虎掌南星生品及不同炮制品指纹图谱和5种核苷类成分含量的比较研究
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篇名: 虎掌南星生品及不同炮制品指纹图谱和5种核苷类成分含量的比较研究
TITLE: Comparative Study on the Fingerprints and Content of 5 Kinds of Nucleosides in Raw Products and Different Processed Products of Pinellia pedatisecta
摘要: 目的:建立虎掌南星生品及不同工艺炮制品的指纹图谱,并测定其中5种核苷类成分的含量,比较生品与不同炮制品之间成分的差异。方法:分别以虎掌南星生品、河南省中药饮片炮制规范制品(简称“《规范》炮制品”)和产地加工炮制一体化新工艺炮制品(简称“一体化新工艺炮制品”)(各12份)为考察样品,采用高效液相色谱(HPLC)法进行分析[色谱柱为SymmetryShieldRP18,流动相为乙腈(A)-0.1%乙酸(B)水溶液(梯度洗脱),流速为0.8mL/min,柱温为30℃,检测波长为270nm,进样量为15µL]。应用《中药色谱指纹图谱相似度评价系统》(2004A版)软件分别建立3种虎掌南星样品的HPLC指纹图谱并进行相似度评价;通过与对照品图谱比较对色谱峰进行指认。同时,对其中5种核苷类成分(腺嘌呤、次黄嘌呤、尿苷、黄嘌呤、肌苷)进行定量分析,并采用SPSS21.0软件对36份样品进行聚类分析。结果:指纹图谱及含量测定的方法学考察结果均符合相关要求。3种样品与其各自对照指纹图谱的相似度均大于0.990。在虎掌南星生品中确定了22个共有峰,在2种炮制品中均确定了16个共有峰(2种炮制品中消失的6个峰相同);指认出了3种样品中均含有的5种核苷类成分,分别为腺嘌呤(3号峰)、次黄嘌呤(7号峰)、尿苷(8号峰)、黄嘌呤(9号峰)、肌苷(11号峰)。含量测定结果显示,2种炮制品中5种核苷类成分的总含量均降低,总含量高低排序为生品>一体化新工艺炮制品>《规范》炮制品。聚类分析结果显示,36份样品可聚为2类,其中生品单独聚为一类,2种炮制品聚为另一类。结论:本研究建立的方法稳定、可行,可用于虎掌南星生品及不同炮制品的质量评价。虎掌南星经炮制后指纹图谱发生了明显变化,5种核苷类成分的总含量均降低,但一体化新工艺炮制品中总含量略高于《规范》炮制品。
ABSTRACT: OBJECTIVE:To establish the fingerprint of raw produc ts and different processed products of Pinellia pedatisecta , to determine the contents of 5 kinds of nucleosides ,and to compare the differences of components between the raw products and processed products. METHODS :P. pedatisecta raw products ,processed products by Processing Standard of Chinese Medicine in Henan Province (called“Standard processed product ”for short )and processed products by new integrated processing technology in the production area (called“new integrated processed product ”for short )were collected as investigation objects (12 batches of each). The determination was performed on SymmetryShield RP 18 column with mobile phase consisted of acetonitrile (A)-0.1% acetic acid aqueous water solution (B)(gradient elution )at the flow rate of 0.8 mL/min,with the column temperature of 30 ℃, the detection wavelength of 270 nm,and the injection volume of 15 µL. HPLC fingerprints of 3 kinds of P. pedatisecta samples were established by using Similarity Evaluation System of TCM Chromatographic Fingerprints (2004 A version ) ,and the similarity of fingerprints was evaluated. The chromatographic peaks were identified by comparing with the reference chromatogram. Five nucleosides (adenine,hypoxanthine,uridine,xanthine,inosine)were quantitatively analyzed. SPSS 21.0 software was used for cluster analysis of 36 batches of samples. RESULTS :The results of fingerprint and content determination met the relevant requirements. The similarity of 3 kinds of sample with their control fingerprint were all greater than 0.990. There were 22 common peaks in the raw products of P. pedatisecta ,and 16 common peaks were identified in the 2 kinds of processed products (the same 6 peaks disappeared from 2 kinds of processed products ). Fivecomponents were identified in 3 kinds of samples ,such as adenine(peak 3),hypoxanthine(peak 7),uridine(peak 8), 1064056472@qq.com xanthine(peak 9)and inosine (peak 11). Results of content determination showed that total contents of 5 kinds of nucleosides in 2 kinds of proc essed products were all· decreased;the contents of them in descending order w as raw product >new i ntegrated processed products >Standard processed products. Results of cluster analysis showed that 36 batches of samples could be clustered into 2 categories,i.e. raw product was clustered into one category and 2 kinds of processed products into other one . CONCLUSIONS :Established method is stable , feasible and suitable for the quality evaluation of raw products and different processed products of P. pedatisecta . Fingerprints have changed significantly and the total content of 5 kinds of nucleosides in P. pedatisecta are all decreased after processing ,but that of new integrated processed products is slightly higher than that of Standard processed products .
期刊: 2020年第31卷第13期
作者: 曹淼淼,朱建光,张振凌,杜莉杰,杨佳宁
AUTHORS: CAO Miaomiao ,ZHU Jianguang ,ZHANG Zhenling ,DU Lijie,YANG Jianing
关键字: 虎掌南星;指纹图谱;含量测定;炮制工艺;核苷类成分;聚类分析
KEYWORDS: Pinellia pedatisecta ;Fingerprint;Content determination ;Processing technology ;Nucleosides;Cluster analysis
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