Solid-phase analytical derivatization as a tool for the quantification of steroid hormones in human urine with HPLC-Q-ToF detection.

A possibility of application of solid-phase analytical derivatization (SPAD) for the quantification of seven steroid hormones (testosterone, dihydrotestosterone, cortisone, cortisol, progesterone, 11α-hydroxyprogesterone, and estrone) in human urine was evaluated. To prepare urine samples for instrumental analysis, SPAD with hydroxylamine was applied after enzymatic hydrolysis of the sample. To achieve high recovery values, extraction and derivatization conditions were optimized. Cartridges packed with end-capped octadecylsilyl silica sorbent provided optimum extraction of target analytes, while the reaction with hydroxylamine in the cartridge was found as a simple and efficient way for the chemical derivatization of steroids. The obtained derivatives were detected by using reversed-phase ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry. The proposed procedure was validated and applied to the analysis of real urine samples to prove the applicability of the proposed method for the routine analysis.

Quantification of cobalt and nickel in urine using inductively coupled plasma atomic emission spectroscopy.

Cobalt and nickel are micronutrients indispensable for the body, therefore, their use with food or as part of vitamin complexes is necessary to maintain health. As a result, trace cobalt and nickel contents are present in human biological fluids - blood and urine. According to the World Anti-Doping Agency prohibited list, they belong to the group of blood doping preparations - erythropoiesis stimulants. Nowadays, methods for their control in biological fluids are being actively developed to establish reasonable allowable contents of these trace elements in human biological fluids. However, in addition to developing highly sensitive methods for the determination of the total content of cobalt and nickel using ICP-MS and ETAAS, the development and comparison of various sample preparation methods that can provide the greatest accuracy, reproducibility and express analysis are also relevant. In the present paper, a comparison of different sample preparation methods - direct analysis, dilution and microwave mineralization of urine samples was shown, the detection and quantification limits were compared, some metrological characteristics that can be achieved using these sample preparation methods were evaluated. The procedure was tested on artificial and real urine samples. Taking the course of vitamin complexes in therapeutic concentrations was shown not to lead to a significant increase in the concentrations of analytes in urine, while taking elevated concentrations (for example, 2-fold) makes it possible to determine them even using ICP-AES. However, even in this case, cobalt and nickel concentrations remain at a relatively low level, not able to lead to a significant increase in erythropoiesis.