A sensitive and reliable method for the detection of lipid peroxidation in biological tissues

被引:27
作者
Anoopkumar-Dukie, S [1 ]
Walker, RB [1 ]
Daya, S [1 ]
机构
[1] Rhodes Univ, Fac Pharm, ZA-6140 Grahamstown, South Africa
关键词
D O I
10.1211/0022357011775299
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
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 C-18 (Waters Spherisorb, 5 mum, 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 (muM) 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.
引用
收藏
页码:263 / 266
页数:4
相关论文
共 13 条
[1]  
DRAPER HH, 1990, METHOD ENZYMOL, V186, P421
[2]  
Fukunaga K, 1998, BIOMED CHROMATOGR, V12, P300, DOI 10.1002/(SICI)1099-0801(199809/10)12:5<300::AID-BMC751>3.0.CO
[3]  
2-#
[4]   THE MEASUREMENT AND MECHANISM OF LIPID-PEROXIDATION IN BIOLOGICAL-SYSTEMS [J].
GUTTERIDGE, JMC ;
HALLIWELL, B .
TRENDS IN BIOCHEMICAL SCIENCES, 1990, 15 (04) :129-135
[5]   Automated high-performance liquid chromatographic separation with spectrofluorometric detection of a malondialdehyde-thiobarbituric acid adduct in plasma [J].
Londero, D ;
LoGreco, P .
JOURNAL OF CHROMATOGRAPHY A, 1996, 729 (1-2) :207-210
[6]  
LOWRY OH, 1951, J BIOL CHEM, V193, P265
[7]   Effect of alpha-tocopherol succinate on free radical and lipid peroxidation levels in BL6 melanoma cells [J].
Ottino, P ;
Duncan, JR .
FREE RADICAL BIOLOGY AND MEDICINE, 1997, 22 (07) :1145-1151
[8]   Melatonin reduces quinolinic acid-induced lipid peroxidation in rat brain homogenate [J].
Southgate, G ;
Daya, S .
METABOLIC BRAIN DISEASE, 1999, 14 (03) :165-171
[9]   Melatonin plays a protective role in quinolinic acid-induced neurotoxicity in the rat hippocampus [J].
Southgate, GS ;
Daya, S ;
Potgieter, B .
JOURNAL OF CHEMICAL NEUROANATOMY, 1998, 14 (3-4) :151-156
[10]   Development and therapeutic potential of kynurenic acid and kynurenine derivatives for neuroprotection [J].
Stone, TW .
TRENDS IN PHARMACOLOGICAL SCIENCES, 2000, 21 (04) :149-154