Quantitative determination of progesterones and corticosteroids in human urine using gas chromatography/mass spectrometry: application to pelvic organ prolapse patients.
A quantitative analytical method using gas chromatography/mass spectrometry (GC/MS) to determine urinary concentrations of eight progesterones and corticosteroids has been developed. After enzymatic hydrolysis with beta-glucouronidase/arylsulfatase, urine samples were extracted by simple one-step solid-phase extraction. Obtained extracts were derivatized with a mixture of N-methyl-N-(trimethylsilyl)trifluoroacetamide/ammonium iodide/dithiothreitol and determined by GC/MS in selected ion monitoring mode to increase the sensitivity. d(4)-Cortisol and d(9)-progesterone were used as internal standards for two different steroid groups. The linear correlation coefficient was in the range of 0.9913 to 0.9998 and recoveries were over 80% for all compounds. Precision and accuracy were in the range of 0.9-18.1 and 84.1-118.7%, respectively. The limit of quantitation (LOQ) was 10 ng/mL for 11-deoxycorticosterone and 21-deoxycortisol and 5 ng/mL for all other analytes. The developed method was successfully applied on pelvic organ prolapsed patients (n = 10, age: 67.9 +/- 4.9) and post-menopausal (n = 10, age: 63.6 +/- 5.5) control women. Urinary levels of most progesterones and corticosteroids except 11-deoxycorticosterone decreased but only that of 17 alpha-hydroxyprogesterone significantly decreased in patients compared with the control groups. Thus, it is concluded that progesterones could be a factor in the pathogenesis of pelvic organ prolapse, and, among them, 17 alpha-hydroxyprogesterone could be a biomarker for pelvic organ prolapse.
Shama
Naz
N
Bioanalysis and Biotransformation Research Center, Korea Institute of Science and Technology, P. O. Box 131, Cheongryang, Seoul, 130-650, South Korea.
Bai
Sang Wook
SW
Chung
Bong Chul
BC
Jung
Byung Hwa
BH
D000305:Adrenal Cortex Hormones / Q000652:urine*; D000368:Aged; D015415:Biomarkers / Q000652:urine; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D017773:Pelvic Floor; D017698:Postmenopause / Q000652:urine*; D011374:Progesterone / Q000652:urine*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016482:Urinalysis / Q000379:methods*; D014596:Uterine Prolapse / Q000175:diagnosis / Q000652:urine*
D000305:Adrenal Cortex Hormones; D015415:Biomarkers; D011374:Progesterone