Determination of Lipid Peroxidation in Desiccated Red Blood Cells

Abstract

Lipid peroxidation of biological membranes is one of the most studied indicators of oxidative stress. Very littleis known about the extent of oxidative damage that occurs during the desiccation of mammalian cells. The objective of this study was to modify and validate a method for the determination of lipid peroxidation in desiccated red blood cells (RBCs). The measurement of malondialdehydes (MDAs), the decomposition products of oxidized polyunsaturated fatty acids, is commonly used as a method for the quantification of lipid peroxidation. The method is based on the reaction of MDAs with thiobarbituric acid, which forms a thiobarbituric acid reactive substances (TBARS) chromophore. Validation of the method to dried RBC specimens included the optimization of an MDA standard curve, spiking studies, measurement of TBARS in lyophilized RBCs, and reexamination of the reagents and processes that are associated with this assay. The recovery of MDA standards, which were lyophilized with RBCs was reasonable (72.96% ؎ 8.85%), as drying and rehydration are associated with sample loss. Significant TBARS concentrations were obtained during lyophilization, dry storage for up to 72 h, and rehydration of RBCs specimens. The efficacy of the antioxidant tert-butyl hydroperoxide in reducing sample autoxidation during the TBARS assay is questionable due to its poor solubility in aqueous solution. Investigation of the interference caused by the 453-nm peak in the TBARS assay revealed that the origin of the peak is, at least in part, related to hydroperoxides. The data shows that the TBARS assay can be applied to desiccated RBC specimens. It also provides first evidence regarding the relationship between oxidative damage and desiccation. The modification of the MDA assay to dried cells will contribute to understanding the mechanism of damage that occurs during dehydration, dry storage and rehydration of dry biomaterials. 165

Red blood cell in vitro quality and function is maintained after S‐303 pathogen inactivation treatment

“…Lipid peroxidation is an indicator of oxidative stress and during RBC storage leads to membrane damage and lysis. 44,45 It has previously been suggested that S-303 treatment of RBCs does not increase oxidative stress. 24 Our results confirm this, as no differences were observed in the lipid peroxidation of RBCs treated with S-303 compared with RT and control units.…”

“…Lipid peroxidation is an indicator of oxidative stress and during RBC storage leads to membrane damage and lysis . It has previously been suggested that S‐303 treatment of RBCs does not increase oxidative stress .…”

Oxidative stress and triglycerides as predictors of subclinical atherosclerosis in prediabetes

“…MDA analysis was based on the protocol of thiobarbituric acid reactive substances. 20 Plasma from EDTA-containing tubes was kept at −80°C for serum 8-OHdG and GSH analysis.…”

Rapidly freeze‐dried human red blood cells for pre‐transfusion alloantibody testing reagents

“…Confocal microscopy technique is accepted as an indication of recovery rate and membrane integrity 17 . Lipid peroxidation is related to the deterioration of biomembranes altering the integrity, permeability and membrane function 15,39,40 . Images showed that FDRBCs stained with DILC preserved their biconcave/spherical morphology and their lipidic integrity (Figure 4b).…”