响应面法优化当归中阿魏酸和藁本内酯提取工艺研究

Response surface methodology optimization for extraction process of ferulic acid and ligustilide from Angelica sinensis

关键词:  提取工艺, 当归, Box-Behnken响应面法, 氢核磁共振

Key words:  extraction process, Angelica sinensis, Box-Behnken response surface method, ¹H-qNMR

摘要:

为优化中药材当归中主要活性成分阿魏酸和藁本内酯同时提取的最佳工艺条件,该文以超声波辅助响应面优化当归中阿魏酸和藁本内酯提取工艺,采用氢核磁共振定量方法同时测定当归中阿魏酸和藁本内酯的含量。优化工艺以阿魏酸和藁本内酯含量的综合评分为考察指标,选取乙醇浓度、提取时间、液料比三个主要因素,采用单因素筛选结合Box-Behnken中心组合设计试验优化提取工艺参数。结果表明:优化的最佳提取工艺条件为乙醇浓度80.87%,液料比13.04 mL·g^-1,提取时间30.14 min,验证试验结果与预测值一致。氢核磁共振定量方法以氘代二甲基亚砜为测试溶剂,吡嗪为内标物,其中吡嗪、阿魏酸和藁本内酯的定量共振峰分别为δ 8.66 ppm、δ 6.37～6.35 ppm和δ 5.55～5.53 ppm。该方法的精密度、稳定性、重复性和加标回收率良好,定量限和检测限较低,能够满足实际分析测试需要。综上所述,利用响应面法对提取工艺条进行优化,实验结果准确可靠,重现性良好,适用于当归中阿魏酸和藁本内酯的同时提取; 氢核磁共振定量方法操作简单、分析速度快、专属性强,可用于当归中阿魏酸和藁本内酯的同时测定。

Abstract:

In order to optimize the optimum technological conditions for the simultaneous extraction of ferulic acid and ligustilide, the extraction process of ferulic acid and ligustilide from A. sinensis were optimized by ultrasonic assisted response surface. Taking the comprehensive score of ferulic acid and ligustilide content as the evaluation index and the ethanol concentration, extraction time and liquid-to-solid ratio were selected as the three main factors, and the contents of ferulic acid and ligustilide in A. sinensis were determined by ¹H-qNMR. Single factor screening combined with Box-Behnken central combination design test was used to optimize the parameters of extraction process. The single-factor screening results showed that a high comprehensive score was obtained under the three conditions of 80% ethanol concentration, liquid-to-solid ratio was 12 mL·g^-1, or 30 min extraction duration. Based on this, the Behnken center combination design test was designed and implemented. The results showed that the influence degree of each experimental factor on the comprehensive score was the ethanol concentration, the liquid-to-solid ratio was the second, and the extraction time was the least. The best optimum extraction conditions were as follows: ethanol concentration was 80.87%, liquid-to-solid ratio was 13.04 mL·g^-1, extraction duration was 30.14 min. The verification test results were consistent with the predicted values. The quantitative resonance peaks of pyrazine, ferulic acid and ligustilide were δ 8.66 ppm, δ 6.37-6.35 ppm and δ 5.55-5.53 ppm, respectively, using DMSO-d₆ as solvent and pyrazine as internal standard substance by ¹H-qNMR. The precision, stability, repeatability and recovery of the method were good, and the limits of quantification and detection were low, which can meet the needs of practical analysis and testing. To sum up, the optimization method of response surface method was used to optimize the extraction process, the experimental results were accurate and reliable, good reproducibility, and suitable for the simultaneous extraction of ferulic acid and ligustilide in A. sinensis. The ¹H-qNMR method had the advantages of simple operation, fast analysis speed and strong specificity, and could be used for the simultaneous determination of ferulic acid and ligustilide in A. sinensis.