响应曲面法优化板蓝根总生物碱含量测定及提取工艺研究

Response Surface Methodology to Optimize the Determination of Total Alkaloids in Radix Isatidis and Study on Extraction Process

目的:应用响应曲面法优化酸性染料比色法测定板蓝根总生物碱的测定方法。方法:在单因素考察实验的结果上,选取缓冲液pH值、缓冲溶液用量和酸性染料用量为3个显著因素进行Box-Benhnken中心组合设计,以板蓝根总生物碱提取率为衡量标准,通过响应曲面法对测定条件进行优化。结果:板蓝根-乙醇-乙酸乙酯(酸碱处理)条件下得出的总生物碱吸光度、含量以及固形物转移率最高,醋酸-醋酸钠缓冲液溶液pH=4.8,缓冲液用量5 mL,溴甲酚绿用量5 mL,三氯甲烷10 mL,萃取1次为板蓝根总生物碱含量测定的最优条件。结论:综合比较三种不同工艺对板蓝根总生物碱的提取效果,板蓝根-乙醇-乙酸乙酯(酸碱处理)效果最好。缓冲溶液的pH值和酸性染料的用量对测定板蓝根总生物碱的含量有显著的影响。

Objective: The acid dye colorimetric method for the determination of total alkaloids in Radix isatidis was optimized by response surface methodology.Methods: Based on the results of the single-factor investigation experiment, three significant factors were selected as the pH value of buffer solution, the amount of buffer solution and the amount of acid dye to carry out the Box-Benhnken center combination design. Assay conditions were optimized.Based on the extraction rate of Radix isatidis total alkaloids, the measurement conditions were optimized by response surface methodology. Results: The absorbance, content and solid transfer rate of total alkaloids obtained under the conditions of Radix isatidis-ethanol-ethyl acetate(acid-base treatment) were the highest, acetic acid-sodium acetate buffer solution pH=4.8, buffer dosage 5 mL, 5 mL of bromocresol green dosage, 10 mL of chloroform, and one extraction were the optimal conditions for the determination of total alkaloids in Radix isatidis. Conclusion: Comprehensively comparing the extraction effects of three different processes on the total alkaloids of Radix isatidis,Radix isatidis-ethanol-ethyl acetate(acid-base treatment) has the best effect. The pH value of the buffer solution and the amount of acid dye had a significant effect on the determination of total alkaloids in Radix isatidis.