目的： 建立一测多评法（QAMS）和外标法（ESM）测定灯盏细辛注射液中 5 个二咖啡酸酯类成分总量和野黄芩苷含量。方法： 采用 Waters Sunfire C18（250 mm×4.6 mm,5 μm）色谱柱,以甲醇-乙腈（30∶70）（A）-0.1% 三氟乙酸水溶液（B）（18∶82）为流动相,二咖啡酸酯检测波长为 327 nm;以 3,5-O-二咖啡酰奎宁酸（1）为内参物,分别建立 3,4-O-二咖啡酰奎宁酸（2）、飞蓬酯乙（3）、4,5-O-二咖啡酰奎宁酸（4）、灯盏细辛酯（5）的相对校正因子（RCF）,计算化合物 2~5 与内参物色谱峰的相对保留时间,测定30批样品的含量,与ESM结果进行比较,验证QAMS 的准确性;野黄芩苷检测波长为335 nm,与3,5-O-二咖啡酰奎宁酸制成混合对照品溶液,采用ESM计算含量,与2020年版《中华人民共和国药典》一部方法（ESM）的结果进行比较,验证本方法的准确性。另取样品,分别在不同 仪器、色谱柱、流动相比例、流速、温度、波长条件下进行全面耐用性考察,采用所建立的方法计算RCF和相对保留时间,测定二咖啡酸酯类成分总量和野黄芩苷的含量,比较结果的RSD,验证所建立方法的实用性。结果： 上述化合物2~5的RCF分别为1.140 7、1.169 1、1.017 2、 1.351 5,相对保留时间分别为 0.92、1.54、1.72、2.11,30批样品QAMS计算的二咖啡酸酯类成分总量与ESM测定结果之间的相对偏差（RAD）均小于5%;30 批样品ESM测得野黄芩苷含量与原方法计算结果之间的 RAD 均小于 5%。结论： 建立的方法可应用于灯盏细辛注射液中5个二咖啡酸酯类成分和野黄芩苷的含量测定。
Objective： To establish determination methods for 5 dicaffeoylquinic acids and scutellarin in Dengzhan Xixin injection by multi-components with single marker method（QAMS）and external standard method（ESM）. Methods： A high-performance liquid chromatography（HPLC）method was developed on a Waters Sunfire C18（250 mm×4.6 mm,5 μm）column with[A（methanol：acetonitrile 30∶70）-B（0.1% trifluoroacetic acid solution）]（18∶82）as mobile phase. The detection wavelength for dicaffeoylquinic acids was set at 327 nm. 3, 5-O-Dicaffeoylquinic acid（1）was chosen as the internal reference substance,the relative correction factors of 4 components（2-5）were determined by multi-point correction method,including 3,4-O-dicaffeoylquinic acid（2）,erigeron B（3）,4,5-O-dicaffeoylquinic acid（4）and erigeroster（5）and the relative retention time of the chromatographic peaks of the components to 3,5-O-dicaffeoylquinic acid were calculated,respectively. The content of compounds 1-5 in 30 batches of Dengzhan Xixin injection samples were determined by relative correction factors,and the relative deviation（RAD）was used as a parameter to compare the results with that of external standard method（ESM）to verify the accuracy of QAMS. In the same system,the detection wavelength of scutellarin was set at 335 nm,and it is formulated with 3,5-O-dicaffeoylquinic acid to make a mixed reference substance,and the content of scutellarin is calculated by external standard method and the relative deviation（RAD） was used as the parameter to compare the method of Chinese Pharmacopoeia（ESM）with that of established method. Durability tests were performed with different chromatographic columns,instruments,levels of different mobile phase proportions,flow rates,temperatures and wavelengths. The relative correction factor,the relative retention time and the results were calculated respectively. The practicability of the established method was verified by comparing the relative standard deviation of the results. Results： The relative correction factors（RCF）of 4 components（2-5）were 1.140 7,1.169 1,1.017 2 and 1.351 5,respectively,and the relative retention values were 0.92,1.54,1.72 and 2.11,respectively. The relative deviation（RAD）of the contents of 5 dicaffeoylquinic acid by QAMs and determined by ESM in 30 batches of Dengzhan Xixin injection were less than 5%. The relative deviation （RAD）of the contents of scutellarin by the established method and by the Chinese Pharmacopoeia method in 30 batches of Dengzhan Xixin injection were less than 5%. Conclusion： The established content determination method can be applied to the determination of 5 dicaffeoylquinic acid and scutellarin in Dengzhan Xixin injection.
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