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-propionohydra zide.
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 microliters 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.
Katayama
M
M
Department of Functional Bioanalysis, Meiji College of Pharmacy, Tokyo, Japan.
Nakane
R
R
Matsuda
Y
Y
Kaneko
S
S
Hara
I
I
Sato
H
H
D019326:17-alpha-Hydroxyprogesterone / Q000097:blood; D000735:Androstenedione / Q000097:blood; D001896:Boron Compounds; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003687:Dehydroepiandrosterone / Q000097:blood; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D008777:Methyltestosterone / Q000097:blood; D011374:Progesterone / Q000097:blood*; D013739:Testosterone / Q000097:blood
C120161:BODIPY FL hydrazide; D001896:Boron Compounds; D005456:Fluorescent Dyes; D013739:Testosterone; D000735:Androstenedione; D003687:Dehydroepiandrosterone; D011374:Progesterone; D019326:17-alpha-Hydroxyprogesterone; D008777:Methyltestosterone