Abstract
Glutathione (GSH) is the most abundant non-protein thiol in biofluids, enabling diverse physiological functions. Among the proposed methods for GSH detection, ultra-high-performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) has the advantages of high sensitivity and efficiency. In this study, a novel analytical method was developed for the determination of GSH using supramolecular solvent (SUPRAS)-based dispersive liquid–liquid microextraction (DLLME) and UHPLC–HRMS. N-Laurylmaleimide was dissolved in tetrahydrofuran, which served three functions: 1) precipitate the proteins present in the biofluid sample, 2) provide a reaction environment for derivatization, and 3) enable the use of SUPRAS as the dispersing agent. Critical parameters were optimized based on single factor testing and response surface methodology. The established method was validated in terms of linearity, accuracy, precision, and successful quantitative analysis of GSH in saliva, urine, and plasma samples. Experimental results showed that SUPRAS as an extraction solvent was particularly suitable for the extraction of GSH from complex matrices. The current study provides a useful tool for accurate measurements of GSH concentrations, which could potentially be used for clinical diagnostics.
Highlights
According to the World Health Organization (WHO), a biomarker refers to any substance, structure, or process that can be measured in the body or its products and influence or predict the incidence of an outcome or disease (IPCS, 2001)
supramolecular solvents (SUPRASs)-based dispersive liquid–liquid microextraction (DLLME) and Ultrahigh-performance liquid chromatography (UHPLC)–high-resolution mass spectrometry (HRMS) were used for the determination of GSH in biofluids
The upper SUPRAS layer was analyzed by UHPLC–HRMS after diluted by methanol
Summary
According to the World Health Organization (WHO), a biomarker refers to any substance, structure, or process that can be measured in the body or its products and influence or predict the incidence of an outcome or disease (IPCS, 2001). GSH-recycling assay (Tietze, 1969; Chowdhury et al, 2013) with commercial assay kits, fluorescence (Cai et al, 2015), electrochemistry (Gao et al, 2016), surface-enhance Raman spectroscopy (SERS) (Saha and Jana, 2013; Zhu et al, 2021), colorimetry (Lee et al, 2018), flow injection analysis (Zitka et al, 2007; Kukoc-Modun et al, 2020), and high-performance liquid chromatography–mass spectrometry (HPLC–MS) (Moore et al, 2013) have been documented Among these methods, HPLC–MS has been a method of choice for the determination of GSH due to its advantages of sensitivity and specificity (Moral et al, 2012; Avula et al, 2013; Moore et al, 2013; Sun et al, 2019). The combination of UHPLC and HRMS has been a powerful tool for the determination of GSH (Vallverdu-Queralt et al, 2015; Zhang et al, 2020b; Zuo et al, 2018)
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