Characterization of Red and Purple-Pericarp Rice (Oryza Sativa L.) Based on Physico-Chemical and Antioxidative Properties of Grains
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Oryza Vol. 54 No. 1, 2017 (57-64) Characterization of red and purple-pericarp rice (Oryza sativa L.) based on physico-chemical and antioxidative properties of grains Priyadarsini Sanghamitra*,TB Bagchi, RP Sah, SG Sharma, Sutapa Sarkar and Nabaneeta Basak ICAR-National Rice Research Institute, Cuttack-753006, Odisha, India *Corresponding author e-mail: [email protected] Received : 18 July 2016 Accepted : 14 March 2017 Published : 19 May 2017 ABSTRACT In the preset study range of variations in physico-chemical, cooking characteristics and antioxidant properties of six pigmented rice (four purple and two red) cultivars from north east India were evaluated. Significant variation (P<0.05) was detected among the cultivars for all the traits evaluated except for volume expansion ratio (VER). All the genotypes had long slender grain. Hulling and milling % for all the genotypes were more than 74 and 61%, respectively. Head rice recovery (HRR) was more than 50% in Manipuriblack, Kalobhat and Assambiroin. The range of amylose content (AC) varied from 2.19 to 24.87% where as Mornodoiga was found with highest AC. All the genotypes except Manipuriblack elongated more than 9mm after cooking. Most of the genotypes were found with soft gel consistency (GC). Similarly, all the genotypes except Assambiroin had water uptake (WU) value ≤ 100ml/100g rice. The concentration of total anthocyanin content (TAC), total phenolic content (TPC) and antioxidant activity (ABTS) differed significantly among the genotypes with highest concentration of these parameters were observed for the purple grain (Mamihunger) whereas no significant difference between the colour groups (red and purple) was observed for total flavonoid content (TFC), gamma- oryzanol and phytic acid content which envisages that value of these parameters depends on genotypes and not on kernel colour. A high correlation of TAC with TPC and ABTS suggest that the major phytochemicals responsible for the tested antioxidant activities are phenolic acids and anthocyanin. Key words: Pigmented rice, antioxidants, grain quality, physico-chemical characteristics Rice (Oryza sativa L.) is consumed as a staple food are very important determinants of cooked rice grain by more than half of the world's population with quality (Ge et al. 2005). The content of amylose in rice approximately 95% of production in Asia (Bhattacharjee is considered the principal determinant of rice quality. et al. 2002). It is the only cereal crop cooked and However, rice varieties with similar amylose content consumed mainly as whole grains, and quality have shown to possess different rice characteristics considerations are much more important than for any on cooking which indicated that secondary differences other food crops (Hossain et al. 2009). Grain quality exist among varieties with similar amylose content. has always been an important consideration in rice Most of the rice crops grown and consumed variety selection and development. It is the primary throughout the world have the white pericarp but there determinant for market place and consumer are many special cultivars of rice known as pigmented acceptability. The Kernel appearance, size, shape, rice characterized by red, black and purple pericarp. nutritional value and cooking characteristics are This pigmentation depends on the kinds of deposition important for judging the quality and preference of rice of phenolic compounds such as anthocyanin and from one group of consumer to another (Kanchana et proanthocyanidin in the aleurone layer of the grain al. 2012). In particular, the cooking and eating qualities r57 r Oryza Vol. 54 No. 1, 2017 (57-64) (Finocchiaro et al. 2010; Pereira- Caro et al. 2013a, laboratory rice huller, Satake, Japan make. The kernels 2013b). Great interest has been shown in the were ground by a grinding machine (Glen mini grinder) polyphenols in rice for their multiple biological activities. and sheaved through 100 mesh size and then stored at These phenolic compounds include ferulic acid and 4°C for further experiments. diferulates, anthocyanins, anthocyanidins and polymeric Anthocyanin content of rice samples was proanthocyanidins (condensed tannins) (Chun et al. measured in UV-V is spectrophotometrically according 2005). Phenolics have free radical scavanging activity to Swain and Hillis (1959) with alcoholic extract. which protect cell against oxidative damage. Gamma-oryzanol content (GOC) was Free radicals have been claimed to play an determined with RP-HPLC. γ -oryzanols extraction by important role in affecting human health by causing HPLC was performed according to Chen et al. (2005) many diseases (e.g., heart diseases, cancer, with simplification. Briefly, 0.5 grams of samples hypertension, diabetes and atherosclerosis). In the past (Brown rice flour) were mixed with 5 ml of HPLC- decade, antioxidants have shown their relevance in the grade isopropanol, vortexed for 2 min at 25°C, prevention of various diseases, in which free radicals centrifuged at 4500 g for 10 min and the supernatant are implicated. This coloured rice are known source of was collected. After 2-3 times repetition, supernatant antioxidant compounds including flavonoid, anthocyanin, fractions were evaporated under hot water bath and phytic acid, proanthocyanidin, tocopherols, tocotrienols, then extracts were dissolved in 5 ml of HPLC-grade γ -oryzanol, and phenolic compounds (Butsat and isopropanol. After filtration through a 0.45 μ m Siriamornpun 2010; Goufo and Trindade 2014) which membrane, 20 μ l aliquots were injected into the column can decrease oxidative stress in vivo, highly effective (C18- Phenomenex Column). It was separated by an in reducing cholesterol levels in the human body and analytical Shimadzu High Performance Liquid exert beneficial effects on human health (Santos-Buelga Chromatography (RP-HPLC) system equipped with an and Scalbert 2000; Ghie and Walton 2007; Lee et al. LC-20AT pump and PDA detector (Shimadzu, Kyoto, 2008). Major anthocyanins such as peonidin, peonidin Japan). The composition of the mobile phase was 35% 3-glucoside and cyanidin 3-glucoside extracted from acetonitrile, 55% methanol and 10% isopropanol and black rice, also reported to exert an inhibitory effect operated in low pressure gradient mode. of cell invasion on various cancer cells (Chen et al. 2006). Total phenolic content (TPC) was determined by modified protocol of Zilic et al. 2011. About 0.3 g of Considering health protecting and promoting brown rice flour sample and 10 ml 70% acetone were effect of pigmented rice, the objective of this study was mixed thoroughly in a centrifuge tube at room to compare physico-chemical, cooking properties and temperature. After centrifugation for 20 minutes at antioxidant potentials of the six pigmented rice cultivars 15000g, aliquots (0.2 ml) of aqueous acetone extracts native to the North-east India viz., Mamihunger, were transferred into test tubes and their volumes made Manipuri black, Chakhao, Kalobhat, Mornodoiga and up to 0.5 ml with distilled water. After addition of the Assambiroin. Folin-Ciocalteu reagent (0.25 ml) and 20% aqueous MATERIALS AND METHODS sodium carbonate solution (1.25 ml), tubes were vortexed. After 40 min, the absorbance was recorded Pigmented paddy varieties were collected from the sub- at 725 nm against a reagent blank. The total phenolic station of NRRI, Gerua, Assam and multiplied in the content of each sample was determined by means of a NRRI experimental field, Cuttack, India. All the paddy calibration curve prepared using catechol and expressed samples were from the recent harvest of Kharif, 2015. as mg catechol equivalents (CE) per gram of brown The agronomic data were collected from the field time rice flour. to time. The paddy samples were sun-dried (moisture up to 12-13%) and cleaned for foreign materials, packed Total flavonoid content was determined in polyethylene bags and kept inside cloth bags, and according to Eberhardt et al. (2000). One gram rice stored at 4°C. The samples were dehulled through grain was extracted in 10 ml of 40% (v/v) ethanol for 30 min at room temperature. The supernatant, after r58 r Properties of red and purple-pericarp rice Sanghamitra et al. centrifugation for 20 min at 15,000g was used in that the blank OD will be 0.453+ 0.002. PA experiments. Briefly, 0.075 ml of 5% NaNO2 was concentration was determined by using the following mixed with 0.5 ml of the sample (ethanolic extract formula: PA% = {(0.463-OD) x 25 V} / (22.05 x M). diluted with 1 ml of water). After 6 min, 0.15 ml of a Here OD is absorbance; V=final volume (ml); 10% AlCl3 solution was added, and the mixture was M=weight of sample (g). allowed to stand for another 5 min. Then, 0.5 ml of 1 M For physical traits, 100 g of rice seeds were NaOH was added, and the volume was made up to 2.5 de-hulled and milled using a standard de-husker and ml with distilled water. The absorbance was measured miller, respectively and the milling, head recovery ratio at 510 nm immediately after mixing, against the blank (HRR), kernel length (KL), kernel breadth (KB) and containing the extraction solvent instead of a sample. length/breadth (L/B) were calculated. For the gel The results are expressed as mg CE (catechine consistency (GC), 100 mg of rice flour was taken in equivalent) per 100g of dry matter. test tube (13 x100 mm), 0.2 ml of ethanol containing ABTS radical scavenging was assayed by 0.03% thymol blue and 2.0 ml of 0.2 N of KOH were modified protocol of Serpen et al. 2008. In this added and kept in boiling water-bath for 8 min, cooled, methodology, both soluble and insoluble fraction of mixed well and kept in ice bath for 20 min. Later, the antioxidant compounds come into contact with the test tubes were laid horizontally on the flat base, graph ABTS radical. The ABTS+ reagent was prepared by paper for one hour and length of gel spreading of those reacting a 7 mmol/l aqueous solution of ABTS with tubes were measured (mm).