Liquid chromatography and supercritical fluid chromatography as alternative techniques to gas chromatography for the rapid screening of anabolic agents in urine

Annual banned‐substance review: analytical approaches in human sports drug testing

“…Also utilizing an orthogonal chromatographic system, Novakova et al . as well as Desfontaine et al . demonstrated the capability of supercritical fluid (SFC) LC interfaced via positive ESI to a QqQ‐based MS/MS system to detect steroidal analytes in human urine.…”

“…In addition, the performance of the assay targeting recently identified long-term metabolites of AAS would be of particular interest to outline the fitness-for-purpose of this alternative to established chromatographic-mass spectrometric test methods. Also utilizing an orthogonal chromatographic system, Novakova et al [54] as well as Desfontaine et al [55] demonstrated the capability of supercritical fluid (SFC) LC interfaced via positive ESI to a QqQ-based MS/MS system to detect steroidal analytes in human urine. In both studies, assays were developed that employed identical instruments consisting of an SFC liquid chromatograph equipped with a so-called high density diol analytical column (100 x 3.0 mm, 1.7 μm particle size) operated with mobile phases composed of CO 2 and methanol/water (49:1, v/v) containing 10 mM ammonium formate.…”

Supercritical fluid chromatography – Mass spectrometry: Recent evolution and current trends

“…This observation was explained by the use of improved ionization sources on the more recent MS instruments, making them more able to handle higher proportion of water [51]. As an example, it was found that, out of 43 anabolic agents tested in human urine, LC provided a sensitivity level equal to 0.1 ng/mL for 98% of the analyzed compounds, while in SFC this percentage was reduced to 76% [52]. A similar result was obtained for vitamin D metabolites, with worse LLOQs in SFC than LC [53].…”

“…The main reason for these negative results is related to the limited injection volume in SFC. Indeed, it is well known that a lower injection volume has to be used in SFC vs. LC, especially when using polar and polar protic solvents such as methanol or water as the injection solvents [53][54][55], which should obviously negatively affect sensitivity. Moreover, different column geometries are generally used in LC and SFC (2.1 mm and 3.0 mm as internal diameters, respectively), which could further increase the dilution factor in SFC and reduce achievable sensitivity [52].…”

“…A second application area that is being under constant development is the implementation of SFC-MS in the forensic and anti-doping control analysis. Indeed, there has been an important number of papers recently released and focusing on several classes of compounds: amphetamines [45,69,70], stimulants and sympathomimetic drugs [51,71,72] or anabolic agents and steroids [44,54,73,74] (Fig. 4).…”