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

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 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.

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

“…Monitoring and screening for CAH patients by measuring levels of 17α‐hydroxyprogesterone (17OHP) or other steroids has become a routine part of many programmes 1,. 2 Numerous methods have been described to determine these steroid hormones, such as fluorimetry,3,, 4 radioimmunoassay (RIA),5–8 and high‐performance liquid chromatography (HPLC) 4,. 9,, 10 Most of these, however, are affected by a degree of interference or cross‐reactivity with other steroids.…”

Miniaturized hollow fiber assisted liquid-phase microextraction and gas chromatography–mass spectrometry for the measurement of progesterone in human serum

“…Therefore, the method enables the precise determination of standards and can be applied to the determination of progesterone in human serum samples. Although, the sensitivity of the miniaturized HF-LPME method is lower than that of LLE method [5][6][7][8][9], the present method is an approach toward realizing green analytical chemistry because the required volume of organic solvent is very small. Moreover, the miniaturized HF-LPME method has excellent cost performance as it requires only a 10-L microsyringe, a porous hollow fiber, and conventional GC-MS.…”

“…* Corresponding author. Several analytical methods for the determination of progesterone in human serum samples have been reported, including liquid chromatography (LC) with fluorescence detection (FD) after derivatization [5], mass spectrometry (MS) [6], and tandem mass spectrometry (MS-MS) [7,8]. On the other hand, gas chromatography-mass spectrometry (GC-MS) with high resolution is useful for the determination of progesterone in human serum sample [9,10].…”

“…The sample preparation method known as liquid-liquid extraction (LLE) [5][6][7][8][9] has been successfully used for the determination of progesterone in human serum samples. However, LLE requires large volumes of organic solvents and additional concentration steps.…”

Determination of progesterone in capsules by high-performance liquid chromatography and UV- spectrophotometry

“…Progesterone is widely used in hormonetherapy and therefore it is necessary to establish a simple and accurate method for its identification and quantitative determination in pharmaceuticals. The literature shows that gas chromatography (GC), gas chromatography-mass spectrometry (GC-MS), [1][2][3][4][5] and high performance liquid chromatography (HPLC) [6][7][8] have been reported for the simultaneous determination of progesterone, 17-hydroxyprogesterone and other 3-keto steroids. However, no single method has been developed for progesterone in single component form.…”