Sensitive HPLC-DMS/MS/MS method coupled with dispersive magnetic solid phase extraction followed by in situ derivatization for the simultaneous determination of multiplexing androgens and 17-hydroxyprogesterone in human serum and its application to patients with polycystic ovarian syndrome.

BACKGROUND AND AIMS: Androgens play important roles in polycystic ovarian syndrome (PCOS). However, measures of androgens based on mass spectrometry (MS) remain complex due to endogenous inferences of isomers or compounds with similar structures. Lack of sensitivity can also affect the accurate quantification of androgens, especially for very low level of 11-oxygenated androgens.

MATERIALS AND

METHODS

We developed a fast and sensitive high-performance liquid chromatography-differential mobility spectrometry tandem mass spectrometry (HPLC-DMS/MS/MS) method for the simultaneous determination of seven androgens and 17-hydroxyprogesterone. Dispersive magnetic solid phase extraction (DMSPE) was conducted with core-shell structured nanoparticles of magnetic graphene oxide (Fe3O4@GO). In situ derivatization was performed using Girard's Reagent P.

RESULTS

Linear ranges of the eight analytes were set in terms of clinical use. Intra- and inter-run precisions were < 16.7 % and 12.9 % for all the analytes and relative error was - 14.7-13.3 % and - 9.3-11.0 %, respectively. Extraction recoveries were 54.0-92.7 % for different analytes. The method was validated and was applied to assay 432 clinical samples.

CONCLUSION

The developed method is green, fast, sensitive and accurate for the determination of endogenous androgens. It can be readily implemented in medical laboratories to provide superior analytical performance over the traditional electrochemiluminescence immunoassay method.