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Analytical and Bioanalytical Chemistry

, Volume 411, Issue 11, pp 2363–2371 | Cite as

Measurement of serum progesterone by isotope dilution liquid chromatography tandem mass spectrometry: a candidate reference method and its application to evaluating immunoassays

  • Yufei Wang
  • Tianjiao Zhang
  • Haijian Zhao
  • Weiyan Zhou
  • Jie Zeng
  • Jiangtao Zhang
  • Chuanbao ZhangEmail author
  • Wenxiang ChenEmail author
Research Paper

Abstract

Measurement of serum progesterone is important in determining ovarian function. Most progesterone measurements are performed with immunoassays and results are often variable. Standardization of the measurements requires a reliable reference method. An isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS) method for the measurement of serum progesterone was developed. Serum samples were spiked with 13C3-progesterone, extracted with a three-step liquid-liquid extraction, and analyzed by LC/MS/MS. A bracketing calibration was used for the analysis and samples were prepared gravimetrically. The developed method showed intra-run and total relative standard deviations (RSDs) of 0.50~0.58% and 0.71~1.33%, respectively. The analytical recoveries were 99.08~101.50%. Measurement results on certified reference materials obtained with this method agreed with the certified values within the stated measurement uncertainties. The method was applied to evaluate immunoassays through split-sample comparisons. A panel of 48 fresh frozen individual samples were measured with the ID-LC/MS/MS method, and six immunoassays and results were compared. Significant calibration biases and sample-specific deviations were observed on some of the immunoassays.

Keywords

Progesterone Reference method ID-LC/MS/MS Immunoassays 

Notes

Acknowledgements

We appreciate the technical support provided by the manufacturers (Abbott, Beckman, Chivd, Mindray, Roche, and Siemens) during the method comparison study.

Funding

This study was supported by grants from the National High Technology Research and Development Program of China (863 Program, No. 2011AA02A102) and The Leaping Development Project of Beijing Biomedical Industry (No. Z161100001816043).

Compliance with ethical standards

The individual serum samples were collected from the leftovers of patient samples at the clinical laboratory of Beijing Hospital, and this study had been reviewed and approved by the Ethics Committee of Beijing Hospital.

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

216_2019_1676_MOESM1_ESM.pdf (301 kb)
ESM 1 (PDF 300 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Yufei Wang
    • 1
    • 2
    • 3
  • Tianjiao Zhang
    • 2
    • 3
  • Haijian Zhao
    • 2
    • 3
  • Weiyan Zhou
    • 2
    • 3
  • Jie Zeng
    • 2
    • 3
  • Jiangtao Zhang
    • 2
    • 3
  • Chuanbao Zhang
    • 1
    • 2
    • 3
    Email author
  • Wenxiang Chen
    • 1
    • 2
    • 3
    Email author
  1. 1.Graduate School of Peking Union Medical College and Chinese Academy of Medical SciencesBeijingChina
  2. 2.National Center for Clinical LaboratoriesBeijing Hospital and National Center of GerontologyBeijingChina
  3. 3.Beijing Engineering Research Center of Laboratory MedicineBeijingChina

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