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The fluorescence and UV absorption spectra were measured with a 850 CRT fluorescence spectrophotometer (Hitachi, Japan) and a Cary 50 probe UV¨Cvis spectrophotometer (Varian, America), respectively. The fluorescence spectrophotometer was also used for the measurement of chemiluminescence spectrum. A Strata TM-X C18 column was used for the extraction of resveratrol. Effects of the reaction parameter on the CL intensity were investigated. The optimum conditions were 1 mol/L H2SO4, 0.025 mmol/L KMnO4, 0.56 mol/L HCHO, 3.5 mL/min of flow rate for P2 and 100 ¦ÌL of sample loop, respectively. Under the selected optimum conditions, the calibration curves for the determination of resveratrol in red wine were established. Under the optimum conditions, the relative chemiluminescence intensity was linear with the concentration of resveratrol from 1.32 ¡Á 10−8 to 1.32 ¡Á 10−5 mol/L. The detection limit was 3.30 ¡Á 10−9 mol/L (3¦Ò). The relative standard deviation for 11 parallel measurement of 1.32 ¡Á 10−5 mol/L resveratrol was 3.8%. The regression equation was ▵I = 10.52 + 0.19 ¡Á 108 C (C is the concentration of resveratrol, mol/L) with a correlation coefficient of 0.9994 (n = 11). The effects of foreign species on the determination of resveratrol were assessed. The tolerable limit of a foreign species was taken if it caused a relative error of less than 5%. The obtained results showed that under the optimized conditions, 1000-fold methanol; 500-fold maltose and fructose; 200-fold citrate, ethanol, lactose, glucose, K+, Na+, Zn2+, NH4+, Ni2+, Cr3+, Ca2+, Al3+, Cl− and 5-fold riboflavin did not affect the determination process. The proposed method was applied to determine resveratrol in red wine and the average concent of resveratrol in the analyzed red wine was 0.409 mg/L. The recovery experiment was also performed and the results obtained are given in Table 1. |
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