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- Title
Influence of isotopically labeled internal standards on quantification of serum/plasma 17α-hydroxyprogesterone (17OHP) by liquid chromatography mass spectrometry.
- Authors
Loh, Tze Ping; Ho, Chung Shun; Hartmann, Michaela F.; Zakaria, Rosita; Lo, Clara Wai Shan; van den Berg, Sjoerd; de Rijke, Yolanda B.; Cooke, Brian R.; Hoad, Kirsten; Graham, Peter; Davies, Stephen R.; Mackay, Lindsey G.; Wudy, Stefan A.; Greaves, Ronda F.
- Abstract
Objectives: Our recent survey of 44 mass spectrometry laboratories across 17 countries identified variation in internal standard (IS) choice for the measurement of serum/plasma 17α-hydroxyprogesterone (17OHP) by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The choice of IS may contribute to inter-method variations. This study evaluated the effect of two common isotopically labeled IS on the quantification of 17OHP by LC-MS/MS. Methods: Three collaborating LC-MS/MS laboratories from Asia, Europe and Australia, who routinely measure serum 17OHP, compared two IS, (1) IsoSciences carbon-13 labeled 17OHP-[2,3,4-13C3], and (2) IsoSciences deuterated 17OHP-[2,2,4,6,6,21,21,21-2H]. This was performed as part of their routine patient runs using their respective laboratory standard operating procedure. Results: The three laboratories measured 99, 89, 95 independent samples, respectively (up to 100 nmol/L) using the 13C- and 2H-labeled IS. The slopes of the Passing-Bablok regression ranged 0.98–1.00 (all 95% confidence interval [CI] estimates included the line of identity), and intercept of <0.1 nmol/L. Average percentage differences of −0.04% to −5.4% were observed between the two IS materials, which were less than the optimal bias specification of 7% determined by biological variation, indicating no clinically significant difference. The results of 12 Royal College of Pathologists of Australasia Quality Assurance Programs (RCPAQAP) proficiency samples (1–40 nmol/L) measured by the laboratories were all within the RCPAQAP analytical performance specifications for both IS. Conclusions: Overall, the comparison between the results of 13C- and 2H-labeled IS for 17OHP showed good agreement, and show no clinically significant bias when incorporated into the LC-MS/MS methods employed in the collaborating laboratories.
- Subjects
LIQUID chromatography-mass spectrometry; MASS spectrometry; LIQUID chromatography; STANDARD operating procedure
- Publication
Clinical Chemistry & Laboratory Medicine, 2020, Vol 58, Issue 10, p1731
- ISSN
1434-6621
- Publication type
Article
- DOI
10.1515/cclm-2020-0318