A sensitive and reliable method for the detection of lipid peroxidation in biological tissues.
A simple, accurate and cost effective method has been designed for the determination of lipid peroxidation in biological tissue samples. The method was a modification and improvement on existing methods available for lipid peroxidation determination. Solid-phase extraction was used to separate the thiobarbituric acid-malondialdehyde complex from thiobarbituric acid-reactive substances and HPLC was performed using a C18 (Waters Spherisorb, 5 microm, 250 x 4.6 mm i.d.) column to achieve isolation of the complex. The procedure was validated with respect to linearity of calibration (0.998), precision, sensitivity and limits of quantitation (1 nmol mL(-1)) and detection (0.5 nmol mL(-1)). Resorcinol was used as an external standard. The method was tested by inducing free radical generation with a known free radical generator, quinolinic acid, in rat brain homogenate. The results showed that the method presented allowed detection of lipid peroxidation products at concentrations in the nanomolar (nM) range compared with the micromolar (microM) range detected by other methods, thus rendering it suitable for use with biological samples. In addition, the modified method allowed for detection of the purified lipid peroxidation products, thus eliminating the possibility of simultaneous detection of impurities that absorb at the same wavelength.
Anoopkumar-Dukie
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Faculty of Pharmacy, Rhodes University, Grahamstown, South Africa.
Walker
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RB
Daya
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D000818:Animals; D001923:Brain Chemistry / Q000187:drug effects*; D007202:Indicators and Reagents; D015227:Lipid Peroxidation / Q000187:drug effects*; D008297:Male; D008315:Malondialdehyde / Q000032:analysis; D009498:Neurotoxins / Q000633:toxicity; D017378:Quinolinic Acid / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis
D007202:Indicators and Reagents; D009498:Neurotoxins; D017392:Thiobarbituric Acid Reactive Substances; D008315:Malondialdehyde; D017378:Quinolinic Acid