Progesterone in the Blood. A Quantitative Method Employing Gas Liquid Chromatography

Abstract

A new method for the measurement of progesterone in the blood has been described in detail. After addition of NaOH to 5 ml plasma, the progesterone was extracted with acetone and then ether. The extract was purified by chromatography through a miniature celite column using trimethylpentane as the mobile phase and 90% methanol as the stationary phase. This step removed a large quantity of pig-mented material and cholesterol. Further purification was achieved by thin layer chromatography in the system chloroform/acetone 9:1. After transfer to a simple dry sample application device, the progesterone was separated and the quantity was measured by means of gas liquid chromatography using 6 ft. XE 60 column. The method has been used to assay samples of plasma from patients at all stages of pregnancy and parturition. Its accuracy and reproducibility have been assessed by multiple measurements of 0.1 [mu]g progesterone added to male pooled plasma. The mean recovery at this level was 78. 6%, SD [plus or minus] 11. 7%, coefficient of variation 14. 8%.