Abstract
Ultra-performance supercritical fluid chromatography–tandem mass spectrometry (UPSFC–MS/MS) is an alternative method for steroid analysis. Continuous development of analytical methodologies for steroid profiling is of major importance in the clinical environment to provide useful and more comprehensive data. The aim of this study was to identify and quantify a large number of endogenous steroids from the four major classes (estrogens, androgens, progestogens and corticosteroids) simultaneously within a short analytical time. This novel UPSFC–MS/MS method with electrospray in positive ionisation (ESI+) mode is robust, selective and present sufficiently high sensitivity to profile nineteen steroids in 50 µL human plasma. Under optimised conditions, nineteen different steroids were separated with high efficiency in the multiple reaction monitoring (MRM) mode. The linearity of the method was good with correlation coefficients (R2) in the range of 0.9983–0.9999 and with calibration range from 0.05–500 ng/mL in human plasma. The intraday and interday precision of the method, as RSD, was less than 15%. The accuracy of the nineteen analytes varied between 80 to 116%. Finally, the novel method was successfully applied for the determination of nineteen steroids within 5 minutes providing the possibility to use it for research as well as routine healthcare practice.
Highlights
The analytical methodologies based on chromatography and tandem mass spectrometryfor the determination of steroids in biological samples have obtained profound consideration in recent past
Separation of nineteen different endogenous steroid hormones and metabolites was successfully achieved within a 5 min run time using an UPSFC–MS/MS method (Fig. 2)
It was reported in a recent review that authors of only 12.5% of all published reports, mentioned simultaneous analysis of 8 to 35 steroid analytes from all four major classes by using gas chromatography (GC)–MS/MS or liquid chromatography (LC)–MS/MS methods[7]
Summary
The analytical methodologies based on chromatography and tandem mass spectrometryfor the determination of steroids in biological samples have obtained profound consideration in recent past. Most of the separation methods of two or more steroids are based on either liquid chromatography (LC) or gas chromatography (GC) coupled to tandem mass spectrometry (MS/MS). These methods offer simultaneous determination of steroids from the four major www.nature.com/scientificreports/. Classes (estrogens, androgens, progestogens and corticosteroids), and provide useful data in the clinical environment[7] These high-tech methods offer tremendous value in obtaining useful structural information on individual steroids and their metabolites[5]. Our method allows for the determination of nineteen endogenous steroid hormone levels in 50 μL plasma, within a few minutes
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