Analysis of 17 ?-hydroxyprogesterone in bloodspots by liquid chromatography tandem mass spectrometry

Salter, Simon J., Cook, Paul, Davies, Justin H. and Armston, Annie E. (2015) Analysis of 17 ?-hydroxyprogesterone in bloodspots by liquid chromatography tandem mass spectrometry. Annals of Clinical Biochemistry, 52 (1), 126-134. (doi:10.1177/0004563214530676). (PMID:24842631)

Abstract

BACKGROUND:

Monitoring of treatment for patients diagnosed with congenital adrenal hyperplasia (CAH) can be performed by measuring the concentration of 17?-hydroxyprogesterone (17OHP) in bloodspots collected on filter papers. A method is described here for measuring 17OHP by liquid chromatography tandem mass spectrometry (LCMSMS).

METHODS:

17OHP was extracted by liquid-liquid extraction and analysed by LCMSMS. The method was validated for sensitivity, specificity, linearity, recovery, ion suppression, precision and bias.

RESULTS:

The standard curve was linear from 0 to 400?nmol/L. Intra-assay %CVs were <10 and inter-assay %CVs were <15 over the range 10-200?nmol/L. Limit of quantitation was 6?nmol/L. No ion suppression was detected. The only interfering compound detected was deoxycorticosterone, an intermediate steroid with the same molecular weight as 17?-hydroxyprogesterone. The method was more accurate and precise than an existing radioimmunoassay. There was poor correlation between the two assays.

CONCLUSIONS:

We have developed a sensitive and specific assay suitable for quantitation of 17OHP in bloodspots. This method performs better than radioimmunoassay and allows smaller samples to be used.

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