Comparison of 7 Published LC-MS/MS Methods for the Simultaneous Measurement of Testosterone, Androstenedione, and Dehydroepiandrosterone in Serum.
BACKGROUND
Recently, LC-MS/MS was stated to be the method of choice to measure sex steroids. Because information on the mutual agreement of LC-MS/MS methods is scarce, we compared 7 published LC-MS/MS methods for the simultaneous measurement of testosterone, androstenedione, and dehydroepiandrosterone (DHEA).


METHODS
We used 7 published LC-MS/MS methods to analyze in duplicate 55 random samples from both men and women. We performed Passing-Bablok regression analysis and calculated Pearson correlation coefficients to assess the agreement of the methods investigated with the median concentration measured by all methods, and we calculated the intraassay CV of each method derived from duplicate results and the CVs between the methods.


RESULTS
Median concentrations of testosterone were 0.22-1.36 nmol/L for women and 8.27-27.98 nmol/L for men. Androstenedione and DHEA concentrations were 0.05-5.53 and 0.58-18.04 nmol/L, respectively. Intraassay CVs were 2.9%-10%, 1.2%-8.8%, 2.7%-13%, and 4.3%-16% for testosterone in women, testosterone in men, androstenedione, and DHEA. Slopes of the regression lines calculated by Passing-Bablok regression analysis were 0.92-1.08, 0.92-1.08, 0.90-1.13, and 0.91-1.41 for all testosterone values, testosterone in women, androstenedione, and DHEA. Intermethod CVs were 14%, 8%, 30%, and 22% for testosterone in women, testosterone in men, androstenedione, and DHEA.


CONCLUSIONS
In general, the LC-MS/MS methods investigated show reasonable agreement. However, some of the assays show differences in standardization, and others show high variation.
Endocrine Laboratory, Department of Clinical Chemistry, VU University Medical Center, Amsterdam, Netherlands;
Rahel M
RM
Büttler
Endocrine Laboratory, Department of Clinical Chemistry, VU University Medical Center, Amsterdam, Netherlands;
Frans
F
Martens
Endocrinology Unit and Center for Applied Biomedical Research, Department of Medical and Surgical Sciences, S. Orsola Malpighi Hospital, University of Bologna, Bologna, Italy;
Flaminia
F
Fanelli
Biocrates Life Sciences AG, Innsbruck, Austria;
Hai T
HT
Pham
Department of Pathology, University of Utah, Salt Lake City, UT; ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT;
Mark M
MM
Kushnir
Laboratory of Clinical Chemistry and Hematology, VieCuri Medical Center Venlo, Netherlands;
Marcel J W
MJ
Janssen
Department of Clinical Biochemistry, University Hospital of South Manchester, Manchester, UK;
Laura
L
Owen
Centre for Endocrinology, Diabetes and Metabolism, School of Clinical and Experimental Medicine, University of Birmingham, UK;
Angela E
AE
Taylor
Department of Growth and Reproduction, Rigshospitalet, Copenhagen University Hospital, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.
Tue
T
Soeborg
Endocrine Laboratory, Department of Clinical Chemistry, VU University Medical Center, Amsterdam, Netherlands;
Marinus A
MA
Blankenstein
Endocrine Laboratory, Department of Clinical Chemistry, VU University Medical Center, Amsterdam, Netherlands; a.heijboer@vumc.nl.
Annemieke C
AC
Heijboer
D000328:Adult; D000735:Androstenedione / Q000097:blood*; D002138:Calibration; D002853:Chromatography, Liquid / Q000592:standards*; D003687:Dehydroepiandrosterone / Q000097:blood*; D005260:Female; D006801:Humans; D008297:Male; D015588:Observer Variation; D012044:Regression Analysis; D015203:Reproducibility of Results; D012737:Sex Factors; D053719:Tandem Mass Spectrometry / Q000592:standards*; D013739:Testosterone / Q000097:blood*
D013739:Testosterone; D000735:Androstenedione; D003687:Dehydroepiandrosterone