High-throughput sample preparation and analysis using 96-well membrane solid-phase extraction and liquid chromatography-tandem mass spectrometry for the determination of steroids in human urine.
A 96-well solid-phase extraction (SPE) system is used to rapidly prepare human urine samples for high-throughput quantitative analysis of two steroids, equilenin and progesterone, by liquid chromatography-tandem mass spectrometry using deuterated estrone as the internal standard. We define high-throughput here as analysis of 384 samples in a 24 h period. A total of 384 samples and standards were extracted by an individual in one day and subsequently analyzed within a 24 h period. The inter- and intratray accuracy and precision obtained over the course of these injections was within 8% coefficient of variation when analyzed by atmospheric pressure chemical ionization mass spectrometry using positive ion detection. A semiautomated sample processing workstation was used to add internal standard and then process 96 samples at a time. The recovery of the analytes from the SPE was approximately 85%. The accuracy and precision obtained was comparable to that ordinarily obtained using manual sample preparation techniques.
Rule
G
G
Cornell University, New York State College of Veterinary Medicine, Ithaca 14850, USA.
Henion
J
J
D001322:Autoanalysis; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D004855:Equilenin / Q000652:urine; D015914:Estrogen Replacement Therapy; D004967:Estrogens / Q000652:urine; D005260:Female; D006801:Humans; D013058:Mass Spectrometry; D011374:Progesterone / Q000652:urine; D011786:Quality Control; D012015:Reference Standards; D013256:Steroids / Q000652:urine*
D004967:Estrogens; D013256:Steroids; D011374:Progesterone; D004855:Equilenin