Determination of Progesterone in Human Peripheral Blood Using Gas-Liquid Chromatography with Electron Capture Detection1

Abstract

A method for the estimation of progesterone in human peripheral blood is described. After converting progesterone to 20[beta]-hydroxy pregn-4-en-3-one, the monochloroacetate derivative of this compound is prepared. Nanogram amounts of the derivative can be estimated using electron capture detection following gas-liquid chromatography. The precision of the method ranges from 10 to 40% for samples containing less than 0.01 [mu]g progesterone, from 10 to 15% for samples containing more than 0.01 [mu]g, and may be as high as 5 to 6% for samples containing 0.1 [mu]g or more. Plasma progesterone values for normal male subjects ranged from <0.015 to 0.048 [mu]g/100 ml (14 individuals, mean: 0.028 [mu]g/100 ml). The progesterone values found in plasma of normal women varied throughout the cycle between <0.015 and 0.11 ug/100 ml (mean: 0.049 [mu]g/100 ml, 10 samples) from the day of onset of menstruation to the 22nd day before onset of menstruation of the next cycle, <0.015-0.42 [mu]g/100 ml (mean: 0.13 [mu]g/100 ml, 10 samples) from day 21 to 15 before onset of menstruation, 0.79-2.06 [mu]g/100 ml (mean: 1.52 [mu]g/100 ml, 12 samples) from day 14 to 8 before onset, of menstruation and 0.10-1.79 [mu]g/100 ml (mean: 0.71 [mu]g/100 ml, 10 samples) from day 7 to 1 before onset of menstruation.