Diagnosis of congenital adrenal hyperplasia by rapid determination of 17α‐hydroxyprogesterone in dried blood spots by gas chromatography/mass spectrometry following microwave‐assisted silylation

Abstract

17alpha-Hydroxyprogesterone (17OHP) is considered to be the biomarker of congential adrenal hyperplasia (CAH). Screening for CAH in newborns by measuring levels of the biomarker of 17OHP has become routine. In the work, a rapid, simple and sensitive technique was developed for the diagnosis of neonatal CAH by the quantitative analysis of 17OHP in neonatal blood spots. The technique was based on microwave-assisted silylation (MAS) followed by gas chromatography/mass spectrometry (GC/MS). In the method, fast derivatization of 17OHP with N,O-bis(trimethylsilyl)trifluoroacetamide was performed by using microwave irradiation, and the trimethylsilyl derivative thus formed was analyzed by GC/MS. The results of the experiment indicate that MAS followed by GC/MS analysis is a rapid, simple and sensitive method for the determination of 17OHP in blood samples. The proposed technique has been shown to have potential as a powerful tool for the rapid diagnosis of neonatal CAH.

Progress in molecular-genetic studies on congenital adrenal hyperplasia due to 11β-hydroxylase deficiency

“…Aldosterone production is low because of low serum potassium and plasma renin. [4][5][6] Molecular-genetic studies of 11 -hydroxylase defi ciency…”

Dried blood spot sampling in combination with LC‐MS/MS for quantitative analysis of small molecules

“…Unfortunately, assay sensitivity and specifi city are still a challenge due to the detection techniques. Gas chromatog-raphy (GC) in combination with mass spectrometric detection (GC-MS) is, in general, more sensitive than the LC-UV or LC-fl uorescence methods; however, GC run times are normally long (Deng et al, 2005). The time-consuming sample preparation (Yoon, 2007) and the necessity of derivatization for many analytes make GC-based methods less suitable for the highthroughput analysis of large sample sizes.…”

Microwave‐assisted silylation followed by gas chromatography/mass spectrometry for rapid determination of ergosterol in cigarettes

“…Three different solvents, ACN, pyridine, and ethyl acetate, have been applied for the derivatization of different components containing hydroxyl or amino groups with BSTFA [21 -23]. In our previous study, ACN was found to have higher polarity than the other two solvents and thus was more prone to absorb microwave energy [18]. Consequently, ACN was selected as the reaction solvent.…”

“…This technique combined with GC/MS was successfully applied to the analysis of 2,5-hexanedione, urinediuretics, and glucocorticoids in human urine, chlorophenols in ash [13 -16]. In our previous studies [17,18], microwave-assisted derivatization was successfully developed for the analysis of amino acids in neonatal blood samples.…”