Nutritional, Phytochemical and Functional Property Evaluation of Composite Flour- useful in Gluten Free Food Products


Akansha, Ekta Singh Chauhan*

Department of Food Science and Nutrition, Banasthali Vidyapith, Rajasthan, India

*Corresponding Author E-mail:



Celiac disease is an antibody-mediated enteropathy that presents permanent intolerance to ingested gluten, for which only treatment is lifelong devotion to a gluten-free diet. The aim of this study was to investigate proximate, phytochemical and antioxidant analysis of gluten-free composite flour. The proximate analysis of composite flour showed moisture: 11.1g/100g, ash: 2.7g/100g, fat: 0.9g/100g, fiber: 4.9g/100g, protein: 19.8g/100g, carbohydrate: 60.6g/100g, vitamin C: 4.5mg/100g, calcium: 120mg/100g iron: 12mg/100g, phosphorus: 283.4mg/100g and zinc: 7.8 mg/100g. Gluten content has been found 0.2g/100g and will be useful for celiac patients.  It also contains some phytochemicals such as flavonoids, saponin, tannin, glycosides and steroids. Some functional properties are assessed namely water absorption capacity (WAC), oil absorption capacity (OAC), emulsion capacity (EC), foaming capacity (FC) and gelation capacity (GC). Therefore, it can be beneficial for celiac disease, anemia, diabetes, cancer and osteoporosis condition along with being nutritionally adequate. Development of food products incorporated with composite flour as a functional food can improve health and prevent various diseases and disorders.


KEYWORDS: Composite flour; chemical composition; gluten-free; celiac disease; functional food.




Composite flour in various researches has been defined as a mixture of wheat and non-wheat flours for the making of leavened breads, pastas and other baked products; or wholly non wheat flour prepared from combinations of flours from roots, cereals, tubers, legumes and other raw materials for making traditional or new products. For organoleptic acceptability and their functional roles of flour components, composite flours is been used widely in production of baked products1. Each component of composite flour is carefully selected in the present study namely teff millet, navy beans and watermelon seeds. It plays a considerable role in contributing towards the nutritional or functional attribute of the product developed.


Teff not only provide nutrients but also have low pasting viscosity and water holding capacity. Being contradictory to this, navy beans have higher foaming stability, oil and water absorption and emulsifying properties. Watermelon seeds are rich in antioxidants and phytochemicals as compared to teff millet and navy beans. It is rich source of essential fatty acid (linoleic acid) with several health benefits2. Prepared composite flour is gluten free along with good nutritional qualities and higher fibre content that can also deal with the health problems. Additionally, gluten free bakery products are generally formulated with flours of rice or maize that have low fibre and protein content. Low dietary fibre content can lead to constipation in patients with celiac disease when they consume such foods3. The major problem related to the gluten-free products is that they are not so good in taste and structure. However, use of composite flour can solve these problems by incorporation of teff millets, navy beans and watermelon seeds. Composite flour contains many functional properties because navy beans have emulsifying and foam stability that leads to increase in the batter’s density and color property of baked products. As per the study, navy beans flour has been found to have highest cake volume similar to wheat flour4. Watermelon seed flour found to be good in overall acceptability such as colour, aroma, crispiness, texture and taste to the baked products5. Thus, as a result, composite flour based baked products will have good physical appearance and texture due to its functional properties. Composite flour will also contain good amount of macro and micronutrients along with bioactive components that possess antioxidant activity.


Thus, the main objective of the research is to evaluate prepared composite flour for its proximate composition, phytochemical and antioxidant activity that can further direct research towards its applications for gluten-free food product development in the market.



Collection of plant material:

The seeds were obtained from the local market of Delhi, India. The seeds were dried at 100ºC in an oven for half an hour. The dried sample of seeds was milled with a mechanical blender and stored in air tight containers in a refrigerator for further analysis.



In this experiment, the used nitric acid and perchloric acid were purchased from Merck, India. For standard calibration of respective elements, we purchased Ca, Fe, P and Zn standard solution (100mg/ml) from Sigma Company Mumbai, India. We prepared the respective desired from the stock solution using lab made double distilled water.


Determination of proximate composition:

Composite flour was taken in a clean, dry and weighed crucible. It was oven dried later on at 1100 C. It was weighed repeatedly until a constant weight was acquired. The crucible was cool down in desiccators every time before weighing. Proximate analysis included the estimation of moisture ash, fat, protein, crude fiber and carbohydrate of seeds. Total ash was estimated by weighing the furnace in incinerated residue at 550ºC for 12 hours. Protein was analyzed by using micro-Kjeldahl distillation method. Carbohydrate content was determined by the difference method.


Determination of minerals:

Chemical estimations were carried out for determining calcium (Ca), iron (Fe), phosphorus (P), and zinc (Zn). The estimation of Ca, Fe, P and Zn was done by using atomic absorption spectrophotometer (AAS) (model VGP 210, Buck Scientific, USA). Vitamin C was estimated by titration method.


The data recorded for respective elements was done in triplicate measurements for its authentication and used for standard deviation calculation.

Phytochemical and Antioxidant screening:

Composite flour was screened for phytochemicals (tannins, steroids, terpenoids, alkaloids, glycosides, carotenoids and flavonoids) according to the procedure as described by6-7. Antioxidant analysis was done on DPPH (diphenylpicryhydrazyl) activity, FRAP (ferric reducing ability of plasma) and nitric oxide scavenging activity8,9,10.


Functional property analysis:

Functional properties were analyzed of composite flour namely water absorption capacity (WAC), oil absorption capacity (OAC), foaming capacity (FC)11 and gelation capacity (GC)12.


Statistical Analysis:

All the results were shown in Mean and Standard Deviation.



Table 1 Proximate composition of composite flour

Nutrients (per 100g)


Moisture (g/100g)


Ash (g/100g)


Fat (g/100g)


Fiber (g/100g)


Protein (g/100g)


Carbohydrate (g/100g)


Vitamin C (mg/100g)


Calcium (mg/100g)


Iron (mg/100g)


Phosphorus (mg/100g)


Zinc (mg/100g)


Gluten (g/100g)



The proximate composition of composite flour is presented in Table 1. The table shows the Mean±SEM (g/100g) of moisture, ash, protein, fat, fiber, carbohydrate, vitamin C, calcium, iron, zinc and phosphorus content. The proximate data revealed that the moisture content (g/100g) was low (11.0±0.1) which was advantageous for prolonging the shelf life of the seeds. On the contrary, slightly decreased amount of moisture was found in other gluten-free composite flour study conducted by13 that is (10.5 g/100g). The ash content of composite flour obtained was (2.7±0.2) which was higher when compared to the study carried out by14 that is (0.3 g/100g). The quantity of ash in any seed sample assumes importance as it determines the nutritionally important minerals. The fat content of composite flour was (0.9±0.1). Similar study was conducted by15 in which data was comparable in fat content with a value (1.4±0.0 g/100g) in raw rice. The fiber content was found (4.9±0.2) in gluten-free composite flour while comparing to the study of16 the fiber content of other gluten free composite flour (50% rice flour and 50% African yam bean flour) was (4.6 g/100g). The protein content of composite flour was (19.8±0.1) which was higher than the observation obtained in other composite flour17 which is (15.3±0.0) respectively. The composite flour was abundant in carbohydrate content (60.5±0.4). Similar study was conducted by18 in which data was comparable in carbohydrate content with a value (60.28 g/100g).


The amount of vitamin C (mg/100 g) present in composite flour was 4.5±0.1. The result suggests that plant seeds are a good source of vitamin C. Similar study was conducted by19 in which data was comparable in vitamin C content with a value (4.1±0.0 g/100g) in composite flour. Calcium content was (120 ±0.3) found in appreciable amount in composite flour. On the contrary, decreased amount was seen in a study conducted by20 that is 52.8±0.3 mg/100g. The amount of phosphorus was (283.4±0.1) found in a good amount. On the contrary, study was conducted by21 lowest amount (20.68±0.01) was found in a sweet potato. The zinc content of composite flour was (7.8±0.1) which was lower than the observation obtained in other composite by22 that is 2.9±0.0 mg/100g. Gluten content in the composite flour found to be 0.2±0.0.  Recent studies have revealed that a limit of 20 mg/kg is admitted23. So, celiac patient can safely consume the products made out of this composite flour.


Phytochemical content:

Table 2 Phytochemical screening of composite flour


Teff millet seeds












Phytochemicals are non-nutritive plant chemicals are also known as phytonutrients. They contain disease preventive properties against chronic diseases such as cardiovascular diseases (CVDs), cancer, diabetes, osteoporosis and vision diseases24. The qualitative phytochemical analysis of the aqueous extracts of teff seed power showed positive results for the presence of flavonoids, saponin, tannin, glycosides and steroid which are shown in Table 2. Epidemiological and animal studies suggest reduction in the risk of chronic diseases associated with oxidative damage with the regular consumption of fruits, vegetables and whole grains.


The prior studies have reported that glycosides and flavonoids show a broad range of biological activities such as antioxidant, antiallergic, antimicrobial and anti-inflammatory25. It has been shown in the studies that flavonoids prevent the development of atherosclerosis because it helps in oxidation of low-density lipoprotein. Saponins have analgesic, anti-nociceptive, antioxidant, antifungal and antiviral effect on cold blooded animals26. It is confirmed from several studies that the tannins exhibit anti-microbial, anti-oxidant and anti-inflammatory properties. The usage of tannins rich food has lots of remedial and beneficial effects on human being and also can be used as drugs to heal the burning injury as it is helpful to stop bleeding from cuts. Tannins have the ability to form a leather resistance on the exposed tissues and helps in protecting the wounds from being affected further27. Steroids were known for their antibacterial activity specifically associated with membrane lipids and cause leakage from liposomes28.


Functional properties analysis:

Table 3 Functional properties analysis of composite flour

Functional properties

Composite flour

Water absorption capacity (%)


Oil absorption capacity (%)


Emulsion capacity (%)


Foaming capacity (%)


Gelation capacity (ºC)



In comparison to other gluten free flours, the water absorption capacity of the composite flour was found (2.0±0.1) higher. Water absorption capacity is an essential functional property required especially for dough handling in food formulations16. Oil absorption capacity is an important functional property retains the flavor of food products that enhances the mouth feel29. The oil absorption capacity of composite flour was (1.5±0.0), which was similar with the findings of30 in yam bean seed flour. This improved mouth feel and flavor retention makes oil absorption an important property in bakery food formulations. Emulsion capacity in composite flour (47.1±0.0) was comparable with the findings of31 in which other composite flour had range between (47.2±2.4; 48.4±2.9 and 48.6±3.74). Foaming capacity was found (7.6±0.1) in a composite flour. On the contrary, slightly increased amount was found in a study conducted by32 that is (12.0). Foaming capacity acts as functional agents in many food formulations including baked products and is desirable attribute for flours that are formulated for the production of a variety of baked products such as cookies, angel cakes, muffins, akara, etc33. So, gluten-free composite based biscuits will help to improved texture, consistency and appearance. Gelation capacity was found (55±0.5) in composite flour. Slightly increased amount was found in a study which is conducted by31 which had range between (56.2±0.5, 59.4±0.1 and 60.5±0.0) in other composite flours at different ratio. In this, starch granules are subjected to heat to form a gel which is important in the manufacturing of baked products.



This composite flour is beneficial for celiac patients as it contains all the essential macro and micro nutrients. This flour is better than what is available in the market, as the existing ones are not sufficient to meet daily requirements of the people suffering from gluten intolerance. They are also rich in phytochemicals which helps in fighting common diseases such as osteoporosis, anemia and constipation which are prevalent in these patients.



The authors are thankful to the Department of Food Science and Nutrition, Banasthali Vidyapith for providing necessary lab facilities.



There is no conflict of interest between the authors.



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Received on 13.02.2019           Modified on 02.03.2019

Accepted on 02.04.2019         © RJPT All right reserved

Research J. Pharm. and Tech. 2019; 12(7):3471-3774.

DOI: 10.5958/0974-360X.2019.00588.2