Analytical methods for determination of glutathione and glutathione disulfide in pharmaceuticals and biological fluids

Abstract

<jats:title>Abstract</jats:title><jats:p>Glutathione is a natural tripeptide that plays a major role in different physiological processes in the human body. Determination of glutathione in different body fluids and tissues is essential for early diagnosis and follow-up of various diseases. The assay of glutathione is problematic because of the high polarity, the limited stability, and the aliphatic structure, which lacks the appropriate chromophore for UV detection. A number of methods have been reported for determination of glutathione using different techniques. High-performance liquid chromatography was employed in both reverse phase and hydrophilic interaction modes. Electrochemical methods exploited the redox activity of glutathione to allow for quantification by different electrodes after chemical modification, including glassy carbon, carbon paste, and nanocomposite electrodes. Capillary zone electrophoresis was used with less need for derivatization which makes it simpler, faster, and more economic. A number of nanosensors and probes have been developed to assay glutathione in biological fluids using semiconductor nanoparticles, quantum dots, genetically engineered green fluorescent probes, and new derivatives of known dye classes. This work is an updated review of the methods of analysis of glutathione and glutathione disulfide in pharmaceuticals and biological fluids with more emphasis on the technical problems and the assay artifacts.</jats:p>

Fluorometric Optimized Determination of Total Glutathione in Erythrocytes

“…In this work, the sample preparation and the derivatization were optimized for direct fluorescence detection of the GSH-OPA isoindole derivative. As reported in the literature, the advantage of OPA-GSH analytical determinations is the higher stability of the fluorescent derivative, with respect to other fluorophores [15,17]. HPLC still represents the best reliable technique that gives precise, accurate and validated results; however, the procedure has a lot of economical/ecological and time-consuming limitations, such as organic solvents consumption and larger chromatographic times.…”

“…The instrument control and data acquisition were carried out using the Millennium 32 software (Milford, Massachusetts, USA). As known from literatur derivatization of GSH and HPLC chromatographic analysis are validated metho total recovery and high stability of the indole fluorescent analyte [15,17].…”

Homogeneous liquid–liquid extraction as an alternative sample preparation technique for biomedical analysis

“…In addition, LLE is non eco-friendly and health-hazardous due to the toxicity of the commonly used extractants [5]. Moreover, LLE has limited efficiency in extraction of highly polar drugs due to the hydrophobic nature of the water immiscible organic solvents used as extractants [6][7][8].…”

Development and Validation of HPLC Method for Simultaneous Estimation of Reduced and Oxidized Glutathione in Bulk Pharmaceutical Formulation

“…Thus, the molar ratio of GSH/GSSG is an important measure of the cellular oxidative stress. The low GSH/GSSG ratio is an important biomarker in a number of human disorders such as cataract, cancer, parkinsonism, diabetes, renal failure, pneumonia, Alzheimer's disease, lead toxicity, cystic fibrosis, dystrophic skin fibroblasts, neurodegenerative disorders, and liver and human immunodeficiency virus infections [12][13][14][15][16]. Determination of GSH is challenging because of the following reasons: (i) GSH can be easily autoxidized to GSSG, which results in a lower GSH/GSSG ratio and the lack of good UV/Vis chromophore or native [17].…”

“…Determination of GSH is challenging because of the following reasons: (i) GSH can be easily autoxidized to GSSG, which results in a lower GSH/GSSG ratio and the lack of good UV/Vis chromophore or native [17]. The number of publications for determination of GSH by different methods have been reported which include HPLC, Electrochemical methods, Capillary electrophoresis, Spectroscopic probes and sensors [14,[18][19][20][21]. But no methods have been reported for simultaneous determination of reduced (GSH) and oxidized glutathione (GSSG) in pharmaceutical formulation.…”