疏血通注射液UPLC指纹图谱及5种核苷类成分含量测定研究

Study on UPLC fingerprint and content determination of 5 nucleosides in Shuxuetong Injection

[目的]建立疏血通注射液超高液相色谱(UPLC)指纹图谱及尿嘧啶、次黄嘌呤、黄嘌呤、肌苷和鸟嘌呤核苷含量测定的方法。[方法]采用ACQUITY UPLC■BEH Amide色谱柱(2.1 mm×100 mm,1.7μm),以0.1%甲酸水-乙腈为流动相进行梯度洗脱,柱温25℃,流速0.3 mL/min,检测波长254 nm。建立33批疏血通注射液的UPLC指纹图谱,采用中药色谱指纹图谱相似度评价软件进行相似度分析,同时测定其中5种核苷类成分的含量。[结果]疏血通注射液指纹图谱及5种核苷类成分含量测定方法的各项方法学考察结果均良好。不同批次疏血通注射液相似度均在0.998以上。33批样品标定了10个共有峰,其中有5个已知峰,分别为尿嘧啶、次黄嘌呤、黄嘌呤、肌苷、鸟嘌呤核苷。该5种核苷类成分的平均含量分别为34.57、153.50、52.76、53.23和14.46μg/mL。[结论]建立的疏血通注射液UPLC指纹图谱及含量测定方法稳定可靠、操作简便,可以为疏血通注射液的质量评价提供参考。

[Objective]To establish a UPLC fingerprint and a determination method of uracil,hypoxanthine,xanthine,inosine and guanine nucleoside in Shuxuetong Injection.[Methods]ACQUITY UPLC■ BEH Amide column(2.1 mm×100 mm,1.7μm)was used for gradient elution with 0.1% formic acid in water and acetonitrile as mobile phase.The column temperature was 25℃,the flow rate was 0.3 mL/min,and the detection wavelength was 254 nm.UPLC fingerprints of 33 batches of Shuxuetong Injection were established.Similarity analysis was conducted by using similarity evaluation software of Chinese medicine chromatographic fingerprint,and the contents of 5 nucleosides in the samples were determined.[Results]The methodology of the fingerprint and content determination methods of 5 nucleosides met the requirements.The similarity of different batches of Shuxuetong Injection was above 0.998.There were 10 common peaks in the fingerprints of 33 batches of samples,among which five components were identified,including uracil,hypoxanthine,xanthine,inosine and guanine nucleoside.The average content of uracil,hypoxanthine,xanthine,inosine and guanine was 34.57,153.50,52.76,53.23 and 14.46μg/mL,respectively.[Conclusion]The established UPLC fingerprint and content determination method for Shuxuetong Injection is reliable and simple to operate,which can provide a certain reference for quality evaluation of Shuxuetong injection.