目的 建立UPLC波长切换法同时测定双黄连注射剂［双黄连注射液、注射用双黄连（冻干）、双黄连粉针剂］中5种特征成分，并对87批样品（注射液63批，冻干剂15批，粉针剂9批）进行测定，对批次间、企业间及剂型间样品结果差异性进行分析。方法 样品经提取后，采用UPLC波长切换法，同时测定了绿原酸、咖啡酸、连翘酯苷A、黄芩苷、连翘苷5种特征成分，色谱柱 ：Waters HSS T3 C₁₈（100 mm×2.1 mm，1.7 μm），以乙腈-0.1%磷酸溶液为流动相进行梯度洗脱，体积流量0.3 mL·min^-1，绿原酸、咖啡酸及连翘酯苷 A检测波长为 324 nm，黄芩苷检测波长为 276 nm，连翘苷检测波长为228 nm；进行专属性、线性关系、精密度、重复性、准确度、稳定性方法学考察；取样品3批，分别按建立的方法、现行质量标准测定方法检测，对比2种方法的检测结果；采用建立的方法对87批样品进行测定。绘制各测定指标结果的频率分布直方图或箱式图，进行注射液不同企业间、批次间各成分含量差异性分析；绘制5个成分及绿原酸和咖啡酸总量的平均含量分布情况雷达图，进行不同剂型之间结果差异性分析；采用 minitab软件对 87批样品测定结果进行主成分分析。结果 建立 UPLC波长切换法经方法学考察符合要求；与现行质量标准测定方法比对，建立的方法检测结果无差异。注射液 2个生产企业样本数相当，绿原酸和咖啡酸总量的均值及离散程度无差异，但不同企业2种成分比例存在差别；黄芩苷、连翘苷及连翘酯苷A含量均值一致，离散程度一致，不同生产企业、不同批次间无显著性差异。雷达图结果显示，冻干剂与粉针剂工艺除干燥方式外完全一致，其绿原酸、咖啡酸、两者总量及黄芩苷含量比例落点几乎重合；连翘苷与连翘酯苷A在粉针剂中含量是冻干剂的近2倍，且批次间差异性大；纵向比较3个剂型，除连翘苷含量相近外，其余成分含量差别均较大。主成分分析结果显示，3个不同剂型被明显区分，现行质量标准控制项目越多的剂型，样品越集中。结论 建立的方法准确可靠，在多样本测定的基础上为标准统一及药品质量标准制定的合理性提供参考。

Objective To establish a UPLC method for simultaneous determination of five components in Shuanghuanglian Injection (injection, lyophilized powder and powder-injection) by switching wavelength, and 87 batches of samples were determined. The differences of results among batches, enterprises and dosage forms were analyzed. Methods The sample were ultrasound-treated with appropriate solvents, UPLC was used to simultaneously determine five components of chlorogenic acid, caffeic acid, forsythiaside A, baicalin and forsythin. Waters HSS T3 C₁₈ (100 mm × 2.1 mm, 1.7 μm) column was used for gradient elution with acetonitrile-0.1% phosphoric acid as mobile phase at a flow rate of 0.3 mL·min^-1. The detection wavelength of chlorogenic acid, caffeic acid and forsythiaside A was 324 nm, baicalin was 276 nm, forsythin was 228 nm. Conduct methodological studies on specificity, linear relationship, precision, repeatability, accuracy, and stability was carry on. Three batches of samples were tested according to the established method and the current quality standard measurement method, and the test results of the two methods were compared. 87 batches of samples were determined. The established method was used to determine 87 batches of samples. Draw a frequency distribution histogram or box chart of the results of each measurement index, and analyze the content differences of various components among different enterprises and batches of injection. The average content distribution of five components and the total amount of chlorogenic acid and caffeic acid was plotted using a radar chart, and the difference in results between different dosage forms was analyzed. The results of 87 batches of samples were analyzed by principal component analysis using minitab software. Results The establishment of UPLC wavelength switching method meets the requirements through methodological investigation. Compared with the current quality standard measurement method, the established method has no difference in detection results. The sample numbers of the two injection production enterprises were similar, and there was no difference in the mean value and dispersion degree of the total amount of chlorogenic acid and caffeic acid, but there were differences in the proportion of components between different enterprises. The mean values of baicalin, forsythrin, and forsythrin A content were consistent, and the degree of dispersion was consistent. There was no significant difference between different manufacturers and different batches. The radar chart results show that the freeze-dried agent and powder injection process are completely consistent except for the drying method, with the total amount of chlorogenic acid, caffeic acid, and baicalin content ratio falling points almost coincident. The content of forsythrin and forsythrin A2 in powder injection was nearly twice as high as that in freeze-dried formulation, and there was significant difference between batches. Longitudinal comparison of the three dosage forms showed that except for the similar content of phillyrin, the content of other components varied significantly. The results of principal component analysis showed that three different dosage forms were clearly distinguished, and the more dosage forms controlled by current quality standards, the more concentrated the samples. Conclusion The method established in this study is accurate and reliable, and provides a basis for the unification of standards and the rationality of drug quality standards on the basis of multi-sample determination.

R284.1