摘要
《中国药典》收载的发酵虫草菌粉产品的质量标准中,规定以鸟苷、腺苷、尿苷的含量作为评价相关产品质量的标准。但除此之外,还有许多其他的核苷类成分对发酵虫草菌粉质量控制的影响尚未被探讨。为探究发酵虫草菌粉及产品质控指标选择的合理性,采用超高效液相色谱-紫外检测法对19批发酵虫草菌粉及产品中9种核苷成分(尿嘧啶、胞苷、鸟嘌呤、尿苷、腺嘌呤、肌苷、鸟苷、胸苷、腺苷)进行了定量分析,建立了发酵虫草菌粉样品的指纹图谱,并结合统计学提供了一种分析指标性成分的方法。通过优化样品的提取方法,选择超声提取法制备19批发酵虫草菌粉及产品的供试液;采用Agilent Eclipse Plus C18色谱柱(150 mm×4.6 mm,3.5μm)进行色谱分析,以甲醇-水为流动相梯度洗脱,对方法的校正曲线、准确度、精密度、重复性和回收率进行了验证。结合对照品指认了指纹图谱中的9个核苷峰,并采用外标一点法测得了各核苷成分的含量。使用化学模式识别对指纹图谱中的共有峰进行分析,聚类分析和主成分分析得到了同样的分类结果:19批样品共分为5类,其中同一发酵虫草菌粉因工艺差异可分为2类,而心肝宝胶囊、百令胶囊、宁心宝胶囊则各单独分为1类。同时,使用主成分分析获得了各样品中的指标性成分,分别为尿苷、鸟苷、腺苷、腺嘌呤、尿嘧啶,并使用聚类分析再次进行确证,验证了指标性成分的合理性。
Currently,guanosine,adenosine,and uridine contents are specified as the quality criteria for related products in the quality standards for fermented Cordyceps powder preparations included in the 2020 edition of Chinese Pharmacopoeia.However,there are many other nucleosides in fermented Cordyceps powder,whose effect on the quality control has not yet been discussed.In this study,an ultra-performance liquid chromatography-ultraviolet detection(UPLC-UV)method was used for the quantitative analysis of 9 nucleosides(uracil,cytidine,guanidine,uridine,adenine,inosine,guanosine,thymidine,and adenosine)in 19 batches of fermented Cordyceps powder samples and products,and the corresponding fingerprints were established.In addition,a method for analyzing the index components was proposed based on statistics.By optimizing the sample extraction method,ultrasound-assisted extraction was selected to process 19 batches of samples.Chromatographic analysis was performed on an Agilent Eclipse Plus C18 column(150 mm×4.6 mm,3.5μm)using methanol and water as the mobile phases under gradient elution.The method was validated based on the calibration curves,accuracy,precision,repeatability,and recovery.The fingerprints of the 19 batches of samples were established,and 16 common peaks were obtained.Among them,nine nucleoside peaks were identified by standards,and their concentrations were determined by the external standard one-point method.Similarity evaluation of the fingerprints was conducted;the similarities of the 19 batches of samples were greater than 0.9.Then,chemical pattern recognition was performed.The same classification results were obtained by hierarchical clustering analysis(HCA)and principal component analysis(PCA).Thus,the samples could be segregated into five classes,and the fermented Cordyceps powders were classified as two types with different fermentation processes.Xinganbao capsules,Bailing capsules and Ningxinbao capsules were each separately classified into one class.This indicated that the chemical recognition pa
作者
曹稳
洪亮
杨明
李绍平
赵静
CAO Wen;HONG Liang;YANG Ming;LI Shaoping;ZHAO Jing(State Key Laboratory for Quality Research of Chinese Medicine, University of Macao, Macao 999078, China;Jiangxi Guoyao Pharmaceutical Co., Ltd., Nanchang 330096, China)
出处
《色谱》
CAS
CSCD
北大核心
2021年第9期1006-1011,共6页
Chinese Journal of Chromatography
基金
国家重点研发计划(2019YFC1711300)
澳门特别行政区科学及科技发展基金(034/2017/A1,0017/2019/AKP)
广东省重点区域研发计划(2020B1111110006)
澳门大学(MYRG2018-00083-ICMS,MYRG2019-00128-ICMS,CPG2021-00009-ICMS).
关键词
指纹图谱
质控指标
主成分分析
聚类分析
发酵虫草菌粉
fingerprints
quality control index
principal component analysis(PCA)
hierarchical clustering analysis
fermented Cordyceps powder
