Uv visible 1800

One hundred microliters of mashed rat’s liver or kidney samples were put into a microtiter plate and added with 100 μL of 0.2% NBT solution. They were incubated at room temperature for 30 min. 50 μL of mixture was obtained and added with 1 mL n,n-dimethylformamide. Afterward, the mixture was centrifuged at 10.000 rpm for 2 min. The absorbance was measured using a spectrophotometer (UV-visible 1800 Shimadzu) at a wavelength of 620 nm. The n,n-dimethylformamide was used as a blank solution. Meanwhile, standard solutions were made from various concentrations of NBT mixed with 1000 mL of n,n-dimethylformamide (Figure-1).

Creatinine level detection was carried out through a colorimetric method using a spectrophotometer (UV-visible 1800 Shimadzu) with a wavelength of 546 nm. The blank solution was prepared based on Integrated Research and Testing Laboratory protocol as follows: 50 μL of distilled water was mixed with 1000 μL of NaOH, incubated for 5 min and mixed with 250 μL of picric acid. The standard solution was made as follows: 50 μL of standard creatinine was added to 1000 μL NaOH, incubated for 5 min and mixed with 250 μL of picric acid, and standard absorbance set as 1. Furthermore, the creatinine level in the rat was detected through blood serum. The blood sample was collected through the orbital sinus and centrifuged at 10.000 rpm for 10 min to obtain the serum. Afterward, 50 μL of blood serum was added with 1000 μL of NaOH, incubated for 5 min and mixed with 250 μL of picric acid. These samples were then measured by the spectrophotometer at a wavelength of 546 nm.