Automated on-line liquid chromatography–photodiode array–mass spectrometry method with dilution line for the determination of bisphenol A and 4-octylphenol in serum

A novel on-line liquid chromatography–photodiode array detection–mass spectrometry (LC–DAD–MS) system was established with restricted-access media (RAM) pre-column and dilution line combined with a column-switching valve. The serum samples were injected directly onto pre-column under diluted condition by dilution line. After elution of proteins in the serum, the analytes were backflushed onto an ODS analytical column using a six-port column-switching device. The influence of the composition of the mobile phase, for instance, organic modifer, ionic strength, pH, dilution times and the rotation time of the switching valve have been investigated using bisphenol A (BPA) and 4-octyphenol (4-OP) as analytes. The evaluations for peak responses and sensitivity were conducted by MS, and proteins were removed by RAM-column with DAD monitoring at 280 nm. The peak shape was improved by adding a dilution line, especially in the case of large volume injection (LVI), which increased the sensitivity of the analysis. The selective and sensitive quantification of BPA and 4-OP in serum sample could be finished within 25 min. The method had linearity in the range 0.1–500 ng/mL with a limit of quantification for BPA and 4-OP of 0.1 and 0.5 ng/mL, respectively. The recoveries were in the range of 80–101% with less than 9.0% RSDs. This on-line LC–MS method demonstrates potential application to evaluating the exposure and risk of BPA and 4-OP in human.