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Antinutrient and Mineral Content of Thirteen Different Varieties of Pearl Millet Locally Grown in Haryana, India

Pearl millet is small seeded grains, mainly produced in tropical and semi-arid region in India and Africa. It is underutilized and lesser known crop with capability of growing in less water areas on about 30 million ha in more than 30 countries of five continents viz., Asia, Africa, North America, South America, and Australia. India is the largest producer of the pearl millet in the world and its cultivation is next to rice, maize and wheat. The major pearl millet growing states in order of area are Rajasthan, Maharashtra, Gujarat, Uttar Pradesh and Haryana and these account for 87% of the total area under cultivation of pearl millet (Rana et al., 2012). In Haryana, production area under this crop during karif2014-15 was 383 hectare with total production of 670 lakh tonnes and average yield of 1749 kgs/ha (Anonymous, 2015). It is cheap and economic crop of dual purpose which is used as food for human, feed for poultry birds and also dry as well as green fodder for cattle. Pearl millet commonly known as bajra and termed as “nutricereal” due to rich source of energy, carbohydrate, protein, fat and ash, besides, it is also a rich source of dietary fiber and minerals (iron, calcium, magnesium, potassium etc) (Abdalla International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 6 Number 5 (2017) pp. 2136-2143 Journal homepage: http://www.ijcmas.com

. It is free from major antinutrients but contains variable amount of phytate and polyphenols and different treatments such as soaking, fermentation, blanching and roasting decrease the level of these antinutrients (Eyzaguirre et al., 2006;Hithamani & Srinivasan, 2014;Lestienne et al., 2005). In developing countries, the commercial processing of these locally grown grains into value-added food and beverage products can be an important driver for economy and to curb malnutrition, therefore, the objective of this study to analyze the antinutrient and mineral content of thirteen pearl millet varieties.

Materials and Methods
Pearl millet varieties viz., composite/desi varieties , improved released variety (HHB-67), released hybrids , white composite variety , white seeded hybrid  and hybrid in pipeline (HHB-265) were procured from the Department of Genetics and Plant Breeding, CCSHAU, Hisar.
The chemicals were procured from Sigma-Aldrich Chemicals Pvt. Ltd. New Delhi (India) and Central Drug House (CDH), New Delhi (India).

Polyphenols
The polyphenols were determined by the method of Swain & Hills (1959). About 500 mg of defatted sample was refluxed with 50 ml methanol containing 1% HCl for 4 hr. The extract was concentrated by evaporating methanol on a boiling water bath and brought its volume to 25 ml with methanolic-HCl. The extract (1.5 ml) was diluted with water to 8.5 ml in 10 ml graduated test tube. The contents were well mixed. Then, 0.5 ml Folin-Denis reagent was added and the tubes were thoroughly shaken. Exactly after 3 min., 1 ml saturated sodium carbonate solution was added and the tubes were thoroughly shaken again and allowed to stand for 1 hr. The absorbance was read at 725 nm using methanolic-HCl as blank. If the solution was cloudy or precipitates appeared, it was centrifuged before readings were taken. A standard curve was plotted by taking 0.5 ml to 4.0 ml of tannic standard solution containing 10 µg to 80 µg of tannic acid.
Where, M = concentration of extract elute obtained from standard graph V = volume made of extract (ml) V1= volume of aliquot (ml) W = weight of sample (g)

Phytic acid
Phytic acid was estimated using method of Haug & Lantzsch (1983). About 1 g finely ground sample was extracted with 25 ml 0.2 N HCl for 3 hr with continuous shaking in a shaker. After proper shaking, it was filtered through Whatmann No.1 filter paper. The volume was made to 25 ml with 0.2 N HCl. Then 0.5 ml of the sample extract was pipetted into a test tube. One ml ferric ammonium sulphate solution was added. The tube was heated in a boiling water bath for 30 min. The contents of the tube were mixed and centrifuged for 30 min. at 3000 rpm. 1 ml supernatant was transferred to another test tube and 1.5 ml bipyridine solution was added. The absorbance at 519 nm against distilled water was measured. For plotting a standard curve, different concentrations i.e. 0.2-1.0 ml of standard sodium phytate solution containing 40-200 µg phytic acid were taken and made to 1.4 ml with water.
Where, M = concentration of extract elute obtained from standard graph V = volume made of extract (ml) V1= volume of aliquot (ml) W = weight of sample (g)

Mineral estimation
Minerals were estimated by using the MP-AES unit (Microwave plasma-atomic emission spectroscopy, Model No. 4100 Agilent Technology). 1 g pearl millet flour was digested with 10 ml of 35% nitric oxide into digestion tubes. Samples were digested according to the ascending order of the temperature: first digested at 80 0C for 20 min., then 120 0C for 30 min., 140 0C for 50 min. and 145 0C for 5 minute. After digestion, samples were made volume up to 100 ml with distilled water. Take 10 ml for running the sample into MP-AES unit which automatically read the sample and give the concentration (ppm) of required mineral estimation.

Polyphenol content
In millet, several phenolic compounds are primarily located in the grain outer layers and can be distinguished such as phenolic acids, flavonoids and condensed polymeric phenols, also known as tannins. Tannins are polymers resulting from the condensation of flavan-3ols (Eyzaguirre et al., 2006), form complex with protein, carbohydrates, and minerals and thus reduce digestibility of these nutrients (Dyke & Rooney, 2006). Phenolics may also contribute colour to pericarp (Mcdonough & Rooney, 1989) which impart undesirable colours in grain products during food processing. Several methods for decreasing the tannins in pearl millet viz., milling/decortication (Lestinne et al., 2005), cooking (Nambiar et al., 2012), fermentation (Eltayeb et al., 2007), germination (Hithamani & Srinivasan, 2014) and chemical treatment (Beta et al., 2000) have been documented. Different varieties of pearl millet studied contained 403.7 to 521 mg polyphenol/100 g (Table 1). Lowest polyphenol content was noticed in WHC-901-445 and highest in HHB-146 variety. No significant difference in polyphenol content was noticed between HHB-146 and HHB-226 varieties. Polyphenol content of other varieties except HHB-67 and HHB-197 was found significantly lower than HHB-226. Variation in polyphenol content (608-788, 741.29 to 767.54 and 502.78 to 658.30 mg/100 g) has been reported by Sharma & Kapoor (1996), Archana et al., (1998) and Sangwan (2005). Perusals of data indicate that White seeded variety has low content of polyphenol than other varieties and this is positively correlated to its colour. Similar reports have been documented which states that dark varieties have high polyphenol content than light (Walter & Marchesan, 2011, Li et al., 2015. This variation in polyphenols may be associated to various factors such as genotypic effect, degree of maturity and environment effect (Radhouane & Fattouch, 2009).
In conclusion it was found that significant variation exist in polyphenol and phytic acid content of thirteen different varieties of pearl millet. Significant amount of different minerals was also present in these varieties of Haryana with good chemical composition as presented in Figure 1. With opting good processing treatments such as malting, roasting, blanching to decrease the level of such antinutrients, can make pearl millet crop better to curb malnutrition problems. As pearl millet is cheap and underutilized crop of Haryana, such knowledge may help the food processors, researchers, and scientist to develop the food products which may lead to diversification in marketing to boost up Indian economy.