Quantitative Detection of Eight Phthalate metabolites in Human Urine Using HPLC-APCI-MS/MS

摘要

Because of the ubiquity of phthalates and their potential role in increasing risk for cancer and reproductive d..rs-function, the need for human exposure assessment stud-ies is urgent In response to this need, we developed a high-throughput, robust, sensitive, accurate, and precise assay for simultaneous measurement of trace levels of eight phthalate metabolites in human urine by HPLC-MS/MS. Human urine samples were processed using enzymatic deconjugation of the glucuronides followed by solid-phase extraction. The eluate was concentrated, and the phthalate metabolites were chromatographically re-solved by reversed-phase HPLC, detected by APCI-tandem mass specirometry, and quantified by isotope dilution. This selective analytical method permits rapid detection (7.7 mm total run time) of eight urinary me-tabolites of the most commonly used phthalates with detection limits in the low nanagram per milliliter range. Assay precision was improved by incorporating 13C4-labeled internal standards for each of the eight analytes, as well as a conjugated internal standard to monitor deconjugation efficiency. This selective, sensitive, and rapid method will help elucidate potential associations (if any) between human exposure to phthalates and adverse health effects.