Tags

Type your tag names separated by a space and hit enter

Development and validation of a UHPLC-HRMS method for the simultaneous determination of the endogenous anabolic androgenic steroids in human serum.
J Chromatogr A. 2019 Nov 06 [Online ahead of print]JC

Abstract

Being performance enhancing hormones, endogenous anabolic androgenic steroids (EAAS) are banned from most competitive sports by the World Anti-doping Agency (WADA). In anti-doping control laboratories, routine assays are mainly performed on urine samples of athletes in and out of competitions. Serum constitutes a promising alternative to urine as it is less subjected to manipulation or contamination that may influence the method sensitivity. The simultaneous determination of EAAS including conjugated metabolites using LC-MS is very challenging due to their contradicting chemical behaviors at the ionization interface of the mass spectrometer. This may prejudice their detection or limit the method sensitivity. Herein, we have addressed these challenges and developed a new method for the simultaneous determination of unconjugated, sulphate- and glucuronide-conjugated EAAS (Androsterone, Etiocholanolone, testosterone, epitestosterone, dihydrotestosterone, dehydroepiandrosterone, androstenedione and 17a-hydroxyprogesterone) in human serum using ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS). The use of mass spectrometric detection in full scan mode facilitated the study of the most versatile adducts for detection and quantitation. A solid phase extraction method was developed for the sample preparation prior to analysis. The method limits of quantitation ranged from 0.006 to 7.904 ng/mL and the recoveries ranged from 70.2% to 96.5%. The method calibration was performed in untreated serum representing realistic matrix composition with correlation coeffecients ranged from 0.9859 to 0.9988. Finally, the serum-levels of the investigated steroids were determined in 4 male and 1 female human subjects to provide estimates of baseline levels based on individual values.

Authors+Show Affiliations

Department of Environmental Science and Analytical Chemistry, Stockholm University, SE-10691 Stockholm, Sweden. Electronic address: hatem.elmongy@aces.su.se.Doping Control Laboratory, Division of Clinical Pharmacology, Karolinska University Hospital, SE-14186 Stockholm, Sweden.Doping Control Laboratory, Division of Clinical Pharmacology, Karolinska University Hospital, SE-14186 Stockholm, Sweden.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31735349

Citation

Elmongy, Hatem, et al. "Development and Validation of a UHPLC-HRMS Method for the Simultaneous Determination of the Endogenous Anabolic Androgenic Steroids in Human Serum." Journal of Chromatography. A, 2019, p. 460686.
Elmongy H, Masquelier M, Ericsson M. Development and validation of a UHPLC-HRMS method for the simultaneous determination of the endogenous anabolic androgenic steroids in human serum. J Chromatogr A. 2019.
Elmongy, H., Masquelier, M., & Ericsson, M. (2019). Development and validation of a UHPLC-HRMS method for the simultaneous determination of the endogenous anabolic androgenic steroids in human serum. Journal of Chromatography. A, 460686. https://doi.org/10.1016/j.chroma.2019.460686
Elmongy H, Masquelier M, Ericsson M. Development and Validation of a UHPLC-HRMS Method for the Simultaneous Determination of the Endogenous Anabolic Androgenic Steroids in Human Serum. J Chromatogr A. 2019 Nov 6;460686. PubMed PMID: 31735349.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development and validation of a UHPLC-HRMS method for the simultaneous determination of the endogenous anabolic androgenic steroids in human serum. AU - Elmongy,Hatem, AU - Masquelier,Michèle, AU - Ericsson,Magnus, Y1 - 2019/11/06/ PY - 2019/07/05/received PY - 2019/09/25/revised PY - 2019/11/05/accepted PY - 2019/11/19/entrez PY - 2019/11/19/pubmed PY - 2019/11/19/medline KW - Chromatography KW - Doping KW - Endogenous Anabolic androgenic steroids KW - Mass spectrometry KW - Solid phase extraction KW - UHPLC SP - 460686 EP - 460686 JF - Journal of chromatography. A JO - J Chromatogr A N2 - Being performance enhancing hormones, endogenous anabolic androgenic steroids (EAAS) are banned from most competitive sports by the World Anti-doping Agency (WADA). In anti-doping control laboratories, routine assays are mainly performed on urine samples of athletes in and out of competitions. Serum constitutes a promising alternative to urine as it is less subjected to manipulation or contamination that may influence the method sensitivity. The simultaneous determination of EAAS including conjugated metabolites using LC-MS is very challenging due to their contradicting chemical behaviors at the ionization interface of the mass spectrometer. This may prejudice their detection or limit the method sensitivity. Herein, we have addressed these challenges and developed a new method for the simultaneous determination of unconjugated, sulphate- and glucuronide-conjugated EAAS (Androsterone, Etiocholanolone, testosterone, epitestosterone, dihydrotestosterone, dehydroepiandrosterone, androstenedione and 17a-hydroxyprogesterone) in human serum using ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS). The use of mass spectrometric detection in full scan mode facilitated the study of the most versatile adducts for detection and quantitation. A solid phase extraction method was developed for the sample preparation prior to analysis. The method limits of quantitation ranged from 0.006 to 7.904 ng/mL and the recoveries ranged from 70.2% to 96.5%. The method calibration was performed in untreated serum representing realistic matrix composition with correlation coeffecients ranged from 0.9859 to 0.9988. Finally, the serum-levels of the investigated steroids were determined in 4 male and 1 female human subjects to provide estimates of baseline levels based on individual values. SN - 1873-3778 UR - https://www.unboundmedicine.com/medline/citation/31735349/Development_and_validation_of_a_UHPLC-HRMS_method_for_the_simultaneous_determination_of_the_endogenous_anabolic_androgenic_steroids_in_human_serum L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9673(19)31115-X DB - PRIME DP - Unbound Medicine ER -
Try the Free App:
Prime PubMed app for iOS iPhone iPad
Prime PubMed app for Android
Prime PubMed is provided
free to individuals by:
Unbound Medicine.