A "dilute-and-shoot" column-switching UHPLC–MS/MS procedure for the rapid determination of branched nonylphenol in human urine: method optimisation and some fundamental aspects of nonylphenol analysis.

Technical grade branched nonylphenol (NP) was determined in human urine by online solid phase extraction–ultra high-performance liquid chromatography–tandem mass spectrometry (SPE–UHPLC–MS/MS). Prior to analysis, urine specimens were simply diluted and enzymatically deconjugated. The run time of the chromatography, including SPE and re-equilibration, was 9 min per injection. The enzymatic cleavage of NP conjugates was optimised with incurred sample material from a human metabolism study: the highest recoveries were obtained with β-glucuronidase from E. coli K 12 in 0.1 M ammonium acetate at pH 6.5, within a minimal hydrolysis time of 30 to 60 min. Using sodium acetate instead of ammonium acetate led to systematically decreased recovery rates. The analytical method was validated regarding its precision (coefficients of variation: 2.9–7.4%), accuracy (relative recovery rates: 93–105%), robustness (relative recovery rates in individual urine matrices: 92–117%), selectivity, and limit of quantification (1.0 μg L−1). Fundamental aspects in the analysis of technical product mixtures such as NP, comprising various isomers and homologues, were considered. Validation results, an exposure scenario and the application of the procedure to real samples, show that it enables a rugged monitoring of NP exposures above, at, and significantly below health-based guidance values, corresponding to daily NP intakes in the low μg kg−1 d−1 range. On the other hand, background levels in non-specifically exposed populations cannot be detected with this method. Hence, while alternative approaches should be pursued for NP analysis at environmental trace level, the speed and simplicity of our method are ideal for high-throughput human biomonitoring in occupational medicine. [ABSTRACT FROM AUTHOR]