Evaluation of CD detection in an HPLC system for analysis of DHEA and related steroids

Abstract

The biological importance of dehydroepiandrosterone (DHEA) is reflected by the fact that DHEA is a crucial precursor of the biosynthesis of the steroidal sex hormones. Simultaneous separation of DHEA, dehydroepiandrosterone sulfate (DHEA-S), pregnenolone, androstenedione and testosterone has been accomplished by reversed-phase ion-pair high-performance liquid chromatography (RP-IP-HPLC) based on isocratic elution applying circular dichroism (CD) detection at 295 nm. Addition of tetrabutylammonium hydrogensulfate to the mobile phase increases the retention of DHEA-S on the C8-silica column by an apparent ion-pairing mechanism without affecting the retention of the other (non-ionic) steroids. CD spectroscopy provides highly selective detection of compounds possessing optically active absorption bands and the separation is even more selective in the higher wavelength range applied. The linearity of the steroid concentration (c, mg mL(-1)) versus peak area was tested in the concentration range of 0.5-2 mg mL(-1) (injected quantities were 10-40 microg). The relative standard deviation (RSD) values for DHEA and DHEA-S indicated a good intra-assay and inter-assay precision of the method.

Circular Dichroism Techniques: Biomolecular and Nanostructural Analyses‐ A Review

“…The capabilities have been applied in several studies (160,161). For instance, Gergely and colleagues have reported the application of HPLC‐CD instrument could provide the selective detection of chiral compounds, and subsequently it allowed the employment of high detection wavelengths, presenting more selective detection with minimal background noise (161).…”

Simultaneous measurement of endogenous cortisol, cortisone, dehydroepiandrosterone, and dehydroepiandrosterone sulfate in nails by use of UPLC–MS–MS

“…Besides the immunoassay techniques mentioned above, hyphenated techniques were validated for the measurement of cortisol and cortisone in human and animal biological matrices with GC-MS and LC-MS [16,17], as well as for the measurement of DHEA and DHEAS [18,19]. LC-MS/MS technology provides the opportunity to accurately determine hormone concentrations even in small amounts of sample.…”

Advances in the analysis of steroid hormone drugs in pharmaceuticals and environmental samples (2004–2010)

“…The successful use of a circular dichroism (CD) detector has been reported in the analysis of prasterone and related steroids [29].…”