Rapid monitoring assay of congenital adrenal hyperplasia with microbore high‐performance liquid chromatography/electrospray ionization tandem mass spectrometry from dried blood spots

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Abstract
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 (µ‐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 (∼12 µL blood). The method described allows for rapid and reliable measurements of 17OHP in dried blood specimens from patients affected by CAH. Copyright © 2001 John Wiley & Sons, Ltd.

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