摘要
目的:建立小续命汤基准样化学成分的定性分析方法,并建立15批小续命汤基准样品指纹图谱,评价其批间质量一致性。方法:根据《古代经典名方关键信息表(25首方剂)》中小续命汤的关键信息,制备小续命汤基准样品。采用超高效液相色谱-四极杆-飞行时间串联质谱法(UPLC-Q-TOF-MS/MS)对小续命汤基准样品进行检测,通过自建数据库、查阅相关文献及与对照品鉴定所得化学成分,并通过与单味药样品及缺单味药阴性样品比对,将各成分进行归属。采用高效液相色谱法(HPLC)建立小续命汤基准样品的指纹图谱,并通过单味药样品及缺单味药阴性样品对共有色谱峰进行药味归属与指认。结果:结合质谱碎片信息、相关文献和数据库检索,从小续命汤基准样品中共鉴定和推测出64个化合物,包括31个黄酮类化合物,8个萜类化合物,12个三萜皂苷类化合物,2个苯酞类化合物,3个苯丙素类化合物,2个姜辣素类化合物,5个生物碱类化合物,1个氰苷类化合物。其中21个源自黄芩,10个源自炒甘草,9个源自人参,8个源自白芍,4个源自防风,3个源自防己,3个源自川芎,2个源自黑顺片,2个源自生姜,1个源自麻黄,1个源自燀苦杏仁。建立的小续命汤基准样品HPLC指纹图谱有23个共有峰,其中峰1、峰2来源于白芍;峰3、峰7来源于防风;峰4、峰8、峰9来源于防己;峰10、峰17、峰18、峰20、峰21来源于炒甘草;峰11~峰16、峰19、峰22来源于黄芩;峰5来源于川芎,峰23来源于生姜;峰6是黄芩和炒甘草共有成分,指认了芍药内酯苷(峰1)、芍药苷(峰2)、升麻素苷(峰3)、5-O-甲基维斯阿米醇苷(峰7)、黄芩苷(峰11)、亥茅酚苷(峰13)、千层纸素A-7-O-β-D-葡萄糖醛酸苷(峰15)、汉黄芩苷(峰16)、甘草酸(峰21)、6-姜辣素(峰23)10个化合物。15批基准样品相似度均>0.999,表明基准样品具有良好的一致性。结论:该研究通过对小续命汤基准样品中的化学成分的鉴定,明确了其物质组成以黄酮类和三萜皂苷类化合物为主,建立的指纹图谱可基本体现小续命汤基准样品的整体化学特征,可为其复方制剂的质量研究提供依据。
Objective:To establish a qualitative analysis method for the chemical constituents of the reference sample of Xiao Xumingtang,and to establish the fingerprint of 15 batches of Xiao Xumingtang,so as to evaluate the quality consistency among batches.Method:According to the key information of Xiao Xumingtang in the Key Information Table of Ancient Famous Classical Formulas(25 Formulas),the reference sample of this formula was prepared,and it was detected by ultra performance liquid chromatographyquadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS).The chemical components were identified by self-constructed database,consulting relevant literature,and comparing with the reference substances,and the components were assigned by comparing with single drug samples and negative samples lacking single drug.The fingerprint of the reference sample of Xiao Xumingtang was established using high performance liquid chromatography(HPLC),and the common peaks were assigned and identified through single drug samples and negative samples lacking single drug.Result:Based on the information of MS fragments,relevant literature,and database retrieval,a total of 64 compounds were identified and inferred from the reference sample of Xiao Xumingtang,including 31 flavonoids,8 terpenoids,12 triterpenoid saponins,2 phthalides,3 phenylpropanoids,2 gingerols,5 alkaloids,and 1 cyanoside.Among them,21 were derived from Scutellariae Radix,10 from stir-fried Glycyrrhizae Radix et Rhizoma,9 from Ginseng Radix et Rhizoma,8 from Paeoniae Radix Alba,4 from Saposhnikoviae Radix,3 from Stephaniae Tetrandrae Radix,3 from Chuanxiong Rhizoma,2 from Aconiti Lateralis Radix Praeparata,2 from Zingiberis Rhizoma Recens,1 from Ephedrae Herba,and 1 from Armeniacae Semen Amarum.The established HPLC fingerprint of the reference sample of Xiao Xumingtang had 23 common peaks,among which,peaks 1 and 2 were derived from Paeoniae Radix Alba,peaks 3 and 7 from Saposhnikoviae Radix,peaks 4,8 and 9 from Stephaniae Tetrandrae Radix,peaks 10,17,18,20 and 21 from stir-fried Glycyrrhizae Radix et Rhizoma,peaks 11-16,19 and 22 from Scutellariae Radix,peak 5 from Chuanxiong Rhizoma,peak 23 from Zingiberis Rhizoma Recens,peak 6 was the common component of stir-fried Glycyrrhizae Radix et Rhizoma and Scutellariae Radix.A total of 10 compounds including albiflorin(peak 1),paeoniflorin(peak 2),cimicifugoside(peak 3),5-O-methylvisammioside(peak 7),baicalin(peak 11),sec-O-glucosylhamaudo(l peak 13),oroxylin A-7-O-β-D-glucuronid(e peak 15),wogonoside(peak 16),glycyrrhizic acid(peak 21)and 6-gingero(l peak 23)were identified.The similarities of 15 batches of reference samples were>0.999,indicating that the reference samples had good consistency.Conclusion:Through the identification of the chemical constituents in the reference sample of Xiao Xumingtang,it is clear that the composition of the samples is mainly composed of flavonoids and triterpenoid saponins.The established fingerprint can basically reflect the overall chemical characteristics of the reference sample of Xiao Xumingtang,which can provide a basis for the quality research of its compound preparations.
作者
田真真
张紫薇
孙建慧
翟张喜
李艳英
TIAN Zhenzhen;ZHANG Ziwei;SUN Jianhui;ZHAI Zhangxi;LI Yanying(Tianjin University of Traditional Chinese Medicine,Tianjing 301617,China;Beijing Increasepharm Co.Ltd.,Beijing 102200,China;Beijing Increasepharm Pharmaceutical Research Co.Ltd.,Beijing 102200,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第17期164-171,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家重点研发计划“政府间国际科技创新合作”重点专项(2021YEE0202100)。
