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Moringa plants: Bioactive compounds and promising applications in food products
Accepted Manuscript Moringa plants: Bioactive compounds and promising applications in food products
S. Saucedo-Pompa, J.A. Torres-Castillo, C. Castro-López, R. Rojas, E.J. Sánchez-Alejo, M. Ngangyo-Heya, G.C.G. MartínezÁvila PII: DOI: Reference:
S0963-9969(18)30430-7 doi:10.1016/j.foodres.2018.05.062 FRIN 7652
To appear in:
Food Research International
Received date: Revised date: Accepted date:
16 December 2017 1 May 2018 24 May 2018
Please cite this article as: S. Saucedo-Pompa, J.A. Torres-Castillo, C. Castro-López, R. Rojas, E.J. Sánchez-Alejo, M. Ngangyo-Heya, G.C.G. Martínez-Ávila , Moringa plants: Bioactive compounds and promising applications in food products. Food Research International (2017), doi:10.1016/j.foodres.2018.05.062
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ACCEPTED MANUSCRIPT Title and Authorship Information Review paper Moringa plants: Bioactive compounds and promising applications in food products Full author names 3
Saucedo-Pompa S., 1
Torres-Castillo J.A., 1 Castro-López C., 1 Rojas R.,
Sánchez-Alejo E.J., 4 Ngangyo-Heya M. and 1 Martínez-Ávila G.C.G.*
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1, 2
Autonomous University of Nuevo León, School of Agronomy. Chemistry and
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1
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Institutional addresses
Biochemistry Laboratory. General Escobedo, 66050, Nuevo León, México Autonomous University of Coahuila, School of Chemistry. Food Science and Technology
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2
Department. Saltillo, 25280, Coahuila, México Autonomous University of Tamaulipas, Institute of Applied Ecology, Gulf Division 356.
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3
Ciudad Victoria, 87019, Tamaulipas, México 4
Autonomous University of Nuevo León, School of Biological Sciences. Department of
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Botany, 66055, San Nicolás de los Garza, Nuevo León, México
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* Corresponding author: e-mail: [email protected] or
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[email protected]
ACCEPTED MANUSCRIPT
Abstract Moringa plants have an extensive range of bioactive compounds that can be obtained from different vegetative structures, such as leaves, seeds, stems and pod husks. These bioactive
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molecules include carbohydrates, phenolic compounds, oils and fatty acids, proteins and
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functional peptides and have great potential to be used in several formulations of food
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products. This report collects recent information concerning bioactive molecules in other species of the Moringaceae family, different from Moringa oleifera. Thus, this document
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aims to describe these bioactive compounds and their functional properties on foodstuffs. In
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addition, more suitable methodologies applied for their extraction and characterization are reviewed. Finally, an overview of patents required to protect Moringa-derived products and
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processes is provided.
Key words: bioactive compounds, Moringa plants, functional applications, extraction
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methods
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Contents 1. Introduction 2. Bioactive compounds of Moringa plants
2.2 Carbohydrates
2.4 Proteins and peptides
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3. Methods for extraction of bioactive compounds
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2.3 Physicochemical properties of oils and fatty acids
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2.1 Phenolic compounds
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4. Functional applications of Moringa plants in food products 5. Food applications of Moringa plants and intellectual protection
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6. Analytical methods for bioactive compound qualification
Conflict of interest
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References
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Acknowledgment
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8. Concluding remarks
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7. Authors viewpoint
Chemical compounds studied in the article Quercetin (PubChem CID: 5280343); Kaempferol (PubChem CID: 5280863); Apigenin (PubChem
CID:
5280443);
Caffeoylquinic
acid
(PubChem
CID:
10155076);
Coumaroylquinic acid (PubChem CID: 6441280); Feruloylquinic acid (PubChem CID:
ACCEPTED MANUSCRIPT 9799386); Raffinose (PubChem CID: 439242); Arabinose (PubChem CID: 66308); Xylose
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(PubChem CID: 644160); Oleic acid (PubChem CID: 445639)
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1. Introduction Moringa plants belong to a mono-generic family called Moringaceae, which includes 13 species of shrubs and trees originating from India and Africa and distributed in many other
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tropical and arid countries (Salaheldeen, Arouaa, Mariod, Chenge, & Abdelrahmanb, 2014; Al_husnan & Alkahtani, 2016). These plants are well known as medicinal, nutritional and
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water purification agents (Salaheldeen et al., 2015). However, some studies have
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demonstrated that bioactive compounds from Moringa plants could be used for the innovation of functional food products and for other industrial food applications (Oyeyinka
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& Oyeyinka, 2018). Figure 1 shows the main bioactive compounds extracted from different
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vegetative structures of Moringa plants. Due to the high quantity and high quality of bioactive compounds from Moringa plants, these plants could be used in several food
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technologies as antimicrobial agents, antioxidant factors, and food fortificants, among other
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nutritional and technological applications (Ogunsina, Radha, & Indrani 2011; Manaois, Morales, & Abilgos-Ramos, 2013; Nkukwana, Muchenje, Masika, et al., 2014; Radha,
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Ogunsina, & Hebina-Babu, 2015; Oyeyinka & Oyeyinka, 2018; Devisetti, Sreerama, &
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Bhattacharya, 2016). For example, Moringa leaf aqueous extracts are good prospects for food application because they can preserve food by inhibiting lipid oxidation and controlling a wide range of pathogenic microorganisms, such as bacteria and fungi, that are important in the food industry (Shah, Bosco, & Mir, 2015; Al_husnan & Alkahtani, 2016). Moreover, the chemical composition and presumed benefits on human health of Moringa oil allow us to consider this plant a nutraceutical food (Sánchez-Machado et al., 2015). Finally, due to the importance of bioactive compounds in different commercial sectors,
ACCEPTED MANUSCRIPT several efforts have recently been made to determine the most appropriate and standardized methods for the extraction and analysis of these compounds from different plant materials and food matrices, including Moringa plants and derived products (Sánchez-Machado et al., 2015; Rodríguez-Pérez, Mendiola, Quirantes-Piné, Ibáñez, & Segura-Carretero, 2016;
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Nouman et al., 2016; Anudeep, Prasanna, Adya, & Radha, 2016). Thus, this review
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summarizes recent knowledge of the bioactive compounds from Moringa plants and their
most suitable methods for the extraction and
characterization of these bioactive
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components.
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potential use in food product formulation. Furthermore, it provides information about the
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2. Bioactive compounds of Moringa plants
Table 1 shows several bioactive compounds obtained from different vegetative structures of
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Moringa plants and the functional activity of these compounds.
2.1 Phenolic compounds
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Several studies have reported the presence and importance of phenolic compounds from
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different Moringa tissues and have described their bioactivity in both in vitro and in vivo tests. Recently, Juhaimi, Ghafoor, Mohamed-Ahmed, Babiker and Özcan (2017) reported that the total phenolic content of M. oleifera young leaves is 22% higher than that of M. peregrina young leaves, which probably makes M. oleifera a better source for obtaining these bioactive compounds. Among the other bioactive molecules, the main phenolic compounds in M. isolariciresinol,
oleifera
leaves include at least 5 lignans (i.e., medioresinol,
secoisolariciresinol and
epipinoresinol glycosides), 26 flavonoids (i.e.,
ACCEPTED MANUSCRIPT quercetin, kaempferol, apigenin, luteolin and myricetin glycosides) and 11 phenolic acids and their derivatives (i.e., caffeoylquinic, feruloylquinic, and coumaroylquinic acids and their isomers) (Rodríguez-Pérez et al., 2015; Castro-López et al., 2017). Flavonoids, secondary metabolites with several metabolic functions, could thus be considered the main
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phenolic compounds in Moringa plants.
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Phenolic compounds differ from each other by their chemical structure and may occur in
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their free form (as aglycones) or, commonly, linked to a sugar moiety (as glycosides) (Makita, Chimuka, Steenkamp, Cukrowska, & Madala, 2016). Nouman et al. (2016)
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reported that twelve flavonoids, including quercetin, kaempferol and apigenin, constitute
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the major class of phenolic compounds in seven cultivars of M. oleifera. The amounts of total flavonoids recorded in the different cultivars analyzed corresponded to 47.0, 30.0 and
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20.0% for quercetin, kaempferol and apigenin derivatives, respectively. These findings
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agree with Rodríguez-Pérez et al. (2015), who reported flavonoids as the predominant group of phenolics in M. oleifera leaves and registered 46, 34 and 7.7% for quercetin,
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kaempferol and apigenin derivatives, respectively. However, it is important to consider that
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phenolic compound yields are strongly dependent not only on the season, the weather conditions and the application of fertilizers but also on the cultivar and genetic variability,
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which could be the most relevant factors for the phytochemical composition of Moringa leaves (Nouman et al., 2016). Two recent studies reported different percentages of flavonoids but almost the same behavior in terms of the flavonoid derivatives present in Moringa plants. Rodríguez-Pérez et al. (2016) reported the highest percentage (43.75%) of quercetin derivatives in M. oleifera leaves, followed by equal percentages (18.75%) of kaempferol, multiforin B and apigenin derivatives. Moreover, Makita et al. (2016) reported seventeen flavonoids with slight differences from the above results. The authors recorded
ACCEPTED MANUSCRIPT 35, 35, 24 and 6% (on average) of quercetin, kaempferol, isorhamnetin, and apigenin derivatives, respectively, in both M. oleifera and M. ovalifolia. These investigations revealed that most of the flavonoids in leaves from different Moringa cultivars are glycosylated with distinct sugar moieties (i.e., rhamnose, sophorose, rutinose, and others
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not identified by the authors) and that the differences depend on the glycosylation
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machinery of each plant (Makita et al., 2016; Nouman et al., 2016; Castro-López et al.,
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2017). Generally, they are glycosylated for storage purposes in different organelles, which increases the bioavailability of these compounds for human consumption (Makita et al.,
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2016; Nouman et al., 2016). In addition, flavonoid glycosides such as rutin have been
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identified as among the most important compounds responsible for antioxidant activity in M. peregrina leaf extracts, which suggests that this property of phenolic compounds still in
spite
glycosylation
2012).
Although
(Dehshahri,
there
Wink,
Afsharypuor,
Asghari,
and
is insufficient information about phenolic
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Mohagheghzadeh,
of
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remains
compounds from M. peregrina, it has been reported that the levels of these compounds
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range from 88 ± 1.1 to 454 ± 16.3 mg per gram of dry leaves depending on the extracting
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solvent (Marwah et al., 2007; Dehshahri et al., 2012; Al-Owaisi, Al-Hadiwi, & Khan, 2014). Flavonoids have been reported in methanolic extracts of M. concanensis
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(Balamurugan & Balakrishnan, 2013). However, it is essential to perform studies aimed to chemically describe and characterize the functional properties of these Moringa species for food applications. Nouman et al. (2016) reported that phenolic acids in the leaves of M. oleifera ranged from 77 to 187 g per gram of dry material depending on the genetic variability of the cultivar. In addition, it has been reported that caffeoylquinic acid and coumaroylquinic acid isomers
ACCEPTED MANUSCRIPT are the main phenolic acids in M. oleifera, representing at least 45.45 and 36.37%, respectively, of the phenolic acids in the leaves of this plant, depending on the extraction method (Amaglo et al., 2010; Rodríguez-Pérez, Quirantes-Piné, Fernández-Gutiérreza, & Segura-Carretero, 2015; Rodríguez-Pérez et al., 2016; Nouman et al., 2016). This result
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agrees with those of the study conducted by Makita et al. (2017), who reported for the first
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time the presence of cis and trans 3-acyl, 4-acyl and 5-acyl caffeoylquinic, p-
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coumaroylquinic and feruloylquinic acids, among other glycosides, in M. ovalifolia. However, in their recent study, Juhaimi et al. (2017) determined that hydroxybenzoic acids
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(gallic acid and p-hydroxybenzoic acid) are the main phenolic acids in both M. oleifera and
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M. peregrina cultivated in Sudan. This suggests that geographic area, climate and soil composition have a great influence on the phenolic composition of Moringa plants.
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On the other hand, Torres-Castillo et al. (2013) associated phenolic contents with
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antioxidant activity in leaves, roots and stems from M. oleifera, with leaves being the organs with the highest amount of these bioactive compounds. Moreover, phenolics have
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been associated with the antimicrobial and antifungal activities of Moringa extracts
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(Tesfay, Magwaza, Mbili, & Mditshwa, 2017; Al_husnan & Alkahtani, 2016), suggesting that the functional properties of Moringa plants could be closely related to their
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biochemical composition, such as phenolic contents. In addition, the presence of pterygospermin and low pH values caused by the addition of M. oleifera leaves to food products have been shown to be suitable for controlling the growth of undesirable microorganisms in food products (Jayawardana, Liyanage, Lalantha, Iddamalgoda, & Weththasinghe, 2015). Regardless of the specific phenolic compounds that are present (flavonoids or phenolic acids), these biomolecules have been associated with other
ACCEPTED MANUSCRIPT functional advantages preventing lipid oxidation and liver damage (Moyo, Oyedemi,
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Masika, & Muchenje, 2012; Shah, Bosco, & Mir, 2015).
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2.2 Carbohydrates
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According to several scientific reports, the content and functionality of carbohydrates from Moringa plants depend on the vegetative structure used for extraction. Abdulkadir, Zawawi,
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and Jahan (2016) ordered the carbohydrate content in different Moringa tissues as follows:
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seeds > stems > pods > leaves.
The present review provides for the first time a description of the different carbohydrates
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reported in M. oleifera that have shown several advantages for human health and food
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technology. It has been reported that carbohydrates constitute at least 38.2 g/100 g of Moringa leaf powder (Sengev, Abu, & Gernah, 2013). Moreover, Moringa leaf powder has
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1.4- and 3-fold more carbohydrates than do soy flour and mushroom powder, respectively,
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which have been used in the formulation of a healthy vegetable soup powder (Farzana, Mohajan, Saha, Hossain, & Haque, 2017). Carbohydrates in plants could be related to
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nonsoluble and soluble dietary fiber, the intake of which helps to prevent or treat several health disorders. Dietary fiber is the indigestible component of the cell wall in plant materials, has antihyperlipidemic and antihypertensive properties, and ranges from 5 to 6 g/100 g of dry weight in Moringa seed flour, depending on the processing method (Ijarotimi, Adeoti, & Ariyo, 2017). However, higher amounts of this fiber were recently reported in M. oleifera leaves, ranging from 18.1 to 21.1 g/100 g of dry weight depending on month in which the plant material is harvested (Cuellar-Nuñez, Luzardo-Ocampo,
ACCEPTED MANUSCRIPT Campos-Vega, Gallegos-Corona, González de Mejía, & Loarca-Piña, 2018). According to the authors, the fiber content is associated with a reduction in colorectal inflammation in mice. Furthermore, Anudeep et al. (2016) characterized soluble dietary fiber with immunomodulatory effects; although no polysaccharide units were reported by these
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authors, 5% neutral sugars such as arabinose and xylose were identified in this fiber from
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defatted seeds of M. oleifera. Similarly, other monosaccharides have been identified in
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Moringa plants; rhamnose has been associated with other bioactive compounds with potential positive effects on human health in different M. oleifera tissues (Amaglo et al.,
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2010). In addition, the oligosaccharide raffinose has been identified as an antinutritional
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sugar in M. oleifera but is important for the fermentation process in probiotic beverage production (Vanajakshi, Vijayendra, Varadaraj, Venkateswaran, & Agrawal, 2015). Since
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fermentation has been reported to improve the essential amino acid, fatty acid, and
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phytochemical compositions of M. oleifera seed flour samples (Ijarotimi, Adeoti, & Ariyo, 2017), this plant has the potential to be used for fermented foodstuff innovations in the food
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industry.
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Moringa gum is a white stem exudate that changes to reddish brown and brownish black after being exposed to the air; it is composed of 41.5, 26.9, 25.9 and 5.6% galactose,
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arabinose, xylose and rhamnose, respectively (Ravi-Varma, Kumar, Reddy et al., 2014; Abhishek & Ahuja, 2017). Moringa gum was identified by Fourier transform-infrared (FTIR) spectroscopy as a polysaccharide with absorption patterns similar to those presented in other samples of polysaccharides (Jarald, Sumati, Edwin et al., 2012). The authors concluded that this polysaccharide has properties similar to those of tragacanth gum, which is a well-established component of many food processes, indicating the functional potential
ACCEPTED MANUSCRIPT of Moringa gum for increasing viscoelastic properties in food formulation (Kurt, Cengiz, & Kahyaoglu, 2016). In a very recent study, three nonhelical polysaccharides with hypoglycemic activity were isolated from M. oleifera leaves (MLP 1, 2, and 3); these polysaccharides presented the
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same monosaccharide composition (xylose, mannose, glucose, galactose, arabinose and
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galacturonic acid) at different molecular ratios, depending on the concentration of the
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extracting solvent (Chen, Zhang, Huang, Fu, & Liu, 2017). As in the case of other polysaccharides, MLPs are conjugated with components such as sulfate radicals and small
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amounts of proteins that might be responsible for additional functional activities such as
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aggregate formation (Ma, Chen, Zhu, & Wang, 2013). Depending on their rheological properties and chemical compositions, these polysaccharides could be used in different
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food processes in which a pseudoplastic flow behavior is needed, as is the case of MLP 1
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and 2, or for the formulation of functional foods with hypoglycemic ability, as is the case of MLP 3, due to their inhibitory effects on -amylase and -glycosylase. These applications
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agree with the suggestion of Muhammad, Asmawi, and Khan (2016) that M. oleifera be
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used as a potential herbal source to treat diabetes mellitus as well as a nutrition and therapeutic agent. Thus, Moringa plants could be considered sources of carbohydrates with
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several promising food applications. However, there is currently limited information on the carbohydrates from Moringa plants, which provides an opportunity for innovative studies and characterization of these functional components.
2.3 Physicochemical properties of oils and fatty acids
ACCEPTED MANUSCRIPT Ben or Behen oil is the commercial name given to Moringa oil, which is commonly obtained from seeds by numerous traditional and emerging methods (Amaglo et al., 2010; Mat Yusoff, Gordon, Ezeh, & Niranjan, 2016; Bhutada, Jadhav, Pinjari, Nemade, & Jainb, 2016; Fakayode & Ajav, 2016; Rodríguez-Pérez et al., 2016). However, chemical
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characterization of fat crude from Moringa has been detailed for other plant structures, such
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as roots, leaves, flowers, and pods (Amaglo et al., 2010). There are strong differences with
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respect to the yield of Moringa oil, depending on the species, solvents and extraction methods applied in each study. Thus, it can be established that the maximum yields ranged
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from 28.6 to 41% of extracted oil under mechanically optimized conditions and using the
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most traditional extraction method (Soxhlet method), respectively, and could reach a production of 3000 kg/ha (Manzoor, Anwar, Iqbal, & Bhanger, 2007; Fakayode & Ajav,
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2016; Bhutada et al., 2016; Rozina, Asif, Ahmad, Zafar, & Ali, 2017). Moreover, it has
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been demonstrated that M. oleifera is a better oil producer than M. peregrina under Mediterranean conditions because it produces significantly higher yields of seeds and does
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so in a more predictable and uniform manner (Vaknin & Mishal, 2017). Thus, as with
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phenolic compounds, the amount of Moringa oil strongly depends on the cultivar and the genetic variability of these plants.
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Moringa seed oils present similar physicochemical parameters than those recorded for other vegetable oils recognized for their nutritional benefits on human health and applications in food formulations (Table 2) (Manzoor et al., 2007; Kharazi, Kenari, Amiri, et al. 2012; Sánchez-Machado et al., 2015; Salaheldeen et al., 2015; Timilsena, Vongsvivut, Adhikari, & Adhikari, 2017). Fatty acids, namely, oleic, palmitic, heptadecanoic, stearic, arachidic, linoleic, linolenic, eicosenoic, and behenic acids, among others, are the main components of Moringa oil (Amaglo et al., 2010; Sánchez-Machado et al., 2015; Bhutada et al., 2016;
ACCEPTED MANUSCRIPT Vaknin & Mishal, 2017). Monounsaturated fatty acids such as palmitoleic (C16:1), oleic (C18:1), and eicosenoic (20:1) acids are present in the largest amounts, at least in six species of Moringa plants (Amaglo et al., 2010; Salaheldeen et al., 2014). According to Bhutada et al. (2016) and Vaknin and Mishal (2017), oleic acid is the most prevalent
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unsaturated acid in M. oleifera (73.5 %) and M. peregrina (74.3 %) seed oils, which can be
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viewed as a healthy alternative to partially hydrogenated vegetable oils. This result
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confirms the outcomes observed by Salaheldeen et al. (2014), Sánchez-Machado et al. (2015), and Hosseinpour, Aghbashlo, Tabatabaei, and Khalife (2016), who reported 76.9,
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71.1 and 83.8% oleic acid among the total unsaturated fatty acids present in oils extracted
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from M. peregrina, M. oleifera and M. concanensis, respectively. Moreover, this trend has been observed in other Moringa species, such as M. ovalifolia, M. stenopetala and M.
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hildebrandtii (Amaglo et al., 2010). Therefore, due to their high unsaturated fatty acid
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content, Moringa seed oils are ideal substitutes for olive oils Gopalakrishnan, Doriya, & Kumar (2016). Moringa seed oils could decrease the risk for high cholesterol and heart
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diseases due to their high monosaturated fatty acid contents (Dollah, Abdulkarim, Ahmad,
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Khoramina, & Ghazali, 2014). In addition, as explained by the authors, these oils have potential for use in high-oleic trans-fat-free baking and could be applied in soft margarine
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formulations due to their physicochemical properties, such as good spreadability at refrigerated temperatures when blended with other vegetable oils. Other bioactive compounds can be found in Moringa oils, such as sterols, carotenoids, terpenoids, phenolics (including flavonoids), and saponins. These compounds are well known for their therapeutic value and application in the food industry, acting as antioxidants, precursors of vitamin A, and antiproliferative agents (Anwar, Latif, Ashraf, & Gilani, 2005; Bhutada et al., 2016; Hernández-Almanza et al., 2016). It was recently
ACCEPTED MANUSCRIPT reported that the refining process of crude oils allows the removal of the majority of unwanted compounds, maintaining the nutritional value and the availability of fatty acids for consumption (Sánchez-Machado et al., 2015). Because refined oil from Moringa seeds maintains a high oleic acid content, is clear and odorless and resists rancidity, this process
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could be coupled with extraction procedures to obtain high-quality Moringa oil, depending
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on future applications (Omosuli, Oloye, & Ibrahim, 2017). Although physicochemical
application
in
edible
formulations
is
available,
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properties of Moringa oils have been well established, limited information on their providing an opportunity for the
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investigation of these functional compounds in food applications.
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2.4 Proteins and peptides
It has been reported that protein contents in Moringa leaves and seeds range from 22 to
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36.7 g protein/100 g of dry weight (Nouman et al., 2016; Gopalakrishnan et al., 2016;
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Vaknin & Mishal, 2017; Makkar and Becker, 1997). These results agree with those previously reported by Amaglo et al. (2010), who recorded the greatest protein content
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(36.6 g protein/100 g of dry weight) in Moringa seeds and contents of 29.4 and 7.8 g
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protein/100 g of dry weight in the leaves and stems, respectively. However, a maximum protein content (37.5 g protein/100 g of dry weight) was recently reported for M. oleifera
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seeds cultivated in Mexico (González-Garza et al., 2017), suggesting that the amount of protein depends not only on the cultivars and species but also on environmental factors. Thus, these protein contents are significantly higher than those described for other seeds widely used in the formulation of food products, such as chia seeds, whose protein content ranged from 17 to 24 g protein/100 g of dry weight (Timilsena et al., 2017). Moreover, M. oleifera seeds have similar storage protein profiles as those recorded for the main legumes, cereals and oilseeds consumed by humans (Agrawal, Shee, & Sharma, 2007). Considering
ACCEPTED MANUSCRIPT the reports reviewed above, Moringa plants can be considered protein sources for the food industry. In addition to information in Moringa plants on alkaline peptides that range from 6 to 16 kDa and lectins with binder properties for water purification (Santos et al., 2009; Bichi,
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2013), the presence of other bioactive compounds with protein nature from different tissues
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of Moringa plants has been reported in recent years. Thus, protein content as well as
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diversity must be considered because this group can include peptides and proteins with several bioactivities and applications in industrial processes and human nutrition, acting as
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antioxidant, antidiabetic, antihypertensive, antimicrobial, and caseinolytic agents (Estrada-
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Salas et al., 2014; Pinto et al., 2015; González-Garza et al. 2017). Recently, GonzálezGarza et al. (2017) reported an increase in the nutraceutical properties such as the
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antioxidant, antihypertensive and antidiabetic activities of proteins from M. oleifera seeds
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after pepsin-trypsin digestion; this increase was associated with the amino acid composition and the broad spectrum of peptides generated by the possible synergistic effect of these
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enzymes.
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Hydrolytic enzymes related to recycling and maintaining physiological processes, including proteinases, amylases, cellulases and invertases, have been reported in the main vegetative
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structures of M. oleifera (Khatun, Absar, & Ashraduzzaman, 2003; Pontual et al., 2012; Satish, Sairam, Ahmed, & Urooj, 2012; Torres-Castillo et al., 2013). Amylases and proteinases are enzymes with potential applications in the food industry. According to Torres-Castillo et al. (2013), the molecular weight of amylases from M. oleifera varies depending on the extraction source, and amylases are generally active at neutral pH, which makes these enzymes good auxiliary additives for the hydrolysis of starches in bakeries to modify the physicochemical properties of the final products. Proteinases have been
ACCEPTED MANUSCRIPT suggested to be added to food products to partially degrade some proteins and modify the physicochemical properties of meat and dairy products to increase digestibility, improve texture, and affect water retention, viscosity and gelling properties (Pontual et al., 2012). Although these proteinases responsible for caseolitic activity are active at acidic pH,
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proteinases from the leaves and roots of M. oleifera have also been related to caseolitic
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activity at alkaline pH and to the hydrolysis of complex proteins from blood, suggesting the
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potential use of these proteinases in the modification and processing of rich-protein food products (Satish et al., 2012).
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Additional hydrolytic enzymes from Moringa have been reported; however, more studies
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are needed to include them in food applications. The most studied hydrolytic enzymes are cellulase and invertase, whose activities are associated with leaf ripening (Khatun, Absar,
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& Ashraduzzaman, 2003); these enzymes could be applied for the modification of texture,
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sweetness or rheological properties of raw materials or food products. Moreover, enzymes involved in oxidative processes in M. oleifera include peroxidases in roots, stems and
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young leaves, while polyphenol oxidases have been characterized from ripe leaves (Khatun,
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Absar, & Ashraduzzaman, 2001; Khatun et al., 2012; Shank, Riyathong, Lee, & Dheeranupattana, 2013). The levels of peroxidase from different vegetative structures vary
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from 13.3 to 167 units per milligram of protein; the callous tissues that derive from these organs present the highest levels of this enzyme (Khatun et al., 2012; Shank et al., 2013). These findings indicate the potential of in vitro techniques to produce this type of enzyme for application in residual peroxide removal from foods and raw materials and in promoting the discoloration of some materials affected by the presence of certain phenolic compounds. Enzymes related to oxidative balance in plants, including catalase, polyphenol oxidase,
peroxidase,
and
glutathione
reductase,
are usually related
to
protection
ACCEPTED MANUSCRIPT mechanisms, and their accumulation is affected under stress. In M. oleifera leaves, the activity of polyphenoloxidase (2.5 units per gram of fresh weight) was higher than that of peroxidase (0.04 units per gram); this enzyme diversity is frequently related to protection mechanisms against oxidative stress (Kar & Mishra, 1976; Meloni et al., 2003; Sravanthi &
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Rao, 2014). Polyphenoloxidase activity has been associated with a 56 kDa protein that is
IP
active at pH 6 and labile to temperatures higher than 50 °C (Khatun, Absar, &
CR
Ashraduzzaman, 2001). The highest concentrations of this enzyme have been reported in ripe M. oleifera leaves, suggesting the possibility of using these leaves as sources of such
US
enzymes and for application for modification of astringency, coloration and oxidation
AN
during food processing.
In addition to enzymes, biofunctional peptides together with nutraceutical properties can
protein
nature,
including
procoagulant,
antihypertensive,
antiinflammatory,
ED
of
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also be incorporated into functional food formulations. The presence of several components
antiproteolytic and antidiabetic properties, with bioactive effects on human health has been
PT
recently reported (Satish et al., 2012; Pereira et al., 2011; González-Garza et al. 2017).
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Thus, the study of these molecules constitutes a starting point for designing functional foods or nutritional supplements with nutraceutical potential from Moringa plants.
AC
Extracts from the roots and leaves of M. oleifera have shown procoagulant and fibrinogenolytic properties related to the presence of several proteolytic activities and thus are considered for potential use as antihemorrhagic agents. It is important to note that because such proteolytic activity has been detected for the hydrolysis of fibrin and casein, these extracts have potential use for modifying textures in dairy and meat products, in addition to the indicated clinical potential (Satish et al., 2012). The Mo-CBP4 protein (M. oleifera chitin binding protein 4) was isolated from Moringa seeds, it decreases pain-related
ACCEPTED MANUSCRIPT abdominal cramps by approximately 50%, and although its mechanism of action still unknown, this protein is resistant to proteolytic digestion and high temperatures and is thus considered to have good potential for designing targeted drugs or providing functional properties for foodstuffs derived from Moringa seeds (Pereira et al., 2011). In addition,
T
active protease inhibitors against trypsin, chymotrypsin, elastase, thrombin, cathepsin B,
IP
and papain have been reported in Moringa roots and leaves; this opens the possibility of
CR
applying these agents for therapeutic purposes as along with agents for decreasing protein hydrolysis in foods, specifically in sea foods and products that require long storage periods
US
at low temperatures (Bijina et al., 2011).
AN
The antihypertensive protein components are other interesting biofunctional molecules that have been reported in Moringa seeds and leaves. These biomolecules have considerable and
M
specific inhibition against the angiotensin-converting enzyme when they are in the native
ED
state; however, products of enzymatic hydrolysis with pepsin and trypsin increased
PT
inhibition to levels comparable to reference drugs (Mansurah et al., 2015).
CE
3. Methods for extraction of bioactive compounds In addition to conventional methods, emerging methodologies have also been investigated
AC
for the extraction of different bioactive phytomolecules. The efficiency of conventional and nonconventional extraction methods depends mainly on some critical points, such as the understanding of the plant matrix, the chemistry of the target bioactive compounds and the stability of these compounds (Azmir et al., 2013). Several methodologies and extraction techniques have been used to release bioactive compounds from Moringa plants. Table 1 summarizes these methods and the best results in terms of yields obtained in several studies.
ACCEPTED MANUSCRIPT Nonconventional or emerging technologies involve techniques that provide high-quality and high-yield extracts and several technical or environmental advantages during extraction processes, such as shorter extraction times and the use of green solvents. Recently, green solvents have been proven to be suitable for the extraction of several phytochemicals
T
(glucosinolates, chlorogenic acids and flavonoids) from M. oleifera leaves (Djande, Piater,
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Steenkamp, Mandala, & Dubery, 2018). The authors reported that with the use of
CR
pressurized hot water and aqueous two-face extraction systems, the same composition of phenolic compounds can be obtained as those recorded when methanol is used as the
US
extracting solvent. Thus, they highlighted the possibility of using green solvents to obtain
AN
bioactive phytochemicals from Moringa plants through the application of simple and environmentally friendly processes.
M
Moreover, specialized equipment has been applied to obtain bioactive compounds with
ED
satisfactory results. According to Rodríguez-Pérez et al. (2015), higher total phenolic contents were obtained from M. oleifera leaves using ultrasound-assisted extraction than
PT
using a conventional method (maceration) independent of the extraction solvent and
CE
concentration. This resulted in a reduction in the duration of the extraction process of 45 min, which could be related to the disruption of plant cell walls with a subsequent increase
new
AC
in solvent penetration to obtain a higher yield of phenolic compounds. This technique offers opportunities
for
food
companies
to
develop
nutraceuticals,
cosmetic
and
pharmaceutical products and food ingredients from Moringa extracts (Rodríguez-Pérez et al. 2015). It was similarly recently reported that microwave-assisted extraction produced more polyphenolic compounds from different plant materials, including Moringa leaves, than did conventional methods. This extraction efficiency was attributed to the powerful shear, high-frequency vibration, high-velocity impaction, and cavitation involved during
ACCEPTED MANUSCRIPT the extraction process (Castro-López et al., 2017). In addition, during microwave-assisted extraction, the high temperature and microwave energy may disrupt the cell wall and release the bioactive compounds into the solvent, in accordance with disruption theory (Kaufmann, Christen, & Veuthey, 2001; Castro-López et al., 2017). This agrees with the
T
findings of Chen et al. (2017), who reported that under optimal conditions (time 70 min,
IP
microwave power 700 W, temperature 70 °C and liquid:solid ratio 35 mL/g), the
CR
experimental yield of MLPs (2.3%) was close to the value predicted (2.9%) for this polysaccharide. Other emerging technologies have also been applied to extract other
US
bioactive compounds from Moringa plants. Three-step downstream processing was
AN
optimized through supercritical fluid extraction, carbon dioxide-expanded ethanol and pressurized hot water extraction to recover a wide range of bioactive compounds, including
M
fatty, amino and organic acids and phenolic compounds from M. oleifera leaves
ED
(Rodríguez-Pérez et al., 2016). These results demonstrated the suitable use of green extraction methods to obtain from M. oleifera leaves different fractions with a varied
PT
composition of phytochemicals for food application.
CE
Alternatively, other green technologies have been successfully applied to obtain Moringa oil using organic solvent-free methodologies. In a recent study, the optimized mechanical
AC
process for oil extraction from M. oleifera was proven to be a clean alternative to Behen oil recovery for the food industry, leaving aside the use of solvents harmful to human health and the environment (Fakayode & Ajav, 2016). Furthermore, the use of a biotechnological procedure for Moringa oil extraction has highlighted the significance of adding enzyme preparations (Neutrase and Celluclast) for greater oil release from M. oleifera seeds, conferring environmental advantages onto these extractions processes compared to those using organic solvents (see Table 1) (Mat Yusoff et al. 2016). Therefore, these green
ACCEPTED MANUSCRIPT technologies
are
emerging
as
suitable
techniques
for the extraction of different
phytochemical molecules with food applications. However, as observed in Table 2, conventional methodologies are still widely applied for the recovery of bioactive compounds from Moringa tissues with good yields and results
T
concerning their bioactivities and functional properties, regardless of the chemical nature of
IP
these compounds. The chemical nature and polarity of the extraction solvent must be
CR
considered for bioactive compound recovery because these factors play an important role in the extraction yield and functional properties of the obtained extracts (Castro-López, Rojas,
US
Sánchez-Alejo, Niño-Medina, & Martínez-Ávila, 2016). As reported by Al-Owaisi, Al-
AN
Hadiwi and Khan (2014), when the polarity of the extracting solvent increases, more phenolic compounds are extracted, which may be related to the polarity given by hydroxyl
M
groups. According to Tesfay et al. (2017), M. oleifera hydroethanolic extracts (50:50,
ED
ethanol:water) are more effective antimicrobial agents than hydromethanolic extracts against three fungal species (Colletotrichum gloeosporioides, Alternaria alternata and
PT
Lasiodiplodia theobromae) that cause postharvest diseases in avocado. These properties
CE
could be related to the presence of bioactive molecules, such as phenolic compounds (flavonoids), alkaloids, carbohydrates and terpenoids, in Moringa plants as identified in
AC
other Moringa species (Al-Owaisi, Al-Hadiwi, & Khan, 2014). In general, nonpolar solvents (hexane, petroleum ether, etc.), mainly hydrocarbons, esters, oils and fatty acids, could be extracted; meanwhile, polar solvents (methanol, ethanol, etc.) allow the extraction of phenolic compounds and organic acids among other polar biomolecules from different structures of Moringa plants. Cleaner technologies have recently been developed for the recovery of bioactive compounds from these plants and have shown good yields and bioactivities; this provides opportunities to use environmentally friendly techniques for the
ACCEPTED MANUSCRIPT extraction of bioactive compounds from Moringa plants (Asare et al., 2012; Fakayode & Ajav, 2016; Al_husnan & Alkahtani, 2016; Mat Yusoff et al., 2016). Therefore, the extraction technique used is very important in the recovery of bioactive compounds from Moringa plants, and research must consider not only the origin but also the extraction
IP
T
solvent.
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4. Functional applications of Moringa plants in food products
Due to their high nutritional and functional characteristics, Moringa plants have potential to
US
be used in several processes and formulations in the food industry, as their bioactive
AN
compounds have demonstrated safe technological advantages in foodstuff development (Table 3). As recently reported, dehydrated Moringa seeds and leaf powder have been used
M
around the world in the formulation of various edible products to obtain fortified or
ED
functional foods (Dachana, Rajiv, Indrani, and Prakashi (2010); Sengev, Abu, & Gernah, 2013; Shah, Bosco, & Mir, 2015; Al_husnan & Alkahtani, 2016; Devisetti, Sreerama, &
PT
Bhattacharya, 2016; Mamta, Dunkwal, Goyal, & Monika, 2017). Sensory parameters such
CE
as color, appearance, aroma, texture, taste and overall acceptability are very important for the food industry because consumer acceptance depends largely on these attributes.
AC
According to Mamta et al. (2017), the addition of a standardized portion of Moringa powder (5 or 7%) allows us to obtain classifications between “liked moderately” and “liked very much” in valued-added Indian foods, making this plant material an option for the formulation of food products. Moreover, Moringa leaves have good functional properties for use in ready-to-eat food products or snacks such as ribbon-shaped toasted products. Due to its high oil absorption capacity, raw Moringa leaf flour can be used in bakery food formulations, while alkali-pretreated Moringa leaf flour could be more suitable for making
ACCEPTED MANUSCRIPT low-fat snack products (Devisetti, Sreerama, & Bhattacharya, 2016). A significant reduction in the contents of antinutritional compounds (e.g., phytic acid and saponins) was observed after the application of hot air (220 °C) for 2 min in the snack-making process. Moreover, it was reported that p-coumaric acid was the most stable phenolic compound
IP
that Moringa leaves could be suitable in foodstuff formulations.
T
after an alkali pretreatment and the above process of heating Moringa flour, which suggests
CR
Furthermore, Moringa powder significantly increases the nutrient contents of several baked products. According to Dachana et al. (2010), the use of dried Moringa leaves (10 %) in
US
cookie formulations significantly increased the protein, iron, calcium and -carotene
AN
contents but maintained sensory qualities such as crumb color, texture, mouthfeel and flavor. These outcomes are in concordance with the results obtained by Sengev, Abu, and
M
Gernah (2013), where the addition of Moringa leaf powder to a wheat bread formulation
ED
significantly increased its nutrimental composition, especially of magnesium, calcium and
PT
-carotene. Moreover, in addition to the mineral content, vitamin A increases in wheat bread when Moringa seed powder is added to the formulation, making Moringa seed
CE
powder a fortified and functional food product that could help maintain proper vision (Bolarinwa, Aruna, & Raji, 2017).
AC
M. oleifera leaves are of special interest for both physicochemical and microbial food preservation. As indicated by Shah, Bosco, and Mir (2015), in addition to improving muscle proteins in raw beef, M. oleifera leaves contain several bioactive substances as phenolic compounds, which can improve the functional properties of muscle proteins and prevent lipid oxidation in raw meat packaged in a high-oxygen-modified atmosphere. Regarding microbial preservation, M. peregrine was identified as a plant that could be a
ACCEPTED MANUSCRIPT source of new antibiotic compounds because its aqueous extracts have shown antimicrobial activity against bacteria, fungi and yeasts at different concentrations. These extracts have greater antibacterial activity against Gram-negative bacterial strains than against Grampositive bacterial strains, which suggests their potential use for inhibiting the growth and
T
film formation of pathogenic food bacteria and therefore maintaining the microbial quality
IP
of several food products (Al_husnan & Alkahtani, 2016). These findings agree with those
CR
reported by Arora and Onsare (2014), who found that the phytochemicals present in M. oleifera pod husks have antimicrobial potential against a wide range of medically important
US
pathogens, including methicillin-resistant Staphylococcus aureus. This antimicrobial effect
of M.
oleifera
had
AN
agrees with Tesfay et al. (2017), who demonstrated that extracts from the leaves and seeds antifungal properties against Colletotrichum, Alternaria and
M
Lasiodiplodia strains when incorporated into an edible coating formulation to extend the
ED
shelf life of avocados. These activities have been related to the biochemical composition of Moringa plants, which includes the presence of the above-described bioactive compounds.
PT
Furthermore, a low-molecular-mass and thermostable protein has been reported in
CE
agricultural biotechnology as a candidate for developing transgenic crops with antifungal properties; however, due to its amino acid contents, it’s potential allergenic effects of this
AC
protein must be investigated (Gifoni et al., 2012; Pinto et al., 2015). According to Timilsena et al. (2017), plant oils have become increasingly popular due to their high proportion of healthy polyunsaturated fatty acids, which the human body is often unable to efficiently synthetize at sufficient levels. Although seeds of Moringa plants are not widely used for extraction, processing or marketing of edible oils, these materials are rich in monounsaturated fatty acids (such as oleic acid) and have high oxidative stability compared to materials containing polyunsaturated fatty acids (Sánchez-Machado et al.,
ACCEPTED MANUSCRIPT 2015), which are promising properties for used in processes related to food making. As noted by Omosuli, Oloye, and Ibrahim (2017), high-oleic-acid vegetable oils, such as Moringa oil, are very stable even in highly demanding applications such as frying because these oils produce less conjugated dienes and trienes during such processes than do
T
polyunsaturated fatty acid-rich oils (Abdulkarim, Long, Lai, Muhammad, & Ghazali,
IP
2007). In addition, M. oleifera seed oil had appropriate physicochemical properties for
CR
commercial use as an edible oil. These results agree with those of Manzoor et al. (2007), who reported that M. concanensis has potential for edible applications and, due to its fatty
US
acid content, could be used to develop nutritionally balanced and highly stable formulations
AN
for the food industry. As previously mentioned, Moringa plants have great potential for use in a wide variety of food products depending on the bioactive compounds and the target
ED
M
foodstuff.
5. Food applications of Moringa plants and intellectual protection
PT
Although several efforts have been made to evaluate the functional properties of Moringa
CE
plants and their bioactive compounds, researchers do not refer to any patent in their published investigations related to these plants (Dou & Kister, 2016). This suggests that
AC
research is conducted with the purpose of simply generating new knowledge and basic investigation because intellectual property concerning the development of food products from Moringa plants and their bioactive compounds is not reflected. According to searches made in the databases WIPO-PATENTSCOPE (World Intellectual Property Organization) and Google Patents (accessed January 12 th , 2018) using the words “Moringa” and “functional foods”, 28 patents were requested around the world in the last 3 years (between 2015 and 2017) related to the main use of Moringa plants for functional
ACCEPTED MANUSCRIPT purposes. Meanwhile, when the words “Moringa” and “food applications” were used, fewer patent records were detected. Of these patents, four corresponded to the same invention describing a method for the preparation of functional extracts from Moringa plants but were registered in different countries. Two more described polymer coatings based on Moringa
T
and neem oil blends to prevent the establishment and proliferation of microorganisms on
IP
cellulosic-rich materials and were similarly registered in different countries. Finally, two
CR
more were related to the use of Moringa extracts with antioxidant activities for the formulation of medicine and health-care food but did not yet include incorporation into
US
food products. Table 4 summarizes the patent records associated with the application of
AN
Moringa plants for the formulation of several food products. In most cases, the vegetative structures of Moringa plants are used in the form of flour or dehydrated powder and, in
M
some cases, are fermented to obtain the target product. However, a specific characterization
ED
or evaluation of bioactive molecules is not mentioned in any patent. In addition, the conditions of preparation, such as drying, humidity, number of steps, concentration of
PT
ingredients, and pH, are not extensively described. Notwithstanding, the benefits to human
CE
health (decreased blood pressure, sleep promotion, mood regulation, stress relief, enhanced immunity, antitumor effects, and others) are reported without indicating evidence or
AC
experimental procedures.
6. Analytical methods for bioactive compound qualification Several recent studies have shown great amounts of bioactive compounds with broad biological activities, such as phenolic compounds, carbohydrates, oils, and fatty acids, as well as proteins and bioactive peptides. Researchers have the opportunity to apply new analytical techniques to facilitate the proper separation and identification of the bioactive
ACCEPTED MANUSCRIPT compounds present in several food samples, including plant materials such as Moringa plants. There are a variety of techniques related to gas and liquid chromatography as well as infrared and mass spectrometry for the adequate separation and identification of different classes of bioactive compounds; these techniques use sophisticated analytical instruments
T
for appropriate identification of such compounds.
IP
FT-IR spectroscopy is a technique used to investigate the composition of diverse food
CR
samples. In a study of carbohydrates present in Moringa plants, the FT-IR spectra displayed typical abortion peaks given by other polysaccharides, which were detected for some
US
functional groups in the successful characterization of gum and polysaccharides from stems
AN
and leaves (Chen et al., 2017; Abhishek & Ahuja, 2017). This result agrees with Baptista et al. (2017), who reported the presence of various functional groups, including amide groups,
M
for the characterization of protein fractions such as albumin and globulin with coagulant
ED
activity from M. oleifera seeds. Moreover, the infrared spectra of Moringa oil (M. peregrina) are similar to those reported for olive, canola and chia oils, with a predominant
PT
peak in the low-wavenumber region (at 1742 cm-1 ) due to C=O carbonyl stretching of ester
CE
functional groups present in lipids and fatty acids. In addition, the intensity of the peak observed in the region of 3004-3010 cm-1 was lower for Moringa oil than for canola and
AC
chia oils, which can be explained by the low polyunsaturated fatty acid contents in Moringa oil (Salaheldeen et al., 2015; Timilsena et al., 2017). FT-IR spectroscopy has some technical advantages over other techniques used for the identification of bioactive molecules because it is a nondestructive and fast method that requires minimal sample preparation (Singh, Andola, Rawat, Pant, & Purohit, 2011). Thus, this technique could be considered a suitable alternative for integral characterization of the bioactive compounds from Moringa plants and derived food products.
ACCEPTED MANUSCRIPT Currently, liquid chromatography coupled with mass spectrophotometry (LC-MS) has been successfully applied for analysis of several plant materials, including Moringa plants and derived products, to characterize a wide number of natural compounds, such as alcohols, nucleosides, phenolics, organic acids, and amino acids (Amaglo et al., 2010; Rodríguez-
T
Pérez et al., 2015; Makita et al., 2016; Nouman et al., 2016; Devisetti, Sreerama, &
IP
Bhattacharya, 2016; Makita et al., 2017). The best published results for M. oleifera leaf
CR
extract characterization have recorded a total of 59 chemical compounds, including amino and organic acids, nucleosides, glucosinolates, lignans, phenolic acids, and flavonoids, as
US
the main bioactive compounds in these extracts (Rodríguez-Pérez et al., 2015). High-
AN
performance liquid chromatography coupled with electrospray ionization quadrupole-time of flight mass spectrometry (HPLC–ESI–QTOF–MS) allows the identification of chemical
M
molecules by interpreting their MS and MS/MS spectra determined by QTOF–MS
ED
compared to scientific literature and open-access mass-spectra databases. These data agree with the published qualitative results, which indicate that this technique can be applied to
PT
identify at least 50 bioactive compounds, such as organic acids, nucleosides glucosinolates,
CE
and phenolic compounds, from M. oleifera leaf optimized extracts (Rodríguez-Pérez et al., 2016). In addition, ultra-HPLC coupled with QTOF–MS was applied in a comparative
AC
analysis of the flavonoid contents of M. oleifera and M. ovalifolia, and the results revealed that these species contained at least 17 glycosylated flavonoids between them (Makita et al., 2016). These findings were comparable to those recently reported by Castro-López et al. (2017), who reported approximately 19 glycosylated flavonoids, among other phenolics, in extracts obtained from M. oleifera leaves using UPLC-ESI-QTOF-MS2 for the identification of these bioactive molecules. Because they provide good levels of peak asymmetry,
efficiency,
and chemical stability, C18 columns could be considerably
ACCEPTED MANUSCRIPT versatile,
high-performance
separation
columns
suitable
for
a
diverse
range
of
phytochemicals. In most cases reviewed, C18 reversed-phase columns have been successfully applied to separate several phenolics and other polar compounds from Moringa plants. In addition, BEH (ethylene-bridged hybrid) C8 and phenyl columns have
T
shown positive effects in the separation of phenolic compounds. Moreover, acidic solutions
IP
(communally formic acid at 0.1 or 0.5%) such as solvent A and acetonitrile (solvent B) at
CR
different gradients are the most used mobile phases in these studies. As aforementioned, the best results for phytochemical characterization from Moringa plants elucidated 59
US
molecules using the following items: Zorbax Eclipse Plus C18 column (1.8 m, 150 mm ×
AN
4.6 mm) (Agilent Technologies, Palo Alto, CA, USA), acidified water (0.5% formic acid, v/v), and acetonitrile. The injection volume in the HPLC was 10 l. The gradient was
M
programmed as follows: 0 min, 5% B; 10 min, 35% B; 65 min, 95% B; 67 min, 5% B; and
ED
finally, a conditioning cycle of 3 min under the initial conditions. The flow rate was set at
PT
0.50 ml/min throughout the gradient, and the injection volume was 10 l (Rodríguez-Pérez et al., 2015). Furthermore, full-screen mass spectra detection is usually carried out in
CE
negative ion mode in a mass range m/z of 50–1500 Da. However, a full scan in positive and negative ion modes is recommended for quantification and identification of glucosinolates,
AC
phenolics, flavonoids, and various other classes of phytochemicals from Moringa plants (Amago et al. 2010). Therefore, LC-MS is an attractive alternative for the identification of several bioactive compounds because this technique could permit discrimination between isomeric compounds depending on the parameters used (Makita et al., 2017). Fatty
acids
from Moringa
plants
after
derivation
to
methyl esters
through
a
transesterification reaction with methanol have been well characterized. For this purpose,
ACCEPTED MANUSCRIPT gas chromatography (GC) equipped with a flame-ionized detector may be considered the most applied analytical tool for the identification and quantification of different fatty acids present in oils and crude fat from different parts of Moringa plants (Amaglo et al., 2010; Nkukwana et al. 2014; Salaheldeen et al., 2015; Sánchez-Machado et al., 2015; Bhutada et
T
al., 2016). However, according to Al_Owaisi, Al-Hadiwi, and Khan (2014), the use of GC-
IP
MS allows the identification of 19, 7 and 6 different bioactive molecules from hexane,
CR
methanol and ethyl acetate extracts, respectively, after ethanoic pre-extraction of M. peregrina leaves. Furthermore, the use of carbon dioxide-expanded ethanol extracts and
US
GC-MS permits the identification of 20 compounds, including ketones and esters
AN
(Rodríguez-Pérez et al., 2016). Thus, LC/MS and GC/MS have become very suitable techniques for identifying bioactive molecules from Moringa plants.
M
However, colorimetric methods are still widely applied by researchers for the quantification
ED
of bioactive compounds from plant materials, including Moringa plants. Although FolinCiocalteu is not a specific method for phenolic determination (Everette et al., 2010), due to
PT
versatility and quickness, which permit analysis of a large number of samples, this
CE
methodology continues to be accepted for scientists to express total phenolic contents in these samples (Al_Owaisi, Al-Hadiwi, & Khan, 2014; Shah, Bosco, & Mir, 2015; Radha,
AC
Ogunsina, & Hebina-Babu, 2015; Rodríguez-Pérez et al., 2016; Juhaimi et al., 2017). Nevertheless, the presence of other reductive nonphenolic compounds with an aromatic ring (e.g., vitamin C and peptides) can produce many interferences in this analysis; therefore, a previous purification step is recommended (Huang, Ou, & Prior, 2005). To determine protein concentration in Moringa plants, Bradford and Lowry assays and Kjeldahl digestion are the most applied methodologies (Table 1) (Pontual et al., 2012; González-Garza et al., 2017; Baptista et al., 2017). Despite being the most used methods,
ACCEPTED MANUSCRIPT Bradford and Lowry assays have some disadvantages, including the quantification of proteins with specific amino acids (tyrosine and tryptophan) involved in color development and limited use in food samples (Sapan, Lundblad, & Price, 1999). In addition, polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE)
T
is a technique used to estimate the molecular weight of proteins and peptides isolated from
IP
Moringa tissues (Pontual et al., 2012; Ullah et al., 2015; González-Garza et al., 2017).
CR
Nevertheless, HPLC shows good results for molar mass distribution through size exclusion in the characterization protein fractions (e.g., globulins, albumin, prolamin, and glutelin)
US
from Moringa seeds (Baptista et al., 2017). Hence, qualification of bioactive compounds
AN
from Moringa plants could be well characterized with the above-described techniques.
M
7. Authors viewpoint
ED
Taken together, these results indicate that bioactive compounds in Moringa plants are a diverse group of molecules that have proven to be good agents against several health
PT
disorders, such as oxidative stress, hypertension, diabetes, hyperlipidemia, and cancer.
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Although numerous studies have been conducted on M. oleifera bioactive compounds, there is not enough information available on the phytochemicals isolated from other species
AC
of the Moringaceae family, which could be considered prospects with promising applications in food products. Considering the vast potential for food application of this family of plants, researchers should focus more on the effect of processing and storing Moringa-based foodstuff on the content and stability of bioactive compounds to gain a better understanding of the role of these compounds in the food matrix and food quality. In addition, more research should also focus on the methods and conditions for structural elucidation of bioactive compounds to identify and isolate new natural bioactive agents
ACCEPTED MANUSCRIPT from Moringa plants, as this could help to address the adverse effects associated with synthetic food
additives.
Although several studies have investigated the functional
properties of the bioactive compounds reviewed above, additional investigation related to digestibility
and
bioavailability
is
needed
to
determine the functionality of these
T
compounds in both in vitro and in vivo systems. In addition, the functional properties
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measured in many of the studies summarized in this review require further validation in all
CR
Moringa species. On the other hand, despite all presented information, clinical trials that confirm the effects of Moringa components, food safety, and legal framework about
US
commercialization are still pendent and were not included because of the orientation of this
AN
review.
M
8. Concluding remarks
ED
This review aimed to highlight the bioactive compounds in Moringa plants and the recent approaches regarding the functional applications and influence of these biocompounds on
PT
functional characteristics in food products. It was observed that the major food products
CE
based on Moringa plants presented high dietary fiber and low fat contents, which suggests that this plant can be used in the formulation of hypocaloric foodstuffs. Thus, it is possible
AC
to develop food products based on Moringa flour with acceptable sensory and nutritional properties when less than 20 g of these material is used, depending on the target foodstuff. Although emerging technologies have been shown to be suitable alternatives for recovering and characterizing bioactive compounds from Moringa plants, conventional methodologies remain valid. Finally, regardless of several promising functional applications in food products, the exploitation of these properties within the industrial field are still scarcely explored.
ACCEPTED MANUSCRIPT Conflicts of interest The authors declare that there are no conflicts of interest
Acknowledgments
T
Saucedo-Pompa thanks Jesús Noel Yáñez Reyes and Fitokimica Industrial de México S.A.
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de C.V. for the facilities given to study his doctorate. Thanks to the Sheltered Agriculture
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Center of the UANL for the photos used in Figure 1.
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References
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Abdulkadir, A. R., Zawawi, D. D., & Jahan, M. S. (2016). Proximate and phytochemical screening of different parts of Moringa oleifera. Russian Agricultural Sciences, 42(1), 34–
M
36.
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Abhishek, R., & Ahuja, M. (2017). Evaluation of carboxymethyl Moringa gum as nanometric carrier. Carbohydrate Polymers, 174, 896–903.
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Agrawal, H., Shee, Ch., & Sharma, A. K. (2007). Isolation of a 66 kDa protein with
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coagulation activity from seeds of Moringa oleifera. Research Journal of Agriculture and Biological Sciences, 3(5), 418–421.
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Al_husnan, L. A., & Alkahtani, M. D. F. (2016). Impact of Moringa aqueous extract on pathogenic bacteria and fungi in vitro. Annals of Agricultural Science, 61(2), 247–250. Al-Owaisi, M., Al-Hadiwi, N., & Khan, S. A. (2014). GC-MS analysis, determination of total phenolics, flavonoid content and free radical scavenging activities of various crude extracts of Moringa peregrina (Forssk.) Fiori leaves. Asian Pacific Journal of Tropical Biomedicine, 4(12), 964–970.
ACCEPTED MANUSCRIPT Amaglo, N. K., Bennett, R. N., Lo Curto, R. B., Rosa, E. A. S., Lo Turco, V., Giuffrida, A., Lo Curto, A., Crea, F., & Timpo, G. M. (2010). Profiling selected phytochemicals and nutrients in different tissues of the multipurpose tree Moringa oleifera L., grown in Ghana. Food Chemistry, 122(1), 1047–1054.
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Anudeep, A., Prasanna, V. K., Adya, S. M., & Radha, Ch. (2016). Characterization of
IP
soluble dietary fiber from Moringa oleifera seeds and its immunomodulatory effects.
CR
International Journal of Biological Macromolecules, 91, 656–662.
Anwar, F., Latif, S., Ashraf, M., & Gilani, A. H. (2005). Moringa oleifera: A food plant
D.
S.,
& Onsare, J. G. (2014). In vitro antimicrobial evaluation and
AN
Arora,
US
with multiple medicinal uses. Phytotherapy Research, 21(1),17–25.
phytoconstituents of Moringa oleifera pod husks. Industrial Crops and Products, 52, 125–
M
135.
ED
Asare, G. A., Gyan, B., Bugyei, K., Adjei, S., Mahama, R., Addo, P., Otu-Nyarko, L., Wiredu, E. K., & Nyarko, A. (2012). Toxicity potentials of the nutraceutical Moringa
PT
oleifera at supra-supplementation levels. Journal of Ethnopharmacology, 139(1), 265–272.
CE
Azmir, J., Zaidul, I. S. M., Rahman, M. M., Sharif K.M., Mohamed, A., Sahena, F., Jahurul, M. H. A., Ghafoor, K., Norulaini N.A.N., & Omar, A. K. M. (2013). Techniques
AC
for extraction of bioactive compounds from plant materials: A review. Journal of Food Engineering, 117(4), 426–436. Balamurugan,
V.,
&
Balakrishnan,
V.
(2013).
Evaluation
of
phytochemical,
Pharmacognostical and antimicrobial activity from the bark of Moringa concanensis Nimmo. International Journal of Current Microbiology and Applied Sciences, 2(4), 117– 125.
ACCEPTED MANUSCRIPT Baptista, A. T. A., Silva, M. O., Gomes, R. G., Bergamasco, R., Vieira, M. F., & Vieira, A. M. S. (2017). Protein fractionation of seeds of Moringa oleifera lam and its application in superficial water treatment. Separation and Purification Technology, 180, 114–124. Bhutada, P. R., Jadhav, A. J., Pinjari, D. V., Nemade, P. R., & Jainb, R. D. (2016). Solvent
T
assisted extraction of oil from Moringa oleifera Lam. seeds. Industrial Crops and Products,
IP
82, 74–80.
CR
Bichi, M. H. (2013). A review of the applications of Moringa oleifera seeds extract in water treatment. Civil and Environmental Research, 3(8), 1–10.
US
Bijina, B., Chellappan, S., Krishna, J. G., Basheer, S. M., Elyas, K. K., Bahkali, A. H., &
AN
Chandrasekaran, M. (2011). Protease inhibitor from Moringa oleifera with potential for use as therapeutic drug and as seafood preservative. Saudi journal of biological sciences, 18(3),
M
273–281.
ED
Bolarinwa, I. F., Aruna, T. E., & Raji, A. O. (2017). Nutritive value and acceptability of bread fortified with moringa seed powder. Journal of the Saudi Society of Agricultural
PT
Sciences, https://doi.org/10.1016/j.jssas.2017.05.002
CE
Castro-López, C., Rojas, R., Sánchez-Alejo, E. J., Niño-Medina, G., & Martínez-Ávila, G. C. G. (2016). Phenolic compounds recovery from grape fruit and by- products: An
AC
Overview of Extraction Methods. In: Grape and Wine Biotechnology (pp. 103–123). Morata, A., & Loira, I., (Eds.), Intech, Croatia. Castro-López, C., Ventura-Sobrevilla, J. M., González-Hernández, M. D., Rojas, R., Ascacio-Valdés, J. A. Aguilar, C. N., & Martínez-Ávila, G.C.G. (2017). Impact of extraction
techniques
on
antioxidant
capacities
and
polyphenol-rich extracts. Food Chemistry, 237, 1139–1148.
phytochemical composition of
ACCEPTED MANUSCRIPT Chen, Ch., Zhang, B., Huang, Q., Fu, X., & Liu, R. H. (2017). Microwave-assisted extraction of polysaccharides from Moringa oleifera Lam. leaves: Characterization and hypoglycemic activity. Industrial Crops and Products, 100, 1–11. Cuellar-Nuñez M.L., Luzardo-Ocampo, I., Campos-Vega, R., Gallegos-Corona, M.A.,
T
González de Mejía, E., & Loarca-Piña, G. (2018). Physicochemical and nutraceutical
IP
properties of Moringa (Moringa oleifera) leaves and their effects in an in vivo AOM/DSS-
CR
induced colorectal carcinogenesis model. Food Research International, 105, 159–168. Dachana, K. B., Rajiv, J., Indrani, D., & Prakashi, J. (2010). Effect of dried Moringa
US
(Moringa oleifera Lam) leaves on rheological, microstructural, nutritional, textural and
AN
organoleptic characteristics of cookies. Journal of Food Quality, 33(5), 660–677. Dehshahri, S., Wink, M., Afsharypuor, S., Asghari, G., & Mohagheghzadeh, A. (2012).
M
Antioxidant activity of methanolic leaf extract of Moringa peregrina (Forssk.) Fiori.
ED
Research in Pharmaceutical Sciences, 7(2), 111–118. Devisetti, R., Sreerama, Y. N., & Bhattacharya, S. (2016). Processing effects on bioactive
PT
components and functional properties of Moringa leaves: development of a snack and
CE
quality evaluation. Journal of Food Science and Technology, 53(1), 647–657. Dou, H., & Kister, J. (2016). Research and development on Moringa Oleifera e
AC
Comparison between academic research and patents. World Patent Information, 47, 21–33. Everette, J. D., Bayart, Q. M., Green, A. M., Abbey, I. A., Wangila, G. W., & Walker, R. (2010). Thorough study of reactivity of various compound classes toward the FolinCiocalteu reagent. Journal of Agricultural and Food Chemistry, 58(14), 8139–8144. Fakayode, O. A., & Ajav, E. A. (2016). Process optimization of mechanical oil expression from Moringa (Moringa oleifera) seeds. Industrial Crops and Products, 90(1), 142–151.
ACCEPTED MANUSCRIPT Farzana, T., Mohajan, S., Saha, T., Hossain, M. N., & Haque, M. Z. (2017). Formulation and nutritional evaluation of a healthy vegetable soup powder supplemented with soy flour, mushroom, and Moringa leaf. Food Science & Nutrition, 5(4), 911–920. Gifoni, J. M., Oliveira, J. T., Oliveira, H. D., Batista, A. B., Pereira, M. L., Gomes, A. S.,
T
Oliveira, H. P., Grangeiro, T. B., & Vasconcelos, I. M. (2012). A novel chitin-binding
IP
protein from Moringa oleifera seed with potential for plant disease control. Peptide
CR
Science, 98(4), 406–415.
González-Garza, N. G., Chuc-Koyoc, J. A., Torres-Castillo, J. A., García-Zambrano, E. A.,
US
Betancur-Ancona, D., Chel-Guerrero, L., & Sinagawa-García, S. R. (2017). Biofunctional
AN
properties of bioactive peptide fractions from protein isolates of Moringa seed (Moringa oleifera). Journal of Food Science and Technology, 54(13), 4268–4276.
M
Google patents https://patents.google.com/. Accessed January 12th 2018
ED
Gopalakrishnan, L., Doriya, K., & Kumar, D. S. (2016). Moringa oleifera: A review on nutritive importance and its medicinal application. Food Science and Human Wellness,
PT
5(2), 49–56.
CE
Hernández-Almanza, A., Montañez, J., Martínez, G., Aguilar-Jiménez, A., ContrerasEsquivel, J. C., & Aguilar, C. N. (2016). Lycopene: Progress in microbial production.
AC
Trends in Food Science & Technology, 56, 142–148. Hosseinpour, S., Aghbashlo, M., Tabatabaei, M., & Khalife, E. (2016). Exact estimation of biodiesel cetane number (CN) from its fatty acid methyl esters (FAMEs) profile using partial least square (PLS) adapted by artificial neural network (ANN). Energy Conversion and Management, 124, 389–398.
ACCEPTED MANUSCRIPT Ijarotimi, O. S., Adeoti, O. A., & Ariyo, O. (2017). Comparative study on nutrient composition,
phytochemical,
and
functional characteristics of raw,
germinated, and
fermented Moringa oleifera seed flour. Food Science & Nutrition, 1(6), 452-–463. Jarald, E. E., Sumati, S., Edwin, S., Ahmad, S., Patni, S., & Daud, A. (2012).
T
Characterization of Moringa oleifera Lam. Gum to Establish it as a Pharmaceutical
IP
Excipient. Indian Journal of Pharmaceutical Education and Research, 46(3), 211–216.
CR
Jayawardana, B. C., Liyanage, R., Lalantha, N., Iddamalgoda, S., & Weththasinghe, P. (2015). Antioxidant and antimicrobial activity of drumstick (Moringa oleifera) leaves in
US
herbal chicken sausages. LWT - Food Science and Technology, 64, 1204–1208.
AN
Juhaimi, F. AL., Ghafoor, K., Mohamed-Ahmed, I. A., Babiker, E. E., & M. M. Özcan. (2017). Comparative study of mineral and oxidative status of Sonchus oleraceus, Moringa and
Moringa
peregrina
Journal
of
Food
Measurement
and
ED
Characterization, 11(4), 1745–1751.
leaves.
M
oleifera
Kar, M., & Mishra, D. (1975). Inorganic pyrophosphatase activity during rice leaf
PT
senescence. Canadian Journal of Botany, 53, 503–511.
CE
Kharazi, S., H., Kenari, R. E., Amiri, Z. R., & Azizkhani, M. (2012). Characterization of iranian virgin olive oil from the roodbar region: A study on zard, mari and phishomi.
AC
Journal of the American Oil Chemists' Society, 89, 1241–1247. Khatun, S., Absar, N., & Ashraduzzaman, M. (2001). Purification, characterization and effect of physico-chemical agents on stability of polyphenoloxidase from sajna (Moringa oleifera L.) Leaves at Mature Stage. Pakistan Journal of Biological Sciences, 4(9), 1129– 1132.
ACCEPTED MANUSCRIPT Khatun, S., Absar, N., & Ashraduzzaman, M. (2003). Changes in physico-compositions and activities of some hydrolytic and oxidative enzymes in the two types of sajna (Moringa oleifera lam) leaves at different maturity levels. Indian Journal of Plant Physiolog, 8, 6–11. Khatun, S., Ashraduzzaman, M., Karim, M. R., Pervin, F., Absar, N., & Rosma, A. (2012). characterization of peroxidase from Moringa
oleifera L. leaves.
T
Purification and
IP
BioResources, 7(3), 3237–3251.
CR
Kurt, A., Cengiz, A., & Kahyaoglu, T. (2016). The effect of gum tragacanth on the rheological properties of salep based ice cream mix. Carbohydrate Polymers, 143, 116–
US
123.
AN
Ma, L., Chen, H., Zhu, W., & Wang, Z. (2013). Effect of different drying methods on physicochemical properties and antioxidant activities of polysaccharides extracted from
M
mushroom Inonotus obliquus. Food Research International, 50, 633–640
ED
Makita, C., Chimuka, L., Cukrowska, E., Steenkamp, P. A., Kandawa-Schutz, M., Ndhlala, A. R., & Madala, N. E. (2017). UPLC-qTOF-MS profiling of pharmacologically important
PT
chlorogenic acids and associated glycosides in Moringa ovalifolia leaf extracts. South
CE
African Journal of Botany, 108, 193–199. Makita, Ch., Chimuka, L., Steenkamp, P., Cukrowska, E., & Madala, E. (2016).
AC
Comparative analyses of flavonoid content in Moringa oleifera and Moringa ovalifolia with the aid of UHPLC-qTOF-MS fingerprinting. South African Journal of Botany, 105, 116–122. Makkar, H. P. S., & Becker K. (1997). Nutrients and antiquality factors in different morphological parts of the Moringa oleifera tree. Journal of Agricultural Science, 128(3), 311–322.
ACCEPTED MANUSCRIPT Mamta, Dunkwal, V., Goyal, M., & Monika. (2017). Organoleptic acceptability of value added products using drumstick leaves powder. Journal of Pharmacognosy and Phytochemistry, 6(4), 1060–1063. Manaois, R. V. Morales, A. V., & Abilgos-Ramos, R. G. (2013). Acceptability, shelf life
T
and nutritional quality of moringa-supplemented rice crackers. Philippine Journal of Crop
IP
Science, 38(2), 1–8.
CR
Mansurah, A. A., Chioma, N. T., Ismaila, M., Abdulmalik, S. A., Williams, C., & Wudil, A. M. (2015). Partial-purification and characterization of angiotensin converting enzyme
AN
Biochemistry Research & Review, 5(1), 39–48.
US
inhibitory proteins from the leaves and seeds of Moringa oleifera. International Journal of
Manzoor, M., Anwar F., Iqbal, T., & Bhanger, M. I. (2007). Physico-chemical
ED
Chemists' Society, 84(5), 413–419.
M
characterization of Moringa concanensis seeds and seed oil. Journal of the American Oil
Marwah, R. G., Fatope, M. O., Mahrooqi, R. A., Varma, G. B. Abadi, A. A., & Al-
PT
Burtamani, S. K. S. (2007). Antioxidant capacity of some edible and wound healing plants
CE
in Oman. Food Chemistry, 101(2), 465–470. Mat-Yusoff, M., Gordon, M. H., Ezeh, O., & Niranjan, K. (2016). Aqueous enzymatic
AC
extraction of Moringa oleifera oil. Food Chemistry, 211, 400–408. Meloni, D. A., Oliva, M. A., Martinez, C. A., & Cambraia, J. (2003). Photosynthesis and activity of superoxide dismutase, peroxidase and glutathione reductase in cotton under salt stress. Environmental and Experimental Botany, 49, 69–76. Moyo, B., Oyedemi, S., Masika, P. J., & Muchenje, V. (2012). Polyphenolic content and antioxidant properties of Moringa oleifera leaf extracts and enzymatic activity of liver from
ACCEPTED MANUSCRIPT goats supplemented with Moringa oleifera leaves/sunflower seed cake. Meat Science, 91(4), 441–447. Muhammad, H. I., Asmawi, M. Z., & Khan, N. A. K. (2016). A review on promising phytochemical, nutritional and glycemic control studies on Moringa oleifera Lam. in
T
tropical and sub-tropical regions. Asian Pacific Journal of Tropical Biomedicine, 6(10),
IP
896–902.
CR
Nkukwana, T. T., Muchenje, T. T., Masika, V., Hoffman, P. J., Dzama, L. C. K., & Descalzo, A. M. (2014). Fatty acid composition and oxidative stability of breast meat from
AN
refrigeration. Food Chemistry, 142, 255–261.
US
broiler chickens supplemented with Moringa oleifera leaf meal over a period of
Nouman, W., Anwar, F., Gull, T., Newton, A., Rosa, E., & Domínguez-Perles, R. (2016).
M
Profiling of polyphenolics, nutrients and antioxidant potential of germplasm’s leaves from
ED
seven cultivars of Moringa oleifera Lam. Industrial Crops and Products, 83, 166–176. Nwakalor, Ch. N. (2014). Sensory evaluation of cookies produced from different blends of
PT
wheat and Moringa oleifera leaf flour. International Journal of Nutrition and Food
CE
Sciences, 3(4), 307–310.
Ogunsina, B. S., Radha, C., & Indrani, D. (2011). Quality characteristics of bread and
AC
cookies enriched with debittered Moringa oleifera seed flour. International Journal of Food Sciences and Nutrition, 62(2), 185–194. Omosuli, S. V., Oloye, D. A., & Ibrahim, T. A. (2017). Effect of drying methods on the physicochemical properties and fatty acid composition of moringa seeds oil. Progress in Food & Nutrition Science, 1, 027–032. Oyeyinka, A. T., & Oyeyinka, S. A. (2018). Moringa oleifera as a food fortificant: Recent trends and prospects. Journal of the Saudi Society of Agricultural Sciences, 17, 127–137.
ACCEPTED MANUSCRIPT PATENTSCOPE-WIPO (World Intellectual Property Organization). https://patentscope.wipo.int/search/es/result.jsf. Accessed January 12th 2018 Pereira, M. L., David de Oliveira, H., Tadeu Abreu de Oliveira, J., Menezes Gifoni, J., de Oliveira Rocha, R., de Oliveira Bezerra de Sousa, D., & Vasconcelos, I. M. (2011).
T
Purification of a chitin-binding protein from Moringa oleifera seeds with potential to
IP
relieve pain and inflammation. Protein and peptide letters, 18(11), 1078–1085.
CR
Pinto, C. E. M., Farias, D. F., Carvalho, A. F. U., Oliveira, J. T. A., Pereira, M. L., Grangeiro, T. B., Freire, J. E. C., Viana, D. A., & Vasconcelos, I. M. (2015). Food safety
US
assessment of an antifungal protein from Moringa oleifera seeds in an agricultural
AN
biotechnology perspective. Food and Chemical Toxicology, 83, 1–9. Pontual, E. V., Carvalho, B. E., Bezerra, R. S., Coelho, L. C., Napoleão, T. H., & Paiva, P.
ED
Chemistry, 135(3), 1848–1854.
M
M. (2012). Caseinolytic and milk-clotting activities from Moringa oleifera flowers. Food
Radha, C., Ogunsina, B. S., & Hebina-Babu, K. T. (2015). Some quality and micro-
PT
structural characteristics of soup enriched with debittered Moringa oleifera seeds flour.
CE
American Journal of Food Science and Technology, 3(5), 145–149. Ravi-Varma, R. N., Kumar, Ch. P., Reddy, A. K., & Raju, P. P. (2014). Evaluation of
AC
Moringa oleifera gum as a sustained release polymer in diclofenac sodium tablet formulation. International Journal of Research in Pharmacy and Chemistry, 4(3), 687–693. Rodríguez-Pérez, C., Mendiola, J. A., Quirantes-Piné, R., Ibáñez E., & Segura-Carretero A. (2016). Green downstream processing using supercritical carbon dioxide, CO 2 -expanded ethanol and pressurized hot water extractions for recovering bioactive compounds from Moringa oleifera leaves. Journal of Supercritical Fluids, 116, 90–100.
ACCEPTED MANUSCRIPT Rodríguez-Pérez, C., Quirantes-Piné, R., Fernández-Gutiérreza, A., & Segura-Carretero, A. (2015). Optimization of extraction method to obtain a phenolic compounds-rich extract from Moringa oleifera Lam leaves. Industrial Crops and Products, 66, 246–254. Rozina., Asif, S., Ahmad, M., Zafar, M., & Ali, N. (2017). Prospects and potential of fatty
T
acid methyl esters of some non-edible seed oils for use as biodiesel in Pakistan. Renewable
IP
and Sustainable Energy Reviews, 74, 687–702.
CR
Salaheldeen, M., Aroua, M. K., Mariod, A. A., Chenge, S. F., Abdelrahman, M. A., & Atabani, A. E. (2015). Physicochemical characterization and thermal behavior of biodiesel
AN
Conversion and Management, 92, 535–542.
US
and biodiesel–diesel blends derived from crude Moringa peregrina seed oil. Energy
Salaheldeen, M., Arouaa, M. K., Mariod, A. A., Chenge, S. F., & Abdelrahmanb, A. M.
ED
Crops and Products, 61, 49–61.
M
(2014). An evaluation of Moringa peregrina seeds as a source for bio-fuel. Industrial
Sánchez-Machado, D. I., López-Cervantes, J., Núñez-Gastélum, J. A., Mora-López, G. S.,
PT
López-Hernández, J., & Paseiro-Losada, P. (2015). Effect of the refining process on
CE
Moringa oleifera seed oil quality. Food Chemistry, 187, 53–57. Santos, A. F., Luz, L. A., Argolo, A. C., Teixeira, J. A., Paiva, P. M., & Coelho, L. C.
AC
(2009). Isolation of a seed coagulant Moringa oleifera lectin. Process biochemistry, 44(4), 504–508.
Sapan, C. V., Lundblad, R. L., & Price, N. C. (1999). Colorimetric protein assay techniques. Biotechnology and Applied Biochemistry, 29, 99–108. Satish, A., Sairam, S., Ahmed, F., & Urooj, A. (2012). Moringa oleifera Lam.: Protease activity against blood coagulation cascade. Pharmacognosy Research, 4(1), 44–9.
ACCEPTED MANUSCRIPT Sengev, A. I., Abu. J. O., & Gernah, D. I. (2013). Effect of Moringa oleifera leaf powder supplementation on some quality characteristics of wheat bread. Food and Nutrition Sciences, 4, 270–275. Shah, M. A., Bosco, S. J. D., & Mir, S. A. (2015). Effect of Moringa oleifera leaf extract
T
on the physicochemical properties of modified atmosphere packaged raw beef. Food
IP
Packaging and Shelf Life, 3, 31–38.
CR
Shank, L. P., Riyathong, T., Lee, V. S., & Dheeranupattana, S. (2013). Peroxidase activity in native and callus culture of Moringa oleifera Lam. Journal of Medical and
US
Bioengineering, 2(3), 163–167.
AN
Sravanthi, J., & Rao, S. G. (2014). Antioxidative studies in Moringa oleifera Lam. Annals of Phytomedicine, 3(2), 101–105.
M
Tesfay, S. Z., Magwaza, L. S., Mbili, N., & Mditshwa, A. (2017). Carboxyl
ED
methylcellulose (CMC) containing Moringa plant extracts as new postharvest organic edible coating for Avocado (Persea americana Mill.) fruit. Scientia Horticulturae, 226,
PT
201–207.
CE
Timilsena, Y. P., Vongsvivut, J., Adhikari, R., & Adhikari, V. (2017). Physicochemical and thermal characteristics of Australian chia seed oil. Food Chemistry, 228, 394–402.
AC
Torres-Castillo, J. A., Sinagawa-García, S. R., Martínez-Ávila, G. C. G., López-Flores, A. B., Sánchez-González, E. I., Aguirre-Arzola, E. V., Torres-Acosta, R. I., Olivares-Sáenz, E., Osorio-Hernández, E., & Gutiérrez-Díez, A. (2013). Moringa oleifera: phytochemical detection, antioxidants, enzymes and antifugal properties. Phyton, 82, 193–202. Ullah, A., Mariutti, R. B., Masood, R., Caruso, I. P., Costa, G. H. G., de Freita, C. M., Santos, C. R., Zanphorlin, L. M., Mutton, M. J. R., Murakami, M. T., & Arni, R. K. (2015).
ACCEPTED MANUSCRIPT Crystal structure of mature 2S albumin from Moringa oleifera seeds. Biochemical and Biophysical Research Communications, 468(1-2), 365–371. Vaknin, Y., & Mishal, A. (2017). The potential of the tropical “miracle tree” Moringa oleifera and its desert relative Moringa peregrina as edible seed-oil and protein crops under
T
Mediterranean conditions. Scientia Horticulturae, 225, 431–437.
IP
Vanajakshi, V., Vijayendra, S. V. N., Varadaraj, M. C., Venkateswaran, G., & Agrawal, R.
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CE
PT
ED
M
AN
US
Food Science and Technology, 63(2), 1268–1273.
CR
(2015). Optimization of a probiotic beverage based on Moringa leaves and beetroot. LWT-
ACCEPTED MANUSCRIPT Table 1. Bioactive compounds from different vegetative structures of Moringa plants, extraction methods and registered yields Moringa species and tissue
Yield
Extraction method
Functional activity/application
S olvent
Reference
Phenolic compounds
M. oleifera stems, leaves and roots M. oleifera leaves
NI
Maceration extraction
Antioxidant activity
Water
T orresCastillo et al. (2013)
5.30 to 47 mg GAE/g dw depending on the solvent mixture 20.3 to 62.4 GAE/g dw depending on the analyzed fraction NI
Ultrasoundassisted extraction
Potential for antioxidant and antiinflammatory properties
Ethanol, Methanol and Acetone
RodríguezPérez et al. (2015)
Supercritical fluids (green and sustainable method)
Antioxidant activity
Sonication
NI
M. oleifera leaves
6.5 g/100 g dw
M. oleifera seeds
10 mg/ml extract
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M. oleifera seeds M. oleifera seeds
22 g/100 g ww 10.9 mg/g dw
M. oleifera kernels
35.1 g/100 g dw
M. oleifera leaves
22.2 to 31.4 g/100 g dw depending on the cultivars* 37.5 g/100 g dw
M. oleifera seeds Oils
8.1, 10.4, 18.5 and 27.5 g/100 g dw, respectively 2.9 g/100 g dw
M. concanensis seeds
37.7 to 40.1 g/100 dw depending on the source
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RodríguezPérez et al. (2016)
Hydromethanolic (70 %)
Nouman et al. (2016)
Antioxidant and reducing power activities
Water
CastroLópez et al. (2017)
Potential use in drug delivery
NI
Varma et al. (2014)
Immunomodulatory effects Potential to enhance human diet
Phosphate buffer
Anudeep et al. (2016) Abdulkadir et al. (2016)
Microwaveassisted extraction Solubilization by agitation
Hypoglycemic activity
Water
Chen et al. (2017)
Coagulation activity
Agrawal et al. (2007)
Solubilization by agitation Solubilization by agitation
Antimicrobial activity
25 mM sodium phosphate buffer, pH 7.5 Deionized water
Digestion by Kjeldahl’s method Solubilization by agitation
NI
Buffer (0.05 M T risHCl, pH 8.0, containing 0.15 M NaCl) NI
Enzymatic antioxidant activity
Buffer phosphate 50 mM pH 7.8
Digestion by Kjeldahl’s method
Antioxidant, antihypertensive and antidiabetic properties
H2 SO4
GonzálezGarcía et al. (2017)
Soxhlet's method
Potential to develop food formulations with high-stability
Hexane
Manzoor et al. (2007)
Agitation/infusion
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Makita et al. (2016)
Microwaveassisted extraction
Mechanical method (injured in stem sites) Enzyme digestion and precipitation AOAC method
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M. oleifera seeds M. oleifera leaves, pods, stem and seeds M. oleifera leaves
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Proteins
M. oleifera stem
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Carbohydrates
77 to 197 mg/g dw depending on the cultivars 2.7 to 15.2 mg GAE/g dw depending on the solid: liquid ratio NI
1) CO2 ; 2) CO2 /Ethanol; 3) Hot water Aqueous methanol (80 %)
Antioxidant potential
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M. oleifera and M. ovalifolia leaves M. oleifera leaves
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M. oleifera leaves
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Compounds
Antifungal activity
NI
Shebek et al. (2015) Pinto et al. (2015) Mat Yusoff et al. (2016) Nouman et al. (2016)
ACCEPTED MANUSCRIPT region M. peregrina kernels M. oleifera seeds
38 g/100 g dw
Soxhlet's method
NI
Sonication
M. oleifera kernels
29 to 40.7 g/100 g dw depending on particle size 11.4 to 28.6 g/100 g ww depending on the processing conditions 11.80 to 41 g/100 g dw depending on the solid: liquid ratio
Solvent extraction
NI
Hexane
Mechanical method (pressure)
NI
NI
Soxhlet's method
Potential as a healthy alternative to partially hydrogenated vegetable oils
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NI= Not identified *Expressed as crude protein after Digestion by Kjeldahl’s method dw= dry weight ww= wet weight
Chloroform:Methanol (3:1)
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M. oleifera seeds
Hexane Hexane
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M. oleifera seeds
Potential to develop frying foods
Salaheldeen et al. (2014) SánchezMachado et al. (2015) Mat Yusoff et al. (2016) Fakayode and Ajav, (2016) Bhutada et al. (2016)
ACCEPTED MANUSCRIPT Table 2. Physicochemical parameters measured in vegetable oils Properties Viscosity (Pa/s) Kinematic Viscosity (mm2 /s)
M. oleifera 42.8 -
M. peregrina 36.18
34.18
M. concanensis -
Salvia hispanica 43.2 -
0.89
0.89
0.89
0.86
-
RI
1.64
-
-
1.46
1.48
FFA
1.29
0.34
0.35
0.34
63.94
77.17
67.73
67
160.62
198.6
187.53
6.5
-
1
-
1.17
-
-
0.11
Conjugated dienes Acid value (mg KOH/g)
SánchezM achado et al. (2015)
Reference
Salaheldee n et al. (2015)
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RI= refractive index FFA= Free fatty acids (%) IV= Iodine value (g I/100 g) SN= Saponification number * values depend on the growing region of the olive tree
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Peroxide value (meq/kg)
179
204 1.97
-
-
1.75
4.33
3.17
+
-
0.70
Salaheldeen et al. (2014)
M anzoor et al. (2007)
1.46 in all samples
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-
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Carotenoids (g/kg)
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SN (mgKOH/g)
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Density (g mL-1 )
Olive oil*
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2.54 Timilsena et al. (2017)
86.3 to 89.4 1.5 to 2.2 2.26 to 2.70 2.35 to 2.40 0.45 to 0.60 Kharazi et al. (2012)
ACCEPTED MANUSCRIPT Table 3. Functional applications of Moringa oleifera in food products
Leaves
2
Bread
Leaves
5
Cookies
Leaves
10 -20
Modified atmosphere packaged raw beef
Leaves
0.3 **
Leaves
20
Vegetable soup powder
Leaves
Bread
Seeds
5
Leaves
5 - 7†
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Ready-to-eat snack
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6 different Indian foods
* Flour to prepare the food product ** Expressed as g/L of moringa leaf extract † Expressed as g/100 of the final product NI= no identified
8.5†
Reference
Protein and minerals
Ogunsina et al. (2011)
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Rice crackers
Related bioactive compounds
NI
Manaois et al. (2013)
Protein, fiber and minerals
Sengev et al. (2013)
NI
Nwakalor et al. (2014)
Phenolic compounds
Shah et al. (2015)
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10 - 20
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Seeds
Improve the nutritional properties with acceptable rheological and sensory characteristics Have good sensory acceptability scores Improve the nutritional composition Products have acceptable sensorial qualities Provide a stabilization of the color and prevent lipid oxidation in raw beef
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Cookies
Functional advantages
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Foodstuff
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Concentration with best results (g/100 g*)
Vegetative structure
Phenolic compounds, saponins and phytic acid Protein, fiber, Improve the vitamin D and nutritional quality and C, and have long shelf life minerals Improve the Protein, fiber, nutritional quality vitamins and keeping sensory minerals attributes Maintain the organoleptic NI acceptability Decrease in antinutritional factors
Devisetti et al. (2016)
Farzana et al. (2017)
Bolarinwa et al. (2017) Mamta et al. (2017)
ACCEPTED MANUSCRIPT
Table 4. Patent records associated with the application of Moringa plants in the formulation of food products Invention core
Formula and preparation method for Moringa health wine
T he invention belongs to the technical field for food processing and discloses a product based on Moringa and other plant materials; it presented several technical advantages and was reported as a good-for-health beverage
Moringa soybean paste and preparation method thereof
T he invention relates to a method for producing fermented soy sauce: (a) Prepare a soy sauce fermented with Moringa. Soak the soybeans for 3 to 4 hours and then boil them. (b) Mix the fermentation product of the Moringa brine extract of step (a) with the fermentation product of step (b). Aging the stage mix in a dark place for 45 to 60 days and evenly mix the fermentation mixture of aged Meju brine extract and Moringa.
Fermented Moringa beverage and method thereof
T he invention provides a beverage of Moringa prepared using a method for retaining Moringa nutrients and the original flavor with good taste. In addition, was reported this as a hypocaloric beverage due to the use of xylitol as a sweetener.
Moringa pepper paste and preparation method thereof
T his invention relates to a method for preparing a Moringa leaf with a moisture content of 4 to 6%. After collecting and washing Moringa leaves, prepare salt water by adding sun salt to clean water to a salt concentration of 17 to 25%. Mix 1.8 to 2.2 kg of Moringa tea from step (A) with 1 liter of alkaline brine and ferment for 45 to 60 days in a sunny place with a mixture of Moringa leaves and salt water. It can be easily ingested and has excellent nutrients and excellent functional ingredients, thus contributing to the national health and functional food industry.
Le av es
Moringa γamino-acid-rich product and its production process and application
T he invention discloses a M. oleifera product rich in γ-amino butyric acid. T his product could be used raw or as an additive for preparing various health foods for decreasing blood pressure, promoting sleeping and other benefits for human healt h.
St e m s an d
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Title of the required patent
V e g et at iv e st r u ct u re in v ol v e d
P u bl ic at io n d at a
C o u n t r y
R oo ts, st e m s, le av es, fl o w er s
20 /0 6/ 20 17
C hi n a
Le av es
05 /0 7/ 20 17
K or e a
Le av es
03 /0 7/ 20 17 19 /0 7/ 20 17
K or e a
31 /0 5/ 20 17
C hi n a
K or e a
ACCEPTED MANUSCRIPT le av es T his invention relates a typical Korean product and its preparation method, including Moringa leaves and pepper powder, to improve the nutritional content. The invention has large amounts of healthy components with features with anticancer action and hyperlipidemia treatment.
Le av es
T he invention relates a product based on fermented rice and Moringa leaves and is prepared through a drying process. T his product was identified as a product with high nutrient contents.
Le av es
T he invention relates the preparation of Moringa soy through an alkaline treatment.
Le av es
Vinegar of Moringa oleifera and manufacturing method thereof Moringa seed rice cake and preparation method thereof
T his invention displays the manufacturing of M. oleifera vinegar and persimmon through five steps, which could be very useful to modern persons and food industries due to the contents of nutrients and functional components.
Le av es
T he invention discloses a rice cake with Moringa seeds, which contain various nutritive ingredients needed by the human body and further have the functions of clearing gut and promoting metabolism.
Moringa seed noodles and preparation method thereof
It describes a method to make noodles from Moringa seed and leaves rich in bioactive compounds with good health care effect and sensory qualities.
Production of Moringa olei fera QQ candy
It discloses a candy formulation with moringa oil and caloric and noncaloric sugars mixed with other ingredients. In addition, this product has a peculiar flavor and health -care functions.
Moringa seed flour and preparation method thereof
T he invention describes a Moringa seed flour mixed with other functional ingredients, which have beneficial effects on the intestines and stomach of the human body, helping digestion.
Se ed s an d le av es Se ed s an d le af Se ed s (o il) Se ed s
Moringa soybe an milk and preparation method thereof
It provides information about Moringa soybean milk capable of supplementing a variety of nutritional elements required by human bodies and the method for its preparation.
Moringa oleifera and blueberry health-care buccal tablet Moringa hango ver-alleviating
T he invention provides a M. oleifera and blueberry health-care buccal tablet. It can serve as a nutritious supplement with has the oxidation resistance, vision fatigue relief, heart function enhancement, immunity improvement, diabetes prevention, and high blood pressure prevention and is simple to produce and industrialize.
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Method for producing a Moringa kimchi seasoning Fermented rice with Moringa and method of making the same Moringa sauce and preparation method thereof
It relates the development of a functional drink capable of accelerating human metabolism, due to the content of moringa leaves.
N E (fr ee ze dr ie d m or in ga ) Le av es
Le av
03 /0 7/ 20 17 22 /1 2/ 20 17 05 /0 7/ 20 17 09 /1 2/ 20 16 23 /1 1/ 20 16
K or e a
16 /1 1/ 20 16
C hi n a
09 /1 1/ 20 16 09 /1 1/ 20 16 21 /0 9/ 20 16
C hi n a
25 /0 3/ 20 15 30 /0
C hi n a
K or e a K or e a K or e a C hi n a
C hi n a C hi n a
C hi
ACCEPTED MANUSCRIPT composition and preparation method thereof
es
9/ 20 15
n a
08 /0 7/ 20 15 29 /0 4/ 20 15 08 /0 4/ 20 15
C hi n a
04 /1 1/ 20 15 06 /0 5/ 20 15
C hi n a
It discloses the invention of a functional cake from Moringa leaves and others plant materials described as a product with a unique taste that is crispy and delicious.
Le av es
Compound Mo ringa oleifera instant granules and preparation method thereof Moringa flower cake and preparation method thereof
It describes a method to obtain granules rich in active ingredients including flavonoids and polysaccharides from M. oleifera leaves and other leaves. T his was described as a functional health food capable of effectively regulating the blood sugar and blood lipid of a human body.
Le av es
It provides information about a healthy Moringa flower cake from the stem and leaves of Moringa plants and other ingredients.
Moringa oleifera biscuit and preparation method thereof
T he invention relates to a biscuit processing method, particularly a method for preparation
Fl o w er an d st e m le af Le av es
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T his invention has ingredients such as fruit pulp (90 grams) and M. oleifera leaf powder (0.2-4 grams) (no additional information was provided).
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Fruit pulp mixed with Moringa oleifera leaf powder
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Moringa flower cake
Le av es
C hi n a C hi n a
G er m a n y
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Graphical abstract
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Highlights
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Moringa is a source of bioactive compounds with relevance for food industry Carbohydrates reported in M. oleifera are reviewed by the first time Bioactive compounds from Moringaceae family different to M. oleifera are reviewed LC and GC-MS are the most suitable methods for identification of bioactive molecules
Graphics Abstract
Figure 1
S. Saucedo-Pompa, J.A. Torres-Castillo, C. Castro-López, R. Rojas, E.J. Sánchez-Alejo, M. Ngangyo-Heya, G.C.G. MartínezÁvila PII: DOI: Reference:
S0963-9969(18)30430-7 doi:10.1016/j.foodres.2018.05.062 FRIN 7652
To appear in:
Food Research International
Received date: Revised date: Accepted date:
16 December 2017 1 May 2018 24 May 2018
Please cite this article as: S. Saucedo-Pompa, J.A. Torres-Castillo, C. Castro-López, R. Rojas, E.J. Sánchez-Alejo, M. Ngangyo-Heya, G.C.G. Martínez-Ávila , Moringa plants: Bioactive compounds and promising applications in food products. Food Research International (2017), doi:10.1016/j.foodres.2018.05.062
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT Title and Authorship Information Review paper Moringa plants: Bioactive compounds and promising applications in food products Full author names 3
Saucedo-Pompa S., 1
Torres-Castillo J.A., 1 Castro-López C., 1 Rojas R.,
Sánchez-Alejo E.J., 4 Ngangyo-Heya M. and 1 Martínez-Ávila G.C.G.*
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1, 2
Autonomous University of Nuevo León, School of Agronomy. Chemistry and
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1
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Institutional addresses
Biochemistry Laboratory. General Escobedo, 66050, Nuevo León, México Autonomous University of Coahuila, School of Chemistry. Food Science and Technology
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2
Department. Saltillo, 25280, Coahuila, México Autonomous University of Tamaulipas, Institute of Applied Ecology, Gulf Division 356.
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3
Ciudad Victoria, 87019, Tamaulipas, México 4
Autonomous University of Nuevo León, School of Biological Sciences. Department of
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Botany, 66055, San Nicolás de los Garza, Nuevo León, México
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* Corresponding author: e-mail: [email protected] or
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[email protected]
ACCEPTED MANUSCRIPT
Abstract Moringa plants have an extensive range of bioactive compounds that can be obtained from different vegetative structures, such as leaves, seeds, stems and pod husks. These bioactive
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molecules include carbohydrates, phenolic compounds, oils and fatty acids, proteins and
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functional peptides and have great potential to be used in several formulations of food
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products. This report collects recent information concerning bioactive molecules in other species of the Moringaceae family, different from Moringa oleifera. Thus, this document
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aims to describe these bioactive compounds and their functional properties on foodstuffs. In
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addition, more suitable methodologies applied for their extraction and characterization are reviewed. Finally, an overview of patents required to protect Moringa-derived products and
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processes is provided.
Key words: bioactive compounds, Moringa plants, functional applications, extraction
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methods
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Contents 1. Introduction 2. Bioactive compounds of Moringa plants
2.2 Carbohydrates
2.4 Proteins and peptides
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3. Methods for extraction of bioactive compounds
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2.3 Physicochemical properties of oils and fatty acids
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2.1 Phenolic compounds
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4. Functional applications of Moringa plants in food products 5. Food applications of Moringa plants and intellectual protection
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6. Analytical methods for bioactive compound qualification
Conflict of interest
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References
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Acknowledgment
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8. Concluding remarks
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7. Authors viewpoint
Chemical compounds studied in the article Quercetin (PubChem CID: 5280343); Kaempferol (PubChem CID: 5280863); Apigenin (PubChem
CID:
5280443);
Caffeoylquinic
acid
(PubChem
CID:
10155076);
Coumaroylquinic acid (PubChem CID: 6441280); Feruloylquinic acid (PubChem CID:
ACCEPTED MANUSCRIPT 9799386); Raffinose (PubChem CID: 439242); Arabinose (PubChem CID: 66308); Xylose
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(PubChem CID: 644160); Oleic acid (PubChem CID: 445639)
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1. Introduction Moringa plants belong to a mono-generic family called Moringaceae, which includes 13 species of shrubs and trees originating from India and Africa and distributed in many other
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tropical and arid countries (Salaheldeen, Arouaa, Mariod, Chenge, & Abdelrahmanb, 2014; Al_husnan & Alkahtani, 2016). These plants are well known as medicinal, nutritional and
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water purification agents (Salaheldeen et al., 2015). However, some studies have
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demonstrated that bioactive compounds from Moringa plants could be used for the innovation of functional food products and for other industrial food applications (Oyeyinka
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& Oyeyinka, 2018). Figure 1 shows the main bioactive compounds extracted from different
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vegetative structures of Moringa plants. Due to the high quantity and high quality of bioactive compounds from Moringa plants, these plants could be used in several food
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technologies as antimicrobial agents, antioxidant factors, and food fortificants, among other
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nutritional and technological applications (Ogunsina, Radha, & Indrani 2011; Manaois, Morales, & Abilgos-Ramos, 2013; Nkukwana, Muchenje, Masika, et al., 2014; Radha,
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Ogunsina, & Hebina-Babu, 2015; Oyeyinka & Oyeyinka, 2018; Devisetti, Sreerama, &
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Bhattacharya, 2016). For example, Moringa leaf aqueous extracts are good prospects for food application because they can preserve food by inhibiting lipid oxidation and controlling a wide range of pathogenic microorganisms, such as bacteria and fungi, that are important in the food industry (Shah, Bosco, & Mir, 2015; Al_husnan & Alkahtani, 2016). Moreover, the chemical composition and presumed benefits on human health of Moringa oil allow us to consider this plant a nutraceutical food (Sánchez-Machado et al., 2015). Finally, due to the importance of bioactive compounds in different commercial sectors,
ACCEPTED MANUSCRIPT several efforts have recently been made to determine the most appropriate and standardized methods for the extraction and analysis of these compounds from different plant materials and food matrices, including Moringa plants and derived products (Sánchez-Machado et al., 2015; Rodríguez-Pérez, Mendiola, Quirantes-Piné, Ibáñez, & Segura-Carretero, 2016;
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Nouman et al., 2016; Anudeep, Prasanna, Adya, & Radha, 2016). Thus, this review
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summarizes recent knowledge of the bioactive compounds from Moringa plants and their
most suitable methods for the extraction and
characterization of these bioactive
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components.
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potential use in food product formulation. Furthermore, it provides information about the
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2. Bioactive compounds of Moringa plants
Table 1 shows several bioactive compounds obtained from different vegetative structures of
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Moringa plants and the functional activity of these compounds.
2.1 Phenolic compounds
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Several studies have reported the presence and importance of phenolic compounds from
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different Moringa tissues and have described their bioactivity in both in vitro and in vivo tests. Recently, Juhaimi, Ghafoor, Mohamed-Ahmed, Babiker and Özcan (2017) reported that the total phenolic content of M. oleifera young leaves is 22% higher than that of M. peregrina young leaves, which probably makes M. oleifera a better source for obtaining these bioactive compounds. Among the other bioactive molecules, the main phenolic compounds in M. isolariciresinol,
oleifera
leaves include at least 5 lignans (i.e., medioresinol,
secoisolariciresinol and
epipinoresinol glycosides), 26 flavonoids (i.e.,
ACCEPTED MANUSCRIPT quercetin, kaempferol, apigenin, luteolin and myricetin glycosides) and 11 phenolic acids and their derivatives (i.e., caffeoylquinic, feruloylquinic, and coumaroylquinic acids and their isomers) (Rodríguez-Pérez et al., 2015; Castro-López et al., 2017). Flavonoids, secondary metabolites with several metabolic functions, could thus be considered the main
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phenolic compounds in Moringa plants.
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Phenolic compounds differ from each other by their chemical structure and may occur in
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their free form (as aglycones) or, commonly, linked to a sugar moiety (as glycosides) (Makita, Chimuka, Steenkamp, Cukrowska, & Madala, 2016). Nouman et al. (2016)
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reported that twelve flavonoids, including quercetin, kaempferol and apigenin, constitute
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the major class of phenolic compounds in seven cultivars of M. oleifera. The amounts of total flavonoids recorded in the different cultivars analyzed corresponded to 47.0, 30.0 and
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20.0% for quercetin, kaempferol and apigenin derivatives, respectively. These findings
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agree with Rodríguez-Pérez et al. (2015), who reported flavonoids as the predominant group of phenolics in M. oleifera leaves and registered 46, 34 and 7.7% for quercetin,
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kaempferol and apigenin derivatives, respectively. However, it is important to consider that
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phenolic compound yields are strongly dependent not only on the season, the weather conditions and the application of fertilizers but also on the cultivar and genetic variability,
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which could be the most relevant factors for the phytochemical composition of Moringa leaves (Nouman et al., 2016). Two recent studies reported different percentages of flavonoids but almost the same behavior in terms of the flavonoid derivatives present in Moringa plants. Rodríguez-Pérez et al. (2016) reported the highest percentage (43.75%) of quercetin derivatives in M. oleifera leaves, followed by equal percentages (18.75%) of kaempferol, multiforin B and apigenin derivatives. Moreover, Makita et al. (2016) reported seventeen flavonoids with slight differences from the above results. The authors recorded
ACCEPTED MANUSCRIPT 35, 35, 24 and 6% (on average) of quercetin, kaempferol, isorhamnetin, and apigenin derivatives, respectively, in both M. oleifera and M. ovalifolia. These investigations revealed that most of the flavonoids in leaves from different Moringa cultivars are glycosylated with distinct sugar moieties (i.e., rhamnose, sophorose, rutinose, and others
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not identified by the authors) and that the differences depend on the glycosylation
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machinery of each plant (Makita et al., 2016; Nouman et al., 2016; Castro-López et al.,
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2017). Generally, they are glycosylated for storage purposes in different organelles, which increases the bioavailability of these compounds for human consumption (Makita et al.,
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2016; Nouman et al., 2016). In addition, flavonoid glycosides such as rutin have been
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identified as among the most important compounds responsible for antioxidant activity in M. peregrina leaf extracts, which suggests that this property of phenolic compounds still in
spite
glycosylation
2012).
Although
(Dehshahri,
there
Wink,
Afsharypuor,
Asghari,
and
is insufficient information about phenolic
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Mohagheghzadeh,
of
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remains
compounds from M. peregrina, it has been reported that the levels of these compounds
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range from 88 ± 1.1 to 454 ± 16.3 mg per gram of dry leaves depending on the extracting
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solvent (Marwah et al., 2007; Dehshahri et al., 2012; Al-Owaisi, Al-Hadiwi, & Khan, 2014). Flavonoids have been reported in methanolic extracts of M. concanensis
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(Balamurugan & Balakrishnan, 2013). However, it is essential to perform studies aimed to chemically describe and characterize the functional properties of these Moringa species for food applications. Nouman et al. (2016) reported that phenolic acids in the leaves of M. oleifera ranged from 77 to 187 g per gram of dry material depending on the genetic variability of the cultivar. In addition, it has been reported that caffeoylquinic acid and coumaroylquinic acid isomers
ACCEPTED MANUSCRIPT are the main phenolic acids in M. oleifera, representing at least 45.45 and 36.37%, respectively, of the phenolic acids in the leaves of this plant, depending on the extraction method (Amaglo et al., 2010; Rodríguez-Pérez, Quirantes-Piné, Fernández-Gutiérreza, & Segura-Carretero, 2015; Rodríguez-Pérez et al., 2016; Nouman et al., 2016). This result
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agrees with those of the study conducted by Makita et al. (2017), who reported for the first
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time the presence of cis and trans 3-acyl, 4-acyl and 5-acyl caffeoylquinic, p-
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coumaroylquinic and feruloylquinic acids, among other glycosides, in M. ovalifolia. However, in their recent study, Juhaimi et al. (2017) determined that hydroxybenzoic acids
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(gallic acid and p-hydroxybenzoic acid) are the main phenolic acids in both M. oleifera and
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M. peregrina cultivated in Sudan. This suggests that geographic area, climate and soil composition have a great influence on the phenolic composition of Moringa plants.
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On the other hand, Torres-Castillo et al. (2013) associated phenolic contents with
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antioxidant activity in leaves, roots and stems from M. oleifera, with leaves being the organs with the highest amount of these bioactive compounds. Moreover, phenolics have
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been associated with the antimicrobial and antifungal activities of Moringa extracts
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(Tesfay, Magwaza, Mbili, & Mditshwa, 2017; Al_husnan & Alkahtani, 2016), suggesting that the functional properties of Moringa plants could be closely related to their
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biochemical composition, such as phenolic contents. In addition, the presence of pterygospermin and low pH values caused by the addition of M. oleifera leaves to food products have been shown to be suitable for controlling the growth of undesirable microorganisms in food products (Jayawardana, Liyanage, Lalantha, Iddamalgoda, & Weththasinghe, 2015). Regardless of the specific phenolic compounds that are present (flavonoids or phenolic acids), these biomolecules have been associated with other
ACCEPTED MANUSCRIPT functional advantages preventing lipid oxidation and liver damage (Moyo, Oyedemi,
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Masika, & Muchenje, 2012; Shah, Bosco, & Mir, 2015).
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2.2 Carbohydrates
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According to several scientific reports, the content and functionality of carbohydrates from Moringa plants depend on the vegetative structure used for extraction. Abdulkadir, Zawawi,
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and Jahan (2016) ordered the carbohydrate content in different Moringa tissues as follows:
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seeds > stems > pods > leaves.
The present review provides for the first time a description of the different carbohydrates
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reported in M. oleifera that have shown several advantages for human health and food
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technology. It has been reported that carbohydrates constitute at least 38.2 g/100 g of Moringa leaf powder (Sengev, Abu, & Gernah, 2013). Moreover, Moringa leaf powder has
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1.4- and 3-fold more carbohydrates than do soy flour and mushroom powder, respectively,
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which have been used in the formulation of a healthy vegetable soup powder (Farzana, Mohajan, Saha, Hossain, & Haque, 2017). Carbohydrates in plants could be related to
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nonsoluble and soluble dietary fiber, the intake of which helps to prevent or treat several health disorders. Dietary fiber is the indigestible component of the cell wall in plant materials, has antihyperlipidemic and antihypertensive properties, and ranges from 5 to 6 g/100 g of dry weight in Moringa seed flour, depending on the processing method (Ijarotimi, Adeoti, & Ariyo, 2017). However, higher amounts of this fiber were recently reported in M. oleifera leaves, ranging from 18.1 to 21.1 g/100 g of dry weight depending on month in which the plant material is harvested (Cuellar-Nuñez, Luzardo-Ocampo,
ACCEPTED MANUSCRIPT Campos-Vega, Gallegos-Corona, González de Mejía, & Loarca-Piña, 2018). According to the authors, the fiber content is associated with a reduction in colorectal inflammation in mice. Furthermore, Anudeep et al. (2016) characterized soluble dietary fiber with immunomodulatory effects; although no polysaccharide units were reported by these
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authors, 5% neutral sugars such as arabinose and xylose were identified in this fiber from
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defatted seeds of M. oleifera. Similarly, other monosaccharides have been identified in
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Moringa plants; rhamnose has been associated with other bioactive compounds with potential positive effects on human health in different M. oleifera tissues (Amaglo et al.,
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2010). In addition, the oligosaccharide raffinose has been identified as an antinutritional
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sugar in M. oleifera but is important for the fermentation process in probiotic beverage production (Vanajakshi, Vijayendra, Varadaraj, Venkateswaran, & Agrawal, 2015). Since
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fermentation has been reported to improve the essential amino acid, fatty acid, and
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phytochemical compositions of M. oleifera seed flour samples (Ijarotimi, Adeoti, & Ariyo, 2017), this plant has the potential to be used for fermented foodstuff innovations in the food
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industry.
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Moringa gum is a white stem exudate that changes to reddish brown and brownish black after being exposed to the air; it is composed of 41.5, 26.9, 25.9 and 5.6% galactose,
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arabinose, xylose and rhamnose, respectively (Ravi-Varma, Kumar, Reddy et al., 2014; Abhishek & Ahuja, 2017). Moringa gum was identified by Fourier transform-infrared (FTIR) spectroscopy as a polysaccharide with absorption patterns similar to those presented in other samples of polysaccharides (Jarald, Sumati, Edwin et al., 2012). The authors concluded that this polysaccharide has properties similar to those of tragacanth gum, which is a well-established component of many food processes, indicating the functional potential
ACCEPTED MANUSCRIPT of Moringa gum for increasing viscoelastic properties in food formulation (Kurt, Cengiz, & Kahyaoglu, 2016). In a very recent study, three nonhelical polysaccharides with hypoglycemic activity were isolated from M. oleifera leaves (MLP 1, 2, and 3); these polysaccharides presented the
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same monosaccharide composition (xylose, mannose, glucose, galactose, arabinose and
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galacturonic acid) at different molecular ratios, depending on the concentration of the
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extracting solvent (Chen, Zhang, Huang, Fu, & Liu, 2017). As in the case of other polysaccharides, MLPs are conjugated with components such as sulfate radicals and small
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amounts of proteins that might be responsible for additional functional activities such as
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aggregate formation (Ma, Chen, Zhu, & Wang, 2013). Depending on their rheological properties and chemical compositions, these polysaccharides could be used in different
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food processes in which a pseudoplastic flow behavior is needed, as is the case of MLP 1
ED
and 2, or for the formulation of functional foods with hypoglycemic ability, as is the case of MLP 3, due to their inhibitory effects on -amylase and -glycosylase. These applications
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agree with the suggestion of Muhammad, Asmawi, and Khan (2016) that M. oleifera be
CE
used as a potential herbal source to treat diabetes mellitus as well as a nutrition and therapeutic agent. Thus, Moringa plants could be considered sources of carbohydrates with
AC
several promising food applications. However, there is currently limited information on the carbohydrates from Moringa plants, which provides an opportunity for innovative studies and characterization of these functional components.
2.3 Physicochemical properties of oils and fatty acids
ACCEPTED MANUSCRIPT Ben or Behen oil is the commercial name given to Moringa oil, which is commonly obtained from seeds by numerous traditional and emerging methods (Amaglo et al., 2010; Mat Yusoff, Gordon, Ezeh, & Niranjan, 2016; Bhutada, Jadhav, Pinjari, Nemade, & Jainb, 2016; Fakayode & Ajav, 2016; Rodríguez-Pérez et al., 2016). However, chemical
T
characterization of fat crude from Moringa has been detailed for other plant structures, such
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as roots, leaves, flowers, and pods (Amaglo et al., 2010). There are strong differences with
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respect to the yield of Moringa oil, depending on the species, solvents and extraction methods applied in each study. Thus, it can be established that the maximum yields ranged
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from 28.6 to 41% of extracted oil under mechanically optimized conditions and using the
AN
most traditional extraction method (Soxhlet method), respectively, and could reach a production of 3000 kg/ha (Manzoor, Anwar, Iqbal, & Bhanger, 2007; Fakayode & Ajav,
M
2016; Bhutada et al., 2016; Rozina, Asif, Ahmad, Zafar, & Ali, 2017). Moreover, it has
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been demonstrated that M. oleifera is a better oil producer than M. peregrina under Mediterranean conditions because it produces significantly higher yields of seeds and does
PT
so in a more predictable and uniform manner (Vaknin & Mishal, 2017). Thus, as with
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phenolic compounds, the amount of Moringa oil strongly depends on the cultivar and the genetic variability of these plants.
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Moringa seed oils present similar physicochemical parameters than those recorded for other vegetable oils recognized for their nutritional benefits on human health and applications in food formulations (Table 2) (Manzoor et al., 2007; Kharazi, Kenari, Amiri, et al. 2012; Sánchez-Machado et al., 2015; Salaheldeen et al., 2015; Timilsena, Vongsvivut, Adhikari, & Adhikari, 2017). Fatty acids, namely, oleic, palmitic, heptadecanoic, stearic, arachidic, linoleic, linolenic, eicosenoic, and behenic acids, among others, are the main components of Moringa oil (Amaglo et al., 2010; Sánchez-Machado et al., 2015; Bhutada et al., 2016;
ACCEPTED MANUSCRIPT Vaknin & Mishal, 2017). Monounsaturated fatty acids such as palmitoleic (C16:1), oleic (C18:1), and eicosenoic (20:1) acids are present in the largest amounts, at least in six species of Moringa plants (Amaglo et al., 2010; Salaheldeen et al., 2014). According to Bhutada et al. (2016) and Vaknin and Mishal (2017), oleic acid is the most prevalent
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unsaturated acid in M. oleifera (73.5 %) and M. peregrina (74.3 %) seed oils, which can be
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viewed as a healthy alternative to partially hydrogenated vegetable oils. This result
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confirms the outcomes observed by Salaheldeen et al. (2014), Sánchez-Machado et al. (2015), and Hosseinpour, Aghbashlo, Tabatabaei, and Khalife (2016), who reported 76.9,
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71.1 and 83.8% oleic acid among the total unsaturated fatty acids present in oils extracted
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from M. peregrina, M. oleifera and M. concanensis, respectively. Moreover, this trend has been observed in other Moringa species, such as M. ovalifolia, M. stenopetala and M.
M
hildebrandtii (Amaglo et al., 2010). Therefore, due to their high unsaturated fatty acid
ED
content, Moringa seed oils are ideal substitutes for olive oils Gopalakrishnan, Doriya, & Kumar (2016). Moringa seed oils could decrease the risk for high cholesterol and heart
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diseases due to their high monosaturated fatty acid contents (Dollah, Abdulkarim, Ahmad,
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Khoramina, & Ghazali, 2014). In addition, as explained by the authors, these oils have potential for use in high-oleic trans-fat-free baking and could be applied in soft margarine
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formulations due to their physicochemical properties, such as good spreadability at refrigerated temperatures when blended with other vegetable oils. Other bioactive compounds can be found in Moringa oils, such as sterols, carotenoids, terpenoids, phenolics (including flavonoids), and saponins. These compounds are well known for their therapeutic value and application in the food industry, acting as antioxidants, precursors of vitamin A, and antiproliferative agents (Anwar, Latif, Ashraf, & Gilani, 2005; Bhutada et al., 2016; Hernández-Almanza et al., 2016). It was recently
ACCEPTED MANUSCRIPT reported that the refining process of crude oils allows the removal of the majority of unwanted compounds, maintaining the nutritional value and the availability of fatty acids for consumption (Sánchez-Machado et al., 2015). Because refined oil from Moringa seeds maintains a high oleic acid content, is clear and odorless and resists rancidity, this process
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could be coupled with extraction procedures to obtain high-quality Moringa oil, depending
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on future applications (Omosuli, Oloye, & Ibrahim, 2017). Although physicochemical
application
in
edible
formulations
is
available,
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properties of Moringa oils have been well established, limited information on their providing an opportunity for the
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investigation of these functional compounds in food applications.
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2.4 Proteins and peptides
It has been reported that protein contents in Moringa leaves and seeds range from 22 to
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36.7 g protein/100 g of dry weight (Nouman et al., 2016; Gopalakrishnan et al., 2016;
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Vaknin & Mishal, 2017; Makkar and Becker, 1997). These results agree with those previously reported by Amaglo et al. (2010), who recorded the greatest protein content
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(36.6 g protein/100 g of dry weight) in Moringa seeds and contents of 29.4 and 7.8 g
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protein/100 g of dry weight in the leaves and stems, respectively. However, a maximum protein content (37.5 g protein/100 g of dry weight) was recently reported for M. oleifera
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seeds cultivated in Mexico (González-Garza et al., 2017), suggesting that the amount of protein depends not only on the cultivars and species but also on environmental factors. Thus, these protein contents are significantly higher than those described for other seeds widely used in the formulation of food products, such as chia seeds, whose protein content ranged from 17 to 24 g protein/100 g of dry weight (Timilsena et al., 2017). Moreover, M. oleifera seeds have similar storage protein profiles as those recorded for the main legumes, cereals and oilseeds consumed by humans (Agrawal, Shee, & Sharma, 2007). Considering
ACCEPTED MANUSCRIPT the reports reviewed above, Moringa plants can be considered protein sources for the food industry. In addition to information in Moringa plants on alkaline peptides that range from 6 to 16 kDa and lectins with binder properties for water purification (Santos et al., 2009; Bichi,
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2013), the presence of other bioactive compounds with protein nature from different tissues
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of Moringa plants has been reported in recent years. Thus, protein content as well as
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diversity must be considered because this group can include peptides and proteins with several bioactivities and applications in industrial processes and human nutrition, acting as
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antioxidant, antidiabetic, antihypertensive, antimicrobial, and caseinolytic agents (Estrada-
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Salas et al., 2014; Pinto et al., 2015; González-Garza et al. 2017). Recently, GonzálezGarza et al. (2017) reported an increase in the nutraceutical properties such as the
M
antioxidant, antihypertensive and antidiabetic activities of proteins from M. oleifera seeds
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after pepsin-trypsin digestion; this increase was associated with the amino acid composition and the broad spectrum of peptides generated by the possible synergistic effect of these
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enzymes.
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Hydrolytic enzymes related to recycling and maintaining physiological processes, including proteinases, amylases, cellulases and invertases, have been reported in the main vegetative
AC
structures of M. oleifera (Khatun, Absar, & Ashraduzzaman, 2003; Pontual et al., 2012; Satish, Sairam, Ahmed, & Urooj, 2012; Torres-Castillo et al., 2013). Amylases and proteinases are enzymes with potential applications in the food industry. According to Torres-Castillo et al. (2013), the molecular weight of amylases from M. oleifera varies depending on the extraction source, and amylases are generally active at neutral pH, which makes these enzymes good auxiliary additives for the hydrolysis of starches in bakeries to modify the physicochemical properties of the final products. Proteinases have been
ACCEPTED MANUSCRIPT suggested to be added to food products to partially degrade some proteins and modify the physicochemical properties of meat and dairy products to increase digestibility, improve texture, and affect water retention, viscosity and gelling properties (Pontual et al., 2012). Although these proteinases responsible for caseolitic activity are active at acidic pH,
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proteinases from the leaves and roots of M. oleifera have also been related to caseolitic
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activity at alkaline pH and to the hydrolysis of complex proteins from blood, suggesting the
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potential use of these proteinases in the modification and processing of rich-protein food products (Satish et al., 2012).
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Additional hydrolytic enzymes from Moringa have been reported; however, more studies
AN
are needed to include them in food applications. The most studied hydrolytic enzymes are cellulase and invertase, whose activities are associated with leaf ripening (Khatun, Absar,
M
& Ashraduzzaman, 2003); these enzymes could be applied for the modification of texture,
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sweetness or rheological properties of raw materials or food products. Moreover, enzymes involved in oxidative processes in M. oleifera include peroxidases in roots, stems and
PT
young leaves, while polyphenol oxidases have been characterized from ripe leaves (Khatun,
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Absar, & Ashraduzzaman, 2001; Khatun et al., 2012; Shank, Riyathong, Lee, & Dheeranupattana, 2013). The levels of peroxidase from different vegetative structures vary
AC
from 13.3 to 167 units per milligram of protein; the callous tissues that derive from these organs present the highest levels of this enzyme (Khatun et al., 2012; Shank et al., 2013). These findings indicate the potential of in vitro techniques to produce this type of enzyme for application in residual peroxide removal from foods and raw materials and in promoting the discoloration of some materials affected by the presence of certain phenolic compounds. Enzymes related to oxidative balance in plants, including catalase, polyphenol oxidase,
peroxidase,
and
glutathione
reductase,
are usually related
to
protection
ACCEPTED MANUSCRIPT mechanisms, and their accumulation is affected under stress. In M. oleifera leaves, the activity of polyphenoloxidase (2.5 units per gram of fresh weight) was higher than that of peroxidase (0.04 units per gram); this enzyme diversity is frequently related to protection mechanisms against oxidative stress (Kar & Mishra, 1976; Meloni et al., 2003; Sravanthi &
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Rao, 2014). Polyphenoloxidase activity has been associated with a 56 kDa protein that is
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active at pH 6 and labile to temperatures higher than 50 °C (Khatun, Absar, &
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Ashraduzzaman, 2001). The highest concentrations of this enzyme have been reported in ripe M. oleifera leaves, suggesting the possibility of using these leaves as sources of such
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enzymes and for application for modification of astringency, coloration and oxidation
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during food processing.
In addition to enzymes, biofunctional peptides together with nutraceutical properties can
protein
nature,
including
procoagulant,
antihypertensive,
antiinflammatory,
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of
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also be incorporated into functional food formulations. The presence of several components
antiproteolytic and antidiabetic properties, with bioactive effects on human health has been
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recently reported (Satish et al., 2012; Pereira et al., 2011; González-Garza et al. 2017).
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Thus, the study of these molecules constitutes a starting point for designing functional foods or nutritional supplements with nutraceutical potential from Moringa plants.
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Extracts from the roots and leaves of M. oleifera have shown procoagulant and fibrinogenolytic properties related to the presence of several proteolytic activities and thus are considered for potential use as antihemorrhagic agents. It is important to note that because such proteolytic activity has been detected for the hydrolysis of fibrin and casein, these extracts have potential use for modifying textures in dairy and meat products, in addition to the indicated clinical potential (Satish et al., 2012). The Mo-CBP4 protein (M. oleifera chitin binding protein 4) was isolated from Moringa seeds, it decreases pain-related
ACCEPTED MANUSCRIPT abdominal cramps by approximately 50%, and although its mechanism of action still unknown, this protein is resistant to proteolytic digestion and high temperatures and is thus considered to have good potential for designing targeted drugs or providing functional properties for foodstuffs derived from Moringa seeds (Pereira et al., 2011). In addition,
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active protease inhibitors against trypsin, chymotrypsin, elastase, thrombin, cathepsin B,
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and papain have been reported in Moringa roots and leaves; this opens the possibility of
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applying these agents for therapeutic purposes as along with agents for decreasing protein hydrolysis in foods, specifically in sea foods and products that require long storage periods
US
at low temperatures (Bijina et al., 2011).
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The antihypertensive protein components are other interesting biofunctional molecules that have been reported in Moringa seeds and leaves. These biomolecules have considerable and
M
specific inhibition against the angiotensin-converting enzyme when they are in the native
ED
state; however, products of enzymatic hydrolysis with pepsin and trypsin increased
PT
inhibition to levels comparable to reference drugs (Mansurah et al., 2015).
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3. Methods for extraction of bioactive compounds In addition to conventional methods, emerging methodologies have also been investigated
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for the extraction of different bioactive phytomolecules. The efficiency of conventional and nonconventional extraction methods depends mainly on some critical points, such as the understanding of the plant matrix, the chemistry of the target bioactive compounds and the stability of these compounds (Azmir et al., 2013). Several methodologies and extraction techniques have been used to release bioactive compounds from Moringa plants. Table 1 summarizes these methods and the best results in terms of yields obtained in several studies.
ACCEPTED MANUSCRIPT Nonconventional or emerging technologies involve techniques that provide high-quality and high-yield extracts and several technical or environmental advantages during extraction processes, such as shorter extraction times and the use of green solvents. Recently, green solvents have been proven to be suitable for the extraction of several phytochemicals
T
(glucosinolates, chlorogenic acids and flavonoids) from M. oleifera leaves (Djande, Piater,
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Steenkamp, Mandala, & Dubery, 2018). The authors reported that with the use of
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pressurized hot water and aqueous two-face extraction systems, the same composition of phenolic compounds can be obtained as those recorded when methanol is used as the
US
extracting solvent. Thus, they highlighted the possibility of using green solvents to obtain
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bioactive phytochemicals from Moringa plants through the application of simple and environmentally friendly processes.
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Moreover, specialized equipment has been applied to obtain bioactive compounds with
ED
satisfactory results. According to Rodríguez-Pérez et al. (2015), higher total phenolic contents were obtained from M. oleifera leaves using ultrasound-assisted extraction than
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using a conventional method (maceration) independent of the extraction solvent and
CE
concentration. This resulted in a reduction in the duration of the extraction process of 45 min, which could be related to the disruption of plant cell walls with a subsequent increase
new
AC
in solvent penetration to obtain a higher yield of phenolic compounds. This technique offers opportunities
for
food
companies
to
develop
nutraceuticals,
cosmetic
and
pharmaceutical products and food ingredients from Moringa extracts (Rodríguez-Pérez et al. 2015). It was similarly recently reported that microwave-assisted extraction produced more polyphenolic compounds from different plant materials, including Moringa leaves, than did conventional methods. This extraction efficiency was attributed to the powerful shear, high-frequency vibration, high-velocity impaction, and cavitation involved during
ACCEPTED MANUSCRIPT the extraction process (Castro-López et al., 2017). In addition, during microwave-assisted extraction, the high temperature and microwave energy may disrupt the cell wall and release the bioactive compounds into the solvent, in accordance with disruption theory (Kaufmann, Christen, & Veuthey, 2001; Castro-López et al., 2017). This agrees with the
T
findings of Chen et al. (2017), who reported that under optimal conditions (time 70 min,
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microwave power 700 W, temperature 70 °C and liquid:solid ratio 35 mL/g), the
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experimental yield of MLPs (2.3%) was close to the value predicted (2.9%) for this polysaccharide. Other emerging technologies have also been applied to extract other
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bioactive compounds from Moringa plants. Three-step downstream processing was
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optimized through supercritical fluid extraction, carbon dioxide-expanded ethanol and pressurized hot water extraction to recover a wide range of bioactive compounds, including
M
fatty, amino and organic acids and phenolic compounds from M. oleifera leaves
ED
(Rodríguez-Pérez et al., 2016). These results demonstrated the suitable use of green extraction methods to obtain from M. oleifera leaves different fractions with a varied
PT
composition of phytochemicals for food application.
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Alternatively, other green technologies have been successfully applied to obtain Moringa oil using organic solvent-free methodologies. In a recent study, the optimized mechanical
AC
process for oil extraction from M. oleifera was proven to be a clean alternative to Behen oil recovery for the food industry, leaving aside the use of solvents harmful to human health and the environment (Fakayode & Ajav, 2016). Furthermore, the use of a biotechnological procedure for Moringa oil extraction has highlighted the significance of adding enzyme preparations (Neutrase and Celluclast) for greater oil release from M. oleifera seeds, conferring environmental advantages onto these extractions processes compared to those using organic solvents (see Table 1) (Mat Yusoff et al. 2016). Therefore, these green
ACCEPTED MANUSCRIPT technologies
are
emerging
as
suitable
techniques
for the extraction of different
phytochemical molecules with food applications. However, as observed in Table 2, conventional methodologies are still widely applied for the recovery of bioactive compounds from Moringa tissues with good yields and results
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concerning their bioactivities and functional properties, regardless of the chemical nature of
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these compounds. The chemical nature and polarity of the extraction solvent must be
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considered for bioactive compound recovery because these factors play an important role in the extraction yield and functional properties of the obtained extracts (Castro-López, Rojas,
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Sánchez-Alejo, Niño-Medina, & Martínez-Ávila, 2016). As reported by Al-Owaisi, Al-
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Hadiwi and Khan (2014), when the polarity of the extracting solvent increases, more phenolic compounds are extracted, which may be related to the polarity given by hydroxyl
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groups. According to Tesfay et al. (2017), M. oleifera hydroethanolic extracts (50:50,
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ethanol:water) are more effective antimicrobial agents than hydromethanolic extracts against three fungal species (Colletotrichum gloeosporioides, Alternaria alternata and
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Lasiodiplodia theobromae) that cause postharvest diseases in avocado. These properties
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could be related to the presence of bioactive molecules, such as phenolic compounds (flavonoids), alkaloids, carbohydrates and terpenoids, in Moringa plants as identified in
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other Moringa species (Al-Owaisi, Al-Hadiwi, & Khan, 2014). In general, nonpolar solvents (hexane, petroleum ether, etc.), mainly hydrocarbons, esters, oils and fatty acids, could be extracted; meanwhile, polar solvents (methanol, ethanol, etc.) allow the extraction of phenolic compounds and organic acids among other polar biomolecules from different structures of Moringa plants. Cleaner technologies have recently been developed for the recovery of bioactive compounds from these plants and have shown good yields and bioactivities; this provides opportunities to use environmentally friendly techniques for the
ACCEPTED MANUSCRIPT extraction of bioactive compounds from Moringa plants (Asare et al., 2012; Fakayode & Ajav, 2016; Al_husnan & Alkahtani, 2016; Mat Yusoff et al., 2016). Therefore, the extraction technique used is very important in the recovery of bioactive compounds from Moringa plants, and research must consider not only the origin but also the extraction
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solvent.
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4. Functional applications of Moringa plants in food products
Due to their high nutritional and functional characteristics, Moringa plants have potential to
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be used in several processes and formulations in the food industry, as their bioactive
AN
compounds have demonstrated safe technological advantages in foodstuff development (Table 3). As recently reported, dehydrated Moringa seeds and leaf powder have been used
M
around the world in the formulation of various edible products to obtain fortified or
ED
functional foods (Dachana, Rajiv, Indrani, and Prakashi (2010); Sengev, Abu, & Gernah, 2013; Shah, Bosco, & Mir, 2015; Al_husnan & Alkahtani, 2016; Devisetti, Sreerama, &
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Bhattacharya, 2016; Mamta, Dunkwal, Goyal, & Monika, 2017). Sensory parameters such
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as color, appearance, aroma, texture, taste and overall acceptability are very important for the food industry because consumer acceptance depends largely on these attributes.
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According to Mamta et al. (2017), the addition of a standardized portion of Moringa powder (5 or 7%) allows us to obtain classifications between “liked moderately” and “liked very much” in valued-added Indian foods, making this plant material an option for the formulation of food products. Moreover, Moringa leaves have good functional properties for use in ready-to-eat food products or snacks such as ribbon-shaped toasted products. Due to its high oil absorption capacity, raw Moringa leaf flour can be used in bakery food formulations, while alkali-pretreated Moringa leaf flour could be more suitable for making
ACCEPTED MANUSCRIPT low-fat snack products (Devisetti, Sreerama, & Bhattacharya, 2016). A significant reduction in the contents of antinutritional compounds (e.g., phytic acid and saponins) was observed after the application of hot air (220 °C) for 2 min in the snack-making process. Moreover, it was reported that p-coumaric acid was the most stable phenolic compound
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that Moringa leaves could be suitable in foodstuff formulations.
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after an alkali pretreatment and the above process of heating Moringa flour, which suggests
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Furthermore, Moringa powder significantly increases the nutrient contents of several baked products. According to Dachana et al. (2010), the use of dried Moringa leaves (10 %) in
US
cookie formulations significantly increased the protein, iron, calcium and -carotene
AN
contents but maintained sensory qualities such as crumb color, texture, mouthfeel and flavor. These outcomes are in concordance with the results obtained by Sengev, Abu, and
M
Gernah (2013), where the addition of Moringa leaf powder to a wheat bread formulation
ED
significantly increased its nutrimental composition, especially of magnesium, calcium and
PT
-carotene. Moreover, in addition to the mineral content, vitamin A increases in wheat bread when Moringa seed powder is added to the formulation, making Moringa seed
CE
powder a fortified and functional food product that could help maintain proper vision (Bolarinwa, Aruna, & Raji, 2017).
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M. oleifera leaves are of special interest for both physicochemical and microbial food preservation. As indicated by Shah, Bosco, and Mir (2015), in addition to improving muscle proteins in raw beef, M. oleifera leaves contain several bioactive substances as phenolic compounds, which can improve the functional properties of muscle proteins and prevent lipid oxidation in raw meat packaged in a high-oxygen-modified atmosphere. Regarding microbial preservation, M. peregrine was identified as a plant that could be a
ACCEPTED MANUSCRIPT source of new antibiotic compounds because its aqueous extracts have shown antimicrobial activity against bacteria, fungi and yeasts at different concentrations. These extracts have greater antibacterial activity against Gram-negative bacterial strains than against Grampositive bacterial strains, which suggests their potential use for inhibiting the growth and
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film formation of pathogenic food bacteria and therefore maintaining the microbial quality
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of several food products (Al_husnan & Alkahtani, 2016). These findings agree with those
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reported by Arora and Onsare (2014), who found that the phytochemicals present in M. oleifera pod husks have antimicrobial potential against a wide range of medically important
US
pathogens, including methicillin-resistant Staphylococcus aureus. This antimicrobial effect
of M.
oleifera
had
AN
agrees with Tesfay et al. (2017), who demonstrated that extracts from the leaves and seeds antifungal properties against Colletotrichum, Alternaria and
M
Lasiodiplodia strains when incorporated into an edible coating formulation to extend the
ED
shelf life of avocados. These activities have been related to the biochemical composition of Moringa plants, which includes the presence of the above-described bioactive compounds.
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Furthermore, a low-molecular-mass and thermostable protein has been reported in
CE
agricultural biotechnology as a candidate for developing transgenic crops with antifungal properties; however, due to its amino acid contents, it’s potential allergenic effects of this
AC
protein must be investigated (Gifoni et al., 2012; Pinto et al., 2015). According to Timilsena et al. (2017), plant oils have become increasingly popular due to their high proportion of healthy polyunsaturated fatty acids, which the human body is often unable to efficiently synthetize at sufficient levels. Although seeds of Moringa plants are not widely used for extraction, processing or marketing of edible oils, these materials are rich in monounsaturated fatty acids (such as oleic acid) and have high oxidative stability compared to materials containing polyunsaturated fatty acids (Sánchez-Machado et al.,
ACCEPTED MANUSCRIPT 2015), which are promising properties for used in processes related to food making. As noted by Omosuli, Oloye, and Ibrahim (2017), high-oleic-acid vegetable oils, such as Moringa oil, are very stable even in highly demanding applications such as frying because these oils produce less conjugated dienes and trienes during such processes than do
T
polyunsaturated fatty acid-rich oils (Abdulkarim, Long, Lai, Muhammad, & Ghazali,
IP
2007). In addition, M. oleifera seed oil had appropriate physicochemical properties for
CR
commercial use as an edible oil. These results agree with those of Manzoor et al. (2007), who reported that M. concanensis has potential for edible applications and, due to its fatty
US
acid content, could be used to develop nutritionally balanced and highly stable formulations
AN
for the food industry. As previously mentioned, Moringa plants have great potential for use in a wide variety of food products depending on the bioactive compounds and the target
ED
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foodstuff.
5. Food applications of Moringa plants and intellectual protection
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Although several efforts have been made to evaluate the functional properties of Moringa
CE
plants and their bioactive compounds, researchers do not refer to any patent in their published investigations related to these plants (Dou & Kister, 2016). This suggests that
AC
research is conducted with the purpose of simply generating new knowledge and basic investigation because intellectual property concerning the development of food products from Moringa plants and their bioactive compounds is not reflected. According to searches made in the databases WIPO-PATENTSCOPE (World Intellectual Property Organization) and Google Patents (accessed January 12 th , 2018) using the words “Moringa” and “functional foods”, 28 patents were requested around the world in the last 3 years (between 2015 and 2017) related to the main use of Moringa plants for functional
ACCEPTED MANUSCRIPT purposes. Meanwhile, when the words “Moringa” and “food applications” were used, fewer patent records were detected. Of these patents, four corresponded to the same invention describing a method for the preparation of functional extracts from Moringa plants but were registered in different countries. Two more described polymer coatings based on Moringa
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and neem oil blends to prevent the establishment and proliferation of microorganisms on
IP
cellulosic-rich materials and were similarly registered in different countries. Finally, two
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more were related to the use of Moringa extracts with antioxidant activities for the formulation of medicine and health-care food but did not yet include incorporation into
US
food products. Table 4 summarizes the patent records associated with the application of
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Moringa plants for the formulation of several food products. In most cases, the vegetative structures of Moringa plants are used in the form of flour or dehydrated powder and, in
M
some cases, are fermented to obtain the target product. However, a specific characterization
ED
or evaluation of bioactive molecules is not mentioned in any patent. In addition, the conditions of preparation, such as drying, humidity, number of steps, concentration of
PT
ingredients, and pH, are not extensively described. Notwithstanding, the benefits to human
CE
health (decreased blood pressure, sleep promotion, mood regulation, stress relief, enhanced immunity, antitumor effects, and others) are reported without indicating evidence or
AC
experimental procedures.
6. Analytical methods for bioactive compound qualification Several recent studies have shown great amounts of bioactive compounds with broad biological activities, such as phenolic compounds, carbohydrates, oils, and fatty acids, as well as proteins and bioactive peptides. Researchers have the opportunity to apply new analytical techniques to facilitate the proper separation and identification of the bioactive
ACCEPTED MANUSCRIPT compounds present in several food samples, including plant materials such as Moringa plants. There are a variety of techniques related to gas and liquid chromatography as well as infrared and mass spectrometry for the adequate separation and identification of different classes of bioactive compounds; these techniques use sophisticated analytical instruments
T
for appropriate identification of such compounds.
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FT-IR spectroscopy is a technique used to investigate the composition of diverse food
CR
samples. In a study of carbohydrates present in Moringa plants, the FT-IR spectra displayed typical abortion peaks given by other polysaccharides, which were detected for some
US
functional groups in the successful characterization of gum and polysaccharides from stems
AN
and leaves (Chen et al., 2017; Abhishek & Ahuja, 2017). This result agrees with Baptista et al. (2017), who reported the presence of various functional groups, including amide groups,
M
for the characterization of protein fractions such as albumin and globulin with coagulant
ED
activity from M. oleifera seeds. Moreover, the infrared spectra of Moringa oil (M. peregrina) are similar to those reported for olive, canola and chia oils, with a predominant
PT
peak in the low-wavenumber region (at 1742 cm-1 ) due to C=O carbonyl stretching of ester
CE
functional groups present in lipids and fatty acids. In addition, the intensity of the peak observed in the region of 3004-3010 cm-1 was lower for Moringa oil than for canola and
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chia oils, which can be explained by the low polyunsaturated fatty acid contents in Moringa oil (Salaheldeen et al., 2015; Timilsena et al., 2017). FT-IR spectroscopy has some technical advantages over other techniques used for the identification of bioactive molecules because it is a nondestructive and fast method that requires minimal sample preparation (Singh, Andola, Rawat, Pant, & Purohit, 2011). Thus, this technique could be considered a suitable alternative for integral characterization of the bioactive compounds from Moringa plants and derived food products.
ACCEPTED MANUSCRIPT Currently, liquid chromatography coupled with mass spectrophotometry (LC-MS) has been successfully applied for analysis of several plant materials, including Moringa plants and derived products, to characterize a wide number of natural compounds, such as alcohols, nucleosides, phenolics, organic acids, and amino acids (Amaglo et al., 2010; Rodríguez-
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Pérez et al., 2015; Makita et al., 2016; Nouman et al., 2016; Devisetti, Sreerama, &
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Bhattacharya, 2016; Makita et al., 2017). The best published results for M. oleifera leaf
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extract characterization have recorded a total of 59 chemical compounds, including amino and organic acids, nucleosides, glucosinolates, lignans, phenolic acids, and flavonoids, as
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the main bioactive compounds in these extracts (Rodríguez-Pérez et al., 2015). High-
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performance liquid chromatography coupled with electrospray ionization quadrupole-time of flight mass spectrometry (HPLC–ESI–QTOF–MS) allows the identification of chemical
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molecules by interpreting their MS and MS/MS spectra determined by QTOF–MS
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compared to scientific literature and open-access mass-spectra databases. These data agree with the published qualitative results, which indicate that this technique can be applied to
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identify at least 50 bioactive compounds, such as organic acids, nucleosides glucosinolates,
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and phenolic compounds, from M. oleifera leaf optimized extracts (Rodríguez-Pérez et al., 2016). In addition, ultra-HPLC coupled with QTOF–MS was applied in a comparative
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analysis of the flavonoid contents of M. oleifera and M. ovalifolia, and the results revealed that these species contained at least 17 glycosylated flavonoids between them (Makita et al., 2016). These findings were comparable to those recently reported by Castro-López et al. (2017), who reported approximately 19 glycosylated flavonoids, among other phenolics, in extracts obtained from M. oleifera leaves using UPLC-ESI-QTOF-MS2 for the identification of these bioactive molecules. Because they provide good levels of peak asymmetry,
efficiency,
and chemical stability, C18 columns could be considerably
ACCEPTED MANUSCRIPT versatile,
high-performance
separation
columns
suitable
for
a
diverse
range
of
phytochemicals. In most cases reviewed, C18 reversed-phase columns have been successfully applied to separate several phenolics and other polar compounds from Moringa plants. In addition, BEH (ethylene-bridged hybrid) C8 and phenyl columns have
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shown positive effects in the separation of phenolic compounds. Moreover, acidic solutions
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(communally formic acid at 0.1 or 0.5%) such as solvent A and acetonitrile (solvent B) at
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different gradients are the most used mobile phases in these studies. As aforementioned, the best results for phytochemical characterization from Moringa plants elucidated 59
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molecules using the following items: Zorbax Eclipse Plus C18 column (1.8 m, 150 mm ×
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4.6 mm) (Agilent Technologies, Palo Alto, CA, USA), acidified water (0.5% formic acid, v/v), and acetonitrile. The injection volume in the HPLC was 10 l. The gradient was
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programmed as follows: 0 min, 5% B; 10 min, 35% B; 65 min, 95% B; 67 min, 5% B; and
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finally, a conditioning cycle of 3 min under the initial conditions. The flow rate was set at
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0.50 ml/min throughout the gradient, and the injection volume was 10 l (Rodríguez-Pérez et al., 2015). Furthermore, full-screen mass spectra detection is usually carried out in
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negative ion mode in a mass range m/z of 50–1500 Da. However, a full scan in positive and negative ion modes is recommended for quantification and identification of glucosinolates,
AC
phenolics, flavonoids, and various other classes of phytochemicals from Moringa plants (Amago et al. 2010). Therefore, LC-MS is an attractive alternative for the identification of several bioactive compounds because this technique could permit discrimination between isomeric compounds depending on the parameters used (Makita et al., 2017). Fatty
acids
from Moringa
plants
after
derivation
to
methyl esters
through
a
transesterification reaction with methanol have been well characterized. For this purpose,
ACCEPTED MANUSCRIPT gas chromatography (GC) equipped with a flame-ionized detector may be considered the most applied analytical tool for the identification and quantification of different fatty acids present in oils and crude fat from different parts of Moringa plants (Amaglo et al., 2010; Nkukwana et al. 2014; Salaheldeen et al., 2015; Sánchez-Machado et al., 2015; Bhutada et
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al., 2016). However, according to Al_Owaisi, Al-Hadiwi, and Khan (2014), the use of GC-
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MS allows the identification of 19, 7 and 6 different bioactive molecules from hexane,
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methanol and ethyl acetate extracts, respectively, after ethanoic pre-extraction of M. peregrina leaves. Furthermore, the use of carbon dioxide-expanded ethanol extracts and
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GC-MS permits the identification of 20 compounds, including ketones and esters
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(Rodríguez-Pérez et al., 2016). Thus, LC/MS and GC/MS have become very suitable techniques for identifying bioactive molecules from Moringa plants.
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However, colorimetric methods are still widely applied by researchers for the quantification
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of bioactive compounds from plant materials, including Moringa plants. Although FolinCiocalteu is not a specific method for phenolic determination (Everette et al., 2010), due to
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versatility and quickness, which permit analysis of a large number of samples, this
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methodology continues to be accepted for scientists to express total phenolic contents in these samples (Al_Owaisi, Al-Hadiwi, & Khan, 2014; Shah, Bosco, & Mir, 2015; Radha,
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Ogunsina, & Hebina-Babu, 2015; Rodríguez-Pérez et al., 2016; Juhaimi et al., 2017). Nevertheless, the presence of other reductive nonphenolic compounds with an aromatic ring (e.g., vitamin C and peptides) can produce many interferences in this analysis; therefore, a previous purification step is recommended (Huang, Ou, & Prior, 2005). To determine protein concentration in Moringa plants, Bradford and Lowry assays and Kjeldahl digestion are the most applied methodologies (Table 1) (Pontual et al., 2012; González-Garza et al., 2017; Baptista et al., 2017). Despite being the most used methods,
ACCEPTED MANUSCRIPT Bradford and Lowry assays have some disadvantages, including the quantification of proteins with specific amino acids (tyrosine and tryptophan) involved in color development and limited use in food samples (Sapan, Lundblad, & Price, 1999). In addition, polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE)
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is a technique used to estimate the molecular weight of proteins and peptides isolated from
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Moringa tissues (Pontual et al., 2012; Ullah et al., 2015; González-Garza et al., 2017).
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Nevertheless, HPLC shows good results for molar mass distribution through size exclusion in the characterization protein fractions (e.g., globulins, albumin, prolamin, and glutelin)
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from Moringa seeds (Baptista et al., 2017). Hence, qualification of bioactive compounds
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from Moringa plants could be well characterized with the above-described techniques.
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7. Authors viewpoint
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Taken together, these results indicate that bioactive compounds in Moringa plants are a diverse group of molecules that have proven to be good agents against several health
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disorders, such as oxidative stress, hypertension, diabetes, hyperlipidemia, and cancer.
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Although numerous studies have been conducted on M. oleifera bioactive compounds, there is not enough information available on the phytochemicals isolated from other species
AC
of the Moringaceae family, which could be considered prospects with promising applications in food products. Considering the vast potential for food application of this family of plants, researchers should focus more on the effect of processing and storing Moringa-based foodstuff on the content and stability of bioactive compounds to gain a better understanding of the role of these compounds in the food matrix and food quality. In addition, more research should also focus on the methods and conditions for structural elucidation of bioactive compounds to identify and isolate new natural bioactive agents
ACCEPTED MANUSCRIPT from Moringa plants, as this could help to address the adverse effects associated with synthetic food
additives.
Although several studies have investigated the functional
properties of the bioactive compounds reviewed above, additional investigation related to digestibility
and
bioavailability
is
needed
to
determine the functionality of these
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compounds in both in vitro and in vivo systems. In addition, the functional properties
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measured in many of the studies summarized in this review require further validation in all
CR
Moringa species. On the other hand, despite all presented information, clinical trials that confirm the effects of Moringa components, food safety, and legal framework about
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commercialization are still pendent and were not included because of the orientation of this
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review.
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8. Concluding remarks
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This review aimed to highlight the bioactive compounds in Moringa plants and the recent approaches regarding the functional applications and influence of these biocompounds on
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functional characteristics in food products. It was observed that the major food products
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based on Moringa plants presented high dietary fiber and low fat contents, which suggests that this plant can be used in the formulation of hypocaloric foodstuffs. Thus, it is possible
AC
to develop food products based on Moringa flour with acceptable sensory and nutritional properties when less than 20 g of these material is used, depending on the target foodstuff. Although emerging technologies have been shown to be suitable alternatives for recovering and characterizing bioactive compounds from Moringa plants, conventional methodologies remain valid. Finally, regardless of several promising functional applications in food products, the exploitation of these properties within the industrial field are still scarcely explored.
ACCEPTED MANUSCRIPT Conflicts of interest The authors declare that there are no conflicts of interest
Acknowledgments
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Saucedo-Pompa thanks Jesús Noel Yáñez Reyes and Fitokimica Industrial de México S.A.
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de C.V. for the facilities given to study his doctorate. Thanks to the Sheltered Agriculture
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Center of the UANL for the photos used in Figure 1.
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References
AN
Abdulkadir, A. R., Zawawi, D. D., & Jahan, M. S. (2016). Proximate and phytochemical screening of different parts of Moringa oleifera. Russian Agricultural Sciences, 42(1), 34–
M
36.
ED
Abhishek, R., & Ahuja, M. (2017). Evaluation of carboxymethyl Moringa gum as nanometric carrier. Carbohydrate Polymers, 174, 896–903.
PT
Agrawal, H., Shee, Ch., & Sharma, A. K. (2007). Isolation of a 66 kDa protein with
CE
coagulation activity from seeds of Moringa oleifera. Research Journal of Agriculture and Biological Sciences, 3(5), 418–421.
AC
Al_husnan, L. A., & Alkahtani, M. D. F. (2016). Impact of Moringa aqueous extract on pathogenic bacteria and fungi in vitro. Annals of Agricultural Science, 61(2), 247–250. Al-Owaisi, M., Al-Hadiwi, N., & Khan, S. A. (2014). GC-MS analysis, determination of total phenolics, flavonoid content and free radical scavenging activities of various crude extracts of Moringa peregrina (Forssk.) Fiori leaves. Asian Pacific Journal of Tropical Biomedicine, 4(12), 964–970.
ACCEPTED MANUSCRIPT Amaglo, N. K., Bennett, R. N., Lo Curto, R. B., Rosa, E. A. S., Lo Turco, V., Giuffrida, A., Lo Curto, A., Crea, F., & Timpo, G. M. (2010). Profiling selected phytochemicals and nutrients in different tissues of the multipurpose tree Moringa oleifera L., grown in Ghana. Food Chemistry, 122(1), 1047–1054.
T
Anudeep, A., Prasanna, V. K., Adya, S. M., & Radha, Ch. (2016). Characterization of
IP
soluble dietary fiber from Moringa oleifera seeds and its immunomodulatory effects.
CR
International Journal of Biological Macromolecules, 91, 656–662.
Anwar, F., Latif, S., Ashraf, M., & Gilani, A. H. (2005). Moringa oleifera: A food plant
D.
S.,
& Onsare, J. G. (2014). In vitro antimicrobial evaluation and
AN
Arora,
US
with multiple medicinal uses. Phytotherapy Research, 21(1),17–25.
phytoconstituents of Moringa oleifera pod husks. Industrial Crops and Products, 52, 125–
M
135.
ED
Asare, G. A., Gyan, B., Bugyei, K., Adjei, S., Mahama, R., Addo, P., Otu-Nyarko, L., Wiredu, E. K., & Nyarko, A. (2012). Toxicity potentials of the nutraceutical Moringa
PT
oleifera at supra-supplementation levels. Journal of Ethnopharmacology, 139(1), 265–272.
CE
Azmir, J., Zaidul, I. S. M., Rahman, M. M., Sharif K.M., Mohamed, A., Sahena, F., Jahurul, M. H. A., Ghafoor, K., Norulaini N.A.N., & Omar, A. K. M. (2013). Techniques
AC
for extraction of bioactive compounds from plant materials: A review. Journal of Food Engineering, 117(4), 426–436. Balamurugan,
V.,
&
Balakrishnan,
V.
(2013).
Evaluation
of
phytochemical,
Pharmacognostical and antimicrobial activity from the bark of Moringa concanensis Nimmo. International Journal of Current Microbiology and Applied Sciences, 2(4), 117– 125.
ACCEPTED MANUSCRIPT Baptista, A. T. A., Silva, M. O., Gomes, R. G., Bergamasco, R., Vieira, M. F., & Vieira, A. M. S. (2017). Protein fractionation of seeds of Moringa oleifera lam and its application in superficial water treatment. Separation and Purification Technology, 180, 114–124. Bhutada, P. R., Jadhav, A. J., Pinjari, D. V., Nemade, P. R., & Jainb, R. D. (2016). Solvent
T
assisted extraction of oil from Moringa oleifera Lam. seeds. Industrial Crops and Products,
IP
82, 74–80.
CR
Bichi, M. H. (2013). A review of the applications of Moringa oleifera seeds extract in water treatment. Civil and Environmental Research, 3(8), 1–10.
US
Bijina, B., Chellappan, S., Krishna, J. G., Basheer, S. M., Elyas, K. K., Bahkali, A. H., &
AN
Chandrasekaran, M. (2011). Protease inhibitor from Moringa oleifera with potential for use as therapeutic drug and as seafood preservative. Saudi journal of biological sciences, 18(3),
M
273–281.
ED
Bolarinwa, I. F., Aruna, T. E., & Raji, A. O. (2017). Nutritive value and acceptability of bread fortified with moringa seed powder. Journal of the Saudi Society of Agricultural
PT
Sciences, https://doi.org/10.1016/j.jssas.2017.05.002
CE
Castro-López, C., Rojas, R., Sánchez-Alejo, E. J., Niño-Medina, G., & Martínez-Ávila, G. C. G. (2016). Phenolic compounds recovery from grape fruit and by- products: An
AC
Overview of Extraction Methods. In: Grape and Wine Biotechnology (pp. 103–123). Morata, A., & Loira, I., (Eds.), Intech, Croatia. Castro-López, C., Ventura-Sobrevilla, J. M., González-Hernández, M. D., Rojas, R., Ascacio-Valdés, J. A. Aguilar, C. N., & Martínez-Ávila, G.C.G. (2017). Impact of extraction
techniques
on
antioxidant
capacities
and
polyphenol-rich extracts. Food Chemistry, 237, 1139–1148.
phytochemical composition of
ACCEPTED MANUSCRIPT Chen, Ch., Zhang, B., Huang, Q., Fu, X., & Liu, R. H. (2017). Microwave-assisted extraction of polysaccharides from Moringa oleifera Lam. leaves: Characterization and hypoglycemic activity. Industrial Crops and Products, 100, 1–11. Cuellar-Nuñez M.L., Luzardo-Ocampo, I., Campos-Vega, R., Gallegos-Corona, M.A.,
T
González de Mejía, E., & Loarca-Piña, G. (2018). Physicochemical and nutraceutical
IP
properties of Moringa (Moringa oleifera) leaves and their effects in an in vivo AOM/DSS-
CR
induced colorectal carcinogenesis model. Food Research International, 105, 159–168. Dachana, K. B., Rajiv, J., Indrani, D., & Prakashi, J. (2010). Effect of dried Moringa
US
(Moringa oleifera Lam) leaves on rheological, microstructural, nutritional, textural and
AN
organoleptic characteristics of cookies. Journal of Food Quality, 33(5), 660–677. Dehshahri, S., Wink, M., Afsharypuor, S., Asghari, G., & Mohagheghzadeh, A. (2012).
M
Antioxidant activity of methanolic leaf extract of Moringa peregrina (Forssk.) Fiori.
ED
Research in Pharmaceutical Sciences, 7(2), 111–118. Devisetti, R., Sreerama, Y. N., & Bhattacharya, S. (2016). Processing effects on bioactive
PT
components and functional properties of Moringa leaves: development of a snack and
CE
quality evaluation. Journal of Food Science and Technology, 53(1), 647–657. Dou, H., & Kister, J. (2016). Research and development on Moringa Oleifera e
AC
Comparison between academic research and patents. World Patent Information, 47, 21–33. Everette, J. D., Bayart, Q. M., Green, A. M., Abbey, I. A., Wangila, G. W., & Walker, R. (2010). Thorough study of reactivity of various compound classes toward the FolinCiocalteu reagent. Journal of Agricultural and Food Chemistry, 58(14), 8139–8144. Fakayode, O. A., & Ajav, E. A. (2016). Process optimization of mechanical oil expression from Moringa (Moringa oleifera) seeds. Industrial Crops and Products, 90(1), 142–151.
ACCEPTED MANUSCRIPT Farzana, T., Mohajan, S., Saha, T., Hossain, M. N., & Haque, M. Z. (2017). Formulation and nutritional evaluation of a healthy vegetable soup powder supplemented with soy flour, mushroom, and Moringa leaf. Food Science & Nutrition, 5(4), 911–920. Gifoni, J. M., Oliveira, J. T., Oliveira, H. D., Batista, A. B., Pereira, M. L., Gomes, A. S.,
T
Oliveira, H. P., Grangeiro, T. B., & Vasconcelos, I. M. (2012). A novel chitin-binding
IP
protein from Moringa oleifera seed with potential for plant disease control. Peptide
CR
Science, 98(4), 406–415.
González-Garza, N. G., Chuc-Koyoc, J. A., Torres-Castillo, J. A., García-Zambrano, E. A.,
US
Betancur-Ancona, D., Chel-Guerrero, L., & Sinagawa-García, S. R. (2017). Biofunctional
AN
properties of bioactive peptide fractions from protein isolates of Moringa seed (Moringa oleifera). Journal of Food Science and Technology, 54(13), 4268–4276.
M
Google patents https://patents.google.com/. Accessed January 12th 2018
ED
Gopalakrishnan, L., Doriya, K., & Kumar, D. S. (2016). Moringa oleifera: A review on nutritive importance and its medicinal application. Food Science and Human Wellness,
PT
5(2), 49–56.
CE
Hernández-Almanza, A., Montañez, J., Martínez, G., Aguilar-Jiménez, A., ContrerasEsquivel, J. C., & Aguilar, C. N. (2016). Lycopene: Progress in microbial production.
AC
Trends in Food Science & Technology, 56, 142–148. Hosseinpour, S., Aghbashlo, M., Tabatabaei, M., & Khalife, E. (2016). Exact estimation of biodiesel cetane number (CN) from its fatty acid methyl esters (FAMEs) profile using partial least square (PLS) adapted by artificial neural network (ANN). Energy Conversion and Management, 124, 389–398.
ACCEPTED MANUSCRIPT Ijarotimi, O. S., Adeoti, O. A., & Ariyo, O. (2017). Comparative study on nutrient composition,
phytochemical,
and
functional characteristics of raw,
germinated, and
fermented Moringa oleifera seed flour. Food Science & Nutrition, 1(6), 452-–463. Jarald, E. E., Sumati, S., Edwin, S., Ahmad, S., Patni, S., & Daud, A. (2012).
T
Characterization of Moringa oleifera Lam. Gum to Establish it as a Pharmaceutical
IP
Excipient. Indian Journal of Pharmaceutical Education and Research, 46(3), 211–216.
CR
Jayawardana, B. C., Liyanage, R., Lalantha, N., Iddamalgoda, S., & Weththasinghe, P. (2015). Antioxidant and antimicrobial activity of drumstick (Moringa oleifera) leaves in
US
herbal chicken sausages. LWT - Food Science and Technology, 64, 1204–1208.
AN
Juhaimi, F. AL., Ghafoor, K., Mohamed-Ahmed, I. A., Babiker, E. E., & M. M. Özcan. (2017). Comparative study of mineral and oxidative status of Sonchus oleraceus, Moringa and
Moringa
peregrina
Journal
of
Food
Measurement
and
ED
Characterization, 11(4), 1745–1751.
leaves.
M
oleifera
Kar, M., & Mishra, D. (1975). Inorganic pyrophosphatase activity during rice leaf
PT
senescence. Canadian Journal of Botany, 53, 503–511.
CE
Kharazi, S., H., Kenari, R. E., Amiri, Z. R., & Azizkhani, M. (2012). Characterization of iranian virgin olive oil from the roodbar region: A study on zard, mari and phishomi.
AC
Journal of the American Oil Chemists' Society, 89, 1241–1247. Khatun, S., Absar, N., & Ashraduzzaman, M. (2001). Purification, characterization and effect of physico-chemical agents on stability of polyphenoloxidase from sajna (Moringa oleifera L.) Leaves at Mature Stage. Pakistan Journal of Biological Sciences, 4(9), 1129– 1132.
ACCEPTED MANUSCRIPT Khatun, S., Absar, N., & Ashraduzzaman, M. (2003). Changes in physico-compositions and activities of some hydrolytic and oxidative enzymes in the two types of sajna (Moringa oleifera lam) leaves at different maturity levels. Indian Journal of Plant Physiolog, 8, 6–11. Khatun, S., Ashraduzzaman, M., Karim, M. R., Pervin, F., Absar, N., & Rosma, A. (2012). characterization of peroxidase from Moringa
oleifera L. leaves.
T
Purification and
IP
BioResources, 7(3), 3237–3251.
CR
Kurt, A., Cengiz, A., & Kahyaoglu, T. (2016). The effect of gum tragacanth on the rheological properties of salep based ice cream mix. Carbohydrate Polymers, 143, 116–
US
123.
AN
Ma, L., Chen, H., Zhu, W., & Wang, Z. (2013). Effect of different drying methods on physicochemical properties and antioxidant activities of polysaccharides extracted from
M
mushroom Inonotus obliquus. Food Research International, 50, 633–640
ED
Makita, C., Chimuka, L., Cukrowska, E., Steenkamp, P. A., Kandawa-Schutz, M., Ndhlala, A. R., & Madala, N. E. (2017). UPLC-qTOF-MS profiling of pharmacologically important
PT
chlorogenic acids and associated glycosides in Moringa ovalifolia leaf extracts. South
CE
African Journal of Botany, 108, 193–199. Makita, Ch., Chimuka, L., Steenkamp, P., Cukrowska, E., & Madala, E. (2016).
AC
Comparative analyses of flavonoid content in Moringa oleifera and Moringa ovalifolia with the aid of UHPLC-qTOF-MS fingerprinting. South African Journal of Botany, 105, 116–122. Makkar, H. P. S., & Becker K. (1997). Nutrients and antiquality factors in different morphological parts of the Moringa oleifera tree. Journal of Agricultural Science, 128(3), 311–322.
ACCEPTED MANUSCRIPT Mamta, Dunkwal, V., Goyal, M., & Monika. (2017). Organoleptic acceptability of value added products using drumstick leaves powder. Journal of Pharmacognosy and Phytochemistry, 6(4), 1060–1063. Manaois, R. V. Morales, A. V., & Abilgos-Ramos, R. G. (2013). Acceptability, shelf life
T
and nutritional quality of moringa-supplemented rice crackers. Philippine Journal of Crop
IP
Science, 38(2), 1–8.
CR
Mansurah, A. A., Chioma, N. T., Ismaila, M., Abdulmalik, S. A., Williams, C., & Wudil, A. M. (2015). Partial-purification and characterization of angiotensin converting enzyme
AN
Biochemistry Research & Review, 5(1), 39–48.
US
inhibitory proteins from the leaves and seeds of Moringa oleifera. International Journal of
Manzoor, M., Anwar F., Iqbal, T., & Bhanger, M. I. (2007). Physico-chemical
ED
Chemists' Society, 84(5), 413–419.
M
characterization of Moringa concanensis seeds and seed oil. Journal of the American Oil
Marwah, R. G., Fatope, M. O., Mahrooqi, R. A., Varma, G. B. Abadi, A. A., & Al-
PT
Burtamani, S. K. S. (2007). Antioxidant capacity of some edible and wound healing plants
CE
in Oman. Food Chemistry, 101(2), 465–470. Mat-Yusoff, M., Gordon, M. H., Ezeh, O., & Niranjan, K. (2016). Aqueous enzymatic
AC
extraction of Moringa oleifera oil. Food Chemistry, 211, 400–408. Meloni, D. A., Oliva, M. A., Martinez, C. A., & Cambraia, J. (2003). Photosynthesis and activity of superoxide dismutase, peroxidase and glutathione reductase in cotton under salt stress. Environmental and Experimental Botany, 49, 69–76. Moyo, B., Oyedemi, S., Masika, P. J., & Muchenje, V. (2012). Polyphenolic content and antioxidant properties of Moringa oleifera leaf extracts and enzymatic activity of liver from
ACCEPTED MANUSCRIPT goats supplemented with Moringa oleifera leaves/sunflower seed cake. Meat Science, 91(4), 441–447. Muhammad, H. I., Asmawi, M. Z., & Khan, N. A. K. (2016). A review on promising phytochemical, nutritional and glycemic control studies on Moringa oleifera Lam. in
T
tropical and sub-tropical regions. Asian Pacific Journal of Tropical Biomedicine, 6(10),
IP
896–902.
CR
Nkukwana, T. T., Muchenje, T. T., Masika, V., Hoffman, P. J., Dzama, L. C. K., & Descalzo, A. M. (2014). Fatty acid composition and oxidative stability of breast meat from
AN
refrigeration. Food Chemistry, 142, 255–261.
US
broiler chickens supplemented with Moringa oleifera leaf meal over a period of
Nouman, W., Anwar, F., Gull, T., Newton, A., Rosa, E., & Domínguez-Perles, R. (2016).
M
Profiling of polyphenolics, nutrients and antioxidant potential of germplasm’s leaves from
ED
seven cultivars of Moringa oleifera Lam. Industrial Crops and Products, 83, 166–176. Nwakalor, Ch. N. (2014). Sensory evaluation of cookies produced from different blends of
PT
wheat and Moringa oleifera leaf flour. International Journal of Nutrition and Food
CE
Sciences, 3(4), 307–310.
Ogunsina, B. S., Radha, C., & Indrani, D. (2011). Quality characteristics of bread and
AC
cookies enriched with debittered Moringa oleifera seed flour. International Journal of Food Sciences and Nutrition, 62(2), 185–194. Omosuli, S. V., Oloye, D. A., & Ibrahim, T. A. (2017). Effect of drying methods on the physicochemical properties and fatty acid composition of moringa seeds oil. Progress in Food & Nutrition Science, 1, 027–032. Oyeyinka, A. T., & Oyeyinka, S. A. (2018). Moringa oleifera as a food fortificant: Recent trends and prospects. Journal of the Saudi Society of Agricultural Sciences, 17, 127–137.
ACCEPTED MANUSCRIPT PATENTSCOPE-WIPO (World Intellectual Property Organization). https://patentscope.wipo.int/search/es/result.jsf. Accessed January 12th 2018 Pereira, M. L., David de Oliveira, H., Tadeu Abreu de Oliveira, J., Menezes Gifoni, J., de Oliveira Rocha, R., de Oliveira Bezerra de Sousa, D., & Vasconcelos, I. M. (2011).
T
Purification of a chitin-binding protein from Moringa oleifera seeds with potential to
IP
relieve pain and inflammation. Protein and peptide letters, 18(11), 1078–1085.
CR
Pinto, C. E. M., Farias, D. F., Carvalho, A. F. U., Oliveira, J. T. A., Pereira, M. L., Grangeiro, T. B., Freire, J. E. C., Viana, D. A., & Vasconcelos, I. M. (2015). Food safety
US
assessment of an antifungal protein from Moringa oleifera seeds in an agricultural
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biotechnology perspective. Food and Chemical Toxicology, 83, 1–9. Pontual, E. V., Carvalho, B. E., Bezerra, R. S., Coelho, L. C., Napoleão, T. H., & Paiva, P.
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Chemistry, 135(3), 1848–1854.
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M. (2012). Caseinolytic and milk-clotting activities from Moringa oleifera flowers. Food
Radha, C., Ogunsina, B. S., & Hebina-Babu, K. T. (2015). Some quality and micro-
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structural characteristics of soup enriched with debittered Moringa oleifera seeds flour.
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American Journal of Food Science and Technology, 3(5), 145–149. Ravi-Varma, R. N., Kumar, Ch. P., Reddy, A. K., & Raju, P. P. (2014). Evaluation of
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Moringa oleifera gum as a sustained release polymer in diclofenac sodium tablet formulation. International Journal of Research in Pharmacy and Chemistry, 4(3), 687–693. Rodríguez-Pérez, C., Mendiola, J. A., Quirantes-Piné, R., Ibáñez E., & Segura-Carretero A. (2016). Green downstream processing using supercritical carbon dioxide, CO 2 -expanded ethanol and pressurized hot water extractions for recovering bioactive compounds from Moringa oleifera leaves. Journal of Supercritical Fluids, 116, 90–100.
ACCEPTED MANUSCRIPT Rodríguez-Pérez, C., Quirantes-Piné, R., Fernández-Gutiérreza, A., & Segura-Carretero, A. (2015). Optimization of extraction method to obtain a phenolic compounds-rich extract from Moringa oleifera Lam leaves. Industrial Crops and Products, 66, 246–254. Rozina., Asif, S., Ahmad, M., Zafar, M., & Ali, N. (2017). Prospects and potential of fatty
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acid methyl esters of some non-edible seed oils for use as biodiesel in Pakistan. Renewable
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and Sustainable Energy Reviews, 74, 687–702.
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Salaheldeen, M., Aroua, M. K., Mariod, A. A., Chenge, S. F., Abdelrahman, M. A., & Atabani, A. E. (2015). Physicochemical characterization and thermal behavior of biodiesel
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Conversion and Management, 92, 535–542.
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and biodiesel–diesel blends derived from crude Moringa peregrina seed oil. Energy
Salaheldeen, M., Arouaa, M. K., Mariod, A. A., Chenge, S. F., & Abdelrahmanb, A. M.
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Crops and Products, 61, 49–61.
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(2014). An evaluation of Moringa peregrina seeds as a source for bio-fuel. Industrial
Sánchez-Machado, D. I., López-Cervantes, J., Núñez-Gastélum, J. A., Mora-López, G. S.,
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López-Hernández, J., & Paseiro-Losada, P. (2015). Effect of the refining process on
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Moringa oleifera seed oil quality. Food Chemistry, 187, 53–57. Santos, A. F., Luz, L. A., Argolo, A. C., Teixeira, J. A., Paiva, P. M., & Coelho, L. C.
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(2009). Isolation of a seed coagulant Moringa oleifera lectin. Process biochemistry, 44(4), 504–508.
Sapan, C. V., Lundblad, R. L., & Price, N. C. (1999). Colorimetric protein assay techniques. Biotechnology and Applied Biochemistry, 29, 99–108. Satish, A., Sairam, S., Ahmed, F., & Urooj, A. (2012). Moringa oleifera Lam.: Protease activity against blood coagulation cascade. Pharmacognosy Research, 4(1), 44–9.
ACCEPTED MANUSCRIPT Sengev, A. I., Abu. J. O., & Gernah, D. I. (2013). Effect of Moringa oleifera leaf powder supplementation on some quality characteristics of wheat bread. Food and Nutrition Sciences, 4, 270–275. Shah, M. A., Bosco, S. J. D., & Mir, S. A. (2015). Effect of Moringa oleifera leaf extract
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on the physicochemical properties of modified atmosphere packaged raw beef. Food
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Packaging and Shelf Life, 3, 31–38.
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Shank, L. P., Riyathong, T., Lee, V. S., & Dheeranupattana, S. (2013). Peroxidase activity in native and callus culture of Moringa oleifera Lam. Journal of Medical and
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Bioengineering, 2(3), 163–167.
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Sravanthi, J., & Rao, S. G. (2014). Antioxidative studies in Moringa oleifera Lam. Annals of Phytomedicine, 3(2), 101–105.
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Tesfay, S. Z., Magwaza, L. S., Mbili, N., & Mditshwa, A. (2017). Carboxyl
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methylcellulose (CMC) containing Moringa plant extracts as new postharvest organic edible coating for Avocado (Persea americana Mill.) fruit. Scientia Horticulturae, 226,
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201–207.
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Timilsena, Y. P., Vongsvivut, J., Adhikari, R., & Adhikari, V. (2017). Physicochemical and thermal characteristics of Australian chia seed oil. Food Chemistry, 228, 394–402.
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Torres-Castillo, J. A., Sinagawa-García, S. R., Martínez-Ávila, G. C. G., López-Flores, A. B., Sánchez-González, E. I., Aguirre-Arzola, E. V., Torres-Acosta, R. I., Olivares-Sáenz, E., Osorio-Hernández, E., & Gutiérrez-Díez, A. (2013). Moringa oleifera: phytochemical detection, antioxidants, enzymes and antifugal properties. Phyton, 82, 193–202. Ullah, A., Mariutti, R. B., Masood, R., Caruso, I. P., Costa, G. H. G., de Freita, C. M., Santos, C. R., Zanphorlin, L. M., Mutton, M. J. R., Murakami, M. T., & Arni, R. K. (2015).
ACCEPTED MANUSCRIPT Crystal structure of mature 2S albumin from Moringa oleifera seeds. Biochemical and Biophysical Research Communications, 468(1-2), 365–371. Vaknin, Y., & Mishal, A. (2017). The potential of the tropical “miracle tree” Moringa oleifera and its desert relative Moringa peregrina as edible seed-oil and protein crops under
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Mediterranean conditions. Scientia Horticulturae, 225, 431–437.
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Vanajakshi, V., Vijayendra, S. V. N., Varadaraj, M. C., Venkateswaran, G., & Agrawal, R.
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(2015). Optimization of a probiotic beverage based on Moringa leaves and beetroot. LWT-
ACCEPTED MANUSCRIPT Table 1. Bioactive compounds from different vegetative structures of Moringa plants, extraction methods and registered yields Moringa species and tissue
Yield
Extraction method
Functional activity/application
S olvent
Reference
Phenolic compounds
M. oleifera stems, leaves and roots M. oleifera leaves
NI
Maceration extraction
Antioxidant activity
Water
T orresCastillo et al. (2013)
5.30 to 47 mg GAE/g dw depending on the solvent mixture 20.3 to 62.4 GAE/g dw depending on the analyzed fraction NI
Ultrasoundassisted extraction
Potential for antioxidant and antiinflammatory properties
Ethanol, Methanol and Acetone
RodríguezPérez et al. (2015)
Supercritical fluids (green and sustainable method)
Antioxidant activity
Sonication
NI
M. oleifera leaves
6.5 g/100 g dw
M. oleifera seeds
10 mg/ml extract
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M. oleifera seeds M. oleifera seeds
22 g/100 g ww 10.9 mg/g dw
M. oleifera kernels
35.1 g/100 g dw
M. oleifera leaves
22.2 to 31.4 g/100 g dw depending on the cultivars* 37.5 g/100 g dw
M. oleifera seeds Oils
8.1, 10.4, 18.5 and 27.5 g/100 g dw, respectively 2.9 g/100 g dw
M. concanensis seeds
37.7 to 40.1 g/100 dw depending on the source
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Hydromethanolic (70 %)
Nouman et al. (2016)
Antioxidant and reducing power activities
Water
CastroLópez et al. (2017)
Potential use in drug delivery
NI
Varma et al. (2014)
Immunomodulatory effects Potential to enhance human diet
Phosphate buffer
Anudeep et al. (2016) Abdulkadir et al. (2016)
Microwaveassisted extraction Solubilization by agitation
Hypoglycemic activity
Water
Chen et al. (2017)
Coagulation activity
Agrawal et al. (2007)
Solubilization by agitation Solubilization by agitation
Antimicrobial activity
25 mM sodium phosphate buffer, pH 7.5 Deionized water
Digestion by Kjeldahl’s method Solubilization by agitation
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Buffer (0.05 M T risHCl, pH 8.0, containing 0.15 M NaCl) NI
Enzymatic antioxidant activity
Buffer phosphate 50 mM pH 7.8
Digestion by Kjeldahl’s method
Antioxidant, antihypertensive and antidiabetic properties
H2 SO4
GonzálezGarcía et al. (2017)
Soxhlet's method
Potential to develop food formulations with high-stability
Hexane
Manzoor et al. (2007)
Agitation/infusion
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Makita et al. (2016)
Microwaveassisted extraction
Mechanical method (injured in stem sites) Enzyme digestion and precipitation AOAC method
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M. oleifera seeds M. oleifera leaves, pods, stem and seeds M. oleifera leaves
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Proteins
M. oleifera stem
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Carbohydrates
77 to 197 mg/g dw depending on the cultivars 2.7 to 15.2 mg GAE/g dw depending on the solid: liquid ratio NI
1) CO2 ; 2) CO2 /Ethanol; 3) Hot water Aqueous methanol (80 %)
Antioxidant potential
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M. oleifera and M. ovalifolia leaves M. oleifera leaves
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M. oleifera leaves
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Compounds
Antifungal activity
NI
Shebek et al. (2015) Pinto et al. (2015) Mat Yusoff et al. (2016) Nouman et al. (2016)
ACCEPTED MANUSCRIPT region M. peregrina kernels M. oleifera seeds
38 g/100 g dw
Soxhlet's method
NI
Sonication
M. oleifera kernels
29 to 40.7 g/100 g dw depending on particle size 11.4 to 28.6 g/100 g ww depending on the processing conditions 11.80 to 41 g/100 g dw depending on the solid: liquid ratio
Solvent extraction
NI
Hexane
Mechanical method (pressure)
NI
NI
Soxhlet's method
Potential as a healthy alternative to partially hydrogenated vegetable oils
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NI= Not identified *Expressed as crude protein after Digestion by Kjeldahl’s method dw= dry weight ww= wet weight
Chloroform:Methanol (3:1)
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M. oleifera seeds
Hexane Hexane
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M. oleifera seeds
Potential to develop frying foods
Salaheldeen et al. (2014) SánchezMachado et al. (2015) Mat Yusoff et al. (2016) Fakayode and Ajav, (2016) Bhutada et al. (2016)
ACCEPTED MANUSCRIPT Table 2. Physicochemical parameters measured in vegetable oils Properties Viscosity (Pa/s) Kinematic Viscosity (mm2 /s)
M. oleifera 42.8 -
M. peregrina 36.18
34.18
M. concanensis -
Salvia hispanica 43.2 -
0.89
0.89
0.89
0.86
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RI
1.64
-
-
1.46
1.48
FFA
1.29
0.34
0.35
0.34
63.94
77.17
67.73
67
160.62
198.6
187.53
6.5
-
1
-
1.17
-
-
0.11
Conjugated dienes Acid value (mg KOH/g)
SánchezM achado et al. (2015)
Reference
Salaheldee n et al. (2015)
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RI= refractive index FFA= Free fatty acids (%) IV= Iodine value (g I/100 g) SN= Saponification number * values depend on the growing region of the olive tree
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Peroxide value (meq/kg)
179
204 1.97
-
-
1.75
4.33
3.17
+
-
0.70
Salaheldeen et al. (2014)
M anzoor et al. (2007)
1.46 in all samples
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Carotenoids (g/kg)
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SN (mgKOH/g)
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IV
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Density (g mL-1 )
Olive oil*
-
2.54 Timilsena et al. (2017)
86.3 to 89.4 1.5 to 2.2 2.26 to 2.70 2.35 to 2.40 0.45 to 0.60 Kharazi et al. (2012)
ACCEPTED MANUSCRIPT Table 3. Functional applications of Moringa oleifera in food products
Leaves
2
Bread
Leaves
5
Cookies
Leaves
10 -20
Modified atmosphere packaged raw beef
Leaves
0.3 **
Leaves
20
Vegetable soup powder
Leaves
Bread
Seeds
5
Leaves
5 - 7†
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Ready-to-eat snack
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6 different Indian foods
* Flour to prepare the food product ** Expressed as g/L of moringa leaf extract † Expressed as g/100 of the final product NI= no identified
8.5†
Reference
Protein and minerals
Ogunsina et al. (2011)
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Rice crackers
Related bioactive compounds
NI
Manaois et al. (2013)
Protein, fiber and minerals
Sengev et al. (2013)
NI
Nwakalor et al. (2014)
Phenolic compounds
Shah et al. (2015)
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10 - 20
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Seeds
Improve the nutritional properties with acceptable rheological and sensory characteristics Have good sensory acceptability scores Improve the nutritional composition Products have acceptable sensorial qualities Provide a stabilization of the color and prevent lipid oxidation in raw beef
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Cookies
Functional advantages
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Foodstuff
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Concentration with best results (g/100 g*)
Vegetative structure
Phenolic compounds, saponins and phytic acid Protein, fiber, Improve the vitamin D and nutritional quality and C, and have long shelf life minerals Improve the Protein, fiber, nutritional quality vitamins and keeping sensory minerals attributes Maintain the organoleptic NI acceptability Decrease in antinutritional factors
Devisetti et al. (2016)
Farzana et al. (2017)
Bolarinwa et al. (2017) Mamta et al. (2017)
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Table 4. Patent records associated with the application of Moringa plants in the formulation of food products Invention core
Formula and preparation method for Moringa health wine
T he invention belongs to the technical field for food processing and discloses a product based on Moringa and other plant materials; it presented several technical advantages and was reported as a good-for-health beverage
Moringa soybean paste and preparation method thereof
T he invention relates to a method for producing fermented soy sauce: (a) Prepare a soy sauce fermented with Moringa. Soak the soybeans for 3 to 4 hours and then boil them. (b) Mix the fermentation product of the Moringa brine extract of step (a) with the fermentation product of step (b). Aging the stage mix in a dark place for 45 to 60 days and evenly mix the fermentation mixture of aged Meju brine extract and Moringa.
Fermented Moringa beverage and method thereof
T he invention provides a beverage of Moringa prepared using a method for retaining Moringa nutrients and the original flavor with good taste. In addition, was reported this as a hypocaloric beverage due to the use of xylitol as a sweetener.
Moringa pepper paste and preparation method thereof
T his invention relates to a method for preparing a Moringa leaf with a moisture content of 4 to 6%. After collecting and washing Moringa leaves, prepare salt water by adding sun salt to clean water to a salt concentration of 17 to 25%. Mix 1.8 to 2.2 kg of Moringa tea from step (A) with 1 liter of alkaline brine and ferment for 45 to 60 days in a sunny place with a mixture of Moringa leaves and salt water. It can be easily ingested and has excellent nutrients and excellent functional ingredients, thus contributing to the national health and functional food industry.
Le av es
Moringa γamino-acid-rich product and its production process and application
T he invention discloses a M. oleifera product rich in γ-amino butyric acid. T his product could be used raw or as an additive for preparing various health foods for decreasing blood pressure, promoting sleeping and other benefits for human healt h.
St e m s an d
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Title of the required patent
V e g et at iv e st r u ct u re in v ol v e d
P u bl ic at io n d at a
C o u n t r y
R oo ts, st e m s, le av es, fl o w er s
20 /0 6/ 20 17
C hi n a
Le av es
05 /0 7/ 20 17
K or e a
Le av es
03 /0 7/ 20 17 19 /0 7/ 20 17
K or e a
31 /0 5/ 20 17
C hi n a
K or e a
ACCEPTED MANUSCRIPT le av es T his invention relates a typical Korean product and its preparation method, including Moringa leaves and pepper powder, to improve the nutritional content. The invention has large amounts of healthy components with features with anticancer action and hyperlipidemia treatment.
Le av es
T he invention relates a product based on fermented rice and Moringa leaves and is prepared through a drying process. T his product was identified as a product with high nutrient contents.
Le av es
T he invention relates the preparation of Moringa soy through an alkaline treatment.
Le av es
Vinegar of Moringa oleifera and manufacturing method thereof Moringa seed rice cake and preparation method thereof
T his invention displays the manufacturing of M. oleifera vinegar and persimmon through five steps, which could be very useful to modern persons and food industries due to the contents of nutrients and functional components.
Le av es
T he invention discloses a rice cake with Moringa seeds, which contain various nutritive ingredients needed by the human body and further have the functions of clearing gut and promoting metabolism.
Moringa seed noodles and preparation method thereof
It describes a method to make noodles from Moringa seed and leaves rich in bioactive compounds with good health care effect and sensory qualities.
Production of Moringa olei fera QQ candy
It discloses a candy formulation with moringa oil and caloric and noncaloric sugars mixed with other ingredients. In addition, this product has a peculiar flavor and health -care functions.
Moringa seed flour and preparation method thereof
T he invention describes a Moringa seed flour mixed with other functional ingredients, which have beneficial effects on the intestines and stomach of the human body, helping digestion.
Se ed s an d le av es Se ed s an d le af Se ed s (o il) Se ed s
Moringa soybe an milk and preparation method thereof
It provides information about Moringa soybean milk capable of supplementing a variety of nutritional elements required by human bodies and the method for its preparation.
Moringa oleifera and blueberry health-care buccal tablet Moringa hango ver-alleviating
T he invention provides a M. oleifera and blueberry health-care buccal tablet. It can serve as a nutritious supplement with has the oxidation resistance, vision fatigue relief, heart function enhancement, immunity improvement, diabetes prevention, and high blood pressure prevention and is simple to produce and industrialize.
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Method for producing a Moringa kimchi seasoning Fermented rice with Moringa and method of making the same Moringa sauce and preparation method thereof
It relates the development of a functional drink capable of accelerating human metabolism, due to the content of moringa leaves.
N E (fr ee ze dr ie d m or in ga ) Le av es
Le av
03 /0 7/ 20 17 22 /1 2/ 20 17 05 /0 7/ 20 17 09 /1 2/ 20 16 23 /1 1/ 20 16
K or e a
16 /1 1/ 20 16
C hi n a
09 /1 1/ 20 16 09 /1 1/ 20 16 21 /0 9/ 20 16
C hi n a
25 /0 3/ 20 15 30 /0
C hi n a
K or e a K or e a K or e a C hi n a
C hi n a C hi n a
C hi
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9/ 20 15
n a
08 /0 7/ 20 15 29 /0 4/ 20 15 08 /0 4/ 20 15
C hi n a
04 /1 1/ 20 15 06 /0 5/ 20 15
C hi n a
It discloses the invention of a functional cake from Moringa leaves and others plant materials described as a product with a unique taste that is crispy and delicious.
Le av es
Compound Mo ringa oleifera instant granules and preparation method thereof Moringa flower cake and preparation method thereof
It describes a method to obtain granules rich in active ingredients including flavonoids and polysaccharides from M. oleifera leaves and other leaves. T his was described as a functional health food capable of effectively regulating the blood sugar and blood lipid of a human body.
Le av es
It provides information about a healthy Moringa flower cake from the stem and leaves of Moringa plants and other ingredients.
Moringa oleifera biscuit and preparation method thereof
T he invention relates to a biscuit processing method, particularly a method for preparation
Fl o w er an d st e m le af Le av es
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T his invention has ingredients such as fruit pulp (90 grams) and M. oleifera leaf powder (0.2-4 grams) (no additional information was provided).
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Fruit pulp mixed with Moringa oleifera leaf powder
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Moringa flower cake
Le av es
C hi n a C hi n a
G er m a n y
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Graphical abstract
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Highlights
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Moringa is a source of bioactive compounds with relevance for food industry Carbohydrates reported in M. oleifera are reviewed by the first time Bioactive compounds from Moringaceae family different to M. oleifera are reviewed LC and GC-MS are the most suitable methods for identification of bioactive molecules
Graphics Abstract
Figure 1