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Analytical performance evaluation and optimization of serum 25(OH)D LC-MS/MS measurement

  • Weiyan Zhou ORCID logo , Meiliang Gong , Yuanli Mao , Xiaofen Yuan , Yuhang Deng , Qianwen Zhang , Wei Guo , Ling Qiu , Xianzhang Huang , Zheng Cao , Jun Xia , Xuhui She , Yulong Cong , Chuanbao Zhang EMAIL logo , Huafen Liu EMAIL logo and Wenxiang Chen EMAIL logo
Published/Copyright: February 19, 2025
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 63 Issue 7

Abstract

Objectives

Measuring serum 25-hydroxyvitamin D is key in clinical labs, but inter-laboratory variations risk diagnostic errors. This study evaluates the performance of current in-house liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods used in top Chinese clinical laboratories and proposes an optimized method for improving serum 25(OH)D measurement accuracy and reliability.

Methods

Seven serum pools with different concentrations of 25(OH)D were prepared and sent to 12 participating laboratories for multiple repeat analysis with their current in-house LC-MS/MS methods and then an optimized LC-MS/MS method. Precision was assessed in terms of coefficient of variance (CV), and trueness was assessed in terms of bias referring to the U.S. National Institute of Standards and Technology (NIST) reference measurement procedure (RMP). The analytical performances of the two methods were compared and evaluated.

Results

Eighty percent and 90 % of the laboratories achieved the defined performance criteria (CV, <12.5 %; mean bias, <8.3 %) with the optimized method for the measurement of 25(OH)D2 and 25(OH)D3, compared with 43 % and 57 % of the laboratories meeting the criteria with their in-house methods, respectively. Precision and trueness improved after applying the optimized method. Although the optimized method didn’t not ensure that all laboratory samples meet the measurement uncertainty (MU) criteria (MU<13.6 %), particularly for low-concentration samples, it significantly reduced the MU compared to the in-house method.

Conclusions

Precision, trueness and MU improved after applying the optimized method. Nonetheless, more efforts are needed to ensure the reliability and accuracy of 25(OH)D measurements in clinical laboratories in China.

Keywords: 25-hydroxyvitamin D2; 25-hydroxyvitamin D3; liquid chromatography-tandem mass spectrometry (LC-MS/MS); quality control; analytical performance comparison
Corresponding authors: Chuanbao Zhang, National Center for Clinical Laboratories and Beijing Engineering Research Center of Laboratory Medicine, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing Hospital, National Center of Gerontology, No 1, Dahua Road, Dongcheng District, Beijing, China, E-mail: ; Huafen Liu, Calibra Scientific, Inc, Hangzhou, China; and Key Laboratory of Digital Technology in Medical Diagnostics of Zhejiang Province, Hangzhou, China, E-mail: ; and Wenxiang Chen, National Center for Clinical Laboratories and Beijing Engineering Research Center of Laboratory Medicine, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing Hospital, National Center of Gerontology, No 1, Dahua Road, Dongcheng District, Beijing, China, E-mail:
Weiyan Zhou and Meiliang Gong contributed equally to this work and share first authorship.

Funding source: National Science and Technology Major Project

Award Identifier / Grant number: 2024ZD0523702

Funding source: National High Level Hospital Clinical Research Funding

Award Identifier / Grant number: BJ-2023-094

Funding source: Capital’s Funds for Health Improvement and Research

Award Identifier / Grant number: 2024-2-4059

Funding source: Shanghai Innovation Medical Device Application Demonstration Project

Award Identifier / Grant number: No.23SHS06200

Acknowledgments

The authors acknowledge participants from the China National Institute of Metrology and By-Health Co., Ltd. for their kind assistance in testing samples during the study.

  1. Research ethics: This study has received permission from the Ethics Committee of Beijing Hospital, with exemption for obtaining informed consent (approval No. 2023BJYYEC-206-02).

  2. Informed consent: Not applicable.

  3. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  4. Use of Large Language Models, AI and Machine Learning Tools: None declared.

  5. Conflict of interest: The authors state no conflict of interest.

  6. Research funding: This work was supported by the National Science and Technology Major Project (2024ZD0523702), the Capital's Funds for Health Improvement and Research (2024-2-4059), the Shanghai Innovation Medical Device Application Demonstration Project (No. 23SHS06200), and the National High Level Hospital Clinical Research Funding (BJ-2023-094).

  7. Data availability: Not applicable.

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/cclm-2024-1416).

Received: 2024-12-05
Accepted: 2025-01-30
Published Online: 2025-02-19
Published in Print: 2025-06-26

© 2025 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2024-1416
Keywords for this article
25-hydroxyvitamin D2; 25-hydroxyvitamin D3; liquid chromatography-tandem mass spectrometry (LC-MS/MS); quality control; analytical performance comparison

Articles in the same Issue

  1. Frontmatter
  2. Editorials
  3. The journey to pre-analytical quality
  4. Manual tilt tube method for prothrombin time: a commentary on contemporary relevance
  5. Reviews
  6. From errors to excellence: the pre-analytical journey to improved quality in diagnostics. A scoping review
  7. Advancements and challenges in high-sensitivity cardiac troponin assays: diagnostic, pathophysiological, and clinical perspectives
  8. Opinion Paper
  9. Is it feasible for European laboratories to use SI units in reporting results?
  10. Perspectives
  11. What does cancer screening have to do with tomato growing?
  12. Computer simulation approaches to evaluate the interaction between analytical performance characteristics and clinical (mis)classification: a complementary tool for setting indirect outcome-based analytical performance specifications
  13. Genetics and Molecular Diagnostics
  14. Artificial base mismatches-mediated PCR (ABM-PCR) for detecting clinically relevant single-base mutations
  15. Candidate Reference Measurement Procedures and Materials
  16. Antiphospholipid IgG Certified Reference Material ERM®-DA477/IFCC: a tool for aPL harmonization?
  17. General Clinical Chemistry and Laboratory Medicine
  18. External quality assessment of the manual tilt tube technique for prothrombin time testing: a report from the IFCC-SSC/ISTH Working Group on the Standardization of PT/INR
  19. Simple steps to achieve harmonisation and standardisation of dried blood spot phenylalanine measurements and facilitate consistent management of patients with phenylketonuria
  20. Inclusion of pyridoxine dependent epilepsy in expanded newborn screening programs by tandem mass spectrometry: set up of first and second tier tests
  21. Analytical performance evaluation and optimization of serum 25(OH)D LC-MS/MS measurement
  22. Towards routine high-throughput analysis of fecal bile acids: validation of an enzymatic cycling method for the quantification of total bile acids in human stool samples on fully automated clinical chemistry analyzers
  23. Analytical and clinical evaluations of Snibe Maglumi® S100B assay
  24. Prevalence and detection of citrate contamination in clinical laboratory
  25. Reference Values and Biological Variations
  26. Temporal dynamics in laboratory medicine: cosinor analysis and real-world data (RWD) approaches to population chronobiology
  27. Establishing sex- and age-related reference intervals of serum glial fibrillary acid protein measured by the fully automated lumipulse system
  28. Hematology and Coagulation
  29. Performance of the automated digital cell image analyzer UIMD PBIA in white blood cell classification: a comparative study with sysmex DI-60
  30. Cancer Diagnostics
  31. Flow-cytometric MRD detection in pediatric T-ALL: a multicenter AIEOP-BFM consensus-based guided standardized approach
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  33. Assessing the clinical applicability of dimensionality reduction algorithms in flow cytometry for hematologic malignancies
  34. Cardiovascular Diseases
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  36. Retraction
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  38. Letters to the Editor
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  40. Reply to “Is this quantitative test fit-for-purpose?”
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  42. The first case of Teclistamab interference with serum electrophoresis and immunofixation
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  46. Congress Abstracts
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