Page 56 - 2021年15期

基于UPLC指纹图谱和一测多评法的虎杖药材质量评价 Δ

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梁 慧 1，2＊ ，潘晓君 ，杨文惠 ，吕渭升 ，罗宇琴 ，吴佳璇 ，魏 梅 ，程学仁 ，兰小勇 ，李振雨 （1.广东
一方制药有限公司技术中心，广东 佛山 528244；2.广东省中药配方颗粒企业重点实验室，广东 佛山
528244；3.中国中药控股有限公司采购中心，广东 佛山 528303）
中图分类号 R284 文献标志码 A 文章编号 1001-0408（2021）15-1842-07
DOI 10.6039/j.issn.1001-0408.2021.15.09
摘 要 目的：建立虎杖药材的超高效液相色谱（UPLC）指纹图谱，并测定药材中4种活性成分的含量，为虎杖药材的质量评价提
供依据。方法：以色谱柱为 Waters BEH C18 （100 mm×2.1 mm，1.7 μm）、流动相为乙腈-0.2％甲酸溶液（梯度洗脱）、流速为 0.4
mL/min、柱温为40 ℃、检测波长为290 nm、进样量为1 μL的色谱条件建立虎杖药材的UPLC指纹图谱，并采用相似度计算、聚类
分析和正交偏最小二乘法判别分析方法对指纹图谱进行评价。以虎杖苷为内参物，测定白藜芦醇、大黄素-8-O-β-D-葡萄糖苷、大
黄素的相对校正因子，建立一测多评法；再利用相对校正因子计算15批虎杖药材中上述4种成分的含量，并将一测多评法与外标
法的结果进行比较。结果：建立了15批虎杖药材的UPLC指纹图谱，确定了12个共有峰，指认出了其中5个成分，分别为虎杖苷、
白藜芦醇、大黄素-8-O-β-D-葡萄糖苷、大黄素和大黄素甲醚。15批虎杖药材指纹图谱的相似度为0.865～0.976；聚类分析将15批
药材分为4类，并呈现一定的产地规律性；正交偏最小二乘法判别分析共确定了7个差异性标志物，差异显著性排序为峰7＞大黄
素-8-O-β-D-葡萄糖苷＞白藜芦醇＞峰 8＞虎杖苷＞峰 1＞峰 10。采用一测多评法测得的 15 批虎杖药材中白藜芦醇、大黄
素-8-O-β-D-葡萄糖苷、大黄素的含量与外标法测得的含量的相对平均偏差均小于5.0％，说明两种方法无明显差异。结论：UPLC
指纹图谱结合一测多评法可便捷、可靠地对虎杖药材进行质量评价；安徽和重庆所产虎杖药材的质量较优。
关键词 虎杖；超高效液相色谱法；指纹图谱；一测多评法；含量测定；质量评价；产地

Quality Evaluation of Polygonum cuspidatum by UPLC Fingerprint and QAMS
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LIANG Hui ，PAN Xiaojun ，YANG Wenhui ，LYU Weisheng ，LUO Yuqin ，WU Jiaxuan ，WEI Mei ，
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CHENG Xueren ，LAN Xiaoyong ，LI Zhenyu （1. Technology Center of Guangdong Yifang Pharmaceutical
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Co.，Ltd.，Guangdong Foshan 528244，China；2. Guangdong Provincial Key Laboratory of TCM Formula
Granule Enterprise，Guangdong Foshan 528244，China；3. Purchasing Center，China Traditional Chinese
Medicine Holdings Co.，Ltd.，Guangdong Foshan 528303，China）
ABSTRACT OBJECTIVE：To establish the UPLC fingerprint of Polygonum cuspidatum，and to determine the contents of four
active ingredients and to provide reference for the quality evaluation of P. cuspidatum. METHODS：The determination was
performed on Waters BEH C18 column（100 mm×2.1 mm，1.7 μm）with mobile phase consisted of acetonitrile-0.2％ formic acid
（gradient elution）at flow rate of 0.4 mL/min. The column temperature was 40 ℃，and detection wavelength was 290 nm. The
sample size was 1 μL. The fingerprints were evaluated by similarity calculation，cluster analysis and orthogonal partial least square
discriminant analysis （OPLS-DA）. Using polydatin as internal standard，relative calibration factors of resveratrol，emodin-8-O-
β-D-glucoside and emodin were determined to develop a method of QAMS. The contents of 4 above components in 15 batches of P.
cuspidatum were calculated by relative calibration factors. The results of QAMS were compared with those of external standard.
RESULTS：UPLC fingerprints of 15 batches of P. cuspidatum were established，and 12 common peaks were confirmed. Five
components were identified，i.e. polydatin，resveratrol，emodin-8-O-β-D-glucoside，emodin，emodin methyl ether. The fingerprint
similarity of 15 batches of P. cuspidatum was in the range of 0.865-0.976. According to cluster analysis，15 batches of P.
cuspidatum were classified into 4 categories，showing certain regularity of origin. Seven markers were identified by OPLS-DA
method. The order of difference significance was peak 7＞emodin-8-O-β-D-glucoside＞resveratrol＞peak 8＞polydatin＞peak 1＞
peak 10. The relative deviation among the contents of resveratrol，emodin-8-O-β-D-glucoside and emodin in 15 batches of P.
cuspidatum determined by QAMS and external standard method was less than 5.0％ ，indicating that there was no significant
difference between the two methods. CONCLUSIONS：UPLC fingerprint combined with QAMS method is convenient and reliable
for the quality evaluation of P. cuspidatum；the quality of P.
Δ 基金项目：广东省省级科技计划项目（No.2018B030323004）；
“广东特支计划”科技创业领军人才项目（No.2017TY04R197） cuspidatum produced in Chongqing and Anhui province is
＊学士。研究方向：中药饮片及中药配方颗粒。电话：0757- better.
85128602。E-mail：773274122@qq.com KEYWORDS Polygonum cuspidatum；UPLC；Fingerprint；
# 通信作者：主管中药师，硕士。研究方向：中药饮片及中药配方 QAMS；Content determination；Quality evaluation；Producing
颗粒。电话：0757-85128602。E-mail：1083656123@qq.com area

·1842 · China Pharmacy 2021 Vol. 32 No. 15 中国药房 2021年第32卷第15期