摘要
目的:建立经典名方阳和汤基准样品的高效液相色谱法(HPLC)指纹图谱,结合化学计量学方法对其质量进行评价,为阳和汤基准样品的质量控制提供参考。方法:采用HPLC建立阳和汤基准样品指纹图谱,使用ZORBAX SB-C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相选择乙腈(A)-0.05%磷酸水溶液(含0.05%三乙胺溶液)(B)梯度洗脱(0~5 min,2%~3%A;5~15 min,3%~5%A;15~65 min,5%~30%A;65~90 min,30%~70%A),流速1.0 mL·min^(-1),柱温35℃,检测波长210、260 nm。采用“中药色谱指纹图谱相似度评价系统”(2012版)并结合聚类分析、主成分分析与偏最小二乘法-判别分析对指纹图谱结果进行分析,评价不同批次阳和汤基准样品的质量差异,并寻找造成质量差异的主要化学成分。结果:建立了阳和汤基准样品的HPLC指纹图谱,确定了13个共有峰,并对各共有峰进行归属,其中2、8号峰来自熟地黄,10、11号峰来自肉桂,1、3~6号峰来自炒白芥子,13号峰来自麻黄,7、9、12号峰来自甘草。通过对照品指认了8个成分,分别为2号峰5-羟甲基糠醛、4号峰芥子碱硫氰酸盐、7号峰甘草苷、8号峰毛蕊花糖苷、10号峰肉桂酸、11号峰桂皮醛、12号峰甘草酸、13号峰盐酸麻黄碱。15批阳和汤基准样品的HPLC指纹图谱与对照指纹图谱的相似度均>0.80;3种化学计量学方法均可将样品分为两类;筛选出了8种主要差异性成分,其中已指认出的有5-羟甲基糠醛、芥子碱硫氰酸盐、毛蕊花糖苷、盐酸麻黄碱。结论:建立的HPLC指纹图谱分析方法灵敏度高、稳定性强,基本体现了阳和汤基准样品的整体化学成分特征,可为该经典名方复方制剂的质量标准建立提供依据。
Objective:To establish a high performance liquid chromatography(HPLC)fingerprint of Yanghetang benchmark sample,and evaluate its quality with chemometric methods,so as to provide a reference for the quality control of this benchmark sample.Method:HPLC was used to establish the fingerprint of Yanghetang benchmark sample with ZORBAX SB-C_(18) column(4.6 mm×250 mm,5μm),the mobile phase was consisted of acetonitrile(A)-0.05%phosphoric acid aqueous solution(containing 0.05%triethylamine solution)(B)for gradient elution(0-5 min,2%-3%A;5-15 min,3%-5%A;15-65 min,5%-30%A;65-90 min,30%-70%A),the flow rate was 1.0 mL·min-1,the column temperature was 35℃,and the detection wavelength was 210,260 nm.Traditional Chinese Medicine(TCM)Chromatographic Fingerprint Similarity Evaluation System(2012 edition)combined with cluster analysis,principal component analysis(PCA)and partial least squares-discriminant analysis(PLS-DA)were used to evaluate the quality differences between different batches of Yanghetang benchmark samples,and to find the main chemical components responsible for the quality differences.Result:HPLC fingerprint of Yanghetang benchmark sample was established,13 common peaks were identified and attributed to each common peak,including peaks 2 and 8 from Rehmanniae Radix Praeparata,peaks 10 and 11 from Cinnamomi Cortex,peaks 1,3-6 from fried Sinapis Semen,peak 13 from Ephedrae Herba,and peaks 7,9,12 from Glycyrrhizae Radix et Rhizoma.Eight of them were identified by comparing with control substance,which were 5-hydroxymethylfurfural(peak 2),sinapine thiocyanate(peak 4),glycyrrhizin(peak 7),verbascoside(peak 8),cinnamic acid(peak 10),cinnamaldehyde(peak 11),glycyrrhizic acid(peak 12)and ephedrine hydrochloride(peak 13).The similarities of the HPLC fingerprints of 15 batches of Yanghetang benchmark samples with the control fingerprint were all greater than 0.80.The three chemometric methods could classify the samples into two categories.Eight differential components were screened out,among which 5-hydroxymethylfurfu
作者
张泽康
王昌海
赵玥瑛
张晴
彭警
杜守颖
白洁
陆洋
ZHANG Zekang;WANG Changhai;ZHAO Yueying;ZHANG Qing;PENG Jing;DU Shouying;BAI Jie;LU Yang(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第10期13-20,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82173989)。
