Rapid monitoring assay of congenital adrenal hyperplasia with microbore high-performance liquid chromatography/electrospray ionization tandem mass spectrometry from dried blood spots.
17-hydroxyprogesterone (17OHP) is the most important plasma parameter for diagnosing and monitoring congenital adrenal hyperplasia (CAH) caused by 21-hydroxylase deficiency. A rapid, simple, and specific method based on microbore high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (micro-HPLC/ESI-MS/MS) was developed to determine the presence of 17OHP on dried filter-paper blood samples from patients with CAH caused by 21-hydroxylase deficiency. 17OHP from dried blood spots formed by the action of Girard reagent P (GirP) turned out to be a water-soluble hydrazone complex. Derivatization with GirP led to higher ESI sensitivity for 17OHP. The LC/MS/MS detection of GirP-derivatized 17OHP (GirP-17OHP) was rapid (<3 min). The method is repeatable and reproducible, with CVs <7% and 12%, respectively. This new method was used for direct determination of 17OHP in dried blood specimens obtained from abnormal children and infants of various ages with a detection limit of 10 ng/mL ( approximately 12 microL blood). The method described allows for rapid and reliable measurements of 17OHP in dried blood specimens from patients affected by CAH.
Lai
C C
CC
Department of Medical Genetics and Medical Research, China Medical College Hospital, Taichung, Taiwan.
Tsai
C H
CH
Tsai
F J
FJ
Lee
C C
CC
Lin
W D
WD
D019326:17-alpha-Hydroxyprogesterone / Q000032:analysis* / Q000097:blood*; D000312:Adrenal Hyperplasia, Congenital / Q000175:diagnosis*; D015415:Biomarkers; D001800:Blood Specimen Collection / Q000379:methods; D002138:Calibration; D002624:Chemistry, Clinical / Q000295:instrumentation / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D015415:Biomarkers; D019326:17-alpha-Hydroxyprogesterone