engleski

Concentration of malondialdehye in human urine samples from area of endemic nephropathy

Endemic neprophaty (EN) is human renal disease of unknown etiology. It affects rural population in Bulgaria, Bosnia and Herzegovina, Croatia, Romania and Serbia. Due to the appearance of EN in rural areas it is considered that some natural compounds should be involved in its etiology. The most plausible theory is that it is caused by some nephrotoxic and carcinogenic mycotoxins. One of the suspected mycotoxins is ochratoxin A that increases the production of reactive oxygen species (ROS) and lipid peroxidation in kidney of experimental animals. The aim of this study was to measure the concentration of malondialdehyde (MDA) as biomarker of lipid peroxidation in human urine samples from endemic region in Croatia and to compare the results with that in controls. In this purpose human urine samples were collected in EN region in Croatia (N=22) and in village where EN is not known (N=15). The concentration of MDA was measured using high performance liquid chromatography (HPLC) as previously described (Drury et al., 1997). Each sample was analyzed in duplicate. All analysis were performed on HPLC with UV-VIS detector Shimadzu (Kyoto, Japan). The mobile phase consisted of 50 mM KH2PO4 and methanol (60:40 v/v). Detector wavelength was set at 532 nm. MDA was separated on C-18 analytical column (125-4 mm) Merck (Darmstad, Germany). Retention time of MDA was around 2.5 minutes and the time of analysis was 10 minutes. For quantification of MDA calibration curve of aqueous standards (0.304, 0.607, 1.518, 3.04 μ M) was used. Calibration curve was linear (r2=0.9994). The concentration of MDA in urine of control group was 0.702 &plusmn ; 0.573 μ M and in urine samples from endemic area 0.557 &plusmn ; 0.295 μ M. Statistical analysis reveled no difference among this two groups indicated that large-scale sample analysis should be performed. Also, some other biomarkers of oxidative stress should be measured. References Drury, J.A., Nycyk, J.A., Cooke, R.W.I. Comparison of urinary and plasma malondialdehyde in preterm infants. Clinica Chimica Acta 263 (1997) 177 – 185

endemic nephropathy; malondialdehyde; mycotoxins; ochratoxin A; reacitve oxigen species

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