Short-column gas chromatography/tandem mass spectrometry for the detection of underivatized anabolic steroids in urine

Abstract

Short-column (3.5 m) gas chromatography (GC)/tandem mass spectrometry (MS/MS) has been investigated for the detection of structurally related, underivatized anabolic steroids in urine. The approach described here demonstrates the ability to rapidly and qualitatively detect underivatized anabolic steroids in spiked urine matrices. In this approach, underivatized steroids are determined using a short-column GC separation, ionized by positive ion chemical ionization, and detected by selected reaction monitoring MS/MS. This approach permits positive identification of underivatized anabolic steroids based on retention time and the production of characteristic product ions. Preliminary detection limits studies in spiked urine samples showed quantitative results between 2 and 40 ng steroid per milliliter of uterine. The potential advantages of this approach compared to present screening methods based on conventional (30 m) GC/MS are its rapidity and selectivity. Reliable qualitative identification can be performed with a short-column GC/MS/MS analysis of less than 6 min with a reduction in sample preparation time due to the elimination of the derivatization step.

General Methods for the Extraction, Purification, and Measurement of Steroids by Chromatography and Mass Spectrometry

“…Short columns can enable faster separation but narrow bore columns can be easily overloaded with sample. Few publications have yet described the benefits for steroids (Rossi et al, 1994).…”

Applications and mechanisms of charge-remote fragmentation

“…The [M À H] À and [M Na] ions of steroid derivatives (bile salts and steroid conjugates) undergo charge-remote fragmentation, providing structural information of the ring systems, including positions of unsaturation, hydroxy, or keto groups (Grif®ths, Zhang & Sjo Èvall, 1993Marschall et al, 1994;Rossi, Johnson & Yost, 1994;Ikeda et al, 1995). Stroobant et al (1995) to occur.…”

A method for determination of one hundred endogenous steroids in human serum by gas chromatography-tandem mass spectrometry.

“…However, the use of GC-MS/MS in the analysis of endogenous steroids has still been limited. Current studies using the GC-MS/MS platform have mostly focused on the quantification of anabolic steroids in the blood of athletes or farmyard animals (Gambelunghe et al 2007, Impens et al 2007, Marcos et al 2002, Raro et al 2016, Rossi et al 1994, Shen et al 2008, Van Vyncht et al 1994, Wong et al 2017, Yamada et al 2008 or on steroid quantifications in wastewaters (Andrasi et al 2013, Kelly 2000, Trinh et al 2011, Zuehlke et al 2005. Blokland et al (2012) simultaneously quantified 47 steroids in the form of unconjugated steroids, glucuronides and sulfates in bovine urine.…”