MSACL 2019 EU Abstract

The Harmoster Initiative: Preliminary Results on the Comparability of Circulating Steroid Measurement Among Ten European Laboratories Using LC-MS/MS

Flaminia Fanelli (1), Marco Cantù (2), Marco Mezzullo (1), Anastasia Temchenko (1), Johanna M Lindner (3), Mirko Peitzsch (4), James M Hawley (5), Stephen Bruce (6), Sieglinde Zelzer (7), Markus Herrmann (7), Mariette T Ackermans (8), Annemieke C Heijboer (8), Jody Van den Ouweland (9), Manfred Rauh (10), Graeme Eisenhofer (4), Brian G Keevil (5), Michael Vogeser (3), Uberto Pagotto (1)
(1) University of Bologna, IT, (2) Ente Ospedaliero Cantonale, Bellinzona, CH, (3) Hospital of the University of Munich (LMU), DE, (4) Technische Universität Dresden, DE, (5) Manchester NHS Foundation Trust, UK, (6) University Hospital of Lausanne (CHUV), CH, (7) Medical University of Graz, AT, (8) Amsterdam UMC, NL, (9) Canisius-Wilhelmina Hospital, Nijmegen, NL, (10) University Hospital, Erlangen, DE

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Flaminia Fanelli (Presenter) University of Bologna

Presenter Bio: Flaminia Fanelli is Junior assistant professor at the Dept of Medical and Surgical Sciences - University of Bologna. She leads the basic science research team of the Endocrinology Unit directed by Prof. Uberto Pagotto. Flaminia has more than 10 years experience in small molecule quantitation, mainly steroids, in human and in vitro fluids by LC-MS/MS. She is involved in national (Ricerca Finalizzata 2017, ga GR-2018-12365398; PRIN 2017 ga 2017ATZ2YK) and EU (ga 22371, 279171, 245009) funded studies focusing on obesity, sex hormone imbalance and adrenal tumors. In 2012 she received the Young Investigators Grant by the Emilia Romagna Region – University Program for the project Development and validation of innovative methods for steroid hormone measurement in serum and saliva (PRUA 1-2012-004). She coauthored 43 peer-reviewed articles (H-index by Scopus: 14).

Relevant Financial Disclosures (within past 24 months) Committee/Board/Advisory Board MSACL Endocrinology Sci Com Discussing Products? NO, presenter will NOT present educational content related to products/services of the ineligible company(ies) listed above. Conflict mitigation NOT REQUIRED.

Background. LC-MS/MS is recommended for circulating steroid measurement, as its intrinsic specificity is supposed to guarantee accurate quantitation. LC-MS/MS also represents a promising opportunity to achieve harmonization of steroid measurements. However, LC-MS/MS assays applied in different laboratories use various pre-analytical and analytical approaches that may influence assay performance and final result. So far, little data have been generated on the comparability of LC-MS/MS steroid measurements.
Objective. To compare the LC-MS/MS measurement of 13 circulating steroids among 10 European centers operating in clinical and research settings.
Methods. Blood from 13 women and 13 men aged 21-70y 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) and plasma (Li-Heparin). Each center received two aliquots of 78 total samples for LC-MS/MS steroid 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), 11deoxycortisol (11S), 17OH-progesterone (17OHP4), androstenedione (A4), testosterone (T) and DHEAS. Most of intra-lab imprecision was <10% for all analytes, with some exceptions <20%. Inter-lab CV% per sample ranged 6.6–31.4% for F, 1.6–16.4% for E, 1.4–34.5% for B, 4.3–39.5% for 11S, 2.3–35.4% for 17OHP4, 2.8–16.9% for A4, 2.4–13.2% for T and 1.0–16.6% for DHEAS. 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. Laboratory mean bias vs all-lab median values ranged -3.5 – 5.8%, -1.9 - 6.5%, -3.2 – 9.1%, -16.5 – 14.6% and -5.0 – 36.9% for DHEAS, E, A4, 11S, and F, respectively. Moreover, mean bias ranged -12.3 – 10.6% and -4.7 – 12.9% for B <10ng/mL and >10ng/mL, respectively, -23.4 - 7.5% and -6.7 - 4.7% for 17OHP4 <1ng/mL and >1ng/mL, respectively, and -2.9 – 6.4% and -8.5 – 8.7% for T <1ng/mL and >1ng/mL, respectively.
Conclusions. Variable intra-lab performances were observed. Inter-lab comparability was good for E, DHEAS, A4 and T, while improvements are needed for F, 11S, B and 17OHP4 measurement. Variability in B and 17OHP4 measurement increased at lower levels, while the opposite was observed for T. The specimen type may have an influence on results. Further analyses are required to define major sources of variability.