Page 59 - 《中国药房》2021年5期

Establishment of UPLC Fingerprint of Poria cocos Aqueous Extract and Study on Its Spectrum-effect
Relationship with Sedative and Hypnotic Effect
WANG Tianhe ，LI Huijun ，ZHANG Dandan ，LUO Xinyao ，XIA Heyuan ，HAN Sijie ，PAN Xiang ，WAN
1
1
1
1
1
1
1
1
2
Ming ，YE Xiaochuan（1. Provincial Key Laboratory of Chinese Medicine Resources and Chinese Medicine
Chemistry，College of Pharmacy，Hubei University of TCM，Wuhan 430065，China；2. Hubei Academy of
TCM/Dept. of Pharmaceutical Administration，Hubei Hospital of TCM，Wuhan 430061，China）
ABSTRACT 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 2 μL. 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. 16
α-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（0.611 5-
0.811 8）；the correlation between 24 peaks and sleep latency was greater than 0.6（0.605 9-0.790 4），except for peaks 14，24 and
2；the correlation of 24 peaks between sleeping rate was greater than 0.6（0.606 4-0.721 6），except for peaks 23，19，17 and 5；
the correlation of 24 peaks between total efficacy was greater than 0.6（0.619 0-0.781 2），except for peaks 2，5，19. The top 10
chromatographic peaks related to the total efficacy were peak 15 （polyporenic acid C），peak 16，peak 8，peak 11，peak 12
（poricoic acid A）， peak 1， peak 7 （16 α-hydroxytrametendicacid）， peak 3， peak 9 （poricoic acid B） and peak 20
（dehydropachymic acid）. CONCLUSIONS：UPLC fingerprint of P. cocos aqueous extract was established and 11 components were
identified. Ten components such as polyporus acid C are closely related to the total efficacy of sedation and hypnosis，which
preliminarily reveal the material basis of the sedative and hypnotic effect of P. cocos.
KEYWORDS Poria cocos；UPLC；Fingerprint；Sedation；Hypnosis；Spectrum-effect relationship；Grey correlation analysis；
Mice

茯苓为多孔菌科真菌茯苓 Poria cocos（Schw.）Wolf 学分析模型，可更全面、多角度地对中药进行评价，常用
的干燥菌核，其药用历史悠久，始载于《五十二病方》，也 于中药质量评价、药效物质基础研究、质量标志物筛选、
收载于历代版本的《中国药典》中；其味甘、淡、平，在《神 中药组方配伍、中药有效部位筛选以及提取纯化工艺优
农本草经》中归为上品，属药食同源中药，久服可安魂养 化等方面 [4－5] 。基于此，本研究先将 10 批不同产地茯苓
神，为九大仙品药草之一，具有利水渗湿、健脾、宁心的 以 水 煎 煮 制 得 水 提 物 ，并 建 立 其 超 高 效 液 相 色 谱
[1]
功效 。茯苓在临床上可单用或与其他中药组成复方， （UPLC）指纹图谱；同时，以阈上剂量戊巴比妥钠协同作
常用于治疗水肿、痰饮、脾虚、便溏泄泻、心神不安、惊悸 用下的小鼠入睡潜伏期和睡眠持续时间，以及阈下剂量
失眠 。茯苓主要含有多糖类、三萜类、蛋白质类、甾体 戊巴比妥钠协同作用下的小鼠入睡率为镇静催眠指标，
[2]
类等化学成分 ，已有文献报道茯苓的多糖部位和乙酸 并结合灰色关联度法进行谱效相关性分析，以期初步揭
[1]
[3]
乙酯部位均具有良好的镇静催眠作用 ，但上述部位的 示茯苓镇静催眠的药效物质基础，为完善茯苓的质量评
成分复杂，具体有哪些成分具有镇静催眠活性尚不 价标准提供依据。
清楚。 1 材料
中药谱效关系研究是将中药化学成分谱与中药药 1.1 主要仪器
效活性有机结合，采用回归分析、相关性分析、灰色关联 本研究所用主要仪器有：Agilent1290型系列四元泵
度分析、主成分分析、人工神经网络、聚类分析等多种数 UPLC 仪（美国 Agilent 公司）、Milli-Q 型超纯水系统（美

中国药房 2021年第32卷第5期 China Pharmacy 2021 Vol. 32 No. 5 ·565 ·