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J Chromatogr B Biomed Sci Appl. 1997 Jan 10;688(1):11-26.

Method for combined analysis of profiles of conjugated progesterone metabolites and bile acids in serum and urine of pregnant women.

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  • 1Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.


A method for analysis of profiles of conjugated progesterone metabolites and bile acids in 10 ml of urine and 1-4 ml of serum from pregnant women is described. Total bile acids and neutral steroids from serum and urine were extracted with octadecylsilane-bonded silica. Groups of conjugates were separated on the lipophilic ion-exchanger triethylaminohydroxypropyl Sephadex LH-20 (TEAP-LH-20). Fractions were divided for steroid or bile acid analyses. Sequences of hydrolysis/solvolysis and separations on TEAP-LH-20 permitted separate analyses of steroid glucuronides, monosulfates and disulfates and bile acid aminoacyl amidates, sulfates, glucuronides and sulfate-glucuronides. Radiolabelled compounds were added at different steps to monitor recoveries and completeness of separation, and hydrolysis/solvolysis of conjugates was monitored by fast-atom bombardment mass spectrometry. The extraction and solvolysis of steroid disulfates in urine were studied in detail, and extraction recoveries were found to be pH-dependent. Following methylation of bile acids, all compounds were analysed by capillary gas chromatography and gas chromatography-mass spectrometry of their trimethylsilyl ether derivatives. Semiquantification of individual compounds in each profile by gas-liquid chromatography had a coefficient of variation of less than 30%. The total analysis required 3 days for serum and 4 days for urine.

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