Page 62 - 《中国药房》2021年第1期

基于 UPLC 指纹图谱与色度值的桑白皮生品和炮制品的鉴别及

炮制终点研究 Δ

黄梦婷 ，潘 玲，邓李红，谢明晏，马永富，魏 梅，程学仁，徐 杰（广东一方制药有限公司/广东省中药配方颗
＊
#
粒企业重点实验室，广东 佛山 528244）

中图分类号 R283 文献标志码 A 文章编号 1001-0408（2021）01-0056-08
DOI 10.6039/j.issn.1001-0408.2021.01.11
摘 要 目的：为桑白皮生品和其炮制品（蜜桑白皮）的鉴别及蜜桑白皮炮制终点的确定提供参考。方法：采用超高效液相色谱
（UPLC）法进行测定。色谱柱为Waters BEH Shield RP C18；流动相为乙腈-0.1％磷酸溶液（梯度洗脱）；流速为0.30 mL/min；柱温为
30 ℃；程序波长为280 nm（0～4 min）、320 nm（4～35 min）。使用《中药色谱指纹图谱相似度评价系统》（2012版）分别建立13批桑
白皮和蜜桑白皮的UPLC指纹图谱以及进行相似度评价，并结合对照品图谱对色谱峰进行指认。测定13批桑白皮和蜜桑白皮粉
末的色度值（L、a、b）并计算其平均总色度值（E），结合偏最小二乘法判别分析（OPLS-DA）法和聚类分析法分析桑白皮炮制前后指
纹图谱和色度值的差异性；同时分析不同炮制时间下蜜桑白皮指纹图谱和色度值的动态变化规律，以确定其炮制终点。结果：桑
白皮炮制前后指纹图谱存在明显差异，13批桑白皮和蜜桑白皮样品的相似度均在0.9以上。桑白皮、蜜桑白皮图谱中分别共标定
了21、23个共有峰，其中峰1、峰2是桑白皮经蜜炙后新产生的化合物；同时，指认了峰2、峰7、峰14、峰19分别为5-羟甲基糠醛、桑
皮苷A、氧化白藜芦醇、桑黄酮G。OPLS-DA分析结果显示，峰1、峰2、峰18、峰20的峰面积/称样量比值以及色度值（L、a、b）是影
响蜜桑白皮炮制前后差异最主要的因素；以E范围75.84～80.88作为蜜桑白皮的炮制终点，确定炮制时间为22～34 min。结论：所
建立的UPLC指纹图谱和色度值的测定方法可用于桑白皮生品和炮制品的鉴别以及蜜桑白皮炮制终点的确定。
关键词 桑白皮；蜜桑白皮；色度值；超高效液相色谱；指纹图谱；最小偏二乘法判别分析；聚类分析

Identification and Study on Processing End-point of Raw and Processed Products of Morus alba Based on
UPLC Fingerprint and Chromaticity
HUANG Mengting，PAN Ling，DENG Lihong，XIE Mingyan，MA Yongfu，WEI Mei，CHENG Xueren，XU Jie
（Guangdong Yifang Pharmaceutical Co.，Ltd./Guangdong Provincial Key Laboratory of Traditional Chinese
Medicine Formula Granule，Guangdong Foshan 528244，China）

ABSTRACT OBJECTIVE：To provide reference for the identification and processing end-point determination of raw Morus alba
and its processed products（honey-processed M. alba）. METHODS：UPLC method was adopted. The determination was performed
on Waters BEH Shield RP C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution（gradient elution）
at the flow rate of 0.30 mL/min. The column temperature was set at 30 ℃. The program wavelengths were set at 280 nm（0-4 min）
and 320 nm（4-35 min）. Similarity Evaluation System for Chromatogram Fingerprint of TCM（2012 edition）was used to establish
UPLC fingerprint and carry out similarity evaluation of 13 batches of M. alba and honey-processed M. alba. The chromatographic
peaks were identified with reference substance fingerprint. The colorimetric value （L，a，b） of 13 batches of M. alba and
honey-processed M. alba powder were determined，and average total colorimetric value（E）was calculated. OPLS-DA and cluster
analysis were adopted to analyze the differences in fingerprints and colorimetric values of M. alba before and after processing. At
the same time，the dynamic change rule of fingerprint and colorimetric value of honey-processed M. alba at different processing
time points were analyzed to determine the processing end-point. RESULTS：There were obvious differences in fingerprints before
and after processing，and the similarity of 13 batches of M. alba and honey-processed M. alba were all higher than 0.9. Totally 21
common peaks were calibrated for M. alba，and 23 common peaks for honey-processed M. alba；peak 1 and peak 2 were newly
produced compounds of honey-processed M. alba. Peak 2，peak 7，peak 14 and peak 19 were identified as 5-hydroxymethylfurfural，
mulberry glucoside A，oxidized resveratrol，mulberry flavonoids G. Results of OPLS-DA showed that the peak area-sample
quantity ratio of peak 1，peak 2，peak 18，peak 20 and the chromaticity values（L，a，b）were the most important factors affecting
the difference of raw and processed products of M. alba. When
Δ 基金项目：广东省科技计划项目（No.2018B030323004）；广东特
the E ranged 75.84-80.88 as the processing end-point of
支计划科技创业领军人才项目（No.2017TY04R197）
honey-processed M. alba，the processing time was determined
＊中药师。研究方向：中药及中药配方颗粒质量标准。电话：
0757-85128604。E-mail：1515183125@qq.com as 22-34 min. CONCLUSIONS： The established UPLC
# 通信作者：主管中药师，硕士。研究方向：中药及中药配方颗粒 fingerprint and colorimetric value determination method can be
质量标准。电话：0757-85128604。E-mail：935394848@qq.com used to identify the raw and processed products of M. alba as

·56 · China Pharmacy 2021 Vol. 32 No. 1 中国药房 2021年第32卷第1期