王不留行炮制前后的UPLC指纹图谱比较及刺桐碱和王不留行黄酮苷的含量测定-中国药房

王不留行炮制前后的UPLC指纹图谱比较及刺桐碱和王不留行黄酮苷的含量测定

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篇名：	王不留行炮制前后的UPLC指纹图谱比较及刺桐碱和王不留行黄酮苷的含量测定
TITLE：	Study on UPLC Fingerprint of Vaccariae Semen before and after Processing and Content Determination of Erythrine and Vaccarin
摘要：	目的：比较王不留行炮制（清炒）前后的指纹图谱差异，并测定其炒制前后刺桐碱、王不留行黄酮苷的含量。方法：采用超高效液相色谱（UPLC）法，色谱柱为YMCTraitC18，流动相为乙腈-水（梯度洗脱），流速为0.35mL/min，检测波长为219nm，柱温为35℃，进样量为1μL。以王不留行黄酮苷为参照，绘制王不留行生品及其炮制品（各17批，编号分别为S1～S17、CS18～CS34）的指纹图谱；采用《中药色谱指纹图谱相似度评价系统（2012版）》进行相似度评价及共有峰指认；采用SPSS20.0软件进行聚类分析、主成分分析和因子分析。采用上述UPLC法测定王不留行生品及其炮制品中刺桐碱、王不留行黄酮苷的含量。结果：17批王不留行生品及其炮制品的UPLC指纹图谱中均有共有峰5个，相似度均大于0.99；共指认了刺桐碱和王不留行黄酮苷等2个共有峰。聚类分析结果显示，S1～S17聚为一类，CS18～CS34聚为一类；主成分分析及因子分析结果显示，第1主成分的方差贡献率为76.418％，刺桐碱、王不留行黄酮苷在第1主成分上有较高载荷（特征值分别为0.976、0.966）。刺桐碱、王不留行黄酮苷检测质量浓度的线性范围分别为6.437～321.832、7.729～386.437μg/mL（r均大于0.999）；检测限、定量限分别为0.085、0.284ng（生品）和0.739、2.465ng（炮制品）；精密度、重复性、稳定性（12h）、耐用性试验的RSD均小于3％（n＝6或n＝5）；平均加样回收率分别为96.42％（RSD＝0.85％，n＝6）、99.13％（RSD＝1.74％，n＝6）。两种成分的含量分别为0.11％～0.20％、0.42％～0.63％（生品）和0.08％～0.11％、0.34％～0.50％（炮制品）。结论：成功建立了王不留行生品及其炮制品的UPLC指纹图谱。王不留行炒制前后的化学成分虽一致性较好，但炒制后刺桐碱、王不留行黄酮苷的含量均有所降低。
ABSTRACT：	OBJECTIVE：Compare the fingerprint difference of Vaccariae Semen before and after processed （stir-fried），and to determine the contents of erythrine and vaccarin before and after stir-fried. METHODS ：UPLC method was adopted. The determination was performed on YMC Trait C 18 column with mobile phase consisted of acetonitrile-water （gradient elution ）at the flow rate of 0.35 mL/min. The detection wavelength was set at 219 nm，and the column temperature was 35 ℃. The sample size was 1 μL. Using vaccarin as reference，the fingerprints of Vaccariae Semen crude product and its processed product （each of 17 batches，S1-S17，CS18-CS34） were drawn. The similarity evaluation and common peak identification were carried out by Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）；cluster analysis ，principle component analysis （PCA）and factor analysis were performed by using SPSS 20.0 software. The contents of erythrine and vaccarin in Vaccariae Semen crude product and its processed product were determined by UPLC. RESULTS ：There were 5 common peaks in UPLC fingerprints of 17 batches of Vaccariae Semen crude product and its processed product. The similarities were all higher than 0.99. Among them ， 2 common peaks were identified ，i.e. erythrine ，vaccarin. Results of cluster analysis showed that S 1-S17 were clustered into one category and CS 18-CS34 were clustered into one category. Results of PCA and factor analysis showed that variance contribution rate of the first principle component was 76.418％；erythrine and vaccarin had higher loading on the first principal component （eigenvalues were 0.976 and 0.966，respectively）. The linear ranges of above 2 components were 6.437-321.832 μg/mL and 7.729-386.437 μg/mL，respectively（r＞0.999）. The limits of detection and quantitation were 0.085，0.284 ng （crude product） and 0.739， 2.465 ng （processed product ）， respectively. RSDs of precision ，reproducibility，stability（12 h）and durability tests were all lower than 3％（n＝6 or n＝5）. E-mail：1083656123@qq.com Average recoveries were 96.42％（RSD＝0.85％，n＝6）and 99.13％（RSD＝1.74％，n＝6）. The contents of the two components were 0.11％-0.20％，0.42％-0.63％（crude product ）and 0.08％-0.11％，0.34％-0.50％（processed product ）. CONCLUSIONS ：UPLC fingerprint of Vaccariae Semen crude product and its processed product are established successfully. Although the chemical constituents in Vaccariae Semen are consistent before and after stir-fried ，the contents of erythrine and vaccarin are all decreased after stir-fried.
期刊：	2020年第31卷第19期
作者：	曹斯琼，吴文平，罗宇琴，马瑞瑞，潘礼业，李国卫，陈向东
AUTHORS：	CAO Siqiong ，WU Wenping ，LUO Yuqin，MA Ruirui，PAN Liye，LI Guowei ，CHEN Xiangdong
关键字：	王不留行；炮制；超高效液相色谱法；指纹图谱；刺桐碱；王不留行黄酮苷；聚类分析；主成分分析
KEYWORDS：	Vaccariae Semen ；Processing；UPLC；Fingerprint；Erythrine；Vaccarin；Cluster analysis ；Principal component
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