A Modified Colorimetric Method for Phytic Acid Analysis in Soybean

Abstract

A quantitative, reproducible, and efficient phytic acid assay procedure is needed to screen breeding populations and support genetic studies in soybeans [Glycine max (L.) Merr.]. The objective of this study was to modify the colorimetric Wade reagent method and compare the accuracy and applicability of this new method in determining seed phytic acid content in soybean with three well‐established phytic acid assay methods: anion exchange column (AEC), high‐performance liquid chromatography (HPLC), and 31P nuclear magnetic resonance (NMR). The CV for repeated measurements of a low phytic acid soybean mutant, CX1834‐1‐6, ranged from 1.8 to 4.2% (n = 5), indicating the results were precise and reproducible. Phytic acid content of 42 soybean genotypes as determined by this method showed a correlation of 93.7 to 96.6% with the measurements by AEC, HPLC, and NMR. According to analysis of covariance, using inorganic P content as a predictor, phytic acid P content in a given sample analyzed by the four assay methods can be estimated with four linear regression models at the α = 0.01 level. Compared with HPLC, AEC, and 31P NMR, this modified colorimetric method is simpler and less expensive for assaying a large number of samples, allowing its effective application in breeding and genetic studies of low phytic acid soybean.

Proximate composition, antioxidant activities and phenolic composition of <em>Cucumis sativus</em> forma hardwickii (Royle) W. J. de Wilde &amp; Duyfjes

“…The phytic acid was measured according to the methods of Gao et al [21]. In brief, 0.5 g of fine powder was mixed with 10 mL 2.4 % HCl and stirred constantly at 220 rpm for 16 h. Centrifuge the mixture at 1000 g at 10°C for 20 min and crude extract was mixed with 1 mL of Wade reagent (0.03 % FeCl 3 .6H 2 O and 0.3 % sulfosalicylic acid in distilled water).…”

Nutritional evaluation of protein isolate from rubber seed in the diet of Labeo rohita : Effects on growth performance, nutrient utilization, whole body composition and metabolic enzymes activity

“…The cyanide content of the RPI was analysed according to alkaline titration method of AOAC (2000), phytic acid and tannin contents were analysed as per the method of Gao et al (2007) and Schanderi (1970), respectively.…”

Evaluation and QTL mapping of phosphorus concentration in soybean seed

“…Upon maturity, seed from each plot was harvested in bulk for analysis. Total phosphorus was determined using the slightly modified procedures of Gao et al (2007) and Zheng et al (2009). Briefly, 5 g of seed from each line were ground using a Foss Cyclotec mill (1093 Sample Mill, Foss, Eden Prairie, MN) equipped with a 1-mm screen.…”