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Determination of natural progesterone levels in bovine serum using gas chromatography coupled with mass spectrometry Cover

Determination of natural progesterone levels in bovine serum using gas chromatography coupled with mass spectrometry

Open Access
|Dec 2025

Abstract

Introduction

Progesterone, a steroidal female sex hormone, can be used for anabolic purposes in cattle. Physiological levels of progesterone in animals are highly variable, and systematic studies of them in various matrices are limited. The purpose of the present study was to apply a developed and validated method routinely used for the determination of the natural hormones 17β-oestradiol and 17β-testosterone in serum to estimate natural cattle progesterone levels. The analytes were taken from serum samples collected from cattle as part of the Polish National Residue Control Plan.

Material and Methods

A total of 1,537 bovine serum samples were analysed: 895 from females and 642 from males. Progesterone was extracted from serum with a mixture of tert-butyl methyl ether and petroleum ether, the extract was derivatised and the analyte was determined by gas chromatography coupled with mass spectrometry.

Results

The basic validation parameters specified for the method, namely repeatability, reproducibility and apparent recovery, indicated it to be useful for the intended purpose. The natural presence of progesterone was detected in 679 samples from females and in 375 samples from males. Progesterone in females was measured in the range of 0.001–496.059 μg L−1 (in 61% of samples it was below 1 μg L−1 and in 32% it was in the range of 1–7 μg L−1, which are the physiological ranges of phases of the reproductive cycle) and progesterone in males was assayed in the range of 0.001–31.792 μg L−1.

Conclusion

The results obtained may serve as a statistical basis for further research.

Language: English
Submitted on: Aug 18, 2025
Accepted on: Dec 10, 2025
Published on: Dec 16, 2025
Published by: National Veterinary Research Institute in Pulawy
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year

© 2025 Iwona Matraszek-Żuchowska, Alicja Kłopot, Paulina Zdonek, Beata Korycińska, Justyna Grzelak, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution 4.0 License.

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