HPLC法同时测定防风中6个主要成分的含量  

Simultaneous determination of six major constituents in the roots of Saposhnikovia divaricata by HPLC

作　　者：赵博[1,2] 杨鑫宝[3,4] 杨秀伟[1] 张连学[2] 刘建勋[3,4] 

机构地区：[1]北京大学天然药物及仿生药物国家重点实验室药学院天然药物学系,北京100191 [2]吉林农业大学中药材学院,长春130118 [3]北京中医药大学,北京100029 [4]中国中医科学院西苑医院,北京100091

出　　处：《药物分析杂志》2013年第3期382-387,共6页Chinese Journal of Pharmaceutical Analysis

基　　金："十二五"国家科技支撑专项(2011BAI07B08)中药材质量评价体系建立及其质量标准示范研究

摘　　要：目的:建立HPLC-DAD法同时测定中药防风中升麻苷、升麻素、5-O-甲基维斯阿米醇苷、印枳树皮苷、防风灵和亥茅酚苷6个主要有效成分的含量。方法:采用正交设计优化提取方法,以提取时间、方法、次数、溶剂比为主要影响因素,以6个分析物的总量为评价指标;分析物的色谱分离用Dikma DiamonsilTMC18色谱柱(250 mm×4.6 mm,5μm)、Dikma Easy Guard C18保护柱(20 mm×4.6 mm,5μm),以甲醇-水为流动相,梯度洗脱,流速为1.0 mL·min-1,检测波长为254 nm,柱温为25℃。结果:确定最佳提取方法为甲醇30倍量回流提取2次,每次2 h。6个分析物的线性范围分别为:升麻苷50～1000 mg·L-1(r=0.9999),升麻素10～200 mg·L-1(r=0.9999),5-O-甲基维斯阿米醇苷20～500 mg·L-1(r=0.9999),印枳树皮苷5～100mg·L-1(r=0.9995),防风灵5～100 mg·L-1(r=0.9991),亥茅酚苷10～200 mg·L-1(r=0.9995);平均加样回收率(n=3)均在95.6%～106.8%,RSD均在1.3%～4.8%。结论:该方法准确,灵敏度高,重复性好,成本低,可用于防风中上述6个主要有效成分的含量测定。Objective： To develop an HPLC -DAD method for simultaneous determination of six main bioactive constituents （prim - 0 - glucosylcimifugin, cimifugin, 4＇ - O -β - D - glucosyl - 5 - O - methylvisamminol, marmesinin, sapodivarin, and sec - 0 - glucosylhamaudol ） in the roots of Saposhnikovia divaricata （ Turcz. ） Schis- chk. （Saposhnikoviae Radix）. Methods：The orthogonal array design was used to optimize the extracting process. The main influential factors of extraction efficiency were extracting time, method, times and solvent ratio. The condi- tions of the extraction were evaluated on the total content of the six analytes. A Dikma DiamonsilTM C18 column （250 mm ×4. 6 mm,5 μm） with a Dikma Easy Guard C18 column（20 mm ×4. 6 mm,5 μm） was used for chromato- graphic separation of the six analytes. The mobile phase, consisting of methanol - water, was programmed for a gradi- ent elution. The flow rate was 1.0 mL ·min- 1, the detection wavelength was at 254 nm, and the column temperature was 25 ℃. Results： The analysis indicated that the optimum conditions of the six analytes were to reflux with 30 fold solvent methanol twice and for 2 hours at each time. The linearity was achieved in the range of 50 - 1000 mg · L-1 （ r = 0. 9999 ） for prim - 0 - glucosylcimifugin, 10 - 200 mg · L-1 （ r = 0. 9999 ） for cimifugin ,20 - 500 mg · L-1 （ r = 0. 9999） for 4＇ - 0 -/3 - D - glucosyl - 5 - 0 - methylvisamminol, 5 - 100 mg~ L- 1 （ r = 0. 9995 ） formarmesinin,5 - 100 mg · L-1 （r =0. 9991 ）for sapodivarin,and 10 -200 mg · L-1 （r =0. 9995） for sec - 0 - glu- cosylhamaudol. The average recoveries（ n = 3 ）were 95.6% -106. 8% with RSD of 1.3% -4. 8%. Conclusion： The method developed is accurate, highly sensitive, well reproducible and low cost, which can be used for the deter- mination of the six above- mentioned bioactive constituents in Saposhnikoviae Radix.

关 键 词：防风 香豆素 色原酮 升麻苷 升麻素 5-O-甲基维斯阿米醇苷 印枳树皮苷 防风灵 亥茅酚苷 高效液相色谱 定量分析 

分 类 号：R917[医药卫生—药物分析学]