Functional and textural properties of a dehulled oat (Avena sativa L) and pea (Pisum sativum) protein isolate cracker

LWT - Food Science and Technology 86 (2017) 418e423

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Functional and textural properties of a dehulled oat (Avena sativa L) and pea (Pisum sativum) protein isolate cracker
n-Martínez a, L.G. Enriquez b, E. Morales-Polanco a, R. Campos-Vega a, M. Gayta ~ a a, * G. Loarca-Pin
noma de Posgrado en Ciencia y Tecnología de los Alimentos, Research and Graduate Studies in Food Science, School of Chemistry, Universidad Auto Quer
etaro, Centro Universitario, Santiago de Queretaro, C.P. 76010, Qro, Mexico b Roquette Food Company Mexico S.A. de C.V., Queretaro, C.P. 76042, Mexico a

a r t i c l e i n f o

a b s t r a c t

Article history: Received 14 March 2017 Received in revised form 2 August 2017 Accepted 4 August 2017 Available online 5 August 2017

In this study, the nutritional, antioxidant and physical properties of a cracker made from dehulled oat flour (Avena sativa L) and pea (Pisum sativum) protein isolate (COP) was investigated. The COP was compared against two commercial crackers, showing a higher nutritional content, emphasizing its high value of protein (24.66 g/100 g cracker), total fiber (18.45 g/100 g cracker) insoluble fiber (13.05 g/100 g cracker), vanillin (0.932 mg/100 g cracker), p-cumaric (0.861 mg/100 g cracker) and avenantramide (1.160 mg/100 g cracker) as well as the low content of lipids (9.07 g/100 g cracker), carbohydrates (62.13 g/100 g cracker), total phenolic compounds (0.42 mgGAE/g cracker) antioxidant capacity DPPH (26.93 mmol eq. Trolox/g cracker) and ABTS (171.61 mmol eq. Trolox/g cracker). Certain differences were also found in textural properties, the COP exhibited lower hardness (19.04 N), and gumminess (4.07 N), and higher values of cohesiveness (0.35), springiness (0.45 mm), and chewiness (0.35). Based on these results, dehulled oat and pea protein isolated crackers have the potential to confer health benefits. © 2017 Published by Elsevier Ltd.

Keywords: Oat Pea protein isolate Cracker Physicochemical Functional and textural properties Texture profile analysis

1. Introduction The manufacturing of processed foods (i.e. snacks) requires the development of new products according to the consumer's preferences and needs, such being an opportunity to incorporate bioactive compounds related to human health benefits as part of the formulations. Cookies and crackers are considered some of the most popular low-moisture baked goods made with soft wheat flour in the United States and Mexico (Kweon, Slade, Levine, & Gannon, 2014). Cookies and crackers have become the most consumed snacks amongst young and adult people due to their low manufacturing cost, convenience, long shelf-life and ability to serve as a vehicle of important nutrients. Furthermore, the consumption of bakery products has been increasing as a result of urbanization and growth of the working female population (Thivani, Mahendran, & Kanimoly, 2016). Nowadays, consumers are more concerned about their health and demand food products that provide health benefits with reduced calories, high protein, dietary fiber content;

* Corresponding author. ~ a). E-mail address: [email protected] (G. Loarca-Pin http://dx.doi.org/10.1016/j.lwt.2017.08.015 0023-6438/© 2017 Published by Elsevier Ltd.

there is also a trend to increase the intake of natural products rather € ckli, Sta €mpfli, than foods that contain synthetic additives (Sto Messner, & Brunner, 2016). Moreover, evidence of diseases such as high blood pressure, diabetes, cardiovascular diseases (CVDs) and among other illnesses due to lifestyle changes, are on the rise (Xavier et al., 2016). The combination of cereals and legumes has been related to the prevention and/or reduction of non-transmissible diseases due to their bioactive compounds such as dietary fiber, phenolic compounds, protein, phytosterols, among others. These compounds have shown antimutagenic, hypocholesterolemic, hypoglycemic and anticarcinogenic properties (Patil, Brennan, Mason, & Brennan, 2016). Besides protein concentrates and isolates have been recently used by the food industry, mostly derived from soybeans, barley, and wheat. However, due to dietary restriction such as allergies and consumer preferences, the food industry is looking for alternate sources of proteins (Toews & Wang, 2013). Pea protein as a concentrate or as an isolate can be an alternative because of its nutritional quality and ability to provide desirable sensory properties such as structure, texture, taste, and color to formulate food products (O'Sullivan, Murray, Flynn, & Norton, 2014). Noteworthy, oat (Avena sativa L.) is the only food recognized as nutraceutical by the U.S. Food and

E. Morales-Polanco et al. / LWT - Food Science and Technology 86 (2017) 418e423

Drugs Administration (FDA) due to its role in the prevention of coronary diseases mainly (Shah, Masoodi, Gani, & Ahmad Ashwar, 2016). According the FDA a functional food is defined as: “… a part of the usual diet to have helpful effects that go beyond elementary nutritional role …” (Martirosyan & Singh, 2015). Functional food can be enriched with ingredients that usually are not present in that particular food. Similarly, the European Food Safety Authority (EFSA) authorizes the labeling of food products with health claims (Mannarino, Ministrini, & Pirro, 2014). Bakery products are considered acceptable diet agents for health and strength (Deepika, 2016). There are some reports related to glutenfree baked products; however, the information on gluten-free crackers made from pea protein and oat flour is rarely found. The information at hand is still limited, particularly regarding the comparative research with commercial products. Knowing the differences between our formulation and the commercially available products provides relevant information about the potential acceptability by the consumer. The aim of this study was to examine physical hardness, resilience, fracturability, cohesiveness, springiness, gumminess, chewiness, physicochemical characteristics, protein, lipids, and carbohydrate and nutritional (TFD, IDF, SDF, RS, antioxidant capacity and free phenolic compounds) properties of an oat (Avena sativa L) and pea protein (Pisum sativum) isolate crackers as a novel alternative functional snack. 2. Materials and methods 2.1. Ingredients and cracker preparation Dehulled oat grain (Avena sativa L.) was obtained from a local market of Queretaro (Mexico). One hundred grams of dehulled oat grain was milled to a fine powder using a coffee grinder (KRUPS GX4100, Mexico) and sieved through a Montinox 40 mesh (0.42 mm) screen, this material was called oat flour (OCF). Commercial pea (Pisum sativum) protein isolate (PPI) Nutralys®F85M res S.A., Lestrem, France). The prepwas supplied by Roquette (Fre aration of the crackers was made based on the standard method AACC (10-50D) (Gaines & Tsen, 1980). To obtain the base formulation, OCF (80%) and PPI (20%) were used as major ingredients in the formulation. The dough was kneaded and sheeted to a uniform thickness and cut into square shapes (10 cm2). The baking was done at 180  C for 10 min in a forced-air convection oven; the oven was equipped with a turbo fan system for heat distribution and the multi-flame system. The characteristics from the oat/pea protein isolate crackers (COP) were compared to two types of wheat crackers: commercial crackers (CC1: carbohydrate: 63 g/100 g cracker, lipids: 20 g/100 g cracker and protein: 7 g/100 g cracker; according to the commercial label) and commercial crackers with reduced lipid content (CC2: carbohydrate: 72 g/100 g cracker, lipids: 6 g/100 g cracker and protein: 11 g/100 g cracker; according to the commercial label). 2.2. Proximate composition AOAC procedures were used to determine moisture (method 925.10), lipid (method 920.39), ash (method 923.03), and nitrogen (method 920.87) contents of the OCF, PPI, COP, CC1 and CC2 samples (AOAC, 2002). The moisture was determined by the method AACC 44e16.01. The nitrogen content was determined by using the Kjeldahl method, with sodium sulfate as a catalyst. The protein content was calculated as nitrogen x 5.83 for OCF and 6.25 for PPI, OCP, CC1 and CC2. The lipid content was obtained from Soxhlet extraction for 6 h with petroleum ether and the ash content was calculated accordingly by the method AACC 30e25.01, 942.05, respectability (AACC, 1995).

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2.3. Nutraceutical composition 2.3.1. Total dietary fiber (TDF) The dietary fiber fractions, containing soluble dietary fractions (SDF) and insoluble dietary fractions (IDF) were determined following AOAC method 991.43 (AOAC, 2002). One gram of each sample (OCF, PPI, COP, CC1, and CC2) was added to 50 mL phosphate buffer pH 6. The samples were placed in a water bath at 100  C, and 0.1 mL of a-amylase solution was added, and incubated for 30 min stirring every 5 min. The samples were rapidly cooled and added with 0.1 mL of protease, and placed in a water bath at 60  C for 30 min. Afterwards, the pH was adjusted at 4, the samples were placed in a water bath at 60  C for 30 min, and 0.3 mL of amyloglucosidase was added. The samples were incubated for 30 min under constant agitation and then diluted with 95% of ethanol (1:4 ratios), then the mixtures were left at room temperature for 24 h. Such samples were filtered at a constant weight and the residues washed three times with 10 mL of distilled water. The residues were placed in an oven at 90  C for 2 h and weighed. The TDF was determined gravimetrically and considered as polysaccharide extract (PE). To quantify IDF, the ethanol was not added. The SDF was calculated by subtracting the IDF proportion from TDF. 2.3.2. Resistant starch (RS) The resistant starch (RS) content was measured following the ~ i, Bravo, and Man ~ as gravimetric method of Saura-Calixto, Gon (1993). The PE (0.1 g) was homogenized with 6 mL of 2 mol/L of KOH and placed in a shaker (Maxi Mix II, Thermolyne type 37600 mixer, San Francisco, Calif., U.S.A.) for 30 min at 25  C under constant agitation. Acetate buffer and 2 mol/L HCl were added and the pH adjusted to 4.75. Subsequently, 60 mL of amyloglucosidase were added and the tube placed in a shaking bath at 60  C for 30 min. The sample was centrifuged (15 min at 3000g) after the incubation. The pellet was re-suspended in 10 mL distilled water and centrifuged twice, freeze dried and weighed. The fraction obtained corresponded to the RS. 2.4. Free phenolic compounds (PCs) 2.4.1. Methanolic extraction of PCs The PCs were extracted according to Cardador-Martinez, Loarca~ a, and Oomah (2002) procedure. Pin 2.4.2. Total PCs The total PCs content was determined by the Folin-Ciocalteu spectrophotometric method adjusted for 96-well plates (Djeridane et al., 2006). The results were expressed as milligrams of gallic acid equivalents per gram of crackers (mg GAE/g cracker). 2.4.3. Analysis of PCs by HPLC-DAD A High-performance liquid chromatography-diode array detection (HPLC-DAD) analysis was conducted on an Agilent 1100 Series HPLC system (Agilent Technologies, Palo Alto, CA, USA) using a Zorbax Eclipse XDB-C18 column (Agilent Technologies, 4.6  250 mm, 5.0 l m). The column was thermostatically controlled at 35  C ± 0.6 and the flow rate was set to 1 mL/min. The mobile phase consisted of two solvents: solvent A (0.1% v/v acetic acid) and solvent B (100% acetonitrile). A linear gradient was used as follows: 90e78.5% of solvent A, held for 2 min, 78.5e76% for 6 min, 76e60% for 2 min, 60e50% for 4 min and 50e90% for 2 min. The detection was performed at 280 nm at 1 s velocity. A volume of 20 mL was injected the samples were analyzed in duplicate. Quantification was carried out using the external standard method with commercial standards of (+)-catechin, rutin, quercetin, vanillin and ellagic, caffeine, p-coumaric, ferulic, gallic, chlorogenic, and sinapic acids.

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2.5. Antioxidant activity

3. Results and discussion

2.5.1. DPPH method The estimation of the Trolox equivalent antioxidant capacity (TEAC) was determined by using the stable radical 1,1-diphenyl-2picrylhydrazyl (DPPH), according to the method reported by Nenadis, Wang, Tsimidou, and Zhang (2004). A total of 20 mL of methanolic extract was mixed with 200 mL of 150 mmol/mmL of DPPH in 80% methanol. The TEAC value was calculated using Trolox as the standard for the calibration curve and expressed as mmol of Trolox equivalents per gram of crackers (mmol eq. Trolox/g cracker).

3.1. Proximate composition

2.5.2. ABTS method The TEAC estimation was performed using the 2,2-azinobis-3ethylbenzothiazoline-6-sulphonic acid (ABTS) assay described by ~ a, Mendoza, Ramos-Go
mez, and Reynoso (2010). A total, Loarca-Pin 20 mL of methanol extraction were mixed with 230 mL of ABTS+ solution. The absorbance was read at 570 nm at room temperature after 0, 4, 10, 30, 60, and 90 min. The TEAC value was calculated using a Trolox calibration curve and expressed as mmol of Trolox equivalents per gram of crackers (mmol eq. Trolox/g cracker). 2.6. Texture profile analysis The texture profile analysis of COP, CC1 and CC2 was done as described by Inglett, Chen, and Liu (2015) using a texture analyzer CT3 (Brookfield, Posgrado en Ciencia y Tecnología de los Alimentos, Research and Graduate Studies in Food Science, School of Chem
noma de Queretaro) equipped with a 5 cm istry, Universidad Auto diameter flat cell. The conditions were carried out at a 1 mm/s velocity pre-and post-test as well as 2 mm/s, 4.5 mm/s and a 5 mm distance, accordingly. The textural parameters, such as hardness (maximum height of the force peak on the first compression cycle), resilience (ratio of work returned by the sample as compressive strain was removed, for the work required for compression), fracturability (the brittleness status during chewing), cohesiveness (ratio of the positive force areas under the first and second compressions), springiness (the contact ratio of the sample during the first compression), gumminess (the product of hardness and cohesiveness), and chewiness (the product of hardness, cohesiveness and springiness) were determined. 2.7. Statistical analysis All measurements were carried out as independent experiments and duplicated or triplicated accordingly. The data were expressed as mean ± standard deviation. The Tukey's test was used to determine significant differences at p < 0.05 for multiple variables. The principal component analyses (PCA) of the significant textural properties of the crackers samples were implemented by using a correlation matrix. A statistical analysis was done with the use of JMP 8.0 (SAS Inst. Inc., Cary, N.C., U.S.A.).

The proximate composition results of raw and cracker flour are presented in Table 1. COP had significantly (p < 0.05) higher protein content (24.66 g/100 g cracker) than CC1 (8.83 g/100 g cracker) and CC2 (14.78 g/100 g cracker). The increase in the protein content in COP flour could be attributed to a blend of the dehulled oat grain (Avena sativa L.) flour and the pea (Pisum sativum) protein isolate. Our results coincide with previous reports of different snacks and cookies made from cereals (wheat) that contained an added legume flour (chickpea, lentil, green and yellow pea) (Cheng & Bhat, 2016; Patil et al., 2016). The high protein content has been related to beneficial health effects decreasing the starch hydrolysis reported in biscuits enriched with bean protein (Zhang, Dhital, & Gidley, 2015). Regarding the lipids content, COP (9.07 g/100 g cracker) had intermediate values compared to CC2 (5.90 g/100 g cracker) and CC1 (13.77 g/100 g cracker), hence, such value is considered as a cracker with lipid reduction (FDA, 2013). A lower lipid content was desirable to increase the shelf-life of the baked products according to Mesías, Holgado, M
arquez-Ruiz, and Morales (2016) in a biscuit made with wheat and chia flour. The carbohydrates content for the COP (62.13 g/100 g cracker) was significantly (p < 0.05) lower than CC1 (72.00 g/100 g cracker) and CC2 (75.81 g/ 100 g cracker). The carbohydrates percentage coincided with the average content of wheat-based cookies (Park, Choi, & Kim, 2015). Besides, low-carbohydrate biscuits have shown to significantly reduce the calories contributed by these foods, providing beneficial health benefits (Aggarwal, Sabikhi, & Sathish Kumar, 2016). 3.2. Nutraceutical analysis Total, insoluble and soluble dietary fraction fiber (TDF, IDF, SDF) and resistant starch (RS) content of the raw and crackers flour as presented in Table 2. COP had significant higher TDF (18.45 g/100 g

Table 2 Total, insoluble and soluble dietary fiber, and resistant starch content of the raw material and the formulation of an oat/pea protein isolate cracker compared to commercially available products. Samples

Total dietary fiber

insoluble fiber

soluble fiber

OCF PPI COP CC1 CC2

16.90 ± 0.52 3.01 ± 0.13 18.45 ± 0.51a 6.25 ± 0.25b 8.59 ± 0.36c

12.33 ± 0.19 0.5 ± 0.002 13.05 ± 0.46a 2.51 ± 0.1b 1.43 ± 0.16b

4.16 0.01 3.60 3.56 2.14

± ± ± ± ±

0.21 0.001 0.18a 0.24a 0.37b

Resistant starch 2.04 ND 1.46 0.94 1.15

± 0.08 ± 0.41a ± 0.28b ± 0.32c

The results are expressed as g/100 g cracker of three independent experiments ± SD (standard deviation). Means with different letters in the same column indicate statistically significant differences among crackers (p < 0.05) from Tukey's test. OCF: Dehulled oat grain flour; PPI: Pea protein isolate; COP: Dehulled oat and pea protein cracker; CC1: Commercial Cracker 1; CC2: Commercial reduced lipids cracker 2; ND: No detected.

Table 1 Proximal composition of the raw material and the formulation of an oat/pea protein isolate cracker compared to commercially available products. Samples

Protein

Lipids

Moisture

OCF PPI COP CC1 CC2

12.58 ± 0.09 87.86 ± 0.43 24.66 ± 0.46a 8.83 ± 0.003b 14.78 ± 0.09c

11.34 ± 0.12 1.00 ± 0.04 9.07 ± 0.05a 13.77 ± 0.04b 5.90 ± 0.07c

8.51 6.78 1.79 5.84 2.56

± ± ± ± ±

0.02 0.05 0.04a 0.01b 0.09c

Ash 1.54 5.72 3.95 5.35 3.49

Carbohydrate ± ± ± ± ±

0.04 0.02 0.01a 0.06b 0.02c

74.43 ± 0.19 5.23 ± 0.02 62.13 ± 0.07a 72.00 ± 0.09b 75.81 ± 0.3c

The results are expressed as g/100 g cracker of three independent experiments ± SD (standard deviation). Different letters in the same column indicate statistically significant differences among crackers (p < 0.05) from Tukey's test. OCF: Dehulled oat grain flour; PPI: Pea protein isolate; COP: Dehulled oat and pea protein cracker; CC1: Commercial Cracker 1; CC2: Commercial reduced lipids cracker 2.

E. Morales-Polanco et al. / LWT - Food Science and Technology 86 (2017) 418e423 Table 3 Total free phenolic compounds and antioxidant capacity (DPPH and ABTS) of the raw material and the formulation of an oat/pea protein isolate cracker compared to commercially available products. Samples

Total phenolic compounds

OCF PPI COP CC1 CC2

0.73 0.47 0.42 0.54 1.47

± ± ± ± ±

0.023 0.051 0.052a 0.033b 0.054c

DPPH 46.40 13.37 26.93 25.28 72.12

ABTS ± ± ± ± ±

3.88 6.66 10.42a 10.26a 5.04b

164.46 148.99 171.61 169.48 298.53

± ± ± ± ±

9.49 9.49 7.45a 9.25a 6.48b

The results of total PCs were expressed as gallic acid equivalent (mg GAE/g cracker) of three independent experiments ± SD (standard deviation). The antioxidant capacity results are expressed as micromoles of trolox equivalent antioxidant capacity (TEAC) (mmol eq. Trolox/g of crackers) of three independent experiments ± SD (standard deviation). Means with different letters in the same column indicate statistically significant differences among crackers (p < 0.05) from Tukey's test. OCF: Dehulled oat grain flour; PPI: Pea protein isolate; COP: Dehulled oat cracker and pea protein; CC1: Commercial Cracker 1; CC2: Commercial reduced lipids cracker 2.

cracker), IDF (13.05 g/100 g cracker) and SDF (3.60 g/100 g cracker) (The rate of soluble and insoluble fiber was 1:3.62) and resistant starch (1.46 g/100 g cracker) than CC1 and CC2 flour. The recommended ratio of soluble and insoluble fiber ranged from 1:4 to 1:3 in biscuits enriched with raspberry fiber, as fiber increases water retention in the human colon influencing the volume and softness of the stool (Gorecka, Pacholek, Dziedzic, & Gorecka, 2015). Fiber intake has been linked to several health benefits concerning human gut, for instance: constipation, colon and breast cancer, hemorrhoids; it also shortens the time of intestine passage. Soluble fiber intake has been associated with a risk reduction of cardiovascular diseases, and also lower blood levels of cholesterol, triglycerides and glucose (Anderson et al., 2009). The fact that the samples showed varied composition of dietary fiber may suggest functional characteristics as well as a possible physiological effect. Our results demonstrate that COP is a good source of dietary fiber and resistant starch.

3.3. Total PCs and antioxidant capacity (DPPH and ABTS) analysis The data of radical scavenging activity and quantification of total PCs are shown in Table 3. Statistical differences were found in all crackers. For the total PCs, the CC2 presented higher mg gallic acid equivalent/g crackers (1.47 mg GAE/g cracker) than CC1 (0.54 mg GAE/g cracker) and COP (0.42 mg GAE/g cracker). Furthermore, the antioxidant capacity measured by the DPPH method in CC2 flours presented higher TEAC values (72.12 mmol eq. Trolox/g cracker) than CC1 (25.28 mmol eq. Trolox/g cracker) and COP (26.93 mmol eq. Trolox/g cracker). The results for the ABTS method exhibited the same behavior in CC2 flours, which demonstrated higher TEAC

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values (298.53 mmol eq. Trolox/g cracker) than CC1 (169.48 mmol eq. Trolox/g cracker) and COP (171.61 mmol eq. Trolox/g cracker). These results may be due to the synthetic antioxidants content in the commercial crackers, particularly higher values presented in CC2 may be explained by the proportion of wheat (bran, flour and wheat germ) (Mazzoncini, Antichi, Silvestri, Ciantelli, & Sgherri, 2015) in these crackers. 3.4. Free phenolic compounds analysis by HPLC-DAD Table 4 shows the profile of free phenolic compounds in the methanolic extract of the different samples analyzed in this study. Some phenolic compounds, predominantly phenolic acids and several flavonoids were quantified. 3.4.1. Phenolic acids Eight phenolic acids were identified on the OCF and COP (Table 4). The most abundant phenolic compound was gallic acid in OCF (5.30 mg/g cracker) and COP (4.240 mg/g cracker), followed by avenanthramide in OCF (1.12 mg/g cracker) and COP (1.160 mg/g cracker), this trend may be due to the content of oat flour in the formulation of COP. The differences might be attributed to the diverse extraction methods, as well as the environmental growing conditions, as suggested by Peterson (2001). Within the CC1 and CC2, the predominant phenolic compound was gallic acid (3.710 and 13.59 mg/g cracker, respectively), followed by ferulic acid (0.900 and 0.966 mg/g cracker, respectively). These results can be explained considering that those phenolic compounds found in the CC1 and CC2 are typically predominant in wheat flour. Similar relı et al. (2016) in biscuits made with sults were reported by Kocadag wheat, barley and colored corn. In general, food processing can also release PCs from its components to which they are bound; it has also been proposed that thermal processing can release bound phenolic that is observed in the processing of cereals and legumes (Duodu, 2014). 3.4.2. Flavonoids Two flavonoids were found in the OCP, COP and CC2 (Table 4) where the most predominant was quercetin in OCF (0.704 mg/g cracker), COP (1.043 mg/g cracker) and CC2 (1.035 mg/g cracker), followed by rutin in OCF (1.281 mg/g cracker), COP (0.685 mg/g cracker) and CC2 (0.528 mg/g cracker). Our results coincide with Luksi c et al. (2016), who reported that during the preparation of buckwheat sourdough bread there was a rutin content decrease. Vogrin ci c et al. (2010) reported that rutin lowering behavior increased within the quercetin content, such result was attributed to the hydrolysis rutin conversion into quercetin during the baking process through the activity of the rutin enzyme on the samples

Table 4 HPLC polyphenol profile of the raw material and the formulation of an oat/pea protein isolate cracker compared to commercially available products. Free phenolic compounds (mg/g)

OCF

PPI

COP

CC1

CC2

Caffeic acid Vanillin p-Coumaric Sinapic acid Pherulic acid Gallic acid Avenantramide Ellagic acid Quercetin Rutin

0.897 ± 0.041 0.791 ± 0.040 0.567 ± 0.011 0.573 ± 0.028 0.872 ± 0.012 5.30 ± 0.013 1.12 ± 0.025 0.954 ± 0.02 0.704 ± 0.048 1.281 ± 0.069

ND 0.08 ± 0.007 0.055 ± 0.0046 ND ND ND ND ND ND ND

0.918 ± 0.03a 0.932 ± 0.063 0.861 ± 0.018a 0.758 ± 0.027a 0.998 ± 0.075a 6.240 ± 0.015a 1.160 ± 0.018a 1.0630 ± 0.05a 1.043 ± 0.031a 0.685 ± 0.087a

ND ND 0.054 0.594 0.900 8.710 0.010 ND ND ND

0.879 ND ND 0.569 0.966 13.59 0.016 0.953 1.035 0.528

± ± ± ± ±

0.082b 0.014a 0.082b 0.031b 0.0052b

± 0.081a

± ± ± ± ± ± ±

0.036a 0.072b 0.028c 0.004b 0.042b 0.0018b 0.0015b

PC's results by HPLC-DAD are expressed as mg/g dry cracker of three independent injections ± SD (standard deviation). Means with different letters in the same line indicate statistically significant differences among crackers (p < 0.05) from Tukey's test. OCF: Dehulled oat grain flour; PPI: Pea protein isolate; COP: Dehulled oat and pea protein cracker; CC1: Commercial Cracker 1; CC2: Commercial reduced lipids cracker 2; ND: No detected.

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Table 5 Texture properties of an oat/pea protein isolate cracker compared to commercially available products. Texture proprieties

COP

CC1

CC2

Hardness (N) Resilience Cohesiveness Springiness (mm) Gumminess (N) Chewiness (mJ)

19.04 ± 4.60a 0.37 ± 0.04a 0.35 ± 0.03a 0.45 ± 0.03a 4.07 ± 0.73a 3.43 ± 0.29 a

21.96 ± 2.84a 0.36 ± 0.02a 0.23 ± 0.03b 0.32 ± 0.04b 7.19 ± 0.62b 1.60 ± 0.21b

26.65 ± 1.81b 0.34 ± 0.03a 0.26 ± 0.02b 0.30 ± 0.05b 36.69 ± 2.81c 2.55 ± 0.22c

Texture profile results of hardness are expressed as Newton (N), springiness millimeter (mm), gumminess Newton (N), Chewiness miliJoule (mJ). Means with different letters in the same line indicate statistically significant differences among crackers (p < 0.05) from Tukey's test. COP: Dehulled oat cracker and pea protein; CC1: Commercial Cracker 1; CC2: Commercial reduced lipids cracker 2. The results are expressed as mean ± SD (standard deviation).

after the water and yeast was added to the flour. 3.5. Textural analysis The texture properties are some of the most important parameters to evaluate the quality of crackers (Siche et al., 2016). Mesías et al. (2016) and Oh, Bae, and Lee (2014), reported that higher protein and insoluble fiber content increased the hardness of biscuits made with wheat and chia flour; however, COP had the lowest hardness (19.04 N) compared to CC1 and CC2 (21.96 N and 26.65 N, respectively) (Table 5). The decreased hardness could be due to the competition of sugar and flour proteins for water, which resulted in lesser gluten formation (Kulthe, Pawar, Kotecha, Chavan, & Bansode, 2014). There were no significant differences in the resilience value of the samples; this may be due to the lack of a homogeneous pore distribution as reported previously (Ahmed & Hussein, 2013). Cohesiveness and springiness of the COP was higher (0.35 mm and 0.45 mm, respectively) than commercial crackers. These results coincide with those reported by Crocket, Ie, and Vodovotz (2011), who added protein to gluten-free products to increase elastic module by cross-linking and to improve flavor quality. The gumminess of the COP (4.07 N) was lower than in commercial crackers. Similar results were reported by Sarabhai et al. (2014), who concluded that the combination of protein concentrates and emulsifiers build up the structure of the cookie dough

similar to gluten protein, improving the textural characteristics of the cookies. The chewiness of the COP was higher (3.43 mJ) than commercial crackers CC1 and CC2 (1.60 mJ and 2.55 mJ, respectively), similar results were reported by Liu, Brennan, Serventi, and Brennan (2016), who suggested that the addition of cereals with low gluten content within biscuits increases the chewiness through the interaction between the fibers and the gluten, meanwhile the starch gelatinization degree decreases. The data were further analyzed by PCA to determine the systematic variation and underlying relationships between physical properties, nutraceutical compounds from all crackers CC1, CC2 and COP tested. The addition of the two main components explained 100% of the total variation. The principal component 1 (PC1) explained 64.9% of the total variation that include: protein, RS, TFD, and IDF being as key elements for nutraceutical properties, at the same time most of these variables were closely located on the same side of the plot explaining the main factors that contribute to the physical characteristics (resilience, cohesiveness and springiness) of COP. Also a lipid content was placed on the opposite side where the crackers CC1 were located. The main component 2 (PC2) explained the reason for the 35.1% of the total variation with the carbohydrates as a principal variable, which affect the physical characteristics such as indicated, hardness, gum and chewiness of CC2 (Fig. 1). Similar results are shown by Ahmad, Pasha, Saeed, and Shahid (2017) which proved that the addition of cereals and legume affect the texture and organoleptic properties. 4. Conclusions We conclude: the cracker formulated in this study included dehulled oat grain (Avena sativa L) flour and pea protein (Pisum sativum) improved the texture properties (hardness, resilience, cohesiveness, springiness, gumminess and chewiness) and certain nutraceutical components such as total dietary fiber, fraction insoluble, soluble fiber, and resistant starch compared to commercial crackers using wheat flour as principal ingredient. Our results provide new strategies to formulate and evaluate crackers that perhaps could be considered as a functional baking process for foods with potential benefits for human health. Acknowledgements Author E. Morales-Polanco was supported by a scholarship from the Consejo Nacional Ciencia y Tecnologia/CONACyT-Mexico (number 249699). English edition by Agustín Ruiz Esparza Y Ballesteros, M.A. References

Fig. 1. Principal component analysis of COP: Dehulled oat grain flour and pea protein isolate cracker (A); CC1: Commercial Cracker 1 (C); CC2: Commercial reduced lipids cracker 2 (-). RS: Resistant starch; TDF: Total dietary fiber; IDF: Insoluble dietary fractions; SDF: Soluble dietary fraction. Biscuits.

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