Evaluation of Simple and Inexpensive High-Throughput Methods for Phytic Acid Determination

High‐throughput/low‐cost/low‐tech methods for phytic acid determination that are sufficiently accurate and reproducible would be of value in plant genetics, crop breeding and in the food and feed industries. Variants of two candidate methods, those described by Vaintraub and Lapteva (Anal Biochem 175:227–24, 1988 ; “VL” methods) and Huang and Lantzsch (J Sci Food Agric 34:1423–1426, 1983 ; “HL” methods), were evaluated. The primary concern with these methods is that, due to interference of matrix constituents including inorganic P, they can overestimate phytic acid and are ineffective at low levels of phytic acid. Twelve seed flours, representing lines of soybean, maize, barley and dry bean, containing a wide range of phytic acid levels, were analyzed by a minimum of eight cooperating laboratories using three variants of the VL method and two variants of the HL method. No method had consistently acceptable (?2.0”) “Horwitz ratios”, a measure of reproducibility, although some treatments approached that. For example, one variant of the VL method when used to assay a soybean flour with a “standard” level of phytic acid had a Horwitz ratio of 2.15. Some variants of the VL method were adequate for analyses of cereal grains regardless of phytic acid level but none accurately measured phytic acid when at low levels in soybean flours. One variant of the HL method in which the 0.2 N HCl extraction media is modified to contain 10% Na₂SO₄, did accurately measure phytic acid levels in both cereal and legume flours regardless of endogenous phytic acid levels or matrix constituents.