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

Analytical and Bioanalytical Chemistry
Yufei WangWenxiang Chen

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 ...Continue Reading

References

Jun 17, 2011·Clinical Chemistry·W Greg MillerCatharine M Sturgeon
Apr 18, 2014·Current Opinion in Endocrinology, Diabetes, and Obesity·Hemamalini KethaRavinder J Singh
Dec 2, 2014·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·María Salomé GachetJürg Gertsch
Jun 25, 2016·The Journal of Steroid Biochemistry and Molecular Biology·Simon TraversEric Pussard

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