Determination of phytic acid in urine by inductively coupled plasma mass spectrometry.

Abstract

An ICPMS method for the determination of phytic acid in human urine based on the total phosphorus measurement of purified extracts of phytic acid is described. Pretreatment of the sample is required to avoid interference in the ICPMS detection from other phosphorus compounds accompanying phytic acid in urine such as phosphate or pyrophosphate. This treatment consists of a simple filtration of the urine sample followed by complete separation of phytic acid from the mentioned phosphorus components using an anion-exchange solid-phase extraction. Separation/recovery conditions, optimized for standards of phytic acid prepared in water and artificial urine, were successfully applied to natural urine samples, resulting in adequate accuracy and precision. Linear range (0.02-0.6 mg of phytic acid L(-)(1)) and limit of detection (5 microg L(-)(1) phytic acid) are adequate for analysis of the usual amounts of phytic acid present in urine. Phosphate, pyrophosphate, and pH of urine samples at concentrations exceeding their normal physiological ranges do not affect the determination of phytic acid. Because of the simplicity, low sample requirement, and relatively high sample throughput (10 to 6 min per sample for runs between 50 and 100 samples, respectively), the present method presents the best alternative to current methods for phytic acid determination in urine. Results also show that the method is adequate for the differentiation of levels of phytic acid excretion from patients suffering from oxalocalcic urolithiasis and healthy controls, suggesting that low phytic acid concentrations in urine lead to elevated risk of oxalocalcic urolithiasis.