血必净注射液超高效液相数字化指纹图谱及4种酚酸类化合物含量测定

目的：采用超高效液相色谱法对不同批次血必净注射液进行指纹图谱的建立及特征成分定量分析，建立血必净注射液的质量控制方法。方法：色谱条件采用ACQUITY UPLC^® HSS T3色谱柱（100 mm×2.1 mm，1.8 μm），以乙腈-0.1%甲酸水为流动相梯度洗脱，流速0.2 mL·min^-1，进样量10 μL，检测波长268.3 nm，柱温40 ℃。对10个批次的血必净注射液进行测定，建立指纹图谱并对其中的4种酚酸类化合物原儿茶酸、原儿茶醛、丹酚酸B、丹酚酸A 进行含量测定。结果：10个批次血必净注射液的指纹图谱相似度均大于0.98，具有良好的相似度，共标定共有峰39个并归属了其中的4个；方法学验证结果表明线性、精密度、重复性、稳定性、加样回收率均符合相关实验要求；血必净注射液中原儿茶酸、原儿茶醛、丹酚酸B、丹酚酸A的平均含量分别为5.26，12.86，2.99，16.37 μg·mL^-1。结论：数字化特征指纹图谱研究并结合主要成分含量测定能够更好、更全面为血必净注射液质量控制提供科学方法和理论基础。

Digital fingerprint of Xuebijing injections by UPLC and content determination of four phenolic acids

OBJECTIVE To establish a fingerprint and perform quantitative analysis of feature ingredients for identification and quality control of different batches of Xuebijing injections by ultra performance liquid chromatography (UPLC).METHODS ACQUITY UPLC^® HSS T3 (100 mm×2.1 mm, 1.8 μm) was used as the separation colum. The analyses were performed under the conditions of gradient elution solvents of acetonitrile and water including 0.1% formic acid. The flow rate was 0.2 mL·min^-1, the injection volume was 10 μL, the detecting wavelength was 268.3 nm and the column temperature was 40℃. Ten batches of Xuebijing injections were determined, the fingerprint was established and the contents of the four phenolic acids were determined, including protocatechuic acid, protocatechualdehyde, salvianolic acid B, and salvianolic acid A.RESULTS According to the established fingerprints of the ten batches of Xuebijing injections, the similarity was above 0.98, 39 common peaks were found and 4 of them were identified. The results of method validation showed that the linearity, precision, repeatability, stability and the recovery rate all met the standards of the relevant experiments. The average contents of protocatechuic acid, protocatechualdehyde, salvianolic acid B and salvianolic acid A in the Xuebijing injections were 5.26, 12.86, 2.99 and 16.37 g·mL^-1.CONCLUSION The combination of digital fingerprints and contents of main compounds can provide scientific methods and theoretical basis for quality control of Xuebijing injections preferably and comprehensively.