Abstract
(1) Antioxidants are involved in body protection mechanisms against reactive oxygen species. Amino acids such as glutathione (GSH) and N-acetylcysteine (NAC) are known to be involved in providing protection against oxidative lethality. A quick and simple method for the determination of NAC and GSH in various biological matrices such as urine, plasma, and homogenates of brain tissues has been developed and described in this work. (2) The assay is based on reversed phase high performance liquid chromatography with spectrofluorimetric detection and on-column derivatization. Butylamine and o-phthaldialdehyde have been used as derivatization reagents. Since o-phthaldialdehyde constitutes a part of the mobile phase, the derivatization reaction and chromatographic separation occur simultaneously. (3) Linearity in the detector response for NAC in human urine was observed in the range of 5–200 nmol mL−1, and NAC and GSH in the brain tissue homogenates were observed in the range of 0.5–5 nmol mL−1 and 0.5–15 nmol mL−1, respectively. Human plasma linearity ranges covered 0.25–5.00 nmol mL−1 and 0.5–15 nmol mL−1 for NAC and GSH, respectively. The LODs for NAC and GSH were 0.01 and 0.02 nmol mL−1 while the LOQs were 0.02 and 0.05 nmol mL−1, respectively. The usefulness of the proposed method was proven through its application to real samples.
Highlights
Thiols belong to a group of organic compounds containing very active and biologically important sulfhydryl group (-SH)
(3) Linearity in the detector response for NAC in human urine was observed in the range of 5–200 nmol mL−1, and NAC and GSH in the brain tissue homogenates were observed in the range of 0.5–5 nmol mL−1 and 0.5–15 nmol mL−1, respectively
The compounds above-mentioned are extensively utilized in procedures dedicated for the control of thiol levels in biological samples and are needed for analyte fluorescence labeling
Summary
Thiols belong to a group of organic compounds containing very active and biologically important sulfhydryl group (-SH). N-acetylcysteine (NAC) plays an essential role in the human body due to its metabolic and medical properties. All of these compounds are widespread in tissues and biological fluids such as plasma and urine [1,2]. Thiols are a substantial cellular component of many key pathways of metabolic processes. They play a significant role as active components of the sulfur cycle as well as take part in the defense mechanisms of the organism against the pathological effects of reactive forms of oxygen and nitrogen [1,3]. It has been proven that changes in some thiol concentrations and their molar ratio can act as potential indicators of cell health and the risk of diseases [2]
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