High-Throughput LC-MS/MS Quantification of Pregnenolone in Human Blood Serum for Clinical Research Purposes

Instrument Type: LCMSMS

Pregnenolone is a biosynthetic precursor to other steroids such as corticosteroids, androgens, and estrogens. Researchers investigating conversion to progesterone or to 17-OH-pregnenolone need to quantify pregnenolone within an analytical range of 0.3 to 1.5 nmol/L in blood serum. Since pregnenolone does not ionize well, derivatization with hydroxyl amine was necessary to reliably achieve the desired detection. We pursued this strategy along with liquid-liquid extraction to develop a simple sample preparation that would provide robust and sensitive quantitation of pregnenolone using ESI.