Method for combined analysis of profiles of conjugated progesterone metabolites and bile acids in serum and urine of pregnant women.
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.
Meng
L J
LJ
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Sjövall
J
J
D001647:Bile Acids and Salts / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000652:urine; D002250:Carbon Radioisotopes; D019826:Cholic Acid; D002793:Cholic Acids / Q000032:analysis / Q000097:blood / Q000737:chemistry / Q000652:urine; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D012739:Gonadal Steroid Hormones / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000652:urine; D006801:Humans; D006868:Hydrolysis; D011247:Pregnancy / Q000097:blood / Q000378:metabolism* / Q000652:urine; D011263:Pregnancy Trimester, Third; D011374:Progesterone / Q000032:analysis / Q000097:blood / Q000737:chemistry / Q000378:metabolism* / Q000652:urine; D015203:Reproducibility of Results; D012822:Silicon Dioxide / Q000737:chemistry; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013739:Testosterone / Q000032:analysis / Q000097:blood / Q000737:chemistry / Q000652:urine; D014316:Tritium
D001647:Bile Acids and Salts; D002250:Carbon Radioisotopes; D002793:Cholic Acids; D012739:Gonadal Steroid Hormones; D014316:Tritium; D013739:Testosterone; D011374:Progesterone; D012822:Silicon Dioxide; D019826:Cholic Acid