Column-switching high-performance liquid chromatography electrospray mass spectrometry coupled with on-line of extraction for the determination of mono- and di-(2-ethylhexyl) phthalate in blood samples

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Abstract
A novel method based on column-switching high-performance liquid chromatography–electrospray mass spectrometry (LC–MS) coupled with an on-line extraction column containing conjugated avidin has been developed for direct injection analysis of di(2-ethylhexyl) phthalate (DEHP) and its metabolite, mono(2-ethylhexyl) phthalate (MEHP), in blood samples. The sample preparation for on-line extraction involved the mixing of blood samples with internal standards, DEHP-d4 and MEHP-d4, in LC glass vials. A linear response was found for column-switching LC–MS when tests were conducted within the validated range of 25 to 1000 ng mL–1 for DEHP and 5 to 1000 ng mL–1 for MEHP, with correlation coefficients (r) greater than 0.999. In addition, the recoveries of DEHP and MEHP from human plasma were calculated by using this method with on-line extraction, yielding recoveries of up to 91.2% (RSD–1 for DEHP, and N.D. is –1 for MEHP). In contrast, high DEHP contamination of commercially available fetal bovine serum samples was found by this method.

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