Login Register Cart 0
Generic placeholder image

Current Pharmaceutical Analysis

Editor-in-Chief

ISSN (Print): 1573-4129
ISSN (Online): 1875-676X

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe
Clinical Trial

Rapid Identification of 44 Steroids in Human Urine Samples using HPLCESI- QTOF-MS

Author(s): Aarti Yadav and Rajesh Dabur *

Volume 18, Issue 4, 2022

Published on: 09 March, 2021

Page: [437 - 448] Pages: 12

DOI: 10.2174/1573412917666210309145639

Price: $65

Abstract

Objectives: Detailed analysis of un-processed and un-derivatized free and conjugated urinary steroids is useful to avoid miscalculations and to diagnose sports doping and adrenal problems, including abnormal steroidogenesis, congenital deficiency of related enzymes, cancer, and other disease conditions. Hence, the present study was conducted to develop a soft ionization method to identify the maximum number of urinary steroids using ultra-performance liquid chromatography coupled with quadrupole time of flight mass spectrometer (HPLC–Q-TOF-MS).

Materials and Methods: HPLC–Q-TOF-MS was carried out for the qualitative detection of steroids and their conjugates in urine samples. The method provides high sensitivity and fast analysis of steroids and their glucuronides without hydrolysis or sample preparation or extraction of steroids.

Results: Using the method, 44 steroids belonging to C-18, C-19, and C-21 classes and their conjugates were resolved and identified using positive and negative modes of ionizations by their characteristic ionization and collision energy induced dissociation behaviors.

Conclusion: The method is time-saving and good to compare samples from different peoples with control or healthy ones as it does not require any kind of pre-treatment or sample processing. It provides a complete picture of steroids metabolism and catabolism. It can be good for doping control or to explore the effects of other drugs. However, in qualitative analysis, one may miss the significant information unless direct methods of steroids analysis to be employed.

Keywords: Steroids, Urine, HPLC, Q-TOF-MS, qualitative analysis, qualitative detection, ionization method.

« Previous
Graphical Abstract

[1]
Markham, J.A. Sex steroids and schizophrenia. Rev. Endocr. Metab. Disord., 2012, 13(3), 187-207.
[http://dx.doi.org/10.1007/s11154-011-9184-2] [PMID: 21603930]
[2]
Lee, A.L.; Ogle, W.O.; Sapolsky, R.M. Stress and depression: possible links to neuron death in the hippocampus. Bipolar Disord., 2002, 4(2), 117-128.
[http://dx.doi.org/10.1034/j.1399-5618.2002.01144.x] [PMID: 12071509]
[3]
Dokras, A. Cardiovascular disease risk in women with PCOS. Steroids, 2013, 78(8), 773-776.
[http://dx.doi.org/10.1016/j.steroids.2013.04.009] [PMID: 23624351]
[4]
Lin, K.T.; Wang, L.H. New dimension of glucocorticoids in cancer treatment. Steroids, 2016, 111, 84-88.
[http://dx.doi.org/10.1016/j.steroids.2016.02.019] [PMID: 26930575]
[5]
Mazzoni, I.; Barroso, O.; Rabin, O. The list of prohibited substances and methods in sport: structure and review process by the world anti-doping agency. J. Anal. Toxicol., 2011, 35(9), 608-612.
[http://dx.doi.org/10.1093/anatox/35.9.608] [PMID: 22080899]
[6]
Schumacher, M.; Guennoun, R.; Mattern, C.; Oudinet, J.P.; Labombarda, F.; De Nicola, A.F.; Liere, P. Analytical challenges for measuring steroid responses to stress, neurodegeneration and injury in the central nervous system. Steroids, 2015, 103, 42-57.
[http://dx.doi.org/10.1016/j.steroids.2015.08.013] [PMID: 26301525]
[7]
Stanczyk, F.Z.; Lee, J.S.; Santen, R.J. Standardization of steroid hormone assays: why, how, and when? Cancer Epidemiol. Biomarkers Prev., 2007, 16(9), 1713-1719.
[http://dx.doi.org/10.1158/1055-9965.EPI-06-0765] [PMID: 17855686]
[8]
Cook-Botelho, J.C.; Bachmann, L.M.; French, D. Steroid hormones; Mass Spectrometry for the Clinical Laboratory, 2017, pp. 205-230.
[9]
Taylor, A.E.; Keevil, B.; Huhtaniemi, I.T. Mass spectrometry and immunoassay: how to measure steroid hormones today and tomorrow. Eur. J. Endocrinol., 2015, 173(2), D1-D12.
[http://dx.doi.org/10.1530/EJE-15-0338] [PMID: 25877990]
[10]
Wang, C.; Gardinali, P.R. Comparison of multiple API techniques for the simultaneous detection of microconstituents in water by on-line SPE-LC-MS/MS. J. Mass Spectrom., 2012, 47(10), 1255-1268.
[http://dx.doi.org/10.1002/jms.3051] [PMID: 23019157]
[11]
Josephs, R.D.; Daireaux, A.; Choteau, T.; Westwood, S.; Wielgosz, R.I. Normal phase-liquid chromatography-tandem mass spectrometry with atmospheric pressure photoionization for the purity assessment of 17β-estradiol. Anal. Bioanal. Chem., 2015, 407(11), 3147-3157.
[http://dx.doi.org/10.1007/s00216-014-8189-6] [PMID: 25245421]
[12]
McDonald, J.G.; Matthew, S.; Auchus, R.J. Steroid profiling by gas chromatography-mass spectrometry and high performance liquid chromatography-mass spectrometry for adrenal diseases. Horm. Cancer, 2011, 2(6), 324-332.
[http://dx.doi.org/10.1007/s12672-011-0099-x] [PMID: 22170384]
[13]
Kushnir, M.M.; Rockwood, A.L.; Bergquist, J. Liquid chromatography-tandem mass spectrometry applications in endocrinology. Mass Spectrom. Rev., 2010, 29(3), 480-502.
[http://dx.doi.org/10.1002/mas.20264] [PMID: 19708015]
[14]
Koal, T.; Schmiederer, D.; Pham-Tuan, H.; Röhring, C.; Rauh, M. Standardized LC-MS/MS based steroid hormone profile-analysis. J. Steroid Biochem. Mol. Biol., 2012, 129(3-5), 129-138.
[http://dx.doi.org/10.1016/j.jsbmb.2011.12.001] [PMID: 22210511]
[15]
Tudela, E.; Deventer, K.; Geldof, L.; Van Eenoo, P. Urinary detection of conjugated and unconjugated anabolic steroids by dilute-and-shoot liquid chromatography-high resolution mass spectrometry. Drug Test. Anal., 2015, 7(2), 95-108.
[http://dx.doi.org/10.1002/dta.1650] [PMID: 24753397]
[16]
Poschner, S.; Zehl, M.; Maier-Salamon, A.; Jäger, W. Simultaneous quantification of estrogens, their precursors and conjugated metabolites in human breast cancer cells by LC-HRMS without derivatization. J. Pharm. Biomed. Anal., 2017, 138, 344-350.
[http://dx.doi.org/10.1016/j.jpba.2017.02.033] [PMID: 28249239]
[17]
Putz, M.; Piper, T.; Casilli, A.; Radler de Aquino Neto, F.; Pigozzo, F.; Thevis, M. Development and validation of a multidimensional gas chromatography/combustion/isotope ratio mass spectrometry-based test method for analyzing urinary steroids in doping controls. Anal. Chim. Acta, 2018, 1030, 105-114.
[http://dx.doi.org/10.1016/j.aca.2018.05.016] [PMID: 30032759]
[18]
Fenske, M. Poor correlation between 6β-hydroxycortisol:cortisol molar ratios and midazolam clearance as measure of hepatic CYP3A activity: a comment. Br. J. Clin. Pharmacol., 2007, 63(5), 632.
[http://dx.doi.org/10.1111/j.1365-2125.2006.02804.x] [PMID: 17094779]
[19]
Massbank. Available from: http://www.massbank.jp/SearchPage. html Accessed on 30-12-2017.
[20]
Massbank. Available from: https://metlin.scripps.edu/landing_page.php?pgcontent=simple_search
[21]
Shackleton, C.H.L.; Chuang, H.; Kim, J.; de la Torre, X.; Segura, J. Electrospray mass spectrometry of testosterone esters: potential for use in doping control. Steroids, 1997, 62(7), 523-529.
[http://dx.doi.org/10.1016/S0039-128X(97)00004-4] [PMID: 9253791]
[22]
Nguyen, P.T.T.T.; Lee, R.S.F.F.; Conley, A.J.; Sneyd, J.; Soboleva, T.K. Variation in 3β-hydroxysteroid dehydrogenase activity and in pregnenolone supply rate can paradoxically alter androstenedione synthesis. J. Steroid Biochem. Mol. Biol., 2012, 128(1-2), 12-20.
[http://dx.doi.org/10.1016/j.jsbmb.2011.10.003] [PMID: 22024430]
[23]
Chico, Y.; Fresnedo, O.; Lacort, M.; Ochoa, B. Effect of estradiol and progesterone on cholesterol 7 α-hydroxylase activity in rats subjected to different feeding conditions. Steroids, 1994, 59(9), 528-535.
[http://dx.doi.org/10.1016/0039-128X(94)90071-X] [PMID: 7846735]
[24]
Mareck, U.; Geyer, H.; Opfermann, G.; Thevis, M.; Schänzer, W. Factors influencing the steroid profile in doping control analysis. J. Mass Spectrom., 2008, 43(7), 877-891.
[http://dx.doi.org/10.1002/jms.1457] [PMID: 18570179]
[25]
Van Renterghem, P.; Van Eenoo, P.; Geyer, H.; Schänzer, W.; Delbeke, F.T. Reference ranges for urinary concentrations and ratios of endogenous steroids, which can be used as markers for steroid misuse, in a Caucasian population of athletes. Steroids, 2010, 75(2), 154-163.
[http://dx.doi.org/10.1016/j.steroids.2009.11.008] [PMID: 19962394]
[26]
Melcangi, R.C.; Panzica, G.; Garcia-Segura, L.M. Neuroactive steroids: focus on human brain. Neuroscience, 2011, 191, 1-5.
[http://dx.doi.org/10.1016/j.neuroscience.2011.06.024] [PMID: 21704130]
[27]
Melcangi, R.C.; Giatti, S.; Garcia-Segura, L.M. Levels and actions of neuroactive steroids in the nervous system under physiological and pathological conditions: Sex-specific features. Neurosci. Biobehav. Rev., 2016, 67, 25-40.
[http://dx.doi.org/10.1016/j.neubiorev.2015.09.023] [PMID: 26657814]
[28]
Gomes, R.L.; Meredith, W.; Snape, C.E.; Sephton, M.A. Analysis of conjugated steroid androgens: deconjugation, derivatization and associated issues. J. Pharm. Biomed. Anal., 2009, 49(5), 1133-1140.
[http://dx.doi.org/10.1016/j.jpba.2009.01.027] [PMID: 19304432]
[29]
Holst, J.P.; Soldin, O.P.; Guo, T.; Soldin, S.J. Steroid hormones: relevance and measurement in the clinical laboratory. Clin. Lab. Med., 2004, 24(1), 105-118.
[http://dx.doi.org/10.1016/j.cll.2004.01.004] [PMID: 15157559]
[30]
Lebesgue, D.; Chevaleyre, V.; Zukin, R.S.; Etgen, A.M. Estradiol rescues neurons from global ischemia-induced cell death: multiple cellular pathways of neuroprotection. Steroids, 2009, 74(7), 555-561.
[http://dx.doi.org/10.1016/j.steroids.2009.01.003] [PMID: 19428444]
[31]
Miller, W.L. Rare defects in adrenal steroidogenesis. Vol. 179. Eur. J. Endocrinol., 2018, 179(3), R125-R141.
[http://dx.doi.org/10.1530/EJE-18-0279] [PMID: 29880708]
[32]
Geyer, H.; Schänzer, W.; Thevis, M. Anabolic agents: recent strategies for their detection and protection from inadvertent doping. Br. J. Sports Med., 2014, 48(10), 820-826.
[http://dx.doi.org/10.1136/bjsports-2014-093526] [PMID: 24632537]
[33]
Handelsman, D.J. Androgen physiology, pharmacology, and abuse. In: Endocrinology: Adult and Pediatric; , 2015, pp. 2368-2393.e16..
[34]
Schiffer, L.; Barnard, L.; Baranowski, E.S.; Gilligan, L.C.; Taylor, A.E.; Arlt, W.; Shackleton, C.H.L.; Storbeck, K.H. Human steroid biosynthesis, metabolism and excretion are differentially reflected by serum and urine steroid metabolomes: A comprehensive review. J. Steroid Biochem. Mol. Biol., 2019, 194, 105439.
[http://dx.doi.org/10.1016/j.jsbmb.2019.105439] [PMID: 31362062]
[35]
Vegeto, E.; Benedusi, V.; Maggi, A. Estrogen anti-inflammatory activity in brain: a therapeutic opportunity for menopause and neurodegenerative diseases. Front. Neuroendocrinol., 2008, 29(4), 507-519.
[http://dx.doi.org/10.1016/j.yfrne.2008.04.001] [PMID: 18522863]
[36]
Deckx, N.; Lee, W.P.; Berneman, Z.N.; Cools, N. Neuroendocrine immunoregulation in multiple sclerosis. Clin. Dev. Immunol., 2013, 2013, 705232.
[http://dx.doi.org/10.1155/2013/705232] [PMID: 24382974]

Rights & Permissions Print Cite
Article Metrics
27
2
© 2024 Bentham Science Publishers | Privacy Policy