超高效液相色谱-二极管阵列-四极杆静电轨道阱质谱法对杆菌肽组分的分析

开发建立了超高效液相色谱-二极管阵列-串联四级杆静电场轨道阱高分辨质谱联用分析杆菌肽组分的方法。采用(Phenomenex C₁₈100 mm×3.0 mm,2.6μm)色谱柱,以0.1%甲酸溶液-0.1%甲酸甲醇乙腈(75+25)溶液为流动相,0.3 mL/min线性梯度洗脱分离,紫外吸收波长为254 nm。采集杆菌肽各组分质谱母离子及其对应子离子,并进行解析,推测组分的结构。在所建立的条件下,检测出9个组分,杆菌肽各组分之间分离良好。利用紫外与质谱归一化法测定含量,两种结果相对偏差在0.01%~3.53%之间。建立的超高效液相色谱-二极管阵列-串联四级杆静电场轨道阱高分辨质谱法能有效分析多肽类药物杆菌肽各组成成分,为杆菌肽的质量控制和工艺优化提供了参考。

Analysis of bacitracin components by ultra high performance liquid chromatography diode array detector coupled with quadrupole orbitrap mass spectrometry

An ultra high performance liquid chromatography diode array detector(DAD)coupled with hybrid quadrupole orbitrap mass spectrometry method was developed for bacitracin components analysis.The UHPLC separation was carried out on a Phenomenon C₁₈(100 mm×3.0 mm,2.6μm)by gradient elution with a mobile phase consisting of 0.1%formic acid aqueous solutionand 0.1%formic acid meoh acetonitrile(75+25)solution at a flow rate of 0.3 mL/min.The.detection wavelength was set at 254 nm.The analysis for bacitracin components were performed with the parent ions and the daughter ions,then estimated their structures.Good resolution of bacitracin components were achieved.Nine bacitracin components were separated and detected.The relative deviation of normalization method by DAD and mass spectrometry was between 0.01%and 3.53%.The established method was effective for the separation and identification of peptide drugs such as bacitracin components and the results were useful for its quality control and process optimization.