Determination of progesterone and 17-hydroxyprogesterone by high performance liquid chromatography after pre-column derivatization with 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a- diaza-s-indacene-3-propionohydrazide
- 1 January 1998
- journal article
- research article
- Published by Royal Society of Chemistry (RSC) in The Analyst
- Vol. 123 (11) , 2339-2342
- https://doi.org/10.1039/a804201b
Abstract
Progesterone, 17-hydroxyprogesterone and four other 3-keto steroids were determined by high performance liquid chromatography with fluorescence detection. Each steroid was derivatized with 4,4-difluoro-5,7-dimethyl-4-bora- 3a,4a-diaza-s-indacene-3-propionohydrazide (BODIPY FL hydrazide) and separated on a Wakosil 5C4 column with acetonitrile–water (7 + 3) as mobile phase. The limits of detection of progesterone, 17-hydroxyprogesterone, dehydroepiandrosterone, androstenedione, testosterone and 17-methyltestosterone were 550–3700 fmol per 10 µl injection (signal-to-noise ratio = 5) serum. The calibration curves were linear up to 1000 ng/ml serum. The proposed method was most sensitive among the available high performance liquid chromatographic methods after fluorescence and chemiluminescence pre-labeling with dansylhydrazine.Keywords
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