金樱子总黄酮的含量测定及酶提工艺优化

为了测定金樱子中的总黄酮含量，并对其纤维素酶辅助提取工艺进行优化，本研究以芦丁为对照品，采用双波长分光光度法，确定测定波长500 nm，参比波长580 nm，对金樱子中的总黄酮含量进行测定；以总黄酮得率为评价指标，考察提取温度、乙醇浓度、纤维素酶用量、液料比等4个因素对总黄酮得率的影响，正交优化纤维素酶辅助提取金樱子总黄酮的工艺。结果表明，在19.90~398.00 μg范围内，芦丁的质量与吸光度差值ΔA呈良好的线性关系（r=0.9981），标准曲线方程为y=0.0008x-0.0047，该方法下测得其得率为87.94 mg/g，平均加样回收率98.23%，相对标准偏差为8.77%；正交试验优化的纤维素酶法辅助提取金樱子总黄酮的最佳工艺条件为：提取温度80℃、乙醇浓度40%、纤维素酶用量0.5%、液料比40：1、提取时间90 min、pH为6.0、提取次数2次，且各因素对金樱子总黄酮提取得率的影响顺序为：提取温度 > 纤维素酶用量 > 液料比 > 乙醇浓度；在该提取条件下，金樱子总黄酮得率为（93.35±3.15）mg/g。因此，纤维素酶法辅助提取金樱子总黄酮可行，得率较高，条件温和。

Content Determination of Total Flavonoids from Rosa laevigata Michx. Fruits and Optimization of Cellulase-assisted Extraction

To determine the content of total flavonoids from Rosa laevigata Michx. fruits, and optimize the process of cellulase-assisted extraction, the content of total flavonoids from Rosa laevigata Michx. fruits were determined with a double-wavelength spectrophotometry with detection wavelength were 500 nm and 580 nm as the reference, using rutin as a standard material, and the process of cellulase-assisted extraction designed by orthogonal test was optimized with the yield of flavonoids as the evaluation index, and the effects of extraction temperature, concentration of ethanol, cellulase dosage, ratio of liquid to material and other 4 factors on the content of total flavonoids were investigated. It showed a good linearity (r=0.9981) between absorbance difference(ΔA) and the rutin mass ranging of 19.90 to 398.00 μg, the standard curve equation was y=0.0008x-0.0047, and the yield of total flavonoids was 87.94 mg/g, the average recovery and RSD were 98.23% and 8.77%, respectively. And the optimumcellulase-assisted extraction conditions of total flavonoids from Rosa laevigata Michx. fruits by orthogonal array design were extraction temperature of 80℃, concentration of ethanol of 40%, cellulase dosage of 0.5%, ratio of liquid to material of 40:1, extraction time of 90 min, pH of 6.0, and extraction numbers of 2. Various factors affecting extracting yield were in a order of extraction temperature > cellulase dosage > ratio of liquid to material > concentration of ethanol. In this condition of extraction, yield of total flavonoids from Rosa laevigata Michx. fruits was (93.35±3.15) mg/g. Therefore, it was suggested that extraction of total flavonoids from Rosa laevigata Michx. fruits by cellulase-assisted extraction was feasible, and the yield of total flavonoids was higher and its conditions was mild.