茯苓水提物UPLC指纹图谱的建立及其镇静催眠作用的谱效关系研究  

Establishment of UPLC Fingerprint of Poria cocos Aqueous Extract and Study on Its Spectrum-effect Relationship with Sedative and Hypnotic Effect

作　　者：王天合 李慧君 张丹丹 罗心遥 夏和元 韩思婕 潘翔 万明 叶晓川 WANG Tianhe;LI Huijun;ZHANG Dandan;LUO Xinyao;XIA Heyuan;HAN Sijie;PAN Xiang;WAN Ming;YE Xiaochuan(Provincial Key Laboratory of Chinese Medicine Resources and Chinese Medicine Chemistry,College of Pharmacy,Hubei University of TCM,Wuhan 430065,China;Hubei Academy of TCM/Dept.of Pharmaceutical Administration,Hubei Hospital of TCM,Wuhan 430061,China)

机构地区：[1]湖北中医药大学药学院中药资源与中药化学省级重点实验室,武汉430065 [2]湖北省中医药研究院/湖北省中医院药事部,武汉430061

出　　处：《中国药房》2021年第5期564-570,共7页China Pharmacy

基　　金：国家重点研发计划中医药现代化研究重点专项(No.2017YFC1703004);湖北省科技计划项目(No.2019ACA121)。

摘　　要：目的:建立茯苓水提物的超高效液相色谱(UPLC)指纹图谱,并考察其与镇静催眠作用的谱效关系。方法:将10批不同产地茯苓经水提后得水提物。采用UPLC法测定该水提物,色谱柱为Waters HSS-C18,流动相为0.1%磷酸溶液-乙腈-甲醇(梯度洗脱),流速为0.4~0.2 mL/min,检测波长为210、242 nm,柱温为40℃,进样量为2μL。采用《中药指纹图谱相似度评价系统》(2012A版)建立10批茯苓水提物的指纹图谱并进行共有峰指认。以小鼠的入睡率、入睡潜伏期和睡眠持续时间为单一药效指标,考察戊巴比妥钠协同作用下10批不同产地茯苓水提物的镇静催眠作用;将单一药效指标和总药效(由单一指标经层次分析法计算而得)进行数据转化后,采用灰色关联度分析法分析茯苓水提物指纹图谱中各共有峰与单一药效及总药效的关联度。结果:10批茯苓水提物共有24个共有峰,指认出11个成分,分别为16-羟基松苓新酸(6号峰)、16α-hydroxytrametendic acid(7号峰)、茯苓酸B(9号峰)、去氢土莫酸(10号峰)、茯苓酸A(12号峰)、猪苓酸C(15号峰)、3-O-乙酰基-16α-羟基松苓新酸(17号峰)、去氢茯苓酸(20号峰)、茯苓酸(21号峰)、松苓新酸(22号峰)、脱氢齿孔酸(24号峰)。经灰色关联度分析发现,24个峰与睡眠持续时间的关联度均大于0.6(0.6115~0.8118),与入睡潜伏期的关联度除14、24、2号峰外,其余均大于0.6(0.6059~0.7904);与入睡率的关联度除23、19、17、5号峰外,其余均大于0.6(0.6064~0.7216);与总药效的关联度除2、5、19号峰外,其余均大于0.6(0.6190~0.7812);与总药效关联度排名前10位的色谱峰分别为15号峰(猪苓酸C)、16号峰、8号峰、11号峰、12号峰(茯苓酸A)、1号峰、7号峰(16α-hydroxytrametendic acid)、3号峰、9号峰(茯苓酸B)、20号峰(去氢茯苓酸)。结论:本研究建立了茯苓水提物的UPLC指纹图谱,指认了11个共有峰成分;猪苓酸C等10种成分与镇静催眠总药效关OBJECTIVE:To establish the UPLC fingerprint of Poria cocos aqueous extract,and to investigate its relationship with sedative and hypnotic effect.METHODS:Ten batches of P.cocos from different areas were extracted with water to obtain the aqueous extract.UPLC method was adopted.The determination was performed on Waters HSS-C18 column with mobile phase consisted of 0.1%phosphoric acid solution-acetonitrile-methanol(gradient elution)at the flow rate of 0.4-0.2 mL/min.The detection wavelengths were set at 210 and 242 nm.The column temperature was 40℃,and sample size was 2μL.The fingerprints of 10 batches of P.cocos aqueous extracts were established by using the Similarity Evaluation System of TCM Fingerprint(2012A version),and the common peaks were identified.The sedative and hypnotic effects of 10 batches of P.cocos aqueous extracts from different areas under the synergistic action of pentobarbital sodium were investigated by taking the sleeping rate,sleep latency and sleep duration of mice as the single efficacy index.After data transformation of single efficacy index and total efficacy(single indexes calculated by analytic hierarchy process),grey correlation analysis was used to analyze the correlation between the common peaks in fingerprint of P.cocos aqueous extract and the single efficacy index and total efficacy.RESULTS:There were 24 common peaks in 10 batches of aqueous extract of P.cocos,and 11 components were identified,i.e.16α-hydroxydehydrotrametenolic acid(peak 6),16α-hydroxytrametendic acid(peak 7),poricoic acid B(peak 9),dehydrotumulosic acid(peak 10),poricoic acid A(peak 12),polyporenic acid C(peak 15),3-O-acetyl-16α-hydroxydehydrotrametenolic acid(peak 17),dehydropachymic acid(peak 20),pachymic acid(peak 21),dehydrotrametenolic acid(peak 22),dehydroeburicoic acid(peak 24).Grey correlation analysis showed,the correlation between 24 peaks and sleep duration was greater than 0.6(0.6115-0.8118);the correlation between 24 peaks and sleep latency was greater than 0.6(0.6059-0.7904),except for peaks 14

关 键 词：茯苓 超高效液相色谱 指纹图谱 镇静 催眠 谱效关系 灰色关联度分析 小鼠 

分 类 号：R284.1[医药卫生—中药学] R285.5[医药卫生—中医学]