研究目的
To develop a robust method for the quantification of progesterone in human serum using isotope dilution-matrix-assisted laser desorption ionization-time of flight mass spectrometry.
研究成果
The 13C3-progesterone-based quantification assay is a robust method for measuring serum progesterone, showing good correlation with conventional methods.
研究不足
The method's robustness and constancy were compared to conventional radioimmunologic assay, but potential interferences from co-extracted compounds were noted.
1：Experimental Design and Method Selection
The study employed isotope dilution mass spectrometry for the quantification of progesterone in human serum, utilizing C4 solid-phase-extraction columns and hexane-based liquid/liquid extraction for sample preparation.
2：Sample Selection and Data Sources
Human serum samples were collected, centrifuged, and adjusted to pH 9.8 ± 0.2. 13C3-Progesterone was added as an internal standard isotope.
3：List of Experimental Equipment and Materials
MALDI-TOF/MS (AutoFlex III Smartbeam with nitrogen laser), SPE Supra-Clean 300 ? C4 columns, 13C3-Progesterone, 2,5-Dihydroxybenzoic acid (DHB).
4：Experimental Procedures and Operational Workflow
Serum samples were prepared, progesterone was extracted using C4 SPE columns, cleaned up with hexane-based liquid/liquid extraction, and analyzed by MALDI-TOF/MS.
5：Data Analysis Methods
The signal at m/z 108.9 of a progesterone fragment was used for quantification, with measurements performed in triplicate.
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