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LC-MS/MS quantitation of plasma progesterone in cattle
R M T Fernandes1, G C Gomes, A M Porcari
1ThoMSon Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas-UNICAMP, Campinas, SP, Brazil.
Theriogenology
|July 30, 2011
Summary
A new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method offers accurate progesterone (P4) quantitation in cattle plasma, outperforming radioimmunoassay (RIA) by reducing interference and eliminating radioactive handling.
Area of Science:
- Veterinary Endocrinology and Reproductive Science
- Analytical Chemistry and Mass Spectrometry
Background:
- Progesterone (P4) quantitation in biological fluids is crucial for reproductive studies.
- Radioimmunoassay (RIA) is common but prone to false positives and interference.
- Liquid chromatography-tandem mass spectrometry (LC-MS/MS) offers higher specificity but is less frequently used for P4.
Purpose of the Study:
- To develop and validate an optimized, rapid, efficient, and cost-effective LC-MS/MS method for P4 quantitation in cattle plasma.
- To compare the performance of the developed LC-MS/MS method against traditional RIA.
- To assess P4 release kinetics from intravaginal P4-releasing devices (IPRDs) in ovariectomized cows.
Main Methods:
- An optimized LC-MS/MS method was developed for direct P4 quantitation in cattle plasma without sample derivatization.
- Plasma P4 levels were measured using both LC-MS/MS and RIA.
- A pairwise study compared P4 quantitation from IPRDs over 192 hours in six ovariectomized cows.
Main Results:
- Both LC-MS/MS and RIA showed similar P4 kinetics (P > 0.05).
- RIA consistently yielded higher P4 values compared to LC-MS/MS (P < 0.05) due to interference and matrix effects.
- The LC-MS/MS method demonstrated limits of detection (LOD) of 0.08 ng/mL and quantitation (LOQ) of 0.25 ng/mL.
Conclusions:
- The optimized LC-MS/MS method provides accurate and reliable P4 quantitation, minimizing false positives and interference common in RIA.
- This method eliminates the need for sample derivatization and avoids radioactive handling, enhancing safety and efficiency.
- The proposed LC-MS/MS technique is a viable alternative and potential gold standard for hormone quantitation in animal reproductive science.
