[关键词]
[摘要]
目的 建立散寒化湿颗粒(Sanhan Huashi Granules,SHG)的HPLC特征图谱,并对全部特征峰进行化学成分指认和药材来源归属,为其整体质量评价提供科学依据。方法 采用Waters Atlantis T3 C18色谱柱(250 mm×4.6 mm,5 μm),以甲醇-0.1%磷酸水溶液为流动相进行梯度洗脱;体积流量为1.0 mL/min;柱温为30 ℃;检测波长为250 nm,建立SHG的HPLC特征图谱。采用UHPLC-QTOF MS、NMR、对照品比对及定向分离技术对10个特征峰进行了准确的化学成分指认并归属处方药味来源,并对20批制剂进行一致性评价。结果 建立了SHG的HPLC特征图谱,10个特征峰得到明确化学成分指认,分别为腺嘌呤(1号峰)、尿苷(2号峰)、鸟苷(3号峰)、肌苷(4号峰)、5-羟甲基糠醛(5号峰)、木兰苷A(6号峰)、佛手酚-5-O-β-D-吡喃葡萄糖苷峰(7号峰)、紫花前胡苷(8号峰)、6'-O-(反式阿魏酰基)-紫花前胡苷(9号峰)、茴香酸对羟基苯乙酯(10号峰),并归属了特征峰处方药味来源,分别为1号峰归属于麻黄、苦杏仁、羌活、广藿香、苍术、白术、焦六神曲、焦麦芽、地龙、徐长卿、葶苈子,2号峰归属于厚朴、羌活、广藿香、佩兰、苍术、白术、地龙、徐长卿、葶苈子,3号峰归属于羌活、广藿香、地龙、徐长卿、葶苈子,4号峰归属于地龙,5号峰归属于焦三仙(焦山楂、焦麦芽、焦六神曲),6号峰归属于厚朴,7~10号峰归属于羌活。20批制剂均符合特征图谱的规定,批间一致性良好。结论 所建立的SHG的HPLC特征图谱方法简便,稳定性、重复性好,初步描绘了SHG的整体化学成分特征,为进一步提升其质量标准奠定了基础,同时也可为其他经典名方质量标准研究提供参考。
[Key word]
[Abstract]
Objective To establish the characteristic chromatogram of Sanhan Huashi Granules (散寒化湿颗粒, SHG) by HPLC, identify the components of all characteristic peaks and the origin of medicinal materials, so as to provide reference for its overall quality control. Methods The separation was developed on Waters Atlantis T3 C18 column (250 mm×4.6 mm, 5 μm) by gradient elution with acetonitrile-0.1% phosphoric acid and the analytes were detected at 250 nm. The column temperature was 30 ℃ and the flow rate was at 1.0 L/min. The 10 characteristic peaks were chemically identified by UHPLC-QTOF MS, NMR, reference substance and directional separation, which were also assigned to individual herbs. Furthermore, the consistency of 20 batches of preparations was evaluated. Results The HPLC characteristic chromatogram of SHG was established. The consistency of 20 batches was good. Ten characteristic peaks [adenine (peak 1), uridine (peak 2), guanosine (peak 3), inosine (peak 4), 5-hydroxymethyl-2-furaldehyde (peak 5), magnoloside A (peak 6), bergaptol-5-O-β-D-glucopyranoside (peak 7), nodakenin (peak 8), 6'-O-feruloylnodakenin (peak 9), 4-hydroxyphenethylanisate (peak10)] were chemically identified and assigned to individual herbs [peak 1 originated from Mahuang (Ephedrae Herba), Kuxingren (Armeniacae Semen Amarum), Qianghuo (Notopterygii Rhizoma et Radix, NRR), Guanghuoxiang (Pogostemonis Herba, PH), Cangzhu (Atractylodis Rhizoma, AR), Baizhu (Atractylodis Macrocephalae Rhizoma, AMR), Jiaoliushenqu (Charred Massa Medicata Fermentata (CMMF), Jiaomaya (Charred Hordei Fructus Germinatus, CHFG), Dilong (Pheretima), Xuchangqing (Cynanchi Paniculati Radix et Rhizoma, CPRR) and Tinglizi (Descurainiae Semen Lepidii Semen, DSLS), peak 2 originated from Houpo (Magnoliae Officinalis Cortex, MOC), NRR, PH, Peilan (Eupatorii Herba), AR, AMR, Pheretima, CPRR and DSLS, peak 3 originated from NRR, PH, Pheretima, CPRR and DSLS, peak 4 originated from Pheretima, peak 5 originated from Charred Crataegi Fructus, CHFG and CMMF, peak 6 originated from MOC, peak 7—10 originated from NRR]. Conclusion The established HPLC characteristic chromatogram method of SHG was simple, stable and repeatable, and the characteristics of the chemical composition of SHG were preliminarily described. It lays a foundation for the promotion of its quality standard, and also provides a reference for the quality standard research of other classical famous prescriptions.
[中图分类号]
[基金项目]
国家重点研发计划“公共安全风险防控与应急技术装备”重点专项—散寒化湿颗粒治疗新冠肺炎的新药创制示范研究(2022YFC0868300)