Abstract
Background:
Mass spectrometry (MS)-based 17-hydroxyprogesterone (17OHP) methods show considerable variation in results in external quality assurance (EQA) programs. An understanding of the current status of MS-based serum/plasma 17OHP quantification is important to facilitate harmonization.
Methods:
A 50-item e-survey related to (1) laboratory characteristics, (2) pre-analytical considerations and (3) analysis of 17OHP was developed and circulated to clinical MS laboratories via professional associations in Asia Pacific, Europe and North America.
Results:
Forty-four laboratories from 17 countries completed the survey. Sample preparation varied between laboratories with protein precipitation and liquid-liquid extraction being the most common processes. Analyte separation was most commonly achieved by liquid chromatography (LC) using a C18 column and mobile phases of water, methanol and formic acid. The ions selected for quantification were 331>97 m/z or 331>109 m/z. Alternative transition ions were used as qualifiers. Twenty-seven of 44 respondents reported preparing their calibrators in-house and variations in material purity and matrix were evident. Nine of 44 laboratories did not participate in an EQA program, and half did not know if their method separated out isobars. The reference intervals, and also their partitioning, reported by the laboratories were highly discrepant, in some cases, by multiple folds.
Conclusions:
Although MS-based methods are similar in many facets, they are highly disparate. Five recommendations have been developed as an outcome of this survey to support the continued improvement of analysis of serum/plasma 17OHP by MS.
Acknowledgments
We wish to thank Professors Leslie Lai, Loralie Langman, Patti Jones, Rachana Santiyanont and all colleagues for assisting in the dissemination of this survey. Thank you to all the participants of this survey for their motivation and enthusiasm in completing this detailed survey. Our thanks are also extended to Drs Lindsey Mackay and Stephen Davies from the National Measurement Institute of Australia for supporting the development of the CRM for 17OHP. Stefan A. Wudy (chair), Michaela Hartmann (member), Yolanda de Rijke (member) and Ronda Greaves (international member) of working group 3 “Harmonisation of Laboratory Assessment” appreciate support from European Cooperation in Science and Technology (COST) Action BM1303 “DSDnet”. Ronda Greaves (chair), Chung Shun Ho (member), Tze Ping Loh (member), Peter Graham (member) and Lisa Jolly (past member) of the Mass Spectrometry Harmonisation Working Group acknowledge the support of the APFCB and the AACB. Finally, we wish to acknowledge the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) for in-kind support to continue this work through the Emerging Technologies Division of the IFCC.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
References
1. Kulle A, Krone N, Holterhus PM, Schuler G, Greaves RF, Juul A, et al. Steroid hormone analysis in diagnosis and treatment of DSD: – Position Paper of EU COST Action BM 1303 “DSDnet. Eur J Endocrinol 2017;176:P1–9.10.1530/EJE-16-0953Search in Google Scholar PubMed PubMed Central
2. Online Mendelian Inheritance in Man. Entry# 201910 – Adrenal Hyperplasia, Congenital due To 21-Hydroxylase deficiency. Available at: http://www.omim.org/entry/201910. Accessed: 18 Dec 2017.Search in Google Scholar
3. Speiser PW, Azziz R, Baskin LS, Ghizzoni L, Hensle TW, Merke DP, et al. Congenital adrenal hyperplasia due to steroid 21-hydroxylase deficiency: an endocrine society clinical practice guideline. J Clin Endocrinol Metab 2010;95:4133–60.10.1210/jc.2009-2631Search in Google Scholar PubMed PubMed Central
4. RCPAQAP Chemical Pathology Programs. Available at: www.rcpaqap.com.au/chempath.www.rcpaqap.com.au/chempath. Accessed: 23 Mar 2017.Search in Google Scholar
5. Midgley P, Azzopardi D, Oates N, Shaw J, Honour J. Virilisation of female preterm infants. Arch Dis Child 1990;65:701–3.10.1136/adc.65.7_Spec_No.701Search in Google Scholar
6. Wong T, Shackleton CH, Covey TR, Ellis G. Identification of the steroids in neonatal plasma that interfere with 17a-hydroxyprogesterone radioimmunoassays. Clin Chem 1992;38:1830–7.10.1093/clinchem/38.9.1830Search in Google Scholar
7. Wudy SA, Wachter UA, Homoki J, Teller WM. 17a-Hydroxyprogesterone, 4-androstenedione, and testosterone profiled by routine stable isotope dilution/gas chromatography-mass spectrometry in plasma of children. Pediatr Res 1995;38:76–80.10.1203/00006450-199507000-00013Search in Google Scholar PubMed
8. Wudy SA, Schuler G, Sánchez-Guijo A, Hartmann MF. The art of measuring steroids: principles and practice of current hormonal steroid analysis. J Steroid Biochem Mol Biol. 2017. pii: S0960-0760(17)30250-9. doi: 10.1016/j.jsbmb.2017.09.003.10.1016/j.jsbmb.2017.09.003Search in Google Scholar PubMed
9. Joint Committee for Traceability in Laboratory Medicine. Available at: www.bipm.org/jctlm. Accessed: 23 Mar 2017.Search in Google Scholar
10. Asia-Pacific Federation of Clinical Biochemistry and Laboratory Medicine (APFCB) Mass Spectrometry Harmonisation Working Group. Available at: http://www.apfcb.org/scientific-activities.html. Accessed: 23 Mar 2017.Search in Google Scholar
11. Australasian Association of Clinical Biochemists (AACB) Harmonisation Committee. Available at: http://www.aacb.asn.au/professionaldevelopment/harmonisation. Accessed: 23 Mar 2017.Search in Google Scholar
12. European Cooperation in Science and Technology (COST) Action BM1303. A Systematic Elucidation on Differences of Sex Development (DSDnet), working group 3 – Harmonisation of Laboratory Assessment. Available at: http://www.dsdnet.eu/wg-3.html. Accessed: 23 Mar 2017.Search in Google Scholar
13. The Association of Clinical Biochemistry and Laboratory Medicine, (ACB). Available at: http://www.acb.org.uk/. Accessed: 24 Oct 2017.Search in Google Scholar
14. American Association of Clinical Chemistry, (AACC). Available at: https://www.aacc.org/. Accessed: 24 Oct 2017.Search in Google Scholar
15. Canadian Society of Clinical Chemists, (CSCC). Available at: https://www.cscc.ca/en/. Accessed: 24 Oct 2017.Search in Google Scholar
16. European Society for Pediatric Endocrinology (ESPE). Available at: https://www.eurospe.org/. Accessed: 24 Oct 2017.Search in Google Scholar
17. Miller WG, Myers GL, Rej R. Why commutability matters. Clin Chem 2006;52:553–4.10.1373/clinchem.2005.063511Search in Google Scholar PubMed
18. Matuszewski BK, Constanzer ML, Chavez-Eng CM. Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS. Anal Chem 2003;75:3019–30.10.1021/ac020361sSearch in Google Scholar PubMed
19. Søeborg T, Frederiksen H, Johannsen TH, Andersson AM, Juul A. Isotope-dilution TurboFlow-LC-MS/MS method for simultaneous quantification of ten steroid metabolites in serum. Clin Chim Acta 2017;468:180–6.10.1016/j.cca.2017.03.002Search in Google Scholar PubMed
20. Clinical and Laboratory Standards Institute. Liquid chromatography-mass spectrometry methods; approved guideline. CLSI document C62-A. Wayne, PA: CLSI, 2014.Search in Google Scholar
21. Tavita N, Greaves RF. Systematic review of serum steroid reference intervals developed using mass spectrometry. Clin Biochem 2017;50:1260–74.10.1016/j.clinbiochem.2017.07.002Search in Google Scholar PubMed
22. Fanelli F, Belluomo I, Di Lallo VD, Cuomo G, De Iasio R, Baccini M, et al. Serum steroid profiling by isotopic dilution-liquid chromatography–mass spectrometry: comparison with current immunoassays and reference intervals in healthy adults. Steroids 2011;76:244–53.10.1016/j.steroids.2010.11.005Search in Google Scholar PubMed
23. Fanelli F, Gambineri A, Belluomo I, Repaci A, Di Lallo VD, Di Dalmazi G, et al. Androgen profiling by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in healthy normal-weight ovulatory and anovulatory late adolescent and young women. J Clin Endocrinol Metab 2013;98:3058–67.10.1210/jc.2013-1381Search in Google Scholar PubMed
24. Greaves RF, Pitkin J, Ho CS, Baglin J, Hunt RW, Zacharin MR. Hormone modeling in preterm neonates: establishment of pituitary and steroid hormone reference intervals. J Clin Endocrinol Metab 2015;100:1097–103.10.1210/jc.2014-3681Search in Google Scholar PubMed
25. Homma K, Hida M, Ikeda KY, Amaga N, Murata M, Hasegawa T. Measurement of serum 17 α-hydroxyprogesterone in newborn infants by stable isotope dilution – gas chromatography/mass spectrometry. Clin Pediatr Endocrinol 2009;18:77–80.10.1297/cpe.18.77Search in Google Scholar PubMed PubMed Central
26. Kulle AE, Welzel M, Holterhus PM, Riepe FG. Implementation of a liquid chromatography tandem mass spectrometry assay for eight adrenal C-21 steroids and pediatric reference data. Horm Res Paediatr 2013;79:22–31.10.1159/000346406Search in Google Scholar PubMed
27. Kushnir MM. Development and performance evaluation of a tandem mass spectrometry assay for 4 adrenal steroids. Clin Chem 2006;52:1559–67.10.1373/clinchem.2006.068445Search in Google Scholar PubMed
28. Kyriakopoulou L, Yazdanpanah M, Colantonio DA, Chan MK, Daly CH, Adeli K. A sensitive and rapid mass spectrometric method for the simultaneous measurement of eight steroid hormones and CALIPER pediatric reference intervals. Clin Biochem 2013;46:642–51.10.1016/j.clinbiochem.2013.01.002Search in Google Scholar PubMed
29. Magnisali P, Dracopoulou M, Mataragas M, Dacou-Voutetakis A, Moutsatsou P. Routine method for the simultaneous quantification of 17alpha-hydroxyprogesterone, testosterone, dehydroepiandrosterone, androstenedione, cortisol, and pregnenolone in human serum of neonates using gas chromatography-mass spectrometry. J Chromatogr A 2008;1206:166–77.10.1016/j.chroma.2008.07.057Search in Google Scholar PubMed
30. Rauh M, Gröschl M, Rascher W, Dörr HG. Automated, fast and sensitive quantification of 17a-hydroxy-progesterone, androstenedione and testosterone by tandem mass spectrometry with on-line extraction. Steroids 2006;71:450–8.10.1016/j.steroids.2006.01.015Search in Google Scholar PubMed
31. Søeborg T, Frederiksen H, Mouritsen A, Johannsen TH, Main KM, Jørgensen N, et al. Sex, age, pubertal development and use of oral contraceptives in relation to serum concentrations of DHEA, DHEAS, 17α-hydroxyprogesterone, δ4-androstenedione, testosterone and their ratios in children, adolescents and young adults. Clin Chim Acta 2014;437:6–13.10.1016/j.cca.2014.06.018Search in Google Scholar PubMed
32. Soldin OP, Sharma H, Husted L, Soldin SJ. Pediatric reference intervals for aldosterone, 17a-hydroxyprogesterone, dehydroepiandrosterone, testosterone and 25-hydroxy vitamin D3 using tandem mass spectrometry. Clin Biochem 2009;42:823–7.10.1016/j.clinbiochem.2009.01.015Search in Google Scholar PubMed PubMed Central
33. Eisenhofer G, Peitzsch M, Kaden D, Langton K, Pamporaki C, Masjkur J, et al. Reference intervals for plasma concentrations of adrenal steroids measured by LC-MS/MS: impact of gender, age, oral contraceptives, body mass index and blood pressure status. Clin Chim Acta 2017;470:115–24.10.1016/j.cca.2017.05.002Search in Google Scholar PubMed PubMed Central
34. Gaudl A, Kratzsch J, Bae YJ, Kiess W, Thiery J, Ceglarek U. Liquid chromatography quadrupole linear ion trap mass spectrometry for quantitative steroid hormone analysis in plasma, urine, saliva and hair. J Chromatogr A 2016;1464:64–71.10.1016/j.chroma.2016.07.087Search in Google Scholar PubMed
35. Sanchez-Guijo A, Hartmann MF, Wudy SA. Introduction to gas chromatography-mass spectrometry. Methods Mol Biol 2013;1065:27–44.10.1007/978-1-62703-616-0_3Search in Google Scholar PubMed
36. Singh RJ. Quantitation of 17-oh-progesterone (ohpg) for diagnosis of congenital adrenal hyperplasia (cah). Methods Mol Biol 2010;603:271–7.10.1007/978-1-60761-459-3_25Search in Google Scholar PubMed
37. Waller K, Henman M, Huynh T, Warner J. 17-hydroxyprogesterone quantification in dried blood spots by high performance liquid chromatography-tandem mass spectroscopy. Int J Pediat Endocrinol 2015;2015:P44.10.1186/1687-9856-2015-S1-P44Search in Google Scholar
38. Aleknaviciute J, Tulen JH, Timmermans M, de Rijke YB, van Rossum EF, de Jong FH, et al. Adrenocorticotropic hormone elicits gonadotropin secretion in premenopausal women. Hum Reprod 2016;31:2360–8.10.1093/humrep/dew190Search in Google Scholar PubMed
39. International Organization for Standardization (ISO). Medical laboratories – Requirements for quality and competence. ISO 15189: 2012.Search in Google Scholar
40. Simeone J, Rainville P. Fast Separation of 20 steroids including isobaric species using UPC2/MS/MS. Technical brief, waters corporation. Available at: http://www.waters.com/webassets/cms/library/docs/720005021en.pdf. Accessed: 30 Aug 2017.Search in Google Scholar
41. Clinical and Laboratory Standards Institute. Characterization and qualification of commutable reference materials for laboratory medicine; approved guideline. CLSI document C53-A. Wayne, PA, USA: CLSI, 2010.Search in Google Scholar
42. Greaves RF. A Guide to harmonisation and standardisation of measurands determined by liquid chromatography – tandem mass spectrometry in routine clinical biochemistry. Clin Biochem Rev 2012;33:123–32.Search in Google Scholar
43. National Measurement Institute. Available at: http://www.measurement.gov.au/ScienceTechnology/Pages/ChemicalMetrologyResearch.aspx. Accessed: 18 Dec 2017.Search in Google Scholar
Supplementary Material:
The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2017-1039).
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