疏血通注射液中游离氨基酸的UPLC指纹图谱研究

Study on UPLC fingerprint of free amino acid in Shuxuetong Injection

目的建立疏血通注射液中游离氨基酸的UPLC指纹图谱分析方法。方法采用异硫氰酸苯酯(PITC)衍生化方法,使用Waters Acquity超高效液相色谱仪,以CORTECS C18柱(150mm×2.1mm,1.6μm),乙腈-0.1%甲酸水进行梯度洗脱,体积流量为0.1mL/min,柱温为25℃,进样量为2μL,检测波长为254nm。结果33批不同批次疏血通注射液中有28个共有峰,其中有15个已知峰,分别为组氨酸(His)、精氨酸(Arg)、丝氨酸(Ser)、甘氨酸(Gly)、谷氨酸(Glu)、天冬氨酸(Asp)、苏氨酸(Thr)、脯氨酸(Pro)、丙氨酸(Ala)、酪氨酸(Tyr)、甲硫氨酸(Met)、缬氨酸(Val)、亮氨酸(Leu)、赖氨酸(Lys)、苯丙氨酸(Phe),不同批次的样品与标准指纹图谱相似度均在0.994以上。结论本实验建立的UPLC指纹图谱分析方法稳定、精密度高,可应用于疏血通注射液的质量控制和评价。

Objective To establish a UPLC fingerprint analysis method for free amino acids in Shuxuetong Injection.Methods Using phenyl isothiocyanate(PITC) derivatization method,using a Waters Acquity ultra performance liquid chromatograph,gradient elution with CORTECS?C18 column(150 mm×2.1 mm,1.6 μm) in acetonitrile-0.1% formic acid water,the flow rate is 0.1 mL/min,column temperature is 25 ℃,the injection volume is 2 μL,and the detection wavelength is 254 nm.Results There are 28 common peaks in 33 batches of different batches of Shuxuetong Injection,including 15 known peaks,namely histidine(His),arginine(Arg),serine(Ser),glycine(Gly),glutamic acid(Glu),aspartic acid(Asp),threonine(Thr),proline(Pro),alanine(Ala),tyrosine(Tyr),methionine(Met),valine(Val),leucine(Leu),lysine(Lys),phenylalanine(Phe),the similarity of samples and standard fingerprints of different batches are above 0.994.Conclusion The UPLC fingerprint analysis method established in this experiment is stable and has high precision.It can be applied to the quality control and evaluation of Shuxuetong Injection.