Journal of Pharmacognosy and Phytochemistry 2020; SP6: 91-97

E-ISSN: 2278-4136 P-ISSN: 2349-8234 International Web-Conference JPP 2020; SP6: 91-97 On

Dr. Namrata Ankush Giri New Trends in Agriculture, Environmental & Biological Sciences for (PhD), Scientist (Food Inclusive Development Technology), Division of Crop (21-22 June, 2020) Utilization, ICAR-Central Tuber Crops Research Institute Sreekariyam P.O., Thiruvananthapuram, , Development and physico-chemical evaluation of protein rich cookies using underutilized Anjudas College of Indigenous Food aungustifolia Technology (Cft-K), Konni, Kerala Dr. Namrata Ankush Giri, Anjudas, Dr. MS Sajeev and Dr. T.

Dr. MS Sajeev Krishnakumar (PhD), Principal Scientist and Head (I/C), Division of Crop Abstract Utilization, ICAR-Central Tuber Curcuma angustifolia is one of the Indian tribal minor tuber crop found growing wild in northeast and Crops Research Institute Sreekariyam P.O., western coastal plains and hills containing highly nutritious and easily digestible starch which is Thiruvananthapuram, Kerala, especially recommended for children. The study was conducted with the objective of utilizing the under India exploited tribal tuber crop Curcuma angustifolia starch by processing it into protein-energy rich functional cookies which is highly suitable for tribal communities. The basic ingredients under this study Dr. T. Krishnakumar were Curcuma angustifolia starch, pearl millet flour and refined wheat flour whereas the functional (PhD), Scientist, Division of ingredients were used as protein sources as whey protein concentrate (WPC), bengal gram flour (BF) and Crop Utilization, ICAR-Central soy flour (SF).The level of Curcuma angustifolia starch was standardized based on experimental trials Tuber Crops Research Institute and incorporated up to 25% in formulation of functional cookies. It is used as source of energy in Sreekariyam P.O., cookies. The level of protein rich functional ingredients were used as 5%, 10% and 15%. Raw materials Thiruvananthapuram, Kerala, were analyzed for chemical properties. Also the produced cookies were evaluated for physical, chemical, India minerals content and sensory characteristics. The results indicated that spread ratio and expansion ratio of functional cookies decreases as the level of functional ingredients increases. Textural characteristics like

breaking hardness of functional cookies were increased as the level of functional ingredients increases.

Results indicated that protein-energy rich functional cookies for tribal people with highest overall sensory acceptable score can be prepared using a whey protein concentrate (15%). Protein rich functional cookies containing 15% whey protein concentrate along with other ingredients was found most sensory acceptable by the panelists and it provides protein 8.24%, fibre 3.33%, carbohydrate 57.3% and energy 481.37Kcal.

Keywords: Protein rich, cookies, Curcuma angustifolia, Physical properties, Nutrirional composition

Introduction

Curcuma species is a minor tuber crop found widely all over south India and many other tropical areas with adequate rainfall. Although highly nutritious and high in medicinal properties, the usage of curcuma species is limited to tribal hamlets and traditional medicinal preparations [1]. Curcuma is reported to be used as a source of an easily digested starch, which is rather similar to that of arrowroot in India, it is utilized on a cottage industry basis for the [2] preparation of invalid and valid foods . Curcuma angustifoliais most commonly found growing wild in India, especially in the northeast and western coastal plains and hills. Such areas include the states of , , , , Orissa, , and Kerala. This species can also be found in Burma, , , and [3]. Correspondence Dr. Namrata Ankush Giri In some part of India Curcuma angustifolia is used as weaning food for babies. The starch (PhD), Scientist (Food obtained from the is given to babies three – four times a day after boiling in water or Technology), Division of Crop milk. This preparation is given as a traditional food for babies. Sometimes it is used as Utilization, ICAR-Central Tuber replacement of mother’s milk. This may be due to the high content of total soluble protein, Crops Research Institute reducing sugar and total free amino acids in this species. The starch of tikhur is used for the Sreekariyam P.O., Thiruvananthapuram, Kerala, preparation of many sweet meals and herbal dishes like halwa, barfi, jalebi etc. It is used India specially during fast. ~ 91 ~

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Farmers also prepare herbal drink “sarbat” through tikhur difference was greater in the case of pasta made with chickpea starch during summer due to its cooling effect [4]. Tikhur is flour [23]. Kaur et al. [24] analyzed the sensory, nutritional and found as a primary ingredient in cakes, fruit preserves, glycemic properties of biscuits from cereal such as oats and biscuits, and puddings [5]. barley and soybean and chickpea combinations for diabetic Tikhur rhizomes are used as appetizer reducing burning patients. Biscuits made from these cereal pulse combinations sensations and stomach pains, removal of stone from kidney, were highly acceptable and were chosen for nutritional useful for ulcer patient[6] and pulp is used for analysis. The results of nutritional analysis showed increased treatment of headache as well as it gives cooling effect[7]. The protein (11.72 g/100 g), crude fiber (1.5 g/100 g) and ash rhizome pulp is a remedy for fever, joint pains and content (4.68 g/100 g) and decreased content of carbohydrates leucorrhoea. The starch obtained from the rhizomes is highly (47 g/100 g) in blend containing refined wheat flour, barley nutritious and easily digestible, therefore, it is recommended and soy flour (25:50:25). for infants, weak children. The starch of tikhur is used for the Considering the health value of Curcuma angustifolia tubers preparation of many sweet meals and herbal dishes like and being underutilized crop coupled with the need to expand halwa, barfi, jalebi etc. It is used specially during fast. the utilization potential of Curcuma angustifolia through the Farmers also prepare herbal drink “sarbat” through tikhur development of value added products, the present study aimed starch during summer due to its cooling effect [4]. The rhizome at the development of functional cookies using Curcuma of the tikhur is light bitter, demulcent, non irritating, nutritive angustifolia starch. and fragrant. Rhizome pulp is used as a remedy for headache, joint pains, jaundice and leucoria [8], essential oil of tikhur Materials and methods rhizome is used against tape worm [9]. a. Materials Cookie type biscuits are widely consumed due to their long Curcuma angustifolia starch, wheat flour, pearl millet flour, shelf life and crisp texture [10, 11]. Cookies are made bengal gram flour, whey protein concentrate, soy flour, traditionally from wheat flour using a number of ingredients refined sugar, hydrogenated fat and baking powder were such as sugar, chocolate chips, peanut butter etc. [12]. procured from the local market of Trivandrum city. Functional cookies have been attempted by various researchers using several types of alternative non-wheat flours b. Methods such as flour, buckwheat flour, cassava flour, quinoa flour etc 1. Preparation of cookies: Composite flours blends were [13-15]. India is one of the largest biscuit producers of the world prepared by using Curcuma angustifolia starch, wheat flour and the bakery industry has been projected to grow annually and pearl millet flour. However the functional ingredients at the rate of 15-17% [16]. Being one of the most acceptable were added as bengal gram flour, whey protein concentrate snack foods by both children and adults cookies/biscuits could and soy flour at the level of 5%, 10% and 15%. The be considered as the best vehicle for nutritional supplements. formulations for the preparation of cookies comprised Whey protein concentrate (WPC) is extensively used as a Curcuma angustifolia starch powder and pearl millet flour functional component, in order to improve the protein content, incorporated at constant rate in the ratio of 1:08. Whereas exclusive of increase in energy value from fat [17]. It is wheat flours was substitute with protein rich ingredients such reported that, it have more biological value and amino acid as whey protein concentrate (WPC), Bengal gram flour (BF) score [18]. WPC is added to different food preparations as a and soy flour (SF) at the level of 5%, 10% and 15% while the best source of protein in order to prevent malnutrition and control cookies were made from 100% refined wheat flour. obesity [19, 20]. Singh et al. [21] reported that, WPC is an The formulation for the preparation of cookies is given in effective ingredient in cake preparation as an egg solids Table 1. Cookies were prepared by creaming methods for replacer useful for people allergic to egg. WPC is having 70% making cookies dough. The ingredients (g) used in proteins and utilized for the preparation of protein rich foods preparation of taro flour based gluten free cookies were gluten and in ice cream, yoghurt, and soft drink etc. [19, 20]. It was free flour blends-100g, powdered sugar-32g, fat-40g, baking reported that, whey ingredients also help to increase batter powder-1g and guar gum-0.5g. Cookies dough was prepared viscosity and crumb structure of muffins and provide tender in a spiral dough mixer and sheeted to a thickness of about texture in biscuits by holding moisture for longer time [22]. 5mm approximately, cut into round shape of 30mm diameter The incorporation of chickpea flour increased the protein, and then transferred to baking tray and baked at 165oC for 20 mineral, fat and indigestible compound content of the pasta, min. Cookies, after attaining room temperature, were packed but total starch content was not affected. Starch hydrolysis in airtight plastic containers and evaluated for different was lower in both types of pasta than in white bread, but the physico- chemical parameters [25, 26].

Table 1: Formulation of protein rich cookies incorporated with Curcuma angustifolia starch

Curcuma angustifolia Wheat flour Pearl millet Whey protein concentrate Bengal gram flour Soy flour Treatments starch (WF) flour (WPC) (BF) (SF) Control -- 100 ------5% WPC 25 50 20 5 -- -- 10% WPC 25 45 20 10 -- -- 15% WPC 25 40 20 15 -- -- 5% BF 25 50 20 -- 5 -- 10% BF 25 45 20 -- 10 -- 15% BF 25 40 20 -- 15 -- 5% SF 25 50 20 -- -- 5 10% SF 25 45 20 -- -- 10 15% SF 25 40 20 -- -- 15

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2. Physical properties Results and discussion Cookies were evaluated for physical characteristics including 1. Physical properties diameter (mm), thickness (mm) and spread ratio. Diameter The physical properties of cookies incorporated with and thickness were measured with the help of digital vernier Curcuma angustifolia starch powder fortified with protein caliper whereas spread ratio of cookies was calculated by sources is mentioned in Table 2. The physical properties of dividing values of diameter (D) by the thickness (T) value protein rich cookies such as diameter, thickness, spread ratio [27].Ten cookies samples were considered for determination of and expansion ratio indicated that the diameter, spread ratio physical properties of cookies. and expansion ratio decreased proportionately with increase in the level of fortification of WPC, BF and SF in cookies 3. Textural properties incorporated with Curcuma angustifolia starch powder (Table The textural properties of cookies were determined using 2). On the contrary, thickness of cookies increased Food texture analyzer [(TA-HDi); Stable Micro System significantly with increase in protein rich ingredients from 5% (U.K)]. Breaking test was used to determine hardness and to 15% addition in cookies. This indicated that replacement of toughness of cookies. The individual cookie was placed over wheat flour with protein sources (WPC, BF and SF) resulted two points of the blade. The blade was attached to the in reduction in lateral expansion of cookies, while it led to crosshead of the instrument. The size of probe used was increased vertical expansion. Increase in thickness of cookies 5mm.The peak force from the resulting curves was considered led to a reduction in the spread ratio (D/T) and when as breaking hardness of the cookie and the area under the peak compared to a spread ratio of 8.26 for control cookies, very force was considered as breaking toughness. Settings were low spread ratio of 3.43 was observed in cookies made from kept for the texture analysis as: Pre test speed-10mm/sec, Test protein sources. Decrease in spread ratio of biscuits has been speed-1mm/sec, Post test speed-1mm/sec, Distance at-50% reported in biscuits made from wheat flour fortified with and Starin-50%. functional additives such as defatted soy flour, cassava- soybean mix, chick pea, bran etc. [13, 31-35]. Singh et al. [36] 4. Proximate composition found that high protein biscuits made from wheat-legume Moisture, protein, fat, ash, crude fiber and minerals (Mn, Cu, blends had low spread ratio. The poor spread ratio of biscuits Zn and Fe) content of different samples of cookies were made from composite flour might be associated with the determined as per standard methods [28]. Minerals content of altered elastic behavior of the dough. Mcwatters [37] attributed cookies was determined by using atomic absorption the spread ratio depression in biscuits made from composite spectrophotometer. Total carbohydrates value was calculated flours to an increase in the relative quantity of hydrophilic by subtracting the total of moisture, protein, fat, crude fiber additives, which compete for water in the dough. and ash content from 100. Total calories were calculated by Breaking hardness of protein rich cookies varied between multiplying protein, carbohydrates and fat content by the 19.33N to 48.73N (Table 2). The fortification of WPC leads factors 4, 4 and 9 respectively. to increase in hardness of cookies as compared to rest of the treatments. WPC fortified cookies showed higher hardness 5. Sensory characteristics even than control when added more than 5%. Lowest Sensory evaluation of cookies was done by group of 15 panel hardness was recorded in cookies fortified with SF. Similar members selected from diverse category of staff at ICAR- results were reported by other researchers also [38, 39]. It might CTCRI. Samples were served with three digit code numbers. be due to the gluten miming properties of WPC in cookie Panelists were instructed about the evaluation procedure. dough. This was probably caused by changes in the Sensory attributes like appearance, colour, aroma, taste, rheological properties of the dough of cookies due to the texture and overall acceptability for all cookies samples were presence of fiber, minerals etc. in pearl millet flour and other evaluated using nine point hedonic scales. The grades for flours. Rheological properties such as water absorption hedonic scale were: like extremely-9, like very much-8, like capacity, dough development time and dough stability time, moderately-7, like slightly-6, neither like nor dislike-5, dislike have been reported to change in whole wheat dough as a slightly-4, dislike moderately-3, dislike very much-2, dislike result of iron and zinc fortification [40]. Singh et al. [35] [29] extremely-1 . obtained hardness value of 26.97 N for biscuits made from refined wheat flour and found that defatted soy flour fortification 6. Statistical analysis enhanced the hardness. Although brittleness is preferred to some Results were expressed as mean of triplicate analyses. A one extent by the consumers, too brittle products could result in way analysis of variance and Duncan’s test were used to packaging problems. However, subsequent sensory evaluation establish the significance of differences among the mean studies reported in the present study showed that cookies with values at the 0.05 significance level. The statistical analyses 15% WPC had high acceptability. were performed using SAS 9.3 software [30].

Table 2: Physical properties of protein rich cookies incorporated with Curcuma angustifolia starch

Treatments Weight (g) Diameter (cm) Thickness (cm) Spread ratio Expansion ratio Hardness (N) Control 11.9± 0.5a 41.85± 0.3a 5.06± 0.3e 8.26± 0.5a 0.97± 0.3a 40.63± 0.53c 5% WPC 11.2± 0.4f 24.2± 0.3c 7± 0.5a 3.46± 0.5g 0.86± 0.3f 35.62±0.39e 10% WPC 11.5± 0.1b 24.5± 0.3b 6.4± 0.5b 3.83± 0.5f 0.87± 0.3e 43.59±0.51b 15% WPC 11.01± 0.1f 24± 0.5c 7± 0.5a 3.43± 0.5g 0.86± 0.5f 48.73±0.83a 5% BF 11.0± 0.5f 22.8± 0.5e 4.6± 0.5 4.96± 0.5c 0.95± 0.5b 17.42±0.34j 10% BF 11.28± 0.3e 22.7± 0.3e 5.6± 0.5d 4.05± 0.3d 0.94± 0.5c 23.67±0.84g 15% BF 11.43± 0.2d 23.6± 0.3d 5.9± 0.3c 4.0± 0.3d 0.90± 0.2d 37.12±0.81d 5% SF 11.23± 0.2f 22.9± 0.5e 4.5± 0.5 5.09± 0.3b 0.95± 0.2b 19.33±0.40i 10% SF 10.99± 0.1g 22.5± 0.2f 5.5± 0.3d 4.09± 0.5d 0.94± 0.1c 29.34±0.44f 15% SF 11.47± 0.2c 23.5± 0.2d 5.9± 0.3c 3.98± 0.5e 0.90± 0.5d 21.59±0.70h * Values are given in mean ± standard deviation of three independent determinations. Values in column followed by the same letter are not significantly different at p ≤ 0.05 as measured by Duncan’s test using SAS ~ 93 ~ Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com

2. Proximate composition and mineral content protein content of gluten free cookies when sweet potato was Proximate composition including moisture, ash, protein, fat, substituted with WPC. They also observed that the high carbohydrates and energy value of protein rich cookies protein level was related with the water binding capacity of incorporated with Curcuma angustifolia starch is presented in soy flour. The increase in level of protein content of cookies Table 3. Moisture content of cookies varied in between 2.86 with fortification of WPC, BF and SF is also represented in to 5.38%. The increase in the level of fortification of protein Fig.1 ingredients from 5 to 15% resulted to increase in the per cent There was somewhat increase in ash and fiber content of of moisture content. The moisture content of protein fortified cookies fortified with protein sources which might be due to cookies found on higher side compared to control. It might be the fortification of WPC, SF and BF in cookies. Giri and due the higher water binding capacity of WPC, BF and SF. Sakhale [26]; Gallagher et al. [43] also reported the biscuits Giri and Sakhale [26] observed that the water content was fortified with whey protein concentrate and casein found high raised from 3.53 to 4.83% and found higher WPC fortified GF in ash content. Control cookies had low crude fiber content cookies. (2.20%) as compared to 2.23-3.44% in the protein fortified Protein content of WPC, BF and SF fortified cookies was cookies. The fat content of protein fortified cookies was significantly superior to that of control. The protein content of observed from 22.03 to 25.66% compared to control. The SF cookies fortified with WPC, BF and SF resulted to increase in fortified cookies had higher fat content as compared to rest of protein content from 6.73% to 10.69%, 8.95% and 9.37% the treatment. Obvious, it has been due to high content of fat respectively. The percent increase in protein content was in SF compared to WF. The significant decrease and increase observed as 58.84, 32.98 and 39.22 with addition of WPC, BF in carbohydrate and calorie content of protein rich cookies and SF respectively as compared to control. Comparable were noted respectively. The increase in protein and fat outcome were reported by Munaza et al. [41] in WPC fortified content leads to decrease in carbohydrate content and increase biscuits. Sathe et al. [42] reported that substitution of wheat in energy value of cookies. These results are in agreements flour with soy flour in crackers leads to increase in protein with Giri et al. [25]. content. Giri and sakhale [26] also found that increase in

Table 3: Proximate composition of protein rich cookies incorporated with Curcuma angustifolia starch

Treatments Moisture (%) Fat (%) Crude fibre (%) Ash (%) Protein (%) Carbohydrate (%) Energy (Kcal/100g) Control 3.29± 0.19d 22.03± 0.08d 2.20± 0.0d 1.17± 0.03e 6.73±0.40e 64.58±0.38a 483.51±0.1e 5% WPC 4.49±0.25b 23.44± 0.29c 2.35± 0.45c 1.14± 0.06e 7.97±0.04d 60.61±0.48c 485.28±0.2c 10% WPC 5.38± 0.47a 24.53± 0.38b 2.42± 0.15b 1.50± 0.03c 8.91±0.00c 57.26±0.51e 485.45±0.3c 15% WPC 5.51± 0.35a 24.55± 0.15b 2.45± 0.36b 1.55± 0.18c 10.69±0.15a 55.25±0.96g 484.71±0.13d 5% BF 2.86± 0.05e 23.07± 0.04c 2.23± 0.21d 1.18±0.08e 7.77±0.03d 62.89±0.35b 490.27±0.5a 10% BF 3.31± 0.44d 23.73± 0.29c 2.56± 0.03b 1.27±0.14d 8.21±0.07c 60.92±0.14c 490.09±0.5a 15% BF 4.44± 0.09b 24.49± 0.14b 3.44± 0.29a 1.33± 0.01d 8.95±0.08c 57.35±0.13e 485.61±0.49c 5% SF 3.32± 0.25d 24.26± 0.21b 2.43± 0.36b 2.55±0.04b 7.50±0.15d 59.94±0.84d 488.1±0.33b 10% SF 3.83± 0.14c 25.56± 0.26a 2.49± 0.28b 3.11±0.11a 8.22±0.02c 56.79±0.14f 490.09±0.5a 15% SF 3.83± 0.12c 25.66± 0.22a 2.53± 0.31b 3.32±0.08a 9.37±0.06b 55.29±0.50g 489.58±0.1a * Values are given in mean ± standard deviation of three independent determinations. Values in column followed by the same letter are not significantly different at p ≤ 0.05 as measured by Duncan’s test using SAS

Fig 1: Protein content of Curcuma angustifolia incorporated cookies fortified with WPC, BF and SF

It was found that minerals such as manganese (Mn), cooper to 4.25 mg/100g respectively which was higher than 0.008 (Cu), iron (Fe) and zinc (Zn) were significantly higher in mg/100g of manganese and 2.3 mg/100g of zinc present in protein fortified cookies as compared to control (Fig. 2). control cookie. Minerals such as magnesium and copper have Copper content ranged from 0.25 to 0.53 mg/100 g in protein been associated with several vital enzyme activities and enriched cookies. Iron content ranged from 1.37 to 3.55 biological functions in the body [44, 45]. The minerals content mg/100g in the protein fortified cookies, when compared to of protein rich cookies fortified with WPC, BF and SF at the only 0.35 mg in the control cookies. Similarly, manganese level of 5 to 15% is graphically represented in Fig. 2. and zinc content ranged from 0.22 to 0.77 mg/100g and 3.34 ~ 94 ~ Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com

Fig 2: Mineral content of protein rich cookies incorporated with Curcuma angustifolia starch

3. Sensory evaluation The records regarding sensorial properties of cookies with SF and BF. Based on panel score, it was investigated incorporated with Curcuma angustifolia starch and fortified that, the cookies with 15% WPC was found most acceptable with WPC, BF and SF is given in Fig.3. Mean sensory scores with regards to overall acceptability. Based on sensorial score for different sensory characteristics of protein enriched of WPC fortified samples, the cookies containing 15% WPC cookies were more than minimum acceptable score of 6 (Fig. was found overall acceptable, considered as standardized. 3). It was observed that cookies were prepared using different Sathe et al. [42] documented parallel findings that, protein rich level of protein sources: WPC, BF and SF at the level of 5 to crackers could be prepared by substituting refined wheat flour 15% were accepted by the panelists. with soy and groundnut flours at 15% level. Singh et al. [35] The data revealed in Fig.3 that cookies with 15% WPC also state that WPC might be fortified up to 15% level in obtained highest scores of texture (8), taste (8.5) and overall cookies without disturbing their overall quality. The acceptability (8.95) as compared to control and other maximum mean score for different sensor y characteristics of treatments. It was found that, the score for cookies fortified cookies developed using 25% Curcuma angustifolia starch, with BF and SF were reduced for texture, mouth feel and taste 40% wheat flour, 20% pearl millet flour and 15% WPC due to beany flavor of SF and particular taste of BF as indicated the commercial scope for manufacturing of good compared to control and WPC fortified cookies. Cookies quality protein rich cookies which can be consumed by people fortified with WPC were more preferred than cookies fortified with protein deficiency and malnutrition problem.

Fig 3: Sensory evolution of protein fortified cookies incorporated with Curcuma angustifolia starch

Conclusions scores for all the sensory parameters, it could be concluded Nutritional and physico-mechanical evaluation of cookies that cookies containing 25% Curcuma angustifolia starch, showed that Curcuma angustifolia starch could be 40% wheat flour, 20% pearl millet flour and 15% WPC was successfully utilized along with other flours such as wheat the best combination for making cookies. The fortification of and pearl millet and fortification with whey protein 15% WPC in cookies resulted in 58.84% rise in proteins of concentrate for the development of protein enriched cookies the cookies. which could be help to combat protein deficiency and malnutrition. The protein rich cookies coupled with the high Acknowledgement content of minerals such as copper, zinc, iron and manganese The authors acknowledge the support and facilities provided could be added advantage for cookies. Based on the high by the Director, ICAR-CTCRI for the study. ~ 95 ~ Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com

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