Abstract
Phytate in plants (inositol phosphates, InsPs) affects mineral bioavailability. However, methods for their quantification may lead to variable results, and some are nonspecific (spectrophotometric techniques). In this study, ion-pair high-performance liquid chromatography (HPLC) was coupled with post-column derivatization to allow fluorescence detection (FLD, λexcitation324/λemission364 nm) of InsPs. The fluorescence derivatization reaction, the main input of this study, was based on a ternary complex among phytate, iron(III), and 1,10-phenanthroline. Phytic acid (InsP6) and three other InsPs (InsP3-5) were analyzed in peanuts and soybean products after extraction in HCl 0.66 M, followed by purification on strong anion exchange cartridges. The novel method, named IP-HPLC-FLD, selectively separated InsP3-6 with linear ranges between 600 and 2000 mg 100 g-1 (R2 > 0.99). The limits of detection and quantification were between 120-180 and 340-540 mg 100 g-1, respectively. As the relative standard deviations were under 10%, the IP-HPLC-FLD method is suitable for phytate analysis.
Published Version
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