Quantification of circulating steroids in individual zebrafish using stacking to achieve nanomolar detection limits with capillary electrophoresis and UV-visible absorbance detection

Abstract

Capillary electrophoresis and UV-visible absorbance detection are used with sample stacking to achieve detection limits ranging from 0.2 to 2 ng/mL (0.8 to 6 nM) for steroids. Stacking is accomplished using negatively charged cyclodextrin steroid-carrier molecules at a discrete pH interface between the reconstituted sample and the separation electrolyte. Steroids are then separated in under 5 min using capillary electrophoresis that incorporates secondary equilibria via sodium dodecyl sulfate and cyclodextrin. The effectiveness of the method for measurements of multiple steroids in limited sample volumes is demonstrated in individual female fish with total circulating blood volumes of 5 μL or less. Steroid recoveries from plasma following a sample processing method developed with commercial extraction cartridges range from 81 to 109 % for 17α,20β-dihydroxy-pregn-4-en-3-one, testosterone, 11-ketotestosterone, estrone, 17β-estradiol, and 17α-ethinyl estradiol. When applied to reproductively active female zebrafish, changes were detected in the levels of circulating steroids as a result of exposure to different solvents and 17β-estradiol.

Optimization of mass spectrometry settings for steroidomic analysis in young and old killifish

“…Notably, in killifish steroidogenesis occurs primarily in gonads, adrenal glands, liver, and brain [22][23][24]. Fish also typically contain high concentrations of steroids, facilitating the study of steroids in vivo [25][26][27].…”

Unexpected differences between planar and column liquid chromatographic retention of 1-acenaphthenol enantiomers controlled by supramolecular interactions involving β-cyclodextrin at subambient temperatures

“…In spite of discovery and extensive studies devoted to other simple (crown ethers, calixarenes, cyclophanes, macrocyclic antibiotics) or more sophisticated, based on mechanically interlocked molecular architecture of macrocyclic compounds (like rotaxanes, catenanes, knotanes), there is still increasing interest in cyclodextrin research [ 14 , 15 ]. This is mainly because of a number of key applications of CDs and extensive usage of macrocyclic oligosaccharides in chemistry (analytical chemistry, separation sciences; e.g., as resolution enhancers, active components of extraction systems) [ 16 – 18 ], micro-electronics and microfluidics devices (components of sensing systems) [ 19 , 20 ], medicine, pharmacy, and cosmetology (drug and bioactive components carriers) [ 21 – 24 ], as well as food industry (aroma carriers, biocomponents extraction, active components of smart food packaging systems) [ 25 – 27 ]. Most recently, there is also an extensive research focusing on their efficient encapsulation properties, working as free molecules or after incorporation within new polysaccharide-based nanomaterials [ 28 , 29 ].…”

Androgens, oestrogens, and progesterone concentrations in wastewater purification processes measured with capillary electrophoresis

“…Naturally, their total steroid amount in the effluent water was much lower than in 459 influent being 126 ng/L. The reason for high concentration of androstenedione may be that 460 progesterone can produce androstenedione that is the precursor in metabolism of testosterone (one of 461 the androgens) and estrogens such as estradiol (Nyakubaya et al 2015) [39]. Otherwise, the removal 462 of the individual steroids were much lower.…”