Abstract
Identifying or designing low phytic acid soybean demands an improved understanding of its dynamics and a need for a reliable and accurate phytate quantification method and hence an improved quantitative technique for accurate and rapid quantification of phytic acid (PA) using high-performance liquid chromatography is proposed in this paper. A rapid PA extraction method utilizing sonication of sample in 0.78 M HCl for 3 min followed by mechanical agitation and separation using strong anion exchange column in a vacuum manifold was optimized. The elution of PA was performed using a RP-C18 column with an isocratic mobile phase [Acetonitrile, 35 mM formic acid and tetrabutylammonium hydroxide (4.8: 5.1: 0.1, v/v/v)]. The modified method was rapid, accurate, precise, and reproducible with relative standard deviation of 1.80 and 3.01 % (n = 10, for 1 mg ml−1) for within and between days respectively with linearity (R2 = 0.999, P < 0.05), low limit of detection (LOD = 7.8 μg ml−1) and limit of quantification (LOQ = 31.25 μg ml−1). PA dynamics was found increased in a linear trend from initial to later developing stages until maturity in both yellow (DS-9814) and black (DS-MM-64) genotypes. The PA content ranged from 2.38–4.72 g 100g−1 in the 20 soybean genotypes screened and the variability in PA content was more in black genotypes (P < 0.0001). The black soybean genotypes JS 76-205 (2.38 g 100g−1), Kalitur (2.50 g 100g−1) and UPSL 652 (2.54 g 100g−1), inherently rich in anthocyanin, contains the lowest PA content and hence can serve as potential genotypes with nutraceutical benefits.
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Abbreviations
- PA:
-
Phytic Acid
- RP:
-
Reverse Phase
- HPLC:
-
High Performance Liquid Chromatography
- P:
-
Phosphorous
- MRP:
-
Multidrug Resistance Associated Protein
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Acknowledgment
The authors sincerely thank Department of Science and Technology, INSPIRE (IF120064) and National Agricultural Science Fund (RNAi 2011), Indian Council of Agricultural Research for providing financial support. The authors also duly acknowledge Dr. Prakash Kumar, Scientist, Indian Agricultural Statistics Research Institute, New Delhi, for data analysis.
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Pandey, V., Krishnan, V., Basak, N. et al. Phytic acid dynamics during seed development and it’s composition in yellow and black Indian soybean (Glycine max L.) genotypes through a modified extraction and HPLC method. J. Plant Biochem. Biotechnol. 25, 367–374 (2016). https://doi.org/10.1007/s13562-015-0348-0
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DOI: https://doi.org/10.1007/s13562-015-0348-0