Quantification of melamine in human urine using cation-exchange based high performance liquid chromatography tandem mass spectrometry

Abstract

Melamine and cyanuric acid have been implicated as adulterants in baby formula in China and pet foods in North America. In China, the effect of melamine or melamine–cyanuric acid adulteration lead to kidney stone development and acute renal failure in thousands of Chinese infants. A selective and sensitive analytical method was developed to measure melamine in human urine in order to evaluate the extent of potential health implications resulting from the consumption of these types of adulterated products in the general US population. This method involves extracting melamine from human urine using cation-exchange solid-phase extraction, chromatographically separating it from its urinary matrix co-extractants on a silica-based, strong-cation exchange analytical column using high performance liquid chromatography, and analysis using positive mode electrospray ionization tandem mass spectrometry. Quantification was performed using modified, matrix-based isotope dilution calibration covering the concentration range of 0.50–100ng/mL. The limit of detection, calculated using replicates of blank and low level spiked samples, was 0.66ng/mL and the relative standard deviations were between 6.89 and 14.9%. The relative recovery of melamine was 101–106%. This method was tested for viability by analyzing samples collected from the general US population. Melamine was detected in 76% of the samples tested, with a geometric mean of 2.37ng/mL, indicating that this method is suitable for reliably detecting background exposures to melamine or other chemicals from which it can be derived.