Analytical Performance of LC-MS/MS Method for Simultaneous Determination of Five Steroids in Serum

Abstract

The accurate measurement of steroid hormones is the fulcrum of modern endocrinology. Our aim was to develop a mass spectrometry coupled with a liquid chromatographic method for 17-hydroxyprogesterone, androstenedione, 11-desoxycortisol, 21-desoxycortisol and cortisol serum quantification simultaneously. The samples were submitted to isotopic internal standards addition, protein precipitation and two dimensional liquid chromatography consisting of trapping column and reverse-phase C18 analytical column following atmospheric pressure chemical ionization and mass spectrometry detection. For all compounds tested functional sensitivity was less than 0.5 ng/mL, precision was less than 15%, recovery ranged from 93% to 120%, linearity ranged from 89% to 111%, and accuracy was considered adequate. In conclusion, we developed a suitable, fast and reliable method for simultaneous routine measurement of 17-hydroxyprogesterone, androstenedione, 11-desoxycortisol, 21-desoxycortisol and cortisol in serum.

Method Development and Validation for Quantification of Cortisol and Cortisone Using Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

“…The presence of endogenous cortisol and cortisone in the serum makes it difficult for method validation analysis. Several studies have used pooled human serum as a blank matrix [19,28]. However, using pooled serum may cause variation in the baseline cortisol and cortisone concentration for different batches of pooled serum.…”

“…Therefore, a highly specific and sensitive analytical tool is needed to quantitate the concentration of the two closely identical molecules such as cortisol and cortisone in biological samples. In this respect, quantitative analysis of low-concentration analytes is possible by using a highly reproducible, selective, and sensitive liquid chromatography-tandem mass spectrometry (LC/MS-MS) methods [19][20][21]. The validated method is useful for investigation of hormone status for the diagnosis and treatment of clinical disorders.…”