Determination of 4-pregnene-3-ones in thymus tissue samples by high performance liquid chromatography

A high performance liquid chromatography method for determining the levels of 4-pregnene-3-ones in tissue containing fat, e.g. in thymus is reported. Following the extraction with chloroform/methanol the fat has been separated from the steroidal-fraction by using disposable extraction-columns. The steroidal-fraction has been analysed in two separation systems, on a RP18-column with a methanol/water gradient and on an amino column with a isopropanol/hexane gradient. The applicability of the described method is demonstrated with calf thymus samples.