You are here

HPLC ve TBA Yöntemi ile Plazma ve Dokularda Total MDA Ölçümü Olgu Sunumu

Measurement of Total Malondialdehyde in Plasma and Tissues by High-Performance Liquid Chromatography and Thiobarbituric Acid Assay

Journal Name:

Publication Year:

Abstract (2. Language): 
Objective: Malondialdehyde, a product of lipid peroxidation, is a good marker for development of oxidative stress. Mateials and Methods: In this study MDA was determined in the tissues and plasma by means of thiobarbituric acid test and directly by reverse phase high-pressure liquid chromatography method after derivatisation with 2,4-dinitrophenylhidrazine. Results: We have found a significant difference between two methods by Mann-Whitney U test (p<0.001). There was the formation of non-lipid-related, malondialdehyde like, TBA-reactive substance that leads to overestimation of the extent of lipid peroxidation. On the contrary, by direct HPLC method, there was a decrease of MDA from plasma and tissue serving as control. Conclusion: Our results indicated that TBA test is not a specific and reliable index of lipid peroxidation. ©2006, Fırat Üniversitesi, Tıp Fakültesi
Abstract (Original Language): 
Amaç: Bir lipit peroksidasyon ürünü olan malondialdehit ölçümünün oksidatif stresin değerlendirilmesinde önemli bir belirteç olduğu bilinmektedir. Gereç ve Yöntem: Çalışmamızda malondialdehit seviyesi karaciğer, böbrek, beyin ve plazmada hem tiyobarbitürik asit yöntemiyle hem de 2,4 dinitrofenilhidrazin ile derivatize edilerek HPLC metoduyla ölçülerek sonuçlar karşılaştırılmıştır. Bulgular: Mann Whitney-U testi ile yapılan istatistik analiz sonucu tiyobarbitirük asit yöntemi malondialdehit miktarı sonuçlarının HPLC yönteminden elde edilen sonuçlara kıyasla önemli derecede yüksek olduğu gözlenmiştir. Doku ve plazma örneklerinde tiyobarbitirük asit yöntemiyle malondialdehit miktarının fazla ölçülmesi lipit olmayan malondialdehide benzer ve tiyobarbitirük asit reaktif maddelerin oluşumu ve interferans vermesiyle açıklanabilmektedir. Buna karşılık direk ve derivatize edilme işleminden sonra uygulanan HPLC yöntemiyle elde edilen MDA sonuçları önemli derecede düşük bulunmuştur. Sonuç: Sonuçlarımız tiyobarbitirük asit testinin, HPLC yöntemine kıyasla gerçek bir lipit peroksidasyon olayının değerlendirilmesinde spesifik ve geçerli olmadığını ortaya koymuştur. ©2006, Fırat Üniversitesi, Tıp Fakültesi
88-92

REFERENCES

References: 

1. Anoopkumar-Dukie S, Walker RB, Daya S. A sensitive and reliable method for the detection of lipid peroxidation in biological tissues. J Pharm Pharmacol 2001; 53: 263-6
2. Cordis GA, Das DK, Riedel W. High-performance liquid chromatographic peak identification of 2, 4-dinitrophenylhydrazine derivatives of lipid peroxidation aldehydes by photodiode array detection. J Chromatogr A 1998; 798: 117-23
3. Templar J, Kon SP, Milligan TP, Newman DJ, Raftery MJ.
Increased plasma malondialdehyde levels in glomerular disease as determined by a fully validated HPLC method. Neprol Dial Transplant 1999; 14: 946-51
4. Slatter DA, Bolton CH, Bailey AJ. The importance of lipid-derived malondialdehyde in diabetes mellitus. Diabetologia 2000;
43: 550-7
5. Pilz J, Meineke I, Gleiter CH. Measurement of free and bound malondialdehyde in plasma by high-performance liquid chromatography as the 2,4-dinitrophenylhydrazine derivative. J
Chromatogr B Biomed Sci Appl 2000; 742: 315-25
6. Dib M, Garrel C, Favier A, Robin V, Desnuelle C. Can
malondialdehyde be used as a biological marker of progression in neurodegenerative disease? J Neurol 2002; 249: 367-74
7. Ceconi C, Cargnoni A, Pasini E, Condorelli E, Curello S, Ferrari R. Evaluation of phospholipid peroxidation as malondialdehyde during myocardial ischemia and reperfusion injury. Am J Physiol 1991; 260: H1057-61
8. Verbunt RJ, Egas JM, Van der Laarse A. Risk of overestimation of free malondialdehyde in perfused rat hearts due to homogenization artifacts. Cardiovasc Res 1996; 31: 603-6
9. Albers DS, Augood SJ, Martin DM, Standaert DG, Vonsattel JP,
Beal MF. Evidence for oxidative stress in the subthalamic nucleus in progressive supranuclear pals. J Neurochem 1999; 73:
881-4
10. Stevens RG, Morris JE, Cordis GA, Anderson LE, Rosenberg DW, Sasser LB. Oxidative damage in colon and mammary tissue of the HFE-knockout mouse. Free Radic Biol Med 2003; 34:
1212-6
11. Cini M and Moretti A. Studies on lipid peroxidation and protein oxidation in the aging brain. Neurobiol Aging 1995; 16: 53-7
12. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;
193: 265-75
13. Yoshioka T, Kawada K, Shimada T, Mori M. Lipid peroxidation in maternal and cord blood and protective mechanism against activated-oxygen toxicity in the blood. Am J Obstet Gynecol
1979; 135: 372-6
14. Uchiyama M, Mihars M. Determination of malondialdehyde precursor in tissues by thiobarbituric acid. Ann Biochem 1978;
86: 271-78
15. Yeo HC, Helbock HJ, Chyu DW, Ames BN. Assay of
malondialdehyde in biological fluids by gas chromatography-mass spectrometry. Anal Biochem 1994; 220: 391-6
16. Lazzarino G, Amorini AM, Fazzina G, Vagnozzi R, Signoretti S, Donzelli S, Di Stasio E, Giardina B, Tavazzi B. Single-sample preparation for simultaneous cellular redox and energy state
determination. Anal Biochem 2003; 322: 51-9
17. Mc Anulty JF, Waller K. The effect of quinacrine on oxidative stress in kidney tissue stored at low temperature after warm ischemic injury. Cryobiology 1999; 39: 197-204

Thank you for copying data from http://www.arastirmax.com