Determination of testosterone and epitestosterone glucuronides in urine by ultra performance liquid chromatography-ion mobility-mass spectrometry

Abstract

UPLC-ion mobility spectrometry separations combined with mass spectrometry (UPLC-IM-MS) and tandem mass spectrometry (UPLC-IM-MS/MS) have been investigated for the simultaneous determination of testosterone and epitestosterone glucuronides in urine. The glucuronide epimers of testosterone and epitestosterone were separated by ion mobility spectrometry prior to mass analysis on the basis of differences in their collision cross sections, which have been measured in nitrogen. Combining ion mobility separation with UPLC/MS enhances the analysis of these low-abundance steroids in urine by selective interrogation of specific retention time, mass-to-charge and mobility regions. Detection limits for the UPLC-IM-MS/MS analysis of TG and ETG were 9.9 ng mL(-1) and 98 ng mL(-1) respectively, equivalent to 0.7 ng mL(-1) and 7.4 ng mL(-1) in urine, with linear dynamic ranges corresponding to 0.7-108 ng mL(-1) and 7.4-147 ng mL(-1) in urine. Repeatability (%RSD) for urine extracts was 0.64% and 2.31% for TG and ETG respectively.

Automated and sensitive determination of four anabolic androgenic steroids in urine by online turbulent flow solid-phase extraction coupled with liquid chromatography–tandem mass spectrometry: A novel approach for clinical monitoring and doping control

“…The LOQs in Milli-Q water were 0.01 ng/mL, which were ten times lower than the reported method by off-line SPE or online SPE analysis [15,19,26,33]. This was attributed to the combination of online SPE with large volume injection and high sensitive MS-MS spectrometry.…”

Formation of multimeric steroid metal adducts and implications for isomer mixture separation by traveling wave ion mobility spectrometry

“…Steroid analysis presents a difficult challenge because of the high number of isomers with very different biological functions . These compounds have been traditionally characterized by gas chromatography (GC) and mass spectrometry (MS) . However, because of limited resolution between some isomers and the requirement for derivatization, the field has moved to other methods including liquid chromatography (LC), nuclear magnetic resonance (NMR), and tandem MS .…”

“…LC was first coupled to high field asymmetric IMS (FAIMS), a spatial separator, for steroid mixture analysis . More recently, LC was coupled to traveling wave IMS (TWIMS), a temporal separator, to analyze steroid glucuronides . In both cases, IMS was used to improve signal to noise ratios and to mobility select the steroids of interest, which increased confidence and sensitivity of the techniques.…”

In vitro evidence of the promoting effect of testosterone in kidney stone disease: A proteomics approach and functional validation

“…Additionally, testosterone can be excreted into the urine [25,26], allowing renal tubular cells to directly expose to high-testosterone environment in male kidney. Altogether, its hormonal and direct effects might be responsible for the risk and high incidence of kidney stone disease in males.…”