Shoichiro Murakawa

The determination of urinary testosterone using thin-layer chromatography and gas chromatography

Abstract A new method has been devised for the relatively rapid and sensitive determination of testosterone in human urine. One fourth of a 24 hour urine sample was incubated with β-glucuronidase after addition of a tracer dose of 1,2–3H-testosterone for the calculation of recovery and extracted with ethyl ether. The crude extract was subjected to thin-layer chromatography (TLC), eluted, acetylated, subjected to TLC again and analyzed by gas chromatography. This method detected as little as 5 μg of testosterone per 24 hour and the rate of recovery was 54.8±0.4% (Mean±S.E.). The value for normal adult men ranged from 41 to 200 μg/day, showing definite decline with advancing age. While the urinary testosterone exhibits a very low level (5>-8 μg/day) in normal adult women, hirsute women excreted almost as much testosterone as normal adult men.