摘要
目的:建立HPLC-DAD法同时测定生药款冬花和蜜炙款冬花中绿原酸、反式咖啡酸、芦丁、金丝桃苷、反式阿魏酸、4,5-O-二咖啡酰基奎宁酸、2,2-二甲基-6-乙酰基苯并二氢吡喃酮、款冬酮和甲基丁酰-3,14-去氢-Z-款冬素酯9个主要成分的含量。方法:采用正交设计优化提取方法,以提取时间、方法、溶剂、溶剂用量为主要影响因素,以9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);以乙腈(A)-0.03%三氟乙酸水溶液(B)为流动相,梯度洗脱,流速为1.0 mL.min-1;检测波长为240 nm;柱温为25℃。结果:确定最佳提取方法为1.00 g生药样品用20 mL 95%乙醇超声提取1 h。绿原酸、反式咖啡酸、芦丁、金丝桃苷、反式阿魏酸、4,5-O-二咖啡酰基奎宁酸、2,2-二甲基-6-乙酰基苯并二氢吡喃酮、款冬酮和甲基丁酰-3,14-去氢-Z-款冬素酯9个分析物的线性范围分别为62.5~2000μg.mL-1(r=0.9999),2.5~80μg.mL-1(r=0.9994),15.62~500μg.mL-1(r=0.9995),25~800μg.mL-1(r=0.9995),50~1600μg.mL-1(r=0.9992),37.5~1200μg.mL-1(r=0.9994),0.195~12.5μg.mL-1(r=0.9991),15.62~500μg.mL-1(r=0.9991),125~4000μg.mL-1(r=0.9995);平均加样回收率均在95.42%~105.0%范围内,RSD均在0.45%~4.5%范围内。结论:该方法准确,灵敏度高,重复性好,成本低,可用于款冬花中9个主要有效成分的含量测定。
Objective: To develop an HPLC - DAD method for simultaneous determination of nine main bioactive constituents including chlorogenic acid, trans - caffeic acid, rutin, hyperoside, trans - ferulic acid, 4,5 - O - dicaf- feoylquinic acid ,2,2 - dimethyl - 6 - acetylchromanonc, tussilagone, and 7β - (3' - ethylcrotonoyloxy ) - 1α - ( 2' - methylbutyryloxy) - 3,14 - dehydro - Z - notonipetranone, in Farfarae Flos ( the dried flower buds of Tussila- go farfara L. ;family Compositae). Methods: Orthogonal array design was used to optimize the extracting process. The main influential factors of extraction efficiency were extracting time, method, as well as volume and ratio of sol- vent. The conditions of the extraction were evaluated by the total content of nine analytes. A Dikma DiamonsilYMc18 column(250 mm ×4.6 mm,5μm) with a Dikma Easy Guard Cls column (20 mm ×4.6 mm,5 μm) was used for chromatographic separation of the nine analytes. The mobile phase, consisting of solvent A (acetonitrile) and solvent B (0.03 % aqueous trifluoroacetic acid) , was programmed as a linear gradient. The flow rate was 1.0 mL min - 1 The absorbance was monitored at a wavelength of 240 nm. The column temperature was 25 ~C. Results: The analy- sis indicated that the optimum conditions for extraction of the nine analytes from the powdered crude drug Farfarae Flos were as follows : dried and pulverized sample ( 1.00 g) was extracted from 20 mL of 95 % aqueous ethanol (20 - fold,v/w) under ultrasonic extractor for 1 h. The linearity was achieved in the range of 62.5 - 2000μg . mL-1 (r =0. 9999) for ehlorogenie acid,2. 580 μg . mL-1 (r =0. 9994) for trans - caffeic acid,15.62 -500μg . mL - 1 ( r = 0. 9995 ) for rutin,25 - 800μg . mL - 1 ( r = 0. 9995 ) for hyperoside,50 - 1600 μg . mL - J ( r = 0. 9992 ) for trans - fenllic acid, 37.5 - 1200μg . mL - 1 ( r = 0. 9994) for 4,5 - O - dicaffeoylquinie acid, 0.195 - 12.5 μg . mL - 1 ( r = 0. 9991 ) for 2,2 - dimethyl - 6 - acetylchromanone, 15.62 - 500 μg.mL - 1 ( r = 0. 9991 ) for tussil- agone,and 125 -4000μg . mL-l(r = 0.9995) for 7β - ( 3' - ethylerotonoyloxy) - 1 α - ( 2' - methylbutyryloxy) - 3,14 - dehydro - Z - notonipetranone. The average recoveries ( n = 3 ) were 95.42% - 105.0% with RSD of O. 45 % - 4.5 %. Conclusion: The method developed is accurate, sensitive, reproducible and cheap cost, which can be used for the determination of the above - mentioned nine bioactive constituents in Farfarae Flos.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2012年第9期1517-1524,1533,共9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
《中国药典》一部标准研究项目(YD-077
YD-078)