Nurten TÜKÖZKAN, Hüsamettin ERDAMAR, Ilgım SEVEN

Measurement of total malondialdehyde in plasma and tissues by high-performance liquid chromatography and thiobarbituric acid assay

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.

Anahtar Kelimeler:

Dokular, Kromatografi, yüksek basınçlı sıvı, Kobaylar, Malondialdehit, Plazma, Tiyobarbitürik asit reaktif maddeleri, Modeller, hayvan

HPLC ve TBA yöntemi ile plazma ve dokularda total MDA ölçümü olgu sunumu

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-lipidrelated, 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.

Keywords:

Tissues, Chromatography, High Pressure Liquid, Guinea Pigs, Malondialdehyde, Plasma, Thiobarbituric Acid Reactive Substances, Models, Animal,

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