Objective To develop a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for simultaneously determining 13 phthalate metabolites in human urine.
Methods Urine samples were prepared by hydrolyzation with beta-glucuronidase and extraction by solid-phase extraction, and then separated on a SB-phenyl RRHD column before applying isotope dilution-HPLC-MS/MS.
Results Good linear relationships were obtained in the range of 0.5 μg/L to 200μg/L with the correlation coefficients above 0.999 for all tested 13 metabolites. The limits of detection were between 0.1μg/L and 0.2 μg/L. The recoveries were in the range of 93.7% to 110.3%, and all relative standard deviations (RSD) were lower than 10%.
Conclusion A sensitive analytical method, based on SPE followed by isotope dilution-HPLC-MS/MS, to quantitatively measure 13 phthalate metabolites in human urine has been developed and validated.
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