Lycopene: A Natural Red Pigment

CHAPTE R 13

Lycopene: A Natural Red Pigment Rahul C. Ranveer Post Graduate Institute of Post Harvest Management, Killa - Roha, Maharashtra, India

1 Introduction Many coloring constituents occur in fruits and vegetables. Anthocyanin pigments are predominant for various colors of plants and plant parts, such as red color in kokum rind (Nayak et al., 2010), plum, cabbage, strawberries, apples, shiso leaves, blueberries, grapes, leaves of the perennial plant (Gordon et al., 2012), bracts of bananas (Gordon et al., 2012), purple sweet potato (Lee et al., 2013), and black rice. Naderi et al. (2012) reported isolation of various pigments, such as betanin from beet root, Bixin/Norbixin from bush seed, capsanthin/capsorubin from paprika, carmine from cochineal pigments, carminic acid from cochineal pigments, chlorophyll from grass, crocin from saffron/gardenia fruits, curcumin from turmeric, and lutein from marigold. Also, some literatures are available on the extraction of lycopene from tomato peel (Ranveer et al., 2013), betanin (anthocyanins) from beet root (Naderi et al., 2012), and bixin/norbixin (carotene) from orange peel (Chedea et al., 2010). Plants are considered as the main pigment producers, however, microorganisms also produce pigments and these natural colorings are pragmatic universally in life. The colorings are observed at every step of life beginning with plant leaves, fruits, and vegetables, plants floras and peel, eyes, animal structures, and in microorganisms. Natural and synthetic colorants are used in various products, such as food, pharmacy, textile, cosmetics, and furniture. Along with improvement in aesthetic value, the natural colorant plays prime role, such as photosynthesis. Without the colorants, photosynthesis will not take place in plants as no life is predominant without chlorophylls and carotenoids. In animals, O2 and CO2 transference could not be possible without hemoglobin or myoglobin. In stress conditions flavonoids and quinines are produced by plants, which are significant for converting light into chemical energy. The melanin safeguards individuals and other animals, these colorants are important for their life cycle in some fungi. Moreover, numerous pigments have been reported for pharmacological activities in many diseases, for example, cancer and cardiovascular diseases, and this has stressed pigments significantly for mankind (Mol et al., 1996; Shahid and Mohammad, 2013; Singh et al., 2014). Natural and Artificial Flavoring Agents and Food Dyes http://dx.doi.org/10.1016/B978-0-12-811518-3.00013-2

427

Copyright © 2018 Elsevier Inc. All rights reserved.

428  Chapter 13 People start linking product qualities with appearance, that is, mainly with color from the beginning, It is mainly factual with meals. In the commencement of the food industry end-users were not conscious about the colorant exploited in food (natural or synthetic), however, in recent years people have expressed their repulsion at artificial colorants when the concepts “artificial colorants” and “diseases” were connected, whereas natural pigments are recognized for pharmacological benefits. On the other hand, only a few colorants of natural origin are allowable for consumption by individuals, and it is very difficult to approve new sources because pigments are listed as food additives by US Food and Drug Administration (USFDA), and have stringent norms for pigments (Calogero et al., 2015; FDA/IFIC, 1993; Rymbai et al., 2011; Zhang and Zhong, 2013).

1.1  Classification of Pigments 1.1.1  On the basis of origin On the basis of origin pigments are classified as inorganic, natural, and synthetic. The pigments obtained from living things, such as animals, plants, and microbes, are considered as natural pigments whereas synthetic pigments are prepared synthetically. Both synthetic and natural pigments are made up of organic compounds, whereas pigments obtained from nature or replicated by synthesis are inorganic compounds (Bauernfeind, 1981). 1.1.2  On the basis of chemical structure of the chromophore Categorization of colorings on the basis of chromophores chemical structure, such as chromophores with conjugated accomplishments consisting of anthocyanins, betalains, carotenoids, caramel, synthetic colorant, and lakes and metal-coordinated porphyrins include chlorophyll, and their isomers and myoglobin (Wong, 1989). 1.1.3  On the basis of structural characteristics of the natural pigments Categorization of natural colorings on their structural characteristics, such as heme and chlorophylls, are derivatives of tetrapyrrole; these derivatives of isoprenoid like iridoids and carotenoids. While purines, pterins, flavins, phenazines, phenoxazines, and betalains are included in N-heterocyclic compounds from tetrapyrroles. Benzopyran derivatives (oxygenated heterocyclic compounds) include anthocyanins and other flavonoid pigments. Benzoquinone, naphthoquinone, anthraquinone are comprised of quinone and melanins (Bauernfeind, 1981). 1.1.4  On the basis of utilization as food additives The pigments are deliberated as food additives and classification completed by the FDA on colors with the necessary authorization that they are man-made and subcategorized as synthetic pigments and lakes. Another cluster of pigments are excused from certification and

Lycopene: A Natural Red Pigment  429 include colorants obtained from natural resources, for example, plants, minerals, or animals, and complements of natural products (Shahid and Mohammad, 2013; Wong, 1989).

2 Lycopene Red-colored fruits and vegetables mainly consist of carotenoids, particularly lycopene. The lycopene in various food and food products is presented in Table 13.1. This natural red pigment is synthesized exclusively by plants and microorganisms (fungi, bacteria, and algae). Lycopene cannot be synthesized either by animals or by humans; therefore they can be exclusively taken from their diet (Rodriguez-Amaya, 2010; Zuorro et al., 2011). Even though the lycopene content of tomatoes is lower than the other commercially grown fruit species, they provide the majority of lycopene in the American diet (Rao and Agarwal, 2000; Srivastava and Srivastava, 2015). Lycopene comprises 83% of the total pigments present in tomatoes (Shi et al., 1999). Lycopene consists of unsaturated hydrocarbon with 11 conjugated and 2 unconjugated double bonds. Lycopene converts into different cis-trans-isomers influenced by sunrays, thermal energy, and chemical reactions (Nguyen and Schwartz, 1999). Many conjugated carbon double bonds give color to the lycopene. Every double bond decreases the energy necessary for electron transition to higher energy states, permitting the particle to absorb visible light of gradually longer wavelengths. The majority of the visible spectrum is absorbed by lycopene and it appears as a red color. In oxidation of lycopene, double bonds present in carbon atoms are broken, separating the compounds into smaller compounds, every one double bonded to an oxygen atom. Even though double bonds between carbon and oxygen are also chromophoric, the much shorter compounds are unable to engross sufficient light to seem as colorful. An analogous effect can be seen if reduction occurs in lycopene; reduction may saturate the lycopene compounds, diminishing its ability to engross light.

Table 13.1: Lycopene content of various food products. Food

Lycopene Content (mg/100 g)

Apricots Chilli Grapefruit Watermelon, fresh Papaya, fresh Grapefruit, pink/red Guava, raw Vegetable juice Tomato

0.005 2.62 3.36 4.10 5.30 3.36 5.40 7.28 3.1–7.74

Source: Singh, P., Goyal, G.K., 2008. Dietary lycopene: its properties and anticarcinogenic effects. Compr. Rev. Food Sci. Food Saf. 7, 255–270.

430  Chapter 13

Figure 13.1: All-trans-Lycopene.

Lycopene is found mainly in tomatoes in all-trans configuration form and the thermally stable form of lycopene is cis-isomers (Zechmeister et al., 1941). Lycopene originates as an isomeric mixture in the blood of human beings, with 60% as cis-isomers out of the total lycopene. The molecular formula of lycopene (C40H56) was initially determined when Willstätter and Escher (1910) presented that lycopene is an isomer of the carotenes. Initially, the chemical structure of lycopene was reported by Karrer et al. (1930), which was afterward confirmed by Kuhn and Grundmann (1932), by categorization of its degraded products following oxidation of chromic acid. The molecular weight of lycopene is 536.85 Da, with the general structure being an aliphatic hydrocarbon with 11 conjugated carbon–carbon double bonds (Fig. 13.1), which imparts a red coloration, as well as fat- and lipid-soluble characteristics. The visible range of light is absorbed by lycopene, and the highest absorption λmax at 472 nm was recorded when lycopene was dissolved in petroleum ether and a differential emanation wavelength of 3078 (Davies, 1976; Moss and Weedon, 1976). As a consequence of the 11 conjugated carbon–carbon double bonds in its backbone, lycopene can tentatively assume 211 or 2048 geometrical configurations (Prasad and Mishra, 2014). All-trans, that is, 5-cis, 9-cis, 13-cis, and 15-cis are some of commonly recognized isomeric forms of lycopene (Fig. 13.2), with the stability of different forms as 5-cis > all-trans > 9-cis > 13-cis > 15-cis > 7-cis > 11-cis, so that the 5-cis form is thermally more stable than the all-trans-isomer (Prasad and Mishra, 2014). A wide variety of antioxidants are present in tomatoes, which includes ascorbic acid, carotenoids, vitamin E, phenolics, and flavonoids (Zuorro et al., 2011). Tomatoes and tomato products are rich in lycopene, a carotenoid, and has created significant attention in recent times, as epidemiological confirmation proposes that it may give protection from cancer and other degenerative illnesses, affected by free radical reactions (Levy et al., 1995; Ranveer et al., 2013). It is well documented that lycopene reduces the occurrence of ischemic heart disease (Zuorro et al., 2011). These positive effects have been first and foremost recognized that the antioxidant potential of tomatoes and tomato-based products,

Lycopene: A Natural Red Pigment  431

Figure 13.2: Lycopene Geometrical Isomers.

as augmented oxidative stress, is an ordinary pathogenic factor for the aforementioned diseases (Ranveer et al., 2013). The previous study reviewed the appearance of lycopene in various portions of tomatoes, such as the skin, the water-insoluble portions, and the fibrous portions, including the fiber and soluble solids (Ranveer et al., 2013; Sharma and Le Maguer, 1996). The results showed that 72%–92% of lycopene was linked with the water-insoluble fractions and skin. This higher amount of lycopene was accumulated more in the peel than other parts. Sharma and Le Maguer (1996) found that tomato extracts, especially skin extracts, contain high amounts of lycopene. Ranveer et al. (2013) reported that lycopene from tomato peels differ from those obtained using whole tomatoes, because of the differences in the chemical composition of the peel and the whole fruit, as well as lycopene is reported to occur in higher concentrations in tomato peel. The peel of tomatoes has the highest total carotenoid concentration, and the locular contents have the highest carotene content. It has been reported that lycopene represents

432  Chapter 13 Table 13.2: Lycopene content of different parts of tomato. S. No.

Sample Name

Lycopene (µg/g)

1 2 3 4

Whole tomato Tomato pulp Peel Industrial waste

83.90 ± 0.79 47.6 ± 0.81 376.17 ± 1.13 176.17 ± 1.09

Source: Ranveer, R.C., Samsher, N.P., Sahoo, A.K., 2013. Effect of different parameters on enzyme-assisted extraction of lycopene from tomato processing waste. Food Bioprod. Process. 91 (4), 370–375.

a substantial proportion of the total carotenoid content of tomato products (Choudhari and Ananthanarayan, 2007). It is estimated as much as 60%–64% of the total carotenoid content consists of lycopene. Considering whole tomatoes, the peel content will be low (5.5%–8.1%), which is the reason for lower lycopene content (Ranveer et al., 2013). Lycopene was found predominantly in the chromoplast of plant tissues. In tomatoes, lycopene biosynthesis increases dramatically during the ripening process, as chloroplast undergoes transformation to chromoplast. Globulous chromoplast containing mainly β-carotene is found in the jelly part of the pericarp while chromoplast is found in the outer part of the pericarp and contains voluminous sheets of lycopene (Choudhari and Ananthanarayan, 2007; Ranveer et al., 2013). The tomato-processing industry waste comprises skin and seeds (approximate in the ratio of 37:63), which lower lycopene content (Machmudah et al., 2012a) as seeds do not contain lycopene (Table 13.2). However, considering the cost of production of lycopene, it can be concluded that the waste of tomatoprocessing industries, in the form of seeds and skin residues, could provide a useful source of lycopene (Ranveer et al., 2013).

3  Extraction of Natural Pigments Several extraction methods have been reported for extraction of pigments from plant origin. Important plant pigments are flavonoids, carotenoids, and chlorophyll. Procedures for extraction of these pigments were reported by various researchers. Anthocyanin from maceration of plant materials that use ethanol, methanol, or n-butanol, acidified with HCL (Lee and Chen, 2002). Cyanidin 3-O-rutinoside and pelargonidin 3-O-rutinoside were significantly extracted from berries Smilax aspera with acidified methanol and purified by chromatographic technique (Longo and Vasapollo, 2006). Giusti and Wrolstad (1996) reported pelargonidin-3-sophoroside-5-glucoside, mono- or diacylated was extracted with cinnamic and malonic acids. Haiyan et al. (2003) reviewed that ethanol was found suitable for extraction of anthocyanin from cherry. Extraction of anthocyanin from various wheat varieties with different pH methods was reported by Jie et al. (2002). Microwave treatment at 624 W for 60 s, together with ultrasonic processing for 40 s, was reported to give optimal extraction of anthocyanin pigment (Jinxing et al., 2003).

Lycopene: A Natural Red Pigment  433 Table 13.3: Lycopene extract methods. Extraction Methods

References

Solvent extraction Hydrostatic pressure processing Enzyme-assisted extraction

Haroon (2014); Colle et al. (2010); Barba et al. (2006) Haroon (2014); Kong et al. (2010); Xi (2006) Choudhari and Ananthanarayan (2007); Zuorro and Lavecchia (2010); Cinar (2005); Ranveer et al. (2013); Ranveer and Sahoo (2015) Haroon (2014); Choksi and Joshi (2007); Tzia and Liadakis (2003); Shi et al. (2009); Vagi et al. (2007) Gutte et al. (2015); Haroon (2014); Eh and Teoh (2012); Chemat et al. (2012, 2004); Rostagno et al. (2003); Li et al. (2004); Mason et al. (2011); Nazir et al. (2009); Zhang et al. (2008); Hromadkova et al. (1999)

SCFE with CO2 Ultrasonic extraction

SCFE, Supercritical fluid extraction.

The literature generally agrees that the stable form of lycopene is found in raw tomatoes, while degradation and isomerization might occur rapidly, even a mild heating treatment during processing or when it is used along with oil or organic solvents (Colle et al., 2010). There are different types of methods for extracting lycopene (Table 13.3).

3.1  Solvent Extraction Extraction using organic solvents is a well-known method that is used in the food industry. Traditionally, organic solvents are used to extract lycopene. Although the method is reliable, it is laborious, cumbersome, and requires the use and disposal of organic solvents (Colle et al., 2010; Haroon, 2014). Carotenoids present in the tomato are fat soluble; common organic solvents used for extraction of lycopene are dichloromethane, hexane, ethanol, acetone, ethyl acetate, petroleum ether, and mixtures of polar or nonpolar solvents in different ratios, such as acetonechloroform (1:2) and hexane-acetone-ethanol (2:1:1) (Barba et al., 2006; Haroon, 2014). The amount of lycopene extracted using hexane/ acetone or hexane/ethanol is higher than when using methanol, dichloromethane, or chloroform (Barba et al., 2006; Haroon, 2014). Organic solvents used for extraction of lycopene are toxic in nature and traceless, which can make it unsuitable for human consumption (Haroon, 2014).

3.2  Hydrostatic Pressure Processing Hydrostatic pressure processing (HPP) with or without high temperature can improve the extraction of lycopene from tomatoes and waste obtained from the tomato-processing industry. Higher yield lycopene and in a shorter time as compared to solvent extract process can be possible by HPP (Xi, 2006). Highly purified lycopene was extracted from tomato by-products using “Extractor Naviglio” (Kong et al., 2010). This extraction method

434  Chapter 13 uses tap water with minimal organic solvent (Haroon, 2014), which did not give any toxic effect.

3.3  Enzyme-Assisted Extraction Cellulase and pectinase enzymes have been reported to substantially improve yields of lycopene from tomato waste. Choudhari and Ananthanarayan (2007) reported that when whole tomatoes were pretreated with optimized conditions of enzyme, the lycopene yield is improved. However, Zuorro and Lavecchia (2010) reported an increased amount of lycopene by 75.6% at 25°C, the extraction was carried in two-step; the first step consists of 5 h enzyme pretreatment, whereas the second step consists of 3 h solvent (hexane or ethyl acetate) extraction. Cellulase and pectinase in different combinations resulted in higher extraction yield of carotenoids from peel of oranges, sweet potato, and carrot (Cinar, 2005). Cellulase act on cellulose, which appear in the primary cell wall below the principal layer of central lamella of the plant cell wall. The primary wall contains a firm structure of cellulose embedded in medium-like gel containing pectic substances, glycoprotein, and hemicellulose. The cellulase enzyme increases cellulose breakdown in glucose, cellobiose, and higher glucose polymers. Pectinase is pectolytic enzyme and hemicellulolytic has the capability to degenerate pectic substances and pectin, further a polymer of 100–200 galacturonic acids, appears in the interior lamella and foremost walls (Choudhari and Ananthanarayan, 2007). Researchers previously documented that lycopene is mostly present in peel parts of the tomato fruit. These enzymes are accomplished to break down the cell wall and unconfined lycopene, which improve the yield.

3.4  Supercritical Fluid Extraction With CO2 Haroon (2014) and Choksi and Joshi (2007) reported that supercritical fluid extraction (SCFE) with carbon dioxide (CO2) gives the best extraction results for lycopene. Many investigators have utilized SCFE for extraction of lycopene from various food samples. During extraction by this method, liquid CO2 (dry ice) is used as the solvent below anticipated pressure or temperature. The food material is packed into the extractor cell. After extraction, the extract is hastened through a control valve (Haroon, 2014; Tzia and Liadakis, 2003). In SCFE CO2 is a perfect solvent for extracting food constituents, because it is nonhazardous, nonflammable, nontoxic, and inexpensive. Lycopene is subtle to light, heat, oxygen, and acids. Isomerization and degradation will be lower as compared to other methods, when SCFE was used for extraction purpose, whereas CO2 lacks polarity and the ability to form specific solvent–solute interactions. Mixing of minute quantities of polar solvent like water, ethanol, methylene chloride, or hexane can improve extractability of CO2. Solubility studies was performed over 50–80°C and a 200–400 bars pressure exhibited that these are suitable working conditions for

Lycopene: A Natural Red Pigment  435 carotenoid extraction (Haroon, 2014; Shi et al., 2009). Response surface methodology can be applicable for maximum extraction of lycopene yield from tomato skin (Haroon, 2014; Vagi et al., 2007). The merchandise obtained by SCFE at 460 bar and 80°C controlled the maximum concentration of carotenoids with 90% of lycopene (Haroon, 2014; Vagi et al., 2007).

3.5  Ultrasonic Extraction Lycopene yield was originated to improve the efficacy of comparatives by sonication (improvement of 26% extraction yield of lycopene in six repetitions in 40 min at 40°C and 70% v/w in the occurrence of ultrasound), lower the extraction temperature, and shorten the total extraction time. Ultrasonication boosted the extraction recovery of trans-lycopene by 75.93% as compared to traditional extraction process, and also minimized the degradation and isomerization of lycopene (Eh and Teoh, 2012). Unadventurous methods of extraction needs were lengthier extraction processes, higher extraction temperatures, and a higher solvent-to-sample ratio for improved efficacy of extraction. Therefore, associated to the enhanced UAE of lycopene, conservative methods are lengthier and less efficient in terms of energy and solvent consumption (Haroon, 2014). The current literature has shown that the ultrasound-assisted extraction can boost the extraction efficacy and concluded acoustic cavitation and some mechanical effects (Chemat et al., 2004, 2012; Rostagno et al., 2003). Sonication cavitation can interrupt cell walls easing solvents to penetrate into the plant material and permitting the intracellular component release. In other mechanical effects produced by ultrasound, it may also be the anxiety of the solvent used for extraction, thus cumulative interaction surface areas between the solvent and beleaguered constituents by allowing superior penetration of solvent into the sample matrix. The ultrasonic-assisted extraction (UAE) is also obligatory shorter extraction times and reduced solvent ingesting (Gutte et al., 2015; Li et al., 2004; Mason et al., 2011; Nazir et al., 2009; Zhang et al., 2008). Whereas, Hromadkova et al. (1999) described that ultrasound-assisted extraction can be performed at a lower temperature, which can avoid thermal destruction to the extracts and minimize the loss of bioactive compounds.

4  Determination of Lycopene Previously, investigators have already described numerous methods to measure and investigate lycopene content in food and biological samples. These comprise ultraviolet– visible (UV–VIS) spectrophotometry (Davis et al., 2003; Fish et al., 2002), liquid chromatography (LC), thin-layer chromatography (TLC), and high-performance liquid chromatography (HPLC) (Britton, 2008; Ishida and Chapman, 2009; Lee and Chen, 2002; Thadikamala et al., 2009).

436  Chapter 13 Various other procedures high-speed countercurrent chromatography (Baldermann et al., 2008), fiber optic visible reflectance spectroscopy (Choudhary et al., 2009), and infrared spectroscopy (De Nardo et al., 2009) are also industrialized. However, they require additional time (usually >20 min) and solvent in order to accomplish departure of the all-Elycopene with its Z isomers. UV–VIS spectrophotometry is more expedient, quicker, and cheaper than HPLC analysis and huge quantities of samples can be administered in a comparatively short time. But, very small amounts of lycopene (less than 1 µg) could not be detected by UV–VIS spectrophotometry, whereas in HPLC 1 µg samples can be detected (Hyman et al., 2004). Though HPLC analysis permits precise quantification of pigments and separation of their isomers, it is difficult to produce continual outcomes and a high level of skill is required (Hyman et al., 2004).

5  Purification Techniques 5.1  Reverse Phase Chromatography Carotenoids are separated by gradient elution normal phase open-column chromatography and analyzed by isocratic-elution reverse phase high-performance liquid chromatography (HPLC) with UV/vis photodiode array detection. The normal-phase elution order of trans pigments was phytoene, phytofluene, β-, zeta-, γ-carotene, and lycopene (Barrie, 1988). An isocratic reversed-phase HPLC assay was developed for monitoring the isomers of α- and β-carotene in vegetables with dark green in color using a mixture of isomers obtained from the light catalyzed oxidation of all-trans α- and β-carotene in the presence of iodine (Nyambaka and Ryley, 1996). Good separation of lycopene from its cis-isomers and other carotenoids has been achieved in vegetable samples on analytical reversed-phase columns with solvent mixtures of methanol and chloroform (94:6; Quackenbush, 1987) and in plasma samples with acetonitrile, methanol, dichloromethane, and water (7:7:2:0.16; Stahl et al., 1992). Researchers on cis-trans-carotenoid isomers have been efficaciously unglued and determined by HPLC using an analytical Ca(OH) 2 column (Chandler and Schwartz, 1987; Koyama et al., 1988; Tsukida et al., 1982). Lesellier et al. (1989) studied the separation of carotenoid stereoisomers at room temperature by the reversed phase HPLC technique using polymeric octadecyl (ODS) phase. Isolation method of lycopene was established by consecutive steps, including petroleum ether extraction, solid-phase extraction using silica cartridges, and supplementary purification with semipreparative HPLC using a Zorbax ODS column. With this method, 6 mg of lycopene was isolated from 10 g of tomato puree (Hakala and Heinonen, 1994). A previous study had conveyed a lycopene production of 775 mg/L by the fermentation of reproduced cultures of Blakeslea trispora grown on optimized GAY (glucose asparagine yeast extract) medium accompanied with vitamin A acetate (1000 ppm) at the commencement of fermentation followed by that of lycopene cyclase inhibitor, piperidine (500 ppm), after 48 h (Choudhari et al., 2009). The focus of the study was to develop a

Lycopene: A Natural Red Pigment  437 simple, novel chromatographic technique for the separation and purification of lycopene from microbial biomass and which could be browbeaten on a commercial scale.

5.2  Thin Layer Chromatography Method A thin line was drawn on the activated TLC plate about 1.5 cm above the bottom. A spot of the extract was placed on the line and allowed to dry. This was followed by a repeated addition of the extract on the same spot. The developing chamber was a beaker containing a mixture of hexane and acetone in the ratio of 3:2. The TLC plate was placed inside the developing chamber and the top was covered. The solvents were allowed to rise on the plate till it reached 1.5 cm close to the top (Rebecca et al., 2014).

5.3  Saponification Method The tomato paste extract was further purified by saponification, referring to method suggested by Zhao et al. (2002). The saponification was carried out in a 40 wt.% KOH solution. Tomato paste (10 g) extract was first dispersed into 2-propanol (1:5, wt./wt.) at 60°C for 1 h under a stream of nitrogen. Then 37 wt.% KOH solution (1:4, v/v) was added and the mixture was stirred at 50°C for 2 h. Finally, the mixture was washed with distilled water to neutrality and then filtered to obtain 0.50 g lycopene crystals.

5.4  Crystallization Method The extracted lycopene was dissolved in dichloromethane/ethanol (1:4) at temperature of 50–60°C, placed in an ice bath for gradual lowering the temperature and then placed in deep freezer overnight to form crystals. The crystals were filtered through Whatman no. 4. filter paper, washed with cold ethanol and dried in freeze-dryer. The crystallization procedure was repeated to obtained crystals with higher level of purity (Ranveer and Sahoo, 2015).

6  Stability of Lycopene 6.1  Effect of Processing on Stability of Lycopene Nguyen and Schwartz (1999) reported that isomerization and oxidation are the main causes of lycopene degradation. In the initial stage of degradation, it consists of reversible isomerization of all-trans-lycopene to less colored, more oxidizable cis-isomers (Boskovic, 1979). Autoxidation of all-trans-lycopene and the cis-isomers occurred parallel to trans-cis-isomerization producing a split of the lycopene molecule into smaller fragments, such as volatile aldehydes and ketones, developing hay or grassy off-flavors (Lovric et al., 1970; Schierle et al., 1997). Ecological influences, such as air, light, and temperature may be very significant for the isomerization and autoxidation of lycopene in tomato products (Anguelova and Warthesen, 2000a,b).

438  Chapter 13

Figure 13.3: Degradation Mechanism of Lycopene.

Lycopene generally undergo two types of changes during processing and storage; isomerization from all-trans to mono- or poly-cis forms and oxidation. During thermal processing of food (cooking, heating, or drying) carotenoids undergo trans-cis-isomerization relative to the time-temperature combination. Degradation of lycopene due to isomerization and oxidation affect the bioavailability and decrease its functionality for health benefits. Due to a highly unsaturated nature they are particularly susceptible to oxidation. Degradation is largely affected by various physical and chemical parameters, such as time, temperature, reaction medium, and environmental conditions. However, the three most important factors that affect the chemical stability of lycopene are heat, light, and oxygen. Studies have shown that isomerization of lycopene from all-trans to mono- or poly-cis form occurs due to changes in the seven conjugated double bonds initially during thermal processing of food followed by degradation. No further trans-cis-isomerization was reported during storage; instead mainly cis-trans-reversion and autooxidation were observed (Lovric et al., 1970) (Fig. 13.3). cistrans-Reisomerization may take place due to the return of the molecule from a high-energy state to stable or ground state as introduction of cis-bonds twists and contracts the molecule making it energy richer and more unstable. Autooxidation of lycopene is irreversible and will lead to fragmentation of the molecule, producing acetone, methylheptenone, laevulinic aldehyde, and probably glycoxal, also, which causes apparent color loss, and typically hay or grass-like odors evolve (Srivastava and Srivastava, 2015). The lycopene stability is around 0.2 g/L at room temperature (Machmudah et al., 2012b).

6.2  Thermal Processing Georgé et al. (2011) studied lycopene stability during high-temperature processing, including cooking, concentration, and dehydration, to prepared tomato products like paste, tomato ketchup, tomato juice. Degradation of lycopene through these methods has typically been recorded and the degree of degradation was reliant on the kind of treatment, the temperature,

Lycopene: A Natural Red Pigment  439 the time, and the presence of oxygen and light. Along with these, the lycopene stability within various storage environments has been explored in various tomato products and even though lycopene losses were noted in all of these, the degradation rate was found to be reliant not only on storage environments, like light, temperature, water activity, and oxygen, but also on the nature of the product (solid or liquid state and microenvironment). Srivastava and Srivastava (2015) and Sharma and Le Maguer (1996) described noteworthy losses of lycopene in serum-free tomato pulp samples, when treated with a high temperature (100°C) in the presence of oxygen, with or without light. Gross (2012) recorded approximately 1%–2% loss in lycopene when tomato juice was heated at 100°C for 7 min. He also observed 57% loss of lycopene, when tomato juice was concentrated using heat. According to the Boskovic (1979), up to 20% reduction in lycopene content in dehydrated tomato products was observed during processing and extended storage. Namitha and Negi (2010) revealed nonsignificant loss of lycopene when tomato halves were dried at 80°C, while significant loss (12%) was recorded at 110°C. Many researchers documented that thermal treatments produced isomerization of lycopene (Schierle et al., 1997; Shi et al., 1999; Stahl and Sies, 1992). Conversely, other investigators have recently specified that general heating does not show any substantial losses of lycopene or a shift in the circulation of cislycopene isomers (Namitha and Negi, 2010; Nguyen and Schwartz, 1999). A great deal of evidence remnants to be congregated on the thermal behavior of lycopene before conclusive solutions can be provided with respect to its stability. Ironically, in the case of lycopene, food processing is in fact a value-added step, in that more lycopene becomes bioavailable following thermal treatment due to the thermally induced rupture of cell walls (Stahl and Sies, 1992; Wojcik et al., 2010). Many researchers have studied the loss of lycopene during heating or drying of tomato products. Nonetheless, the lack of documentary evidence is available on the effect of heating the lycopene content throughout spray-drying of tomato concentrate. Few researchers are only concerned with stability of lycopene during storage of dried tomato powder manufactured with various drying methods (Anguelova and Warthesen, 2000a,b; Lovric et al., 1970; Purkayastha et al., 2013). The lycopene content of some processed tomato products are presented in Table 13.4. Table 13.4: Lycopene content of some processed tomato products. Tomato Products

Lycopene Content (µg/g of Weight)

Cooked tomato Tomato sauces Tomato juice Tomato paste Tomato powder Ketchup Tomato soup

37 62 50.0–116.0 54.0–500.0 1126.3–1264.9 99.0–134.4 79.9

Source: Singh, P., Goyal, G.K., 2008. Dietary lycopene: its properties and anticarcinogenic effects. Compr. Rev. Food Sci. Food Saf. 7, 255–270.

440  Chapter 13

6.3 Ultrasonication Use of ultrasound has been deliberated in food processing, particularly in washing, drying, homogenizing, and sanitization of foodstuffs and in the extraction of constituents from fruit and vegetables (Cucheval and Chow, 2008; Fernandes et al., 2008; Riera et al., 2004; Rodrigues et al., 2008; Tiwari et al., 2009). The drying rate can be increased by the application of ultrasound, extractability, superior homogenizing of milk and juices, but it may also affect the quality of food products. In food and food products various changes like flavor, color, viscosity, and chemical composition have been documented. Most vicissitudes are connected to the formation of free radicals throughout ultrasound application and to the restricted increase in temperature, resulting in bubble collapse while sonocavitation (GarciaNoguera et al., 2010; Pingret et al., 2013). Substantial loss of lycopene was noted at 60°C and ultrasound power densities ranging from 1000 to 3000 W/L. When the sample treated with ultrasound power density of 2000 W/L, highest degradation of lycopene (51%) was noted, whereas lesser degradation was noted at higher and lower power density. Low power densities (55–63 W/L) did not show any consequence on lycopene levels at the same temperature. The rise in lycopene content at high ultrasound power densities might be connected to the intensification in the bioavailability of lycopene. Ultrasound processing may improve the bioavailability of lycopene by breaking down cell walls, which weakens the bond between lycopene and the fruit tissue (Srivastava and Srivastava, 2015).

6.4 Light Heat and light are the two major factors that highly affect the food quality in processing and material handling procedure. Lycopene content in tomato-based food products can be considered as quality index. Shi et al. (2008) studied the effect of visible light exposure of different intensities provided by an adjustable daylight lamp in a controlled environmental chamber at 5 ± 1°C for 2–12 days. According to this study no significant difference was found for total and all-trans-lycopene during light exposure. The cis-isomer lycopene reduction was statistically significant and the percent changes were 30% lower than the untreated samples. Thus, it can be said that cis-isomer lycopene is less stable than alltrans-lycopene under the light exposure. Lycopene stability was studied using standard lycopene liquefied in hexane and illuminating at 2000–3000 lx at 25°C for 6 days (Lee and Chen, 2002). As there was increase in illumination time, all-trans-lycopene was decreased. There were 94% losses reported in total lycopene after a 144 h exposure time. There was an increasing trend observed at the beginning of 5-cis-lycopene level, whereas after 2 h of illumination a decreasing trend was recorded. Parallel observation was recorded for 9-cis, 13-cis, and 15-cis-lycopene, which showed that isomerization and degradation of lycopene and its cis-isomers might progress concurrently. An improved level of mono-cis-lycopene was observed possibly due to the change of all-trans-lycopene to mono-cis-lycopene after which

Lycopene: A Natural Red Pigment  441 a reduction could occur due to the change of mono-cis form to another cis-form through intermediate all-trans-lycopene or undergo degradation. Shi et al. (2002) studied the effect of irradiation on canola oil at 2010 (outdoor), 900, 650, and 140 (indoor) µmol/m2/s for 1–6 days and found a decrease in the amount of total and trans-lycopene and an increase in the amount of cis-isomers. In this experiment the results showed that irradiation caused more loss in total lycopene than heating treatments at 25, 100, and 180°C while the rate of formation of cisisomers was lower in both the experiments.

6.5 Oxygen Oxygen atoms with unpaired electrons exert tremendous power on other atoms’ electrons and may tear other molecules apart in order to get one. The loss of an electron is called oxidation, because whenever an electron is lost, it is usually to oxygen. As discussed earlier, heat, light, and oxygen are three potent factors leading to the degradation of carotenoids and loss of their biological activity, which is very important from a health point of view. Wong and Bohart (1957) studied the stability of lycopene in different packaging environments and found that air-packed tomato juice powder retained the lowest lycopene levels compared with CO −2 , N −2 , or vacuum-packed samples. According to Sharma and Le Maguer (1996), vacuum and dark storage combinations gave the lowest lycopene loss in tomato pulp. Ax et al. (2003) studied the stability of lycopene by dissolving it in the oil phase of oil-in-water emulsions, under oxygen saturation or oxygen-free conditions. At 25°C, about 25% of lycopene was degraded within 30 h in the oxygen-removed emulsions, whereas a lycopene loss of about 80% was found for oxygen saturation. Lycopene destabilization was about 3 times higher in the presence of oxygen than under inert conditions. Several authors studied the effect of oxygen removal to a certain extent and complete exclusion of oxygen on the stability of lycopene. Removal of oxygen in water by flushing the system with nitrogen produced no encourageable results. However, complete exclusion of oxygen using enzymes, such as glucose oxidase, led to significant improvement in lycopene stability (Ribeiro et al., 2003). Nitrogen or argon head space can also be provided to improve the stability of lycopene by keeping the exposure to atmospheric oxygen to a minimum (Nguyen and Schwartz, 1999).

6.6 Dehydration Lycopene is important not only because of the color it imparts but also because of the recognized health benefits associated with its presence. Degradation of lycopene not only affects the attractive color of the products but also their nutritive value. Dehydration is an important and commonly used method, which increases the shelf life, and the product can be stored for longer periods. Dehydration is defined as “the application of heat under controlled conditions to remove the majority of water normally present in the food by evaporation.”

442  Chapter 13 Loss of lycopene in tomato pulp during spray-drying with constant feed rate, feed temperature and pressure with different flow rates of drying air and air inlet temperature of the compressed air was studied by Goula et al. (2006). The results revealed that the kinetics of lycopene degradation follows a first-order reaction with a reaction rate constant dependent on product moisture content, in addition to temperature. Loss of lycopene was recorded in ranging from 8.07% to 20.93%, when spray-drying of tomato pulp was performed (Goula and Adamopoulos, 2005). Data analysis of the experiment produced associations among lycopene retention and the adjustable working circumstances. Lycopene loss increased with an increase in air inlet temperature and compressed air flow rate. The extent of a degradation reaction in a food during drying is mainly controlled by its temperature and for all the reaction types, the rate constant is a function of temperature as controlled by the apparent energy of activation. (Goula and Adamopoulos, 2005). The decrease in lycopene content reported here was due to an actual degradation of lycopene, rather than to a progressive conversion from the all-translycopene to a less strongly colored, less intensely absorbing cis-form. The rate of dehydration is predisposed by temperature, as well as by moisture content or in other arguments; it is affected by water activity, light, and dissolved oxygen. Cernisev (2010) studied the influence of product moisture and temperature on the color change due to nonenzymatic browning in tomato quarters during drying at various temperatures in the range of 50–90°C. Tomato samples, which were dried at 80 and 90°C and had the shortest drying time, exhibited sensory quality attributes that were strongly modified and showed signs of deterioration, while tomatoes dried at 50 and 60°C had intensive red color without signs of browning. Although the sample dried to final moisture at 50°C received the highest rating for appearance, it was not significantly different from that dried at 60°C. Tomatoes dried to final moisture at low-temperature conditions were rated superior in sensory quality attributes to those dried at high temperature. Chou and Chua (2001) also explained that reduced quality of food products because of browning effects is mainly due to the thermal effect of the drying. It was previously suggested that the first stage of lycopene degradation during drying and storage of tomato powders is the reversible isomerization of all-trans-lycopene to less colored, more oxidizable cis-isomers (Boskovic, 1979). Anguelova and Warthesen (2000a,b) also found that cis-lycopene isomers, while present at low levels in raw tomatoes, may have increased during spray-drying of the powders. A study by Shi et al. (1999) showed that cis-isomers were not detected in the fresh tomato samples, but a significant increase was found in the cis-isomers with simultaneous decrease in the all-trans-isomers during the osmotic-vacuum-drying, vacuum-drying, and air-drying dehydration to produce dehydrated tomato samples. According to Nguyen and Schwartz (1999) dehydration of tomatoes at mild temperatures does not usually cause significant losses in total lycopene content, but the conversion of trans- to cis-isomers always occurs in the dehydrated products. Demiray et al. (2013) studied the effect of partial dehydration to obtain a product possessing final moisture content of 25% using a forced air oven at 40, 60, and 80°C for different lengths of treatment. The highest value of lycopene was found in the dehydrated sample at 80°C

Lycopene: A Natural Red Pigment  443 (76.4 mg/100 g dry matter), while the lowest value was found at 40°C (58.5 mg/100 g dry matter), confirming that in these samples lycopene was damaged by the length of the drying process. Nevertheless, investigation performed by Shi et al. (1999) indicated substantial loss in lycopene content in the dehydration of tomato products. Different processing methods like cooking, cooling, and canning do not usually cause large changes in total lycopene content, but maximum lycopene can be rehabilitated from the all-trans-form into the cis-isomer. According to the conclusions of Wojcik et al. (2010) and Stahl and Sies (1992), lycopene is constant throughout heating and industrial treatment, and it also recovers bioavailability of lycopene.

7  Health Benefits of Lycopene Recently, carotenoids particularly lycopene were proven for possible anticancer properties not only the scientific literature but also epidemiological data. The strong documentation related to evidence is found in different organizations like US National Research Council of the Academy of Sciences (1989), the NCI (1987), and the World Cancer Research Fund, the American Institute for Cancer Research (1997), and these organizations have been suggested to increase consumption of citrus fruits, cruciferous vegetables, green and yellow vegetables, and fruits and vegetables with higher vitamins A and C content, which possibly reduces cancer risk. UKDoH (1999) and the WHO (1990) made similar commendations. Among all carotenoids, one of the furthermost effective antioxidants found was lycopene (Di Mascio et al., 1989; Kong et al., 2010; Miller et al., 1996; Woodall et al., 1997), it possess 10 times higher singlet-oxygen-quenching capability than α-tocopherol and double than that of β-carotene (Di Mascio et al., 1989). It has engrossed consideration owing to its physicochemical and biological properties, particularly associated with its natural antioxidant properties. Cumulative scientific documented reports support the role of lycopene as a micronutrient with significant health benefits because it seems to deliver a protective effect in contradiction to a broad range of epithelial cancers. In the last decade the studies on lycopene and tomatoes has become the hottest topic for nutraceutical and food researchers (Shi and Le Maguer, 2000). Lycopene has increased attention in the past few years as new documentary proof that its protective effect, in contradiction of degenerative diseases, predisposed by free-radical reactions, such as cancer and coronary heart disease (Story et al., 2010). Lycopene was found to be more effective in inhibition of human cancer cell propagation than α and β-carotenes (Levy et al., 1995). Gullett et al. (2010) showed that consumption of fresh tomatoes gives a protective effect against the risk of cancer of the digestive track. In another case study conducted by Colditz et al. (1985), high consumption of tomatoes in the elderly American population had a reduction in mortality from cancer at all sites by 50%. In colon cancer patients, lycopene was also reported to reduce insulin-like growth factor-I levels. High serum levels of insulin-like growth factor-I are connected with a higher risk of colon and other types of cancer, which was also supported by different epidemiological studies (Walfisch et al., 2007).

444  Chapter 13 The health-promoting roles related to biochemical mechanisms of lycopene is not fully understood; however, it possesses the antioxidant properties (Rao and Agarwal, 1999), which has been exposed to be effective protecting from oxidative damage to DNA, protein, and lipids, is assumed to be strongly accountable. Modulation of cell–cell communication is another activity of lycopene (Zhang et al., 1991), embarrassment of cell propagation (Levy et al., 1995), and confrontation to bacterial contamination may also be involved. Current literatures propose that chronic diseases, including cancer and cardiovascular disease, are related with inflammation and coagulation. Jorge (2001) and Zimmermann et al. (1999) have suggested that inflammatory pathways induce cardiovascular diseases, such as atherosclerosis and other coronary syndromes. The useful properties of certain rehabilitations, like 3-hydroxyl-3-methylglutaryl coenzyme A reductase inhibitors and angiotensin converting enzyme inhibitors, have been accredited in part to the embarrassment of inflammation. Yaping et al. (2003) assessed the antiinflammatory and anticoagulant activities of lycopene using rat studies. Lycopene was provided in the form of oleoresin. To study the antiinflammatory activity, croton oil-induced mouse ear edema model was used, whereas to study the anticoagulant properties the glass slide method was used. A lycopene dose for 4 days decreases swelling of the treated ear with efficacy similar to that of amoxicillin, which is a well-known inflammatory agent. Along with these, coagulation time was also increased by the lycopene. These outcomes recommended the health-promoting roles of lycopene with its antiinflammatory and anticoagulant activities. Scolastici et al. (2007) investigated the antigenotoxic/antimutagenic effects of lycopene in Chinese hamster ovary cells (CHO) treated with hydrogen peroxide, methylmethane sulfonate (MMS), or 4-nitroquinoline-1-oxide (4-NQO). Different concentrations, that is, 10, 25, and 50 µM of 95% lycopene (97%), were verified under three different protocols: earlier, concurrently, and after mutagens treatment. To determine the damage level in DNA, comet and cytokinesis-block micronucleus methods were used. Results exhibited that lycopene lowered the occurrence of micronucleated cells persuaded by the three mutagens. Though, this chemopreventive activity was reliant on the concentrations and treatment calendars used. Analogous results were noted in the comet assay, although some developments of primary DNA injury were noticed when the carotenoid was administered after the mutagens. Their discoveries inveterate the chemopreventive activity of lycopene, and exhibited that this effect ensues under different mechanisms.

7.1  Colorectal Cancer Colorectal adenomas, that is, where a polyp is a precursor for most colorectal cancer, blood levels of lycopene were 35% lower as compared to when the person subjected without polyps (Erhardt et al., 2003). β-Carotene inclined to 25.5% less in blood levels, but this difference was nonsignificant according to investigators. Finally the researchers concluded that only little concentration of plasma lycopene (less than 70 µg/L) and smoking augmented

Lycopene: A Natural Red Pigment  445 the probability of colorectal adenomas, but the intensification in risk was moderately considerable: lower concentration of lycopene augmented risk by 230% whereas in smoking it increased by 302%.

7.2  Prostate Cancer The role of nourishment and nutritional supplements in the growth and advancement of prostate cancer characterizes gradually recurrent topic of conversation (Barber and Barber, 2002). Consciousness among relations between tomato products, lycopene, and health outcomes are gradually increased among the public and the biomedical community. Researchers from various disciplines starting with epidemiology, clinical medicine, nutrition, agriculture, and molecular and cell biology have been reported in their reports about providing fascinating data signifying that the carotenoid particularly lycopene and tomato products might be responsible for prevention of cancer, lowering the chances of cardiovascular disease, and preventive the morbidity or mortality of other chronic diseases (Miller et al., 2002). Carotenoids might be reacting with free radicals of oxygen, whichever transfer of the electron, which is unpaired by keeping carotenoid in an enthusiastic in triplet state, the additional energy existence degenerate as heat, or by “bleaching” of the carotenoid. The former leaves the carotenoid intact and therefore is able to be involved in numerous cycles of free-radical scavenging, and the latter results in decomposition of the carotenoid. Providentially, it is the previous that preponderates, and the efficacy of this procedure appears to be connected with number of double bonds present in the carotenoid structure. Attention has been increased in lycopene, in specific, as it has a greater amount of double bonds and thus has been originated to be the greatest powerful scavenger of free radicals of oxygen all the carotenoids (Miller et al., 1996; Rao et al., 2003). Lycopene not only possesses oxygen-free radicals scavenge activity, but also it reacts with reactive other oxygen species that is, nitrogen dioxide and hydrogen peroxide (Bohm et al., 1995; Woodall et al., 1997) and thus it protect cells from oxidative damage. Fascinatingly, efficiency of lycopene in scavenging for nitrogen dioxide was twice than that of β-carotene (Bohm et al., 1995; Tinkler et al., 1994; Woodall et al., 1997). Lycopene has also been confirmed to have additional conceivable anticancer belongings predominantly with respect to modulation of intercellular communication and modifications in intracellular gesturing pathways (Stahl and Sies, 1996). These comprise directive in intercellular opening of joints (Zhang et al., 1992), an upsurge in cellular discrepancy (Bankson et al., 1991), and modifications in phosphorylation of certain regulatory proteins (Matsushima-Nishiwaki, 1995). It is recognized that concerning the role or certainly the importance of these effects in vivo; though, lycopene has been established to be expressively more efficient than any carotene in constraining insulin-like growth factor type 1 encouraged propagation of a number of cancer cell lines (Levy et al., 1995) and reduced in incidence of both unprompted and chemically persuaded mammary tumors in animal models (Nagasawa et al., 1997; Sharoni

446  Chapter 13 et al., 1997). In prostate cancer, specifically, a study has established embarrassment of cell line propagation in the presence of physiological concentrations of lycopene along with αtocopherol (Pastori et al., 1998). Substantial amounts of lycopene are found in the prostate glands of human beings, and many current reviews recommend that in humans with a high amount of blood lycopene involve a subordinate chance of prostate carcinoma (Clinton, 1999). In a Harvard Health Experts Continuation Study, suggested interrelationships among different carotenoids intake, retinal, fruits and vegetables, and the probability of prostate cancer (Giovannucci et al., 1995). They also concluded that intake of fresh tomatoes, tomato sauce, and pizza, those are major sources of dietary lycopene, and predominately reduce the occurrence of prostate cancer. Earlier studies recommended that the accrued human epidemiological confirmation designated that nourishments extraordinary in tomatoes might decrease the chances of emergent cervical, colon, esophageal, rectal, and stomach cancers (Batieha et al., 1993; Bjelke, 1974; CookMozaffari et al., 1979; Giovannucci, 2002; Potischman et al., 1994; Ramon et al., 1993; Tajima and Tominaga, 1985). Pizza has been constructively correlated to lower the chances of prostate cancer in North America. Revealing evidences are accessible on sex hormone-related cancer sites. Silvano et al. (2006) revealed the intake of pizza on the chances of breast, ovarian, and prostate cancers by analyzing reports of three hospital-based case-control studies performed in Italy between 1991 and 2002. The study encompassed with 2569 women with breast cancer, 1031 with ovarian cancer, 1294 men with prostate cancer, and a total of 4864 controls. These were compared with nonpizza consumers, the multivariate odds ratios for eaters were 0.97 [95% confidence interval (CI) 0.86–1.10] for breast, 1.06 (95% CI 0.89–1.26) for ovarian, and 1.04 (95% CI 0.88–1.23) for prostate cancer. Consistent approximations for unvarying consumers (more than 1 portion per week) were 0.92 (95% CI 0.78–1.08), 1.00 (95% CI 0.80–1.25), and 1.12 (95% CI 0.88–1.43), respectively. The consequences do not demonstrate a related role of pizza on the risk of sex hormone-related cancers. The alteration with designated studies from North America recommends that nutritional and existent associates of pizza eating vary among dissimilar inhabitants and community. Tomatoes have been shown to be obliging in decreasing the risk of prostate cancer. A 14-month study conducted by Boileau et al. (2003) underscores the importance of a healthy, whole-food, tomato-rich diet in the prevention of prostate cancer. When the rats nourished with lycopene-enrich food and treated with N-methyl-nitrosourea (a carcinogen) and testosterone to encourage prostate cancer had analogous risk of death from prostate cancer as that of rats supplemented with normal food. Whereas, when rats fed with whole tomato powder there were 26% less probability to die by prostate cancer. The study concluded that 80% of the group fed with normal diet and 72% of the rats fed lycopene had capitulated to prostate cancer, whereas only 62% of the rats nourished with whole tomato powder died.

Lycopene: A Natural Red Pigment  447 Many scientists showed that tomatoes contain not only the lycopene but also contain various protective phytochemicals, and recommended that the lycopene occurred in human prostate tissue and the blood of animals and humans who remain free of prostate cancer might designate acquaintance to higher amounts of lycopene, as well as other constituents working in collaboration with it. Etminan et al. (2004) conducted 21 metaanalysis studies and endorses that consumption of tomatoes, specifically cooked tomatoes, gives safeguards toward prostate cancer. After the compilation of data from 21 studies, an 11% reduction in chances of prostate cancer was observed in those who consumed the highest amount of raw tomatoes, whereas a 19% reduction was recorded in those who consumed cooked tomato products. Though the epidemiological confirmation of the role of lycopene in prevention of cancer is convincing, this role remains to be confirmed. There are limited human interference trials examining the significance of lycopene in reduction of cancer risk. Various scientists have explored the possessions of tomato or tomato product (lycopene) supplementation on oxidative damage to lipids, proteins, and DNA (Agarwal and Rao, 1998; Pool-Zobel et al., 1997; Rao and Agarwal, 1998). A primary report has specified that tomato extract supplementation in the form of oleoresin pills lowers the levels of prostate-specific antigen in patients with prostate cancer (Kucuk et al., 2002).

7.3  Pancreatic Cancer Pancreatic cancer is considered one of the deadliest cancers and developments so quickly that an individual with the disease who is contributing to studies die even earlier than they were interviewed can be accomplished—so the reimbursements noted in the following study of a nourishment rich in tomatoes and tomato-based products are particularly substantial. Nkondjock et al. (2005) carried their study in Canada in which 462 persons with pancreatic cancer were age- and gender-matched with 4721 persons free from disease. Afterward, tuning for age, province, body mass index (BMI), smoking, educational achievement, nutritional folate, and total caloric consumption, the data indicated that the people consuming the lycopene, a carotenoid providing mostly by tomatoes, had a 31% decrease in their risk of pancreatic cancer. Among nonsmoker persons and with nourishments richest in β-carotene or total carotenoids reduced the chances of pancreatic cancer by 43 and 42%, respectively. The investigators recognized the exclusive mechanism through which lycopene safeguards against cancer: triggering cancer-preventive phase II enzymes. When the investigators nurtured breast and liver cancer cells with lycopene, the carotenoid triggered the manufacture and activity of the phase II detoxification enzymes [NAD(P)H: quinone oxidoreductase (NQ01) and glutamylcysteine synthetase (GCS)]. Lycopene ramped up manufacture and activity of these protective enzymes by causing the appearance of a correspondent gene called luciferase that then triggered the “antioxidant response element” in other genetic factors that encode

448  Chapter 13 the enzymes, thus producing the genes to direct amplified enzyme production. In relation to other carotenoids, comprising β-carotene, astaxanthin, and phytoene, did not show this effect. Because much epidemiological indication designates that lycopene acts significantly with other phytochemicals to give tomatoes their protecting belongings, and new research has shown that consumption of tomato products is more effective than consumption of lycopene alone in preventing cancer, the investigators revealed that other carotenoids encourage phase II enzymes via different pathways from that used by lycopene.

7.4  Coronary Heart Diseases Tomato lycopene may also provide benefits for cardiac diseases. Epidemiological studies have also reinforced the assumption that eating heat-processed tomatoes might decrease the chances of coronary heart diseases as the lycopene delays inactively with DNA damage due to oxidation and low-density lipoproteins (Clinton, 1998; Diaz et al., 1997; Gester, 1997; Hadley et al., 2003; Ojima et al., 1993; Weisburger, 1998). Lycopene’s capability to work as an antioxidant and free-radical scavenger that are frequently connected with carcinogenesis is possibly a key to the mechanism for its beneficiary belongings on human health (Khachik et al., 1995). Investigators recommended that along with its converse connotation with various cancers, a high intake of lycopene may play a role in cardiac disease preclusion. The study began with the 39,876 middle-aged and older women who were free from cardiac disease and cancer. Throughout the 7 years of the study, women who had eaten lycopene-rich foods (tomato-based products, including tomatoes, tomato juice, tomato sauce, and pizza) for 7–10 times each week were found to have a 29% fewer chance of CVD as compared to women consuming less than 1.5 servings of tomato products per week. Women who consumed more than 2 servings weekly with the oil-based tomato products, predominantly tomato sauce and pizza, showed better results, a 34% lower risk of CVD. Other research performed in Europe also proposed that liking raw tomatoes or tomato products like sauce or paste many times in a week is a delightful way to protect the cardiac system. Visioli et al. (2003) described that when a group of 12 healthy women consumed tomato products, which were fed with 8 mg of lycopene every day for a 3 week period, their LDL cholesterol was reduced by a considerable amount vulnerable to oxidation of free radicals, the first stage in the construction of atherosclerotic commemoration development and a main risk factor for cardiac disease. Lipophilic constituents restricted in tomatoes can avoid cardiac diseases by controlling the atherogenic progressions in vascular endothelium arbitrated by oxidized low-density lipoproteins (LDLs). Balestrieri et al. (2004) explored that lycopene in connotation with α-tocopherol or tomato lipophilic extracts improves acyl-platelet-activating influence by biosynthesis in endothelial cells throughout oxidative stress. Lycopene provides protection in contradiction to oxidative stress and was also exemplified when human skin was exposed to UV light. Lycopene was originating to be especially

Lycopene: A Natural Red Pigment  449 demolished compared to β-carotene, signifying a more active or defensive role (Ribayo-Mercado et al., 1995). In a multicenter case-control study, the correlation among antioxidant status and critical myocardial infarction was assessed (Kohlmeier et al., 1997). Subjects were enlisted from 10 European nations to exploit the erraticism in contact within the research. Adipose tissue antioxidant levels, which are pointers of long-term acquaintance than blood antioxidant levels, were used as indicators of antioxidant status. Surgery specimens of adipose tissue were taken unswervingly after the infarction and were analyzed for numerous carotenoids. After alteration for a range of dietary variables, individually lycopene levels were found to be defensive. A study from Johns Hopkins University, Baltimore, showed that smokers with low levels of circulating carotenoids were at increased risk for succeeding myocardial infarction (Handelman et al., 1996). Inferior blood lycopene levels were correspondingly initiated with greater chances for and death from coronary artery disease in a populace study associating Lithuanian and Swedish cohorts with dissimilar rates of death from coronary artery disease (Kristenson et al., 1997).

References Agarwal, S., Rao, A.V., 1998. Tomato lycopene and low-density lipoprotein oxidation: a human dietary intervention study. Lipids 33 (10), 981–984. Anguelova, T., Warthesen, J., 2000a. Degradation of lycopene, α-carotene, and β-carotene during lipid peroxidation. J. Food Sci. 65 (1), 71–75. Anguelova, T., Warthesen, J., 2000b. Lycopene stability in tomato powders. J. Food Sci. 65 (1), 67–70. Ax, K., Mayer-Miebach, E., Link, B., Schuchmann, H., Schubert, H., 2003. Stability of lycopene in oil-in-water emulsions. Eng. Life Sci. 4, 199–201. Baldermann, S., Ropeter, K., Kohler, N., Fleischmann, P., 2008. Isolation of all-trans lycopene by high-speed counter-current chromatography using a temperature-controlled solvent system. J. Chromatogr. A 1192 (1), 191–193. Balestrieri, M.L., Prisco, R.D., Nicolaus, B., Pari, P., Schiano Moriello, V., Strazzullo, G., Iorio, E.L., Servillo, L., Balestrieri, C., 2004. Lycopene in association with α-tocopherol or tomato lipophilic extracts enhances acyl-platelet-activating factor biosynthesis in endothelial cells during oxidative stress. Free Radic. Biol. Med. 36 (8), 1058–1067. Bankson, D.D., Countryman, C.J., Collins, S.J., 1991. Potentiation of the retinoic acid-induced differentiation of HL-60 cells by lycopene. Am. J. Clin. Nutr. 53 (Suppl.), 13–17. Barba, A.I.O., Hurtado, M.C., Mata, M.C.S., Ruiz, V.F., de Tejada, M.L.S., 2006. Application of a UV-vis detection-HPLC method for a rapid determination of lycopene and beta-carotene in vegetables. Food Chem. 95 (2), 328–336. Barber, N.J., Barber, J., 2002. Lycopene and prostate cancer. Prostate Cancer Prostatic Dis. 5, 6–12. Barrie, T., 1988. Analytical and preparative chromatography of tomato paste carotenoids. J. Food Sci. 53 (3), 954–959. Batieha, A.M., Armenian, H.K., Norkus, E.P., Morris, J.S., Spate, V.E., Comstock, G.W., 1993. Serum micronutrients and subsequent risk of cervical cancer in a population-based nested case-control study. Cancer Epidermiol. Biomark. Prev. 2 (4), 335–339. Bauernfeind, J.C., 1981. Natural food colors. In: Bauernfeind, J.C. (Ed.), Carotenoids as Colorants and Vitamin A Precursors. Academic Press, New York, NY, pp. 1–45. Bjelke, F., 1974. Case-control study in Norway. Scand. J. Gastroenterol. 9 (Suppl. 31), 42–48. Bohm, F., Tinkler, J.H., Truscott, T.G., 1995. Carotenoids protect against cell membrane damage by the nitrogen dioxide radical. Nat. Med. 1, 98–99.

450  Chapter 13 Boileau, T.W., Liao, Z., Kim, S., Lemeshow, S., Erdman, Jr., J.W., Clinton, S.K., 2003. Prostate carcinogenesis in N-methyl-N-nitrosourea (NMU)-testosterone-treated rats fed tomato powder, lycopene, or energy-restricted diets. J. Natl. Cancer Inst. 95 (21), 1578–1586. Boskovic, M.A., 1979. Fate of lycopene in dehydrated tomato products: carotenoid isomerization in food system. J. Food Sci. 44, 84–86. Britton, G., 2008. TLC of carotenoids. In: Waksmundzka-Hajnos, M., Sherma, J., Kowalska, T. (Eds.), Thin Layer Chromatography in Phytochemistry. CRC Press, New York, NY, pp. 543–573. Calogero, G., Bartolotta, A., Di Marco, G., Di Carlo, A., Bonaccorso, F., 2015. Vegetable-based dye-sensitized solar cells. Chem. Soc. Rev. 44 (10), 3244–3294. Cernisev, S., 2010. Effect on conventional and multistage drying processing on non-enzymatic browning in tomato. J. Food Eng. 96, 114–118. Chandler, L.A., Schwartz, S.J., 1987. HPLC-separation of cis-trans isomers in fresh and processed fruits and vegetables. J. Food Sci. 52, 669–672. Chedea, V.S., Kefalas, P., Socaciu, C., 2010. Patterns of carotenoid pigments extracted from two orange peel wastes (Valencia and navel var.). J. Food Biochem. 34, 101–110. Chemat, F., Grondin, I., Costes, P., 2004. High power ultrasound effects on lipid oxidation of refined sunflower oil. Ultrason. Sonochem. 11, 281–285. Chemat, F., Vian, M.A., Cravotto, G., 2012. Green extraction of natural products: concept and principles. Int. J. Mol. Sci. 13, 8615–8627. Choksi, P.M., Joshi, V.Y., 2007. A review on lycopene: extraction, purification, stability and applications. Int. J. Food Prop. 10 (2), 289–298. Chou, S.K., Chua, K.J., 2001. New hybrid drying technologies for heat sensitive foodstuffs. Trends Food Sci. Technol. 12, 359–369. Choudhari, S.M., Ananthanarayan, L., 2007. Enzyme aided extraction of lycopene from tomato tissues. Food Chem. 102, 77–81. Choudhari, S.M., Ananthanarayan, L., Singhal, R.S., 2009. Purification of lycopene by reverse phase chromatography. Food Bioproc. Technol. 2, 391–399. Choudhary, R., Bowser, T.J., Weckler, P., Maness, N.O., McGlynn, W., 2009. Rapid estimation of lycopene concentration in watermelon and tomato puree by fiber optic visible reflectance spectroscopy. Postharv. Biol. Technol. 52 (1), 103–109. Cinar, I., 2005. Effects of cellulase and pectinase concentrations on the colour yield of enzyme extracted plant carotenoids. Proc. Biochem. 40, 945–949. Clinton, S.K., 1998. Lycopene: chemistry, biology, and implications for human health and disease. Nutr. Rev. 56 (2), 35–51. Clinton, S.K., 1999. The dietary antioxidant network and prostate carcinoma. Cancer 86 (9), 1629–1630. Colditz, G.A., Branch, L.G., Lipnick, R.J., 1985. Increased green and yellow vegetable intake and lowered cancer deaths in an elderly population. Am. J. Clin. Nutr. 41 (1), 32–36. Colle, I., Lemmens, L., Van Buggenhout, S., Van Loey, A., Hendrickx, M., 2010. The effect of thermal processing on the degradation, isomerization and bioaccessibility of lycopene in tomato pulp. J. Food Sci. 75, C753–C759. Cook-Mozaffari, P.J., Azordegan, F., Day, N.E., 1979. Esophageal cancer studies in the Caspian litoral of Iran: results of a case-control study. Br. J. Cancer 39 (5), 292–309. Cucheval, A., Chow, R.C.Y., 2008. A study on the emulsion of oil by power ultrasound. Ultrason. Sonochem. 15, 916–992. Davies, B.H., 1976. Carotenoids. Goodwin, T.W. (Ed.), Chemistry and Biochemistry of Plant Pigments, vol. 2, second ed. Academic Press, New York, NY, pp. 38–165. Davis, A.R., Fish, W.W., Perkins-Veazie, P., 2003. A rapid spectrophotometric method for analyzing lycopene content in tomato and tomato products. Postharv. Biol. Technol. 28 (3), 425–430. De Nardo, T., Shiroma-Kian, C., Halim, Y., Francis, D., Rodriguez-Saona, L.E., 2009. Rapid and simultaneous determination of lycopene and b-carotene contents in tomato juice by infrared spectroscopy. J. Agric. Food Chem. 57, 1105–1112.

Lycopene: A Natural Red Pigment  451 Demiray, E., Tulek, Y., Yilmaz, Y., 2013. Degradation kinetics of lycopene. β-Carotene and ascorbic acid in tomatoes during hot air drying. LWT Food Sci. Technol. 50 (1), 172–176. Di Mascio, P., Kaiser, S., Sies, H., 1989. Lycopene as the most effective biological carotenoid singlet oxygen quencher. Arch. Biochem. Biophys. 274 (2), 532–538. Diaz, M.N., Frei, B., Vita, J.A., Keaney, J.F., 1997. Antioxidants and atherosclerotic heart disease. N. Engl. J. Med. 337 (12), 408–416. Eh, A.L.S., Teoh, S.G., 2012. Novel modified ultrasonication technique for the extraction of lycopene from tomatoes. Ultrason. Sonochem. 19 (1), 151–159. Erhardt, J.G., Meisner, C., Bode, J.C., Bode, C., 2003. Lycopene, β-carotene, and colorectal adenomas. Am. J. Clin. Nutr. 78 (6), 1219–1224. Etminan, M., Takkouche, B., Caamano-Isorna, F., 2004. The role of tomato products and lycopene in the prevention of prostate cancer: a meta-analysis of observational studies. Cancer Epidemiol. Biomark. Prev. 13 (3), 340–345. FDA/IFIC, 1993. Regulation of color additives. FDA/IFIC brochure. Food and Drug Administration, Washington, DC. Fernandes, F.A.N., Gallão, M.I., Rodrigues, S., 2008. Effect of osmotic dehydration and ultrasound pre-treatment on cell structure: melon dehydration. LWT Food Sci. Technol. 41, 604–610. Fish, W.W., Perkins-Veazie, P., Collins, J.K., 2002. A Quantitative assay for lycopene that utilizes reduced volumes of organic solvents. J. Food Comp. Anal. 15, 309–317. Garcia-Noguera, J., Oliveira, F.I.P., Gallão, M.I., Weller, C.L., Rodrigues, S., Fernandes, F.A.N., 2010. Ultrasound-assisted osmotic dehydration of strawberries: effect of pre-treatment time and ultrasonic frequency. Drying Technol. 28, 294–303. Georgé, S., Tourniaire, F., Gautier, H., Goupy, P., Rock, E., Caris-Veyrat, C., 2011. Changes in the contents of carotenoids, phenolic compounds and vitamin C during technical processing and lyophilisation of red and yellow tomatoes. Food Chem. 124 (4), 1603–1611. Gester, H., 1997. The potential role of lycopene for human health. J. Am. Coll. Nutr. 16 (2), 109–126. Giovannucci, E., 2002. A review of epidemiologic studies of tomatoes, lycopene, and prostate cancer. Exp. Biol. Med. 227 (56), 852–859. Giovannucci, E.L., Ascherio, A., Rimm, E.B., Stampfer, M.J., Colditz, G.A., Willett, W.C., 1995. Intake of carotenoids and retinol in relationship to risk of prostate cancer. J. Natl. Cancer Inst. 87 (23), 1767–1776. Giusti, M.M., Wrolstad, R.E., 1996. Radish anthocyanin extract as a natural red colorant for maraschino cherries. J. Food Sci. 61, 688–694. Gordon, A., Cruz, A.P.G., Cabral, L.M.C., de Freitas, S.C., Taxi, C.M.A.D., Donangelo, C.M., de Andrade Mattietto, R., Friedrich, M., da Matta, V.M., Marx, F., 2012. Chemical characterization and evaluation of antioxidant properties of Açaí fruits (Euterpe oleraceae Mart.) during ripening. Food Chem. 133 (2), 256–263. Goula, A.M., Adamopoulos, K.G., 2005. Stability of lycopene during spray drying of tomato pulp. LWT–Food Sci. Technol. 38 (5), 479–487. Goula, A.M., Nikas, V.A., Chatzitakis, P.C., Adampoulos, K.G., 2006. Prediction of lycopene degradation during drying process of tomato pulp. J. Food Eng. 74, 27–46. Gross, J., 2012. Pigments in Vegetables: Chlorophylls and Carotenoids. Springer Science and Business Media, New York, NY. Gullett, N.P., Amin, A.R., Bayraktar, S., Pezzuto, J.M., Shin, D.M., Khuri, F.R., Aggarwal, B.B., Surh, Y.J., Kucuk, O., 2010. Cancer prevention with natural compounds. Semin. Oncol. 37 (3), 258–281. Gutte, K.B., Sahoo, A.K., Ranveer, R.C., 2015. Effect of ultrasonic treatment on extraction and Ω-3 fatty acid of flaxseed oil. OCL 22 (6), D606. Hadley, C.W., Clinton, S.K., Schwartz, S.J., 2003. The consumption of processed tomato products enhances plasma lycopene concentrations in association with reduced lipoprotein sensitivity to oxidative damage. J. Nutr. 133, 727–732. Haiyan, G., Lei, Z., Xiaosong, H., Liying, S., 2003. Study on the extraction and stability of cherry pigment. Food Ferment. Ind. 29 (4), 54–57.

452  Chapter 13 Hakala, S.H., Heinonen, M., 1994. Chromatographic purification of natural lycopene. J. Agric. Food Chem. 42, 1314–1316. Handelman, G.J., Parker, L., Cross, C.E., 1996. Destruction of tocopherols, carotenoids and retinol in human plasma by cigarette smoke. Am. J. Clin. Nutr. 63 (4), 559–565. Haroon, S., 2014. Extraction of lycopene from tomato paste and its immobilization for controlled release. Master’s thesis submitted to The University of Waikato, New Zealand. Hromadkova, Z., Kovacikova, J., Ebringerov’a, A., 1999. Study of the classical and ultrasound-assisted extraction of the corn cob xylan. Ind. Crops Prod. 9, 101–109. Hyman, J.R., Gaus, J., Foolad, M.R., 2004. A rapid and accurate method for estimating tomato lycopene content by measuring chromaticity values of fruit puree. J. Am. Soc. Hortic. Sci. 129 (5), 717–723. Ishida, B.K., Chapman, M.H., 2009. Carotenoid extraction from plants using a novel, environmentally friendly solvent. J. Agric. Food Chem. 57 (3), 1051–1059. Jie, Y., Huimi, G., Meizhen, C., 2002. Study on the physicochemical properties and extraction process of red pigment from He Dong black wheat. Food Ferment. Ind. 28 (11), 12–16. Jinxing, C., Xiufeng, L., Zhaomeng, L., Cuiping, A., 2003. Study on extraction technology of strawberry pigments and its physicochemical properties. Food Ferment. Ind. 29 (5), 69–73. Jorge, P., 2001. Inflammatory pathways in atherosclerosis and acute coronary syndromes. Am. J. Cardiol. 88 (Suppl.), 11K–15K. Karrer, P., Helfenstein, A., Wehri, H., Wettstein, A., 1930. Pflanzenfarbstoffe. XXV. Leber die konstitution des lycopins und carotins. Acta 14, 154–162. Khachik, F., Beecher, G.R., Smith, Jr., J.C., 1995. Lutein, lycopene, and their oxidative metabolites in chemoprevention of cancer. J. Cell Biochem. 22 (4), 236–246. Kohlmeier, L., Clarke, J.D., Gomez-Gracia, E., Martin, B.C., Steck, S.E., Kardinaal, A.F., Ringstad, J., Thamm, M., Masaev, V., Riemersma, R., Martin-Moreno, J.M., Huttunen, J.K., Kok, F.J., 1997. Lycopene and myocardial infarction risk in the EURAMIC study. Am. J. Epidemiol. 146 (2), 618–626. Kong, K.W., Khoo, H.E., Prasad, K.N., Ismail, A., Tan, C.P., Rajab, N.F., 2010. Revealing the power of the natural red pigment lycopene. Molecules 15 (2), 959–987. Koyama, Y., Hosomi, M., Miyata, A., Hashimoto, H., Reames, S.A., 1988. Supplementary and revised assignment of the 7,9-9,9′, 13,13′-, 9,13′-di-cis and 9,9′,13-tri-cis isomers of β-carotene in high performance liquid chromatography using a column of calcium hydroxide. J. Chromatogr. 439, 417–422. Kristenson, M., Zieden, B., Kucinskiene, Z., Elinder, L.S., Bergdahl, B., Elwing, B., 1997. Antioxidant state and mortality from coronary heart disease in Lithuanian and Swedish men: concomitant cross-sectional study of men aged 50. Br. Med. J. 314 (11), 629–633. Kucuk, O., Sarkar, F.H., Djuric, Z., Sakr, W., Pollak, M.N., Khachik, F., Banerjee, M., Bertram, J.S., Wood, Jr., D.P., 2002. Effects of lycopene supplementation in patients with localized prostate cancer. Exp. Biol. Med. 227, 881–885. Kuhn, R., Grundmann, C., 1932. Die konstitution des Lycopins. Ber. Deutsch. Chem. Ges. 65, 1880–1889. Lee, M.T., Chen, B.H., 2002. Stability of lycopene during heating and illumination in a model system. Food Chem. 78, 425–432. Lee, M.J., Park, J.S., Choi, D.S., Jung, M.Y., 2013. Characterization and quantitation of anthocyanins in purplefleshed sweet potatoes cultivated in Korea by HPLC-DAD and HPLC-ESI-QTOF-MS/MS. J. Agric. Food Chem. 61 (12), 3148–3158. Lesellier, E., Marty, C., Berset, C., Tchapla, A., 1989. Optimization of the isocratic non aqueous reverse phase (NARP) HPLC separation of trans/cis-α- and β-carotenes. J. High Resolut. Chromatogr. 12, 447–454. Levy, J., Bosin, E., Feldman, B., Giat, Y., Miinster, A., Danilenko, M., Sharoni, Y., 1995. Lycopene is a more potent inhibitor of human cancer cell proliferation than either alpha-carotene or beta-carotene. Nut. Cancer 24 (3), 257–266. Li, H., Pordesimo, L., Weiss, J., 2004. High intensity ultrasound assisted extraction of oil from soybeans. Food Res. Int. 37, 731–738. Longo, L., Vasapollo, G., 2006. Extraction and identification of anthocyanins from Smilax aspera L. berries. Food Chem. 94, 226–231.

Lycopene: A Natural Red Pigment  453 Lovric, T., Sablek, Z., Boskovic, M., 1970. Cis-trans isomerisation of lycopene and colour stability of foam-mat dried tomato powder during storage. J. Sci. Food Agric. 21, 641–647. Machmudah, S., Winardi, S., Sasaki, M., Goto, M., Kusumoto, N., Hayakawa, K., 2012a. Lycopene extraction from tomato peels by-product containing tomato seed using supercritical carbon dioxide. J. Food Eng. 108 (2), 290–296. Machmudah, S., Zakaria, Winardi, S., Sasaki, M., Goto, M., Kusumoto, N., Hayakawa, K., 2012b. Lycopene extraction from tomato peels by-product containing tomato seed using supercritical carbon dioxide. J. Food Eng. 108, 290–296. Mason, T.J., Chemat, F., Vinatoru, M., 2011. The extraction of natural products using ultrasound or microwaves. Curr. Org. Chem. 15, 237–247. Matsushima-Nishiwaki, R., 1995. Suppression by carotenoids of microcystin-induced morphological changes in mouse hepatocytes. Lipids 30 (4), 1029–1034. Miller, E.C., Hadley, C.W., Schwartz, S.J., Erdman, Jr., J.W., Boileau, T.W., Clinton, S.K., 2002. Lycopene, tomato products, and prostate cancer prevention: have we established causality? Pure Appl. Chem. 74 (8), 1435–1441. Miller, N.J., Sampson, J., Candeiar, L.P., Bramley, P.M., Rice-Evans, C.A., 1996. Antioxidant activities of carotenes and xanthophylls. FEBS Lett. 384 (3), 240–246. Mol, J., Jenkins, G., Schäfer, E., Weiss, D., 1996. Signal perception, transduction and gene expression involved in anthocyanin biosynthesis. Crit. Rev. Plant Sci. 155 (6), 525–557. Moss, G.P., Weedon, B.C.L., 1976. Chemistry of the carotenoids. Goodwin, T.W. (Ed.), Chemistry and Biochemistry of Plant Pigments, vol. 1, second ed. Academic Press, New York, NY, pp. 149–224. Naderi, N., Ghazali, H.M., Hussin, A.S.M., Amid, M., Manap, M.Y.A., 2012. Characterization and quantification of dragon fruit (Hylocereus polyrhizus) betacyanin pigments extracted by two procedures. Pertanika J. Trop. Agric. Sci. 35 (1), 33–40. Nagasawa, H., Mitamura, T., Sakamoto, S., Yamamoto, K., 1997. Effects of lycopene on spontaneous mammary tumor development in SHN virgin mice. Anticancer Res. 15 (1), 118–123. Namitha, K., Negi, P.S., 2010. Chemistry and biotechnology of carotenoids. Crit. Rev. Food Sci. Nutr. 50 (8), 728–760. Nayak, C.A., Srinivas, P., Rastogi, N.K., 2010. Characterisation of anthocyanins from Garcinia indica Choisy. Food Chem. 118, 719–724. Nazir, N., Ramli, N., Mangunwidjaja, D., 2009. Extraction, trans-esterification and process control in from Jatropha curcas. Eur. J. Lipid Sci. Technol. 111, 1185–1200. Nguyen, M.L., Schwartz, S.J., 1999. Lyopene: chemical and biological properties. Food Technol. 53 (2), 38–45. Nkondjock, A., Ghadirian, P., Johnson, K.C., Krewski, D., 2005. Dietary intake of lycopene is associated with reduced pancreatic cancer risk. J. Nutr. 135 (3), 592–597. Nyambaka, H., Ryley, J., 1996. An isocratic reversed-phase HPLC separation of the stereoisomers of the provitamin A carotenoids (α- and β-carotene) in dark green vegetables. Food Chem. 55 (1), 63–72. Ojima, F., Sakamoto, H., Ishiguro, Y., Terao, J., 1993. Consumption of carotenoids in photosensitized oxidation of human plasma and plasma low-density lipoprotein. Free Radic. Biol. Med. 15 (4), 377–384. Pastori, M., Pfander, H., Boscoboinik, D., Azzi, A., 1998. Lycopene in association with α-tocopherol inhibits at physiological concentrations proliferation of prostate cancer cells. Biochem. Biophys. Res. Commun. 250 (4), 582–585. Pingret, D., Tixier, A.S.F., Chemat, F., 2013. Degradation during application of ultrasound in food processing: a review. Food Control 31, 593–606. Pool-Zobel, B.L., Bub, A., Muller, H., Wollowski, I., Rechkemmer, G., 1997. Consumption of vegetables reduces genetic damage in humans: first result of a human intervention trial with carotenoid-rich foods. Carcinogenesis 18 (9), 1847–1850. Potischman, N., Hoover, R.N., Brinton, L.A., Swanson, C.A., Herrero, R., Tenorio, F., de Britton, R.C., Gartan, E., Reeves, W.C., 1994. The relations between cervical cancer and serological markers of nutritional status. Nutr. Cancer 21 (2), 193–201.

454  Chapter 13 Prasad, A.K., Mishra, P.C., 2014. Modeling the mechanism of action of lycopene as a hydroxyl radical scavenger. J. Mol. Model. 20 (5), 1–10. Purkayastha, M.D., Nath, A., Deka, B.C., Mahanta, C.L., 2013. Thin layer drying of tomato slices. J. Food Sci. Technol. 50 (4), 642–653. Quackenbush, F.W., 1987. Reverse phase HPLC separation of cis and trans-carotenoids and its application to β-carotenes in food materials. J. Liq. Chromatogr. 10, 643–653. Ramon, J.M., Serra, L., Como, C., Oromi, J., 1993. Dietary factors and gastric cancer risk. Cancer 71 (6), 1731–1735. Ranveer, R.C., Sahoo, A.K., 2015. Effects of cellulase and pectinase combinations on the recovery of lycopene from tomato processing industry waste. Agro Food Ind. Hi Tech 26 (1), 46–49. Ranveer, R.C., Samsher, N.P., Sahoo, A.K., 2013. Effect of different parameters on enzyme-assisted extraction of lycopene from tomato processing waste. Food Bioprod. Process. 91 (4), 370–375. Rao, A.V., Agarwal, S., 1998. Bioavailability and in vivo antioxidant properties of lycopene from tomato products and their possible role in the prevention of cancer. Nutr. Cancer 31 (3), 199–203. Rao, A.V., Agarwal, S., 1999. Role of lycopene as antioxidant carotenoid in the prevention of chronic diseases: a review. Nutr. Res. 19 (2), 305–323. Rao, A.V., Agarwal, S., 2000. Role of antioxidant lycopene in cancer and heart disease. J. Am. College Nutr. 19 (3), 563–569. Rao, L.G., Guns, E., Rao, A.V., 2003. Lycopene: its role in human health and disease. Agro Food Ind. Hi Tech 7–8, 25–30. Rebecca, L.J., Sharmila, S., Das, M.P., Seshiah, C., 2014. Extraction and purification of carotenoids from vegetables. J. Chem. Pharm. Res. 6 (4), 594–598. Ribayo-Mercado, J.D., Garmyn, M., Gilchrest, B.A., Russel, R.M., 1995. Skin lycopene is destroyed preferentially over β-carotene during ultraviolet irradiation in humans. J. Nutr. 125 (7), 1854–1859. Ribeiro, H.S., Ax, K., Schubert, H., 2003. Stability of lycopene emulsions in food systems. J. Food Sci. 68, 2730–2734. Riera, E., Golás, Y., Blanco, A., Gallego, A., Blasco, M., Mulet, A., 2004. Mass transfer enhancement in supercritical fluids extraction by means of power ultrasound. Ultrason. Sonochem. 11, 241–244. Rodrigues, S., Pinto, G.A.S., Fernandes, F.A.N., 2008. Optimization of ultrasound extraction of phenolic compounds from coconut (Cocos nucifera) shell powder by response surface methodology. Ultrason. Sonochem. 15, 95–100. Rodriguez-Amaya, D.B., 2010. Quantitative analysis, in vitro assessment of bioavailability and antioxidant activity of food carotenoids: a review. J. Food Comp. Anal. 23 (7), 726–740. Rostagno, M.A., Palma, M., Barroso, C.G., 2003. Ultrasound assisted extraction of soy isoflavones. J. Chromatogr. A 1012, 119–128. Rymbai, H., Sharma, R.R., Srivastav, M., 2011. Biocolorants and its implications in health and food industry: a review. Int. J. Pharm. Tech. Res. 3 (4), 2228–2244. Schierle, J., Bretzel, W., Buhler, J., Faccin, N., Hess, D., Steiner, K., Schuep, W., 1997. Content and isomeric ratio of lycopene in food and human blood plasma. Food Chem. 59, 459–465. Scolastici, C., Alves de Lima, R.O., Barbisan, L.F., Ferreira, A.L., Ribeiro, D.A., Salvadori, D.M.F., 2007. Lycopene activity against chemically induced DNA damage in Chinese hamster ovary cells. Toxicol. Vitro 21, 840–845. Shahid, M., Mohammad, F., 2013. Recent advancements in natural dye applications: a review. J. Clean. Prod. 53, 310–331. Sharma, S.K., Le Maguer, M., 1996. Lycopene in tomatoes and tomato pulp fractions. J. Food Sci. 2, 107–113. Sharoni, Y., Giron, E., Rise, M., Levy, J., 1997. Effects of lycopene-enriched oleoresin on 7,12-dimetheyl-benz [a] anthracene-induced rat mammary tumors. Cancer Detect. Prev. 21 (2), 118–123. Shi, J., Dai, Y., Kakuda, Y., Mittal, G., Xue, S.J., 2008. Effect of heating and exposure to light on the stability of lycopene in tomato puree. Food Control 19 (5), 514–520. Shi, J., Khatri, M., Xue, S.J., Mittal, G.S., Ma, Y., Li, D., 2009. Solubility of lycopene in supercritical CO2 fluid as affected by temperature and pressure. Sep. Purif. Technol. 66 (2), 322–328.

Lycopene: A Natural Red Pigment  455 Shi, J., Le Maguer, M., 2000. Lycopene in tomatoes: chemical and physical properties affected by food processing. Crit. Rev. Food Sci. Nutr. 40 (1), 1–42. Shi, J., Le Maguer, M., Kakuda, Y., Liptay, A., Niekamp, F., 1999. Lycopene degradation and isomerization in tomato dehydration. Food Res. Int. 32, 15–21. Shi, J., Wu, Y., Bryan, M., Maguer, L.M., 2002. Oxidation and isomerization of lycopene under thermal treatment and light irradiation in food processing. Nutraceut. Food 7, 179–183. Silvano, G., Renato, T., Cristina, B., Eva, N., Maurizio, M., Silvia, F., Attilio, G., Carlo, L.V., 2006. Pizza consumption and the risk of breast, ovarian and prostate cancer. Eur. J. Cancer Prev. 15 (1), 74–76. Singh, M., Kaur, M., Silakari, O., 2014. Flavones: an important scaffold for medicinal chemistry. Eur. J. Med. Chem. 84, 206–239. Srivastava, S., Srivastava, A.K., 2015. Lycopene; chemistry, biosynthesis, metabolism and degradation under various abiotic parameters. J. Food. Sci. Technol. 52 (1), 41–53. Stahl, W., Schwartz, W., Sunqvist, A.R., Sies, H., 1992. Cis-trans isomers of lycopene and β-carotene in human serum and tissues. Arch. Biochem. Biophys. 294, 173–177. Stahl, W., Sies, H., 1992. Uptake of lycopene and its geometrical isomers is greater from heat-processed than from unprocessed tomato juice in humans. J. Nutr. 122, 2161–2166. Stahl, W., Sies, H., 1996. Lycopene: a biologically important carotenoid for human? Arch. Biochem. Biophys. 336, 1–9. Story, E.N., Kopec, R.E., Schwartz, S.J., Harris, G.K., 2010. An update on the health effects of tomato lycopene. Annu. Rev. Food Sci. Technol. 1, 189–2010. Tajima, K., Tominaga, S., 1985. Dietary habits and gastrointestinal cancers: a comparative case control study of stomach and large intestinal cancer in Nagoya. Jpn. J. Cancer Res. 76 (3), 705–716. Thadikamala, S., Doradla, U., Kushala, H., Patsamatla, L.D.S., Devarapalli, K., Pullasi, B., 2009. HPLC method for the determination of lycopene in crude oleoresin extracts. Asian J. Chem. 21 (1), 139–148. Tinkler, J.H., Boehm, F., Schalch, W., Truscott, T.G., 1994. Dietary carotenoids protect human cells from damage. J. Photochem. Photobiol. 26, 283–285. Tiwari, B.K., Muthukumarappan, K., O’Donnell, C.P., Cullen, P.J., 2009. Inactivation kinetics of pectin methylesterase and cloud retention in sonicated orange juice. Innov. Food Sci. Emerg. Technol. 10, 166–171. Tsukida, K., Saiki, K., Takii, T., Koyama, Y., 1982. Separation and determination of cis-trans-β-carotenes by highperformance liquid chromatography. J. Chromatogr. 245, 359–364. Tzia, C., Liadakis, G., 2003. Extraction Optimization in Food Engineering. Marcell Dekker, New York, NY. UKDoH, 1999. Committee on Medical Aspects of Food and Nutrition Policy (COMA) Annual Report, Department of Health, UK. Vagi, E., Simandi, B., Vasarhelyine, K.P., Daood, H., Kery, A., Doleschall, F., Nagy, B., 2007. Supercritical carbon dioxide extraction of carotenoids, tocopherols and sitosterols from industrial tomato by-products. J. Supercrit. Fluids 40 (2), 218–226. Visioli, F., Riso, P., Grande, S., Galli, C., Porrini, M., 2003. Protective activity of tomato products on in vivo markers of lipid oxidation. Eur. J. Nutr. 42 (1), 201–206. Walfisch, S., Walfisch, Y., Kirilov, E., Linde, N., Mintentag, H., Agbaria, R., Sharoni, Y., Levy, J., 2007. Tomato lycopene extract supplementation decreases insulin-like growth factor-I levels in colon cancer patients. Eur. J. Cancer Prev. 16 (4), 298–303. Weisburger, J.H., 1998. Evaluation of the evidence on the role of tomato products in disease prevention. Proc. Soc. Exp. Biol. Med. 218 (2), 140–143. WHO (World Health Organization), 1990. Diet, nutrition and the prevention of chronic disease. Technical Report Series 797. World Health Organization, Geneva. Willstätter, R., Escher, H.H.Z., 1910. Lycopene extracted from tomato. Physiol. Chem. 76, 214–225. Wojcik, M., Burzynska-Pedziwiatr, I., Wozniak, L.A., 2010. A review of natural and synthetic antioxidants important for health and longevity. Curr. Med. Chem. 17 (28), 3262–3288. Wong, D.W.S., 1989. Colorants. Mechanism and Theory in Food Chemistry. AVI, Westport, CT, pp. 147–187. Wong, F., Bohart, S.G., 1957. Observations on the color of vacuum dried tomato juice powder during storage. Food Technol. 5, 293–296.

456  Chapter 13 Woodall, A.A., Lee, S.W.M., Weesie, R.J., Jackson, M.J., Britton, G., 1997. Oxidation of carotenoids by free radicals: relationship between structure and reactivity. Biochim. Biophys. Acta 1336 (1), 33–42. Xi, J., 2006. Effect of high pressure processing on the extraction of lycopene in tomato paste waste. Chem. Eng. Technol. 29 (6), 736–739. Yaping, Z., Wenli, Y.U., Weile, H.U., Ying, Y., 2003. Anti-inflammatory and anti-coagulant activities of lycopene in mice. Nutr. Res. 23, 1591–1595. Zechmeister, L., LeRosen, A.L., Went, F.W., Pauling, L., 1941. Prolycopene, a naturally occurring stereoisomer of lycopene. Proc. Natl. Acad. Sci. USA 27, 468–474. Zhang, L.X., Cooney, R.V., Bertram, J.S., 1991. Carotenoids enhance gap junctional communication and inhibit lipid peroxidation in C3H/10T1/2 cells: relationship to their cancer chemopreventive action. Carcinogenisis 12, 2109–2114. Zhang, L.X., Cooney, R.V., Bertram, J.S., 1992. Carotenoids upregulate conexin 43 gene expression independent of their provitamin A or antioxidant properties. Cancer Res. 52, 5707–5712. Zhang, Z.S., Wang, L.J., Li, D., 2008. Ultrasound assisted extraction of oil from flax seed. Sep. Purif. Technol. 62, 192–198. Zhang, Y., Zhong, Q., 2013. Encapsulation of bixin in sodium caseinate to deliver the colorant in transparent dispersions. Food Hydrocoll. 33 (1), 1–9. Zhao, Y., Qian, S., Yu, W., Xue, Z., Shen, H., Yao, S., Wang, D., 2002. Antioxidant activity of lycopene extracted from tomato paste towards trichloromethyl peroxyl radical CCl3O2. Food Chem. 77, 209–212. Zimmermann, J., Herrlinger, S., Metzger, T., Wanner, C., 1999. Inflammation enhances cardiovascular risk and mortality in hemodialysis patients. Kidney Int. 55, 648–658. Zuorro, A., Fidaleo, M., Lavecchia, R., 2011. Enzyme-assisted extraction of lycopene from tomato processing waste. Enzyme Microbiol. Technol. 49, 567–573. Zuorro, A., Lavecchia, R., 2010. Mild enzymatic method for the extraction of lycopene from tomato paste. Biotechnol. Biotechnol. Equip. 24, 1854–1857.

Further Reading Chiu, Y.T., Chiu, C.P., Chien, J.T., Ho, G.H., Yang, J., Chen, B.H., 2007. Encapsulation of lycopene extract from tomato pulp waste with gelatin and poly(-glutamic acid) as carrier. J. Agric. Food Chem. 55 (13), 5123–5130. Noble, A.C., 1975. Investigation of the color changes in heat concentrated tomato pulp. J. Agric. Food Chem. 23, 48–49. Pohar, K.S., Gong, M.C., Bahnson, R., Miller, E.C., Clinton, S.K., 2003. Tomatoes, lycopene and prostate cancer: a clinician’s guide for counselling those at risk for prostate cancer. World J. Urol. 21, 9–14.