LC-ESI/MS/MS method for rapid screening and confirmation of 44 exogenous anabolic steroids in human urine

Abstract

A sensitive and rapid method based on liquid chromatography-triple-quadrupole tandem mass spectrometry (LC-MS/MS) with electrospray ionization (ESI) has been developed and validated for the screening and confirmation of 44 exogenous anabolic steroids (29 parent steroids and 15 metabolites) in human urine. The method involves an enzymatic hydrolysis, liquid-liquid extraction, and detection by LC-MS/MS. A triple-quadrupole mass spectrometer was operated in positive ESI mode with selected reaction monitoring (SRM) mode for the screening and product ion scan mode for the confirmation. The protonated molecular ions were used as precursor ions for the SRM analysis and product ion scan. The intraday and interday precisions of the target analytes at concentrations of the minimum required performance levels for the screening were 2-14% and 2-15%, respectively. The limits of detection for the screening and confirmation method were 0.1-10 ng/mL and 0.2-10 ng/mL, respectively, for 44 steroids. This method was successfully applied to analysis of urine samples from suspected anabolic steroid abusers.

Sensitivity of GC‐EI/MS, GC‐EI/MS/MS, LC‐ESI/MS/MS, LC‐Ag+CIS/MS/MS, and GC‐ESI/MS/MS for analysis of anabolic steroids in doping control

“…Except for the injection mode, the conditions were identical for the GC‐based methods. The LC conditions were based on papers published previously by our group . All targeted analytes and their abbreviation are listed in Table .…”

“…Sample preparation was based on our previous reports . Urine (2 mL) samples were loaded onto a PAD‐1 column that had been washed with 2 mL of DW.…”

“…In addition, new analytical approaches using GC combined with atmospheric pressure ionization (API) sources, such as atmospheric‐pressure chemical ionization (APCI) and atmospheric‐pressure photoionization (APPI), have been recently reported . Liquid chromatography‐electrospray ionization/tandem mass spectrometry (LC‐ESI/MS/MS) has also been used as a complementary analytical tool for general GC‐MS procedures . Many other approaches based on the ionization of steroids using ESI, APCI, APPI, and CIS have been investigated by several research groups .…”

Urinary detection of conjugated and unconjugated anabolic steroids by dilute‐and‐shoot liquid chromatography‐high resolution mass spectrometry

“…[4] In general, analytical methods to detect AAS include hydrolysis, extraction and detection by liquid chromatography-mass spectrometry (LC-MS) [5] or by gas chromatography-mass spectrometry (GC-MS). Phase-I reactions include i.a.…”

High‐temperature high‐performance liquid chromatography on a porous graphitized carbon column coupled to an Orbitrap mass spectrometer with atmospheric pressure photoionization for screening exogenous anabolic steroids in human urine

“…Furthermore, because this technique does not allow for direct detection of steroid metabolites, there is need for a derivatization step, which increases the analysis time. Diverse analytical methods using liquid chromatography/mass spectrometry (LC/MS) with electrospray ionization (ESI) have been developed recently as analytical tools complementary to general GC/MS procedures . Recently, an LC/MS/MS method with metal ion coordination ionspray was developed for the detection of 84 anabolic steroids and metabolites .…”