苦参总黄酮抗菌提取物中7种黄酮成分的结构鉴定及含量测定研究  

Isolation of Antibacterial Activity Constituents of Total Flavonoids from Sophora Flavescens and Simultaneous Determination of Seven Flavonoids by HPLC

作　　者：杨志欣[1] 李霞[1] 刘明玉 张文君[3] 单柏松 宋春晓 邓伟哲[4] YANG Zhi-xin LI Xia LIU Ming-yu ZHANG Wen-jun SHAN Bai-song SONG Chun-xiao DENG Wei- zhe(College of Pharmacy, Heilongjiang University of Chinese Medicine, Harbin 150040, China The Second Hospital of Har- bin, Harbin 150056, China Pharmaceutical Research Institute, Harbin University of Commerce, Harbin 150076, China No. 211 Hospital of PLA, Harbin 150080, China)

机构地区：[1]黑龙江中医药大学药学院,哈尔滨150040 [2]哈尔滨市第二医院,哈尔滨150056 [3]哈尔滨商业大学药学院,哈尔滨150076 [4]中国人民解放军第211医院,哈尔滨150080

出　　处：《中国药学杂志》2016年第24期2169-2174,共6页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金面上资助项目(81274091);黑龙江省自然科学基金资助项目(H2016057);黑龙江省自然科学基金资助项目(D200744);国家教育部春晖计划资助项目(Z2008-1-15016);黑龙江中医药大学"优秀创新人才支持计划"资助项目(2012)

摘　　要：目的明确苦参总黄酮抗菌活性的化学成分，建立高效液相色谱法同时测定7种黄酮成分的方法。方法采用硅胶柱、ODS柱色谱等方法进行分离纯化，综合理化性质和波谱学数据进行结构鉴定。HPLC同时测定7种黄酮成分，采用DikmaC18色谱枉，甲醇-水梯度洗脱，检测波长280nm。结果从苦参总黄酮抗菌提取物中分离鉴定7个化合物，分别为苦参啶（1）、槐属二氢黄酮G（2）、苦参酮（3）、异脱水淫羊藿素（4）、异黄腐醇（5）、芒柄花素（6）和三叶豆紫檀苷（7）。上述7种成分依次在质量浓度8．70—87．00、44．25～442．50、128．10～1281．00、9．40～94．00、48．40～484．00、14．20-142．00、25．70—257．00μg·mL-1内线性关系良好，r值均大于0．9990，加样回收率均符合要求。其中，苦参酮、异黄腐醇、槐属二氢黄酮G含量相对高，分别占提取物的（16．24±0．02）％、（7．60±0．02）％、（6．59±0．03）％。结论苦参总黄酮抗菌作用的主要化学成分可能是苦参酮、异黄腐醇、槐属二氢黄酮G等。所建立的HPLC测定法简便、准确，可用于同时测定苦参总黄酮中7种成分。OBJECTIVE To study the chemical constituents of the total flavonoids from Sophoraflavescens and establish a method for simultaneous determination of seven compounds. METHODS The compounds were isolated by chromatography on silica gel and ODS column and their structures were elucidated by spectroscopic analysis. The samples were analyzed on a Dikma C18 column （4. 6 mmx 250 mm,5 μm） ; gradient elution was performed using mobile phase composed of methanol （A）and water （B） ; the detection was carried out using a photodiode array detector at 280 nm. RESULTS Seven compounds were isolated and their structures were i- dentified as kuratidine （ 1 ）, sophoraflavanone G （ 2 ）, kurarinone （ 3 ）, isoanhydroicaritin （4）, isoxanthohumol （ 5 ）, formononetin （6） , and trifolirhizin （7）. The calibration curve was linear within 8.70 - 87.00, 44.25 - 442. 50, 128.10 - 1 281.00, 9. 40 - 94. 00, 48. 40 -484. 00, 14. 20 - 142. 00, and 25.70 -257.00 μg mL-1 for kuraridine, sophoraflavanone G, kurarinotae, isoanhyd- roicaritin, isoxanthohumol, formononetin, and trifolirhizin, respectively （r 〉 0. 999 0） , and the extraction recoveries varied from 95% to 105%. CONCLUSION The main chemical components contributing to antibacterial activity of total flavonoids may be sophorafla- vanone G, kurarinone, and isoxanthohumol. The method is simple, rapid, accurate, and can be used simultaneously to determine the contents of the seven active ingredients.

关 键 词：苦参总黄酮 苦参酮 槐属二氢黄酮G 抗菌活性 高效液相色谱法 分离纯化 结构鉴定 

分 类 号：R284[医药卫生—中药学]