【作者】 李斌；

【导师】 杜方麓；

【作者基本信息】 湖南中医药大学 ， 中药学， 2007， 硕士

【摘要】 目的:研究溶剂法分离纯化红车轴草中总黄酮的工艺,将红车轴草提取物总黄酮含量由17.29%提高到50%以上。方法:①采用高效液相色谱法测量红车轴草粗提物中芒柄花素和鹰嘴豆芽素的含量,以ODS C₁₈分析柱分离测定,以甲醇-水（60∶40）为流动相,以260nm为检测波长,以外标法测定。②70%乙醇对粗提物进行提取,以除去脂溶性的杂质和色素。用乙酸乙酯对醇提干膏进行提取。然后用碱水溶液对乙酸乙酯提取液进行萃取,以分离黄酮类成分和其他成分,采用正交试验对醇提条件,乙酸乙酯提取条件及碱水萃取条件进行优化。结果:①高效液相色谱法测得红车轴草粗提物总黄酮含量为17.29%,芒柄花素和鹰嘴豆芽素在0.13μg-1.2μg范围内呈良好的线性关系,二者峰面积（Y）对进样量（X）的直线回归方程分别为:鹰嘴豆芽素Y=36.93X-0.101（r=0.9998）;芒柄花素Y=34.40X-0.105（r=0.9999）。②乙醇提取最优条件为:70%的乙醇在80℃温度下提取3次,每次溶剂用量（mL）分别为粗提物重量（g）的40倍、20倍、20倍,每次提取1.5h;乙酸乙酯提取的最佳工艺为:乙酸乙酯用量（mL）为醇提浸膏重量（g）的30倍、20倍、10倍,在80℃温度下提取3次,每次1.5h;碱水萃取的最佳条件为:乙酸乙酯液浓缩至2/3体积,用5%Na₂CO₃溶液萃取3次,每次用量相当于乙酸乙酯提取液原体积的1.6倍,碱水液盐酸调pH值5～6。结论:溶剂法分离纯化红车轴草粗提物总黄酮,工艺简单,符合工厂生产实际,可以生产出总黄酮含量大于50%的产品,总黄酮的转移率为65.67%。更多还原

【Abstract】 Objective: To establish isolation and purification technical process for total flavonoids of Trifolium pretense L. to heighten the content of total flavonoids from 17.29% to 50%.Method:①HPLC was used to determine the content of formonone and biochanin in crude extract of Trifolium pretense L. The chromatographic column was ODS C₁₈ column, the mobile phase consisted of methanol and water （ 60∶40 ）, the eluate was detected at 260nm, formonone and biochanin was determinated in external standard method.②70% ethanol were used to extract the crude extract, and then use acetoacetate to extract the extractant of ethanol, then base solution was used to extract the acetoacetate , separate the base solution , add hydrochloric acid to adjust the pH 5～6 to precipitate the flavonoids. Each process was optimized by orthogonal experiment design. Results:①The total flavonoids content was 17.29%, formonone and biochanin was good linear in the range of 0.13μg -1.2μg, the regression equation of each one was: formonone Y=36.93X- 0.101 （r=0.9998） biochanin Y=34.40X - 0.105 （ r=0.9999 ） .②The optimum ethanol extraction process was that adding 40,20,20 times of 70% ethanol for extracting three times （1.5 hour each time） at 80℃; The optimum acetoacetate extraction process was that 30,20,10 times of acetoacetate, extracting temperature was 80℃and extracted three times ,1.5 hours at a time; The optimum alkali solution extraction process was that acetoacetate solution was concentrated to 2/3 volume and adding 5% Na₂CO₃ solution extracted for 3 times ,ratio of acetoacetate （mL） to 5% Na₂CO₃ solution （mL） was 1-1.6 one time.Conclusion: The technical process of isolation and purification total flavonoids of Trifolium pretense L. was suitable to modern industrialization and it can produce the product with total flavonoids higher than 50%.更多还原