Determination of glutathione in biological material by flow-injection analysis using an enzymatic recycling reaction

Redegeld, Frank A.; Opstal, M.A.J. van; Houdkamp, E.; Bennekom, W.P. van

doi:10.1016/0003-2697(88)90048-6

Cited by 56 publications

(18 citation statements)

References 17 publications

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“…It has been shown that GSH reductase affects the background signal by the slow reduction of DTNB [37]. Reducing agents, such as ascorbic acid, mercaptoethanol, dithiothreitol and cysteine, or thiol reactive compounds could interfere with the DTNB assay as well [38]. Decreased GSH levels could be due to the leakage and/or oxidation to GSSG (oxidized GSH).…”

Section: Discussionmentioning

confidence: 99%

Influence of Pre-Storage Irradiation on the Oxidative Stress Markers, Membrane Integrity, Size and Shape of the Cold Stored Red Blood Cells

Antosik

Czubak

Gajek

et al. 2015

Transfus Med Hemother

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Background: To investigate the extent of oxidative damage and changes in morphology of manually isolated red blood cells (RBCs) from whole blood, cold stored (up to 20 days) in polystyrene tubes and subjected to pre-storage irradiation (50 Gy) and to compare the properties of SAGM-preserved RBCs stored under experimental conditions (polystyrene tubes) with RBCs from standard blood bag storage. Methods: The percentage of hemolysis as well as the extracellular activity of LDH, thiobarbituric acid-reactive substances, reduced glutathione (GSH), and total antioxidant capacity (TAC) were measured. Changes in the topology of RBC membrane, shape, and size were evaluated by flow cytometry and judged against microscopy images. Results: Irradiation caused significant LDH release as well as increased hemolysis and lipid peroxidation, GSH depletion, and reduction of TAC. Prolonged storage of irradiated RBCs resulted in phosphatidylserine exposure on the cell surface. By day 20, approximately 60% of RBCs displayed non-discoid shape. We did not notice significant differences in percentage of altered cells and cell volume between RBCs exposed to irradiation and those not exposed. Conclusion: Irradiation of RBC transfusion units with a dose of 50 Gy should be avoided. For research purposes such as studying the role of antioxidants, storage of small volumes of RBCs derived from the same donor would be more useful, cheaper, and blood-saving.

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Section: Discussionmentioning

confidence: 99%

Influence of Pre-Storage Irradiation on the Oxidative Stress Markers, Membrane Integrity, Size and Shape of the Cold Stored Red Blood Cells

Antosik

Czubak

Gajek

et al. 2015

Transfus Med Hemother

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“…Reduced and oxidized vitamin C was determined using a modified HPLC method [ 19,20]. Red blood cells were washed twice in PBS (500 g, 10 min) and immediately prepared for spectrophotomctric flow injection analysis of reduced and oxidized glutathione [21]. Methemoglobin con centration was calculated from measurements with a Coming 2500 CO oximeter spectrophotometer (CibaCorning, Medfiled, Mass., USA) in blood drawn from the left ventricle.…”

Section: Assessment O F Redox Status {Ant ¡Oxidative Capaci Ty and LImentioning

confidence: 99%

Inhalation of Nitric Oxide: Effect on Cerebral Hemodynamics and Activity, and Antioxidant Status in the Newborn Lamb

et al. 1996

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Ventilation with nitric oxide (NO) is increasingly being used to treat pulmonary hypertension in the newborn. In the brain, NO has vasoactive properties and is involved in neurotransmission. However, the effect of inhaled NO on the cerebral blood flow (CBF) and on the cerebral activity is not known. Furthermore, there is little information on the influence of this free radical gas on the redox status in pulmonary vessels. We therefore investigated the effect of inhaled NO (2–60 ppm) on CBF, cerebral activity and redox status in blood effluent from the pulmonary circulation in 6 ventilated newborn lambs before and during group B streptococci (GBS)-induced pulmonary hypertension. Blood pressure in the pulmonary artery (P_ap) and aorta (P_ao), carotid artery blood flow (Q_car) to assess changes in CBF, and electrocortical activity were measured. Blood gases, indices of free radical status and methemoglobin were determined in blood samples obtained from the left ventricle. Inhalation of NO, before and during GBS-induced pulmonary hypertension, decreased P_ap and P_CO2 and increased P_O2. Multiple linear regression revealed that Q_car was positively related to P_CO2, but not to inhaled NO or P_O2 before or during GBS conditions. Electrocortical activity and indices of antioxidative capacity and lipid peroxidation did not change significantly. Methemoglobin was not detected. In conclusion, inhalation of NO (up to 60 ppm) lowered Pap without directly affecting CBF, electrocortical activity, and redox status in the pulmonary vessels. CBF, however, can indirectly be influenced by NO-mediated changes in P_CO2·

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“…Erythrocyte GSH (RBC GSH) serves as a convenient biological reserve in which to measure GSH status, and several valid measurement methods are available, with enzymatic recycling being the most sensitive. [15][16][17][18] Importantly, the reactive nature of GSH causes challenges in sample stability and necessitates correct sample collection methods. 19 The bioavailability of orally administered cysteine is believed to be the rate-limiting step for the synthesis of glutathione, 10 although the amino acids glutamate/ glutamine are also integral to GSH synthesis.…”

Section: Introductionmentioning

confidence: 99%

Effects of Oral Glutathione Supplementation on Systemic Oxidative Stress Biomarkers in Human Volunteers

Allen

Bradley

2011

The Journal of Alternative and Complementary Medicine

135

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Background: The tripeptide glutathione (GSH) is the most abundant free radical scavenger synthesized endogenously in humans. Increasing mechanistic, clinical, and epidemiological evidence demonstrates that GSH status is significant in acute and chronic diseases. Despite ease of delivery, little controlled clinical research data exist evaluating the effects of oral GSH supplementation. Objectives: The study objectives were to determine the effect of oral GSH supplementation on biomarkers of systemic oxidative stress in human volunteers. Design: This was a randomized, double-blind, placebo-controlled clinical trial. Setting/location: The study was conducted at Bastyr University Research Institute, Kenmore, WA and the Bastyr Center for Natural Health, Seattle, WA. Subjects: Forty (40) adult volunteers without acute or chronic disease participated in this study. Intervention: Oral GSH supplementation (500 mg twice daily) was given to the volunteers for 4 weeks. Outcome measures: Primary outcome measures included change in creatinine-standardized, urinary F2-isoprostanes (F2-isoP) and urinary 8-hydroxy-2¢-deoxyguanosine (8-OHdG). Changes in erythrocyte GSH concentrations, including total reduced glutathione (GSH), oxidized glutathione (GSSG), and their ratio (GSH:GSSG) were also measured by tandem liquid chromatography/mass spectrometry. Analysis of variance was used to evaluate differences between groups. Results: There were no differences in oxidative stress biomarkers between treatment groups at baseline. Thirtynine (39) participants completed the study per protocol. Changes in creatinine standardized F2-isoP (ng/mg creatinine) (0.0 -0.1 versus 0.0 -0.1, p = 0.38) and 8-OHdG (lg/g creatinine) (-0.2 -3.3 versus 1.0 -3.2, p = 0.27) were nonsignificant between groups at week 4. Total reduced, oxidized, and ratio measures of GSH status were also unchanged. Conclusions: No significant changes were observed in biomarkers of oxidative stress, including glutathione status, in this clinical trial of oral glutathione supplementation in healthy adults.

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Determination of glutathione in biological material by flow-injection analysis using an enzymatic recycling reaction

Cited by 56 publications

References 17 publications

Influence of Pre-Storage Irradiation on the Oxidative Stress Markers, Membrane Integrity, Size and Shape of the Cold Stored Red Blood Cells

Influence of Pre-Storage Irradiation on the Oxidative Stress Markers, Membrane Integrity, Size and Shape of the Cold Stored Red Blood Cells

Inhalation of Nitric Oxide: Effect on Cerebral Hemodynamics and Activity, and Antioxidant Status in the Newborn Lamb

Effects of Oral Glutathione Supplementation on Systemic Oxidative Stress Biomarkers in Human Volunteers

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