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

Hamad, Amal E.; Elshahawy, Mohamed; Negm, Amr; Mansour, Fotouh R.

doi:10.1515/revac-2019-0019

Cited by 33 publications

(19 citation statements)

References 144 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Fluorometric Optimized Determination of Total Glutathione in Erythrocytes

et al. 2021

View full text Add to dashboard Cite

Glutathione is a tripeptide natural product characterized by a non-canonical peptide bond with an amide moiety linking the nitrogen of cysteine to the γ-carboxyl of glutamate, and is found ubiquitously in nature, in animals, plants and microorganisms. One of the most abundant biological matrices is represented by erythrocytes, being glutathione the only sulfur-containing mechanism for the red blood cell oxidative protection. Several analytical methods for glutathione determination from different samples are described in the literature and most of these methods are based on the use of high-performance liquid chromatography. HPLC equipment is not available in all the biochemical laboratories, and, moreover, displays lot of economic and ecological limitations, including organic solvent consumption and time-consuming analysis. Here, an organic-free high-throughput fluorometric methodology for the analysis of total glutathione in erythrocytes is reported, avoiding the use of time-consuming and not-sustainable techniques.

show abstract

Section: Discussionmentioning

confidence: 99%

“…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].…”

Section: Hplc Glutathione Determinationmentioning

confidence: 99%

Fluorometric Optimized Determination of Total Glutathione in Erythrocytes

et al. 2021

View full text Add to dashboard Cite

show abstract

“…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].…”

Section: Introductionmentioning

confidence: 99%

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

Hammad

Abdallah

Bedair

et al. 2021

J of Separation Science

Self Cite

View full text Add to dashboard Cite

Liquid-liquid extraction is a widely used technique of sample preparation in biomedical analysis. In spite of the high pre-concentration capacities of liquidliquid extraction, it suffers from a number of limitations including time and effort consumption, large organic solvent utilization, and poor performance in highly polar analytes. Homogeneous liquid-liquid extraction is an alternative sample preparation technique that overcomes some drawbacks of conventional liquid-liquid extraction, and allows employing greener organic solvents in sample treatment. In homogeneous liquid-liquid extraction, a homogeneous phase is formed between the aqueous sample and the water-miscible extractant, followed by chemically or physically induced phase separation. To form the homogeneous phase, aqueous samples are mixed with water-miscible organic solvents, waterimmiscible solvents/cosolvents, surfactants, or smart polymers. Then, phase separation is induced chemically (adding salt, sugar, or buffer) or physically (changing temperature or pH). This mode is rapid, sustainable, and cost-effective in comparison with other sample preparation techniques. Moreover, homogeneous liquid-liquid extraction is more suitable for the extraction of delicate macromolecules such as enzymes, hormones, and proteins and it is more compatible with liquid chromatography with tandem mass spectrometry, which is a vital technique in metabolomics and proteomics. In this review, the principle, types, applications, automation, and technical aspects of homogeneous liquid-liquid extraction are discussed.

show abstract

“…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].…”

Section: Introductionmentioning

confidence: 99%

“…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.…”

Section: Introductionmentioning

confidence: 99%

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

Singh¹,

MJ²,

Anchliya³

2021

AustinJAnalPharmChem

View full text Add to dashboard Cite

The objective of this study was the development, optimization, and validation of a RP-HPLC method for the quantification of reduced glutathione (GSH) and oxidized glutathione (GSSG) in pharmaceutical formulations The separation utilized a C18 column at room temperature with absorption wavelength 210nm. The mobile phase was an isocratic flow of a 95:5 (v/v) mixture of 25mM phosphate buffer (pH 2.7) and methanol with flow rate at 1.0 mL/min. Validation of the method assessed with the methods ability in seven categories: linearity, range, limit of detection, limit of quantification, accuracy, precision, and selectivity. The method show an acceptable degree of linearity with r²=0.9994 and 0.999 over a concentration range of 10-200 μg/mL for GSH and GSSG respectively. The detection limit and quantification limit for GSH 20.7μg/mL and 69.24μg/mL and for GSSG 17.22μg/mL and 57.42μg/mL respectively. The percent recovery of the method was 99.98-100.93 %. Following validation, the method was employed in the determination of glutathione in pharmaceutical formulations in the form of a liposome. The proposed method offers a simple, accurate, and inexpensive way to quantify reduced glutathione.

show abstract

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

Cited by 33 publications

References 144 publications

Fluorometric Optimized Determination of Total Glutathione in Erythrocytes

Fluorometric Optimized Determination of Total Glutathione in Erythrocytes

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

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

Contact Info

Product

Resources

About