Page 75 - 《中国药房》2021年第9期

西南地区有柄石韦UPLC指纹图谱的建立及酚酸类成分的测定 Δ

＊
#
邱韵静 ，索彩仙，潘礼业，何民友，陈向东，李国卫（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 R282 文献标志码 A 文章编号 1001-0408（2021）09-1093-07
DOI 10.6039/j.issn.1001-0408.2021.09.12

摘 要 目的：建立西南地区有柄石韦的超高效液相色谱（UPLC）指纹图谱，并测定其中4种酚酸类成分（新绿原酸、咖啡酸、绿原
酸、隐绿原酸）的含量。方法：采用Waters Cortecs T3 C18 （100 mm×2.1 mm，1.6 μm）色谱柱，以甲醇-0.1％磷酸溶液为流动相进行梯
度洗脱，流速为0.35 mL/min，检测波长为326 nm，柱温为30 ℃，进样量为1 μL。以UPLC法结合《中药色谱指纹图谱相似度评价
系统》（2012版）建立有柄石韦的UPLC指纹图谱，采用SPSS 20.0软件对数据进行聚类分析和主成分分析，并利用外标法测定来自
西南地区20批有柄石韦中4种酚酸类成分的含量。结果：有柄石韦的UPLC指纹图谱中确定了9个共有峰，指认出峰1、峰3、峰4、
峰 5、峰 9 分别为新绿原酸、咖啡酸、绿原酸、隐绿原酸和异绿原酸 C；不同批次有柄石韦中各色谱峰相对保留时间的 RSD 为 0～
0.68％，相对峰面积的RSD为0～62.35％；20批药材指纹图谱与对照图谱的相似度均不小于0.990。聚类分析结果显示，不同产地
的有柄石韦可聚为3类；主成分分析结果表明，不同产地的有柄石韦质量存在一定差异。新绿原酸、咖啡酸、绿原酸、隐绿原酸检
2
测浓度的线性范围分别为 0.61～61.41、0.18～17.60、2.00～200.11、0.62～61.51 μg/mL（R 均大于 0.999 9），精密度、重复性和稳定
性试验的RSD均小于2.00％，加样回收率为96.23％～98.17％（RSD为0.96％～2.28％，n＝6）；20批样品中，上述4种酚酸类成分
的含量分别为0.385 3～1.891 9、0.018 0～0.129 5、2.569 5～10.676 0、0.563 5～1.860 5 mg/g。结论：所建UPLC指纹图谱较全面地
反映了有柄石韦的主要化学成分。酚酸类成分可作为有柄石韦质量的主要评定指标；西南地区有柄石韦药材的质量高低排序为
重庆产药材＞四川产药材＞贵州产药材。
关键词 有柄石韦；指纹图谱；新绿原酸；咖啡酸；绿原酸；隐绿原酸；超高效液相色谱法；聚类分析；主成分分析

Establishment of UPLC Fingerprint of Pyrrosia petiolosa from Southwest China and Content
Determination of Phenolic Acids Component
QIU Yunjing，SUO Caixian，PAN Liye，HE Minyou，CHEN Xiangdong，LI Guowei（Guangdong Yifang
Pharmaceutical Co.，Ltd./Key Laboratory of Guangdong TCM Formula Granule Enterprise，Guangdong Foshan
528244，China）

ABSTRACT OBJECTIVE：To establish a UPLC fingerprint of Pyrrosia petiolosa from southwest China，and to determine the
contents of 4 kinds of phenolic acids （neochlorogenic acid， caffeic acid， chlorogenic acid and cryptochlorogenic acid）.
METHODS：The determination was performed on Waters Cortecs T3 C18 column（100 mm×2.1 mm，1.6 μm）with mobile phase
consisted of methanol-0.1％ phosphoric acid（gradient elution）at the flow rate of 0.35 mL/min. The detection wavelength was set
at 326 nm. The column temperature was 30 ℃，and injection volume was 1 μL. UPLC method was used to establish the UPLC
fingerprint of P. petiolosa in combination with the Similarity Evaluation System of TCM Chromatographic Fingerprints （2012
edition）. Cluster analysis and principle component analysis（PCA）were performed by using SPSS 20.0 software. The contents of 4
kinds of phenolic acids in 20 batches of P. petiolosa were determined by external standard method. RESULTS：There were 9
common peaks for the UPLC fingerprint of P. petiolosa. Peaks 1，3，4，5 and 9 were identified as neochlorogenic acid，caffeic
acid，chlorogenic acid，cryptochlorogenic acid and isochlorogenic acid C，respectively. RSDs of the relative retention time of each
peak in different batches of P. petiolosa were 0-0.68％，and the RSDs of the relative peak area were 0-62.35％. The similarities
between the fingerprint of 20 batches of medicinal materials and the control chromatogram were not less than 0.990. The result of
cluster analysis showed that P. petiolosa from different regions could be sorted into three species. Results of PCA showed the
differences among P. petiolosa from different regions. The linear range of neochlorogenic acid，caffeic acid，chlorogenic acid and
cryptochlorogenic acid were 0.61-61.41， 0.18-17.60， 2.00-200.11， 0.62-61.51 μ g/mL （R ＞0.999 9）. RSDs of precision，
2
reproducibility and stability tests were all lower than 2.00％ .
Δ 基金项目：广东省省级科技计划项目（No.2018B030323004）
The recoveries were 96.23％-98.17％（RSD＝0.96％-2.28％，
＊学士。研究方向：中药配方颗粒质量标准。E-mail：365485924
n＝6）. Among 20 batches of samples，the contents of above 4
@qq.com
# 通信作者：主管中药师，硕士。研究方向：中药配方颗粒质量标 kinds of phenolic acids were 0.385 3-1.891 9，0.018 0-0.129 5，
准。E-mail：452048107@qq.com 2.569 5-10.676 0，0.563.5-1.860 5 mg/g. CONCLUSIONS：

中国药房 2021年第32卷第9期 China Pharmacy 2021 Vol. 32 No. 9 ·1093 ·