ECE2019 Poster Presentations Interdisciplinary Endocrinology 2 (37 abstracts)
Harmonization status among ten European laboratories using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for steroid measurements in serum: preliminary results from the HarmoSter consortium
Flaminia Fanelli 1 , Marco Mezzullo 1 , Anastasia Temchenko 1 , Marco Cantù 2 , Johanna M Lindner 3 , Mirko Peitzsch 4 , James M Hawley 5 , Stephen Bruce 6 , Sieglinde Zelzer 7 , Markus Herrmann 7 , Annemieke C Heijboer 8 , Jody Van den Ouweland 9 , Graeme Eisenhofer 4 , Brian G Keevil 5 , Manfred Rauh 10 , Michael Vogeser 3 & Uberto Pagotto 1
1Endocrinology Unit, Center for Applied Biomedical Research, Department of Medical and Surgical Sciences, University of Bologna, Bologna, Italy; 2Laboratory of Clinical Biochemistry and Pharmacology, Department of Laboratory Medicine EOLAB, Ente Ospedaliero Cantonale, Bellinzona, Switzerland; 3Institute of Laboratory Medicine, Hospital of the University of Munich (LMU), Munich, Germany; 4 Institute of Clinical Chemistry and Laboratory Medicine, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany; 5Department of Clinical Biochemistry, University Hospital South Manchester, Manchester NHS Foundation Trust, Manchester, United Kingdom; 6Clinical Chemistry Laboratory, University Hospital of Lausanne, Lausanne, Switzerland; 7Clinical Institute for Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Graz, Austria; 8Endocrine Laboratory, Department of Clinical Chemistry, VU Medical Center - Academic Medical Center University Hospital, Amsterdam, Netherlands; 9Department of Clinical Chemistry, Canisius-Wilhelmina Hospital, Nijmegen, Netherlands; 10Department of Pediatrics and Adolescent Medicine, University Hospital, Erlangen, Germany.
Background: LC-MS/MS is replacing immunoassays for serum steroid measurement in clinical labs worldwide. The intrinsic high specificity of LC-MS/MS is supposed to guarantee accurate quantitation, however, differences in pre-analytical and analytical procedures, and overall performance may influence the result. A few studies so far investigated the reproducibility among different LC-MS/MS assays for sex steroids. No data are available for adrenal steroids.
Aim: To compare the LC-MS/MS measurement of 13 serum steroids among 10 European Centers.
Methods: Blood from 13 women and 13 men aged 21–70 y was collected in fasting state, at 8:00–9:00 am, after 10 min saline infusion in three different vacuum tubes to obtain serum (gel separator, GS and beads-clot-activator) and plasma (Li-Heparin) (78 total samples). Aliquots were sent to each lab for LC-MS/MS measurement according to the lab’s own procedure. Intra-lab duplicate measurement CV%, inter-lab CV% and Bland-Altman analyses were performed.
Results: Preliminary results from six labs were obtained on cortisol (F), cortisone (E), corticosterone (B), 11 deoxycortisol (11S), 17OH-progesterone (17OHP4), androstenedione (A4), testosterone (T) and DHEAS. Intra-lab CV% ranged 3.1–9.9 (F), 2.7–13.8 (E), 3.3–8.0 (B), 3.8–14.4 (11S), 5.8–18.1 (17OHP4<1 ng/ml) 2.0–9.9 (17OHP4>1 ng/ml), 4.2–8.8 (A4), 4.3–7.6 (T) and 4.1–11.4 (DHEAS). Median inter-lab CV% were 5.0, 5.1, 6.8. 7.3, 8.9, 14.0 and 15.5 for DHEAS, E, T, A4, B, 11S and F, respectively, and 13.2 and 5.9 for 17OHP4 <1 ng/ml and >1 ng/ml, respectively. Cases showing inter-lab CV>15% were 0 for T; 1 (1.3%) for E and DHEAS; 3 (3.8%) for A4; 13 (16.7%) for B; 25 (32.1%) for 17OHP4; 36 (46.2%) for 11S and 42 (53.8%) for F. Among GS-sera, cases increased to 18 (69.2%) for both 11S and F. Results from each lab were compared to all-lab median values. Mean bias ranged −3.5 – 5.8%, -1.9 - 6.5%, −3.2 – 9.1%, -10.6 – 11.2%, −16.5 – 14.6% and −5.0 – 36.9% for DHEAS, E, A4, B, 11S, and F, respectively. Moreover, mean bias ranged −23.4 −7.5% and −6.7 − 4.7% for 17OHP4<1 ng/ml and >1 ng/ml, respectively, and −2.9 – 6.4% and −8.5 – 8.7% for T<1 ng/ml and >1 ng/ml, respectively.
Conclusions: Variable intra-lab performances were observed. Inter-lab reproducibility was good for E, B, DHEAS, A4 and T, but poor for F and 11S. The specimen type may influence the results. 17OHP4 reproducibility was higher at high than low levels, while the opposite was unexpectedly found for T. Further analyses will help to define and correct major sources of variability.
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Endocrine Abstracts
ISSN 1470-3947 (print) | ISSN 1479-6848 (online)
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