【摘要】 目的:使用超高效液相色谱-四极杆串联飞行时间质谱法（UPLC-Q TOF MS法）对国内外3家企业溶菌酶的一级结构进行分析研究。方法:采用BEH300 C₄（50 mm×2.1 mm, 1.7μm）色谱柱和BEH300 C₁₈（150 mm×2.1 mm, 1.7μm）色谱柱，以0.1%甲酸/水溶液为流动相A,0.1%甲酸/乙腈溶液为流动相B,梯度洗脱，流速0.2 mL·min^-1;采用Q TOF MS的电喷雾正离子模式，相对分子质量测定时采用MS模式进行扫描，氨基酸序列和二硫键连接方式测定时采用MS^E模式进行扫描。结果:3种来源溶菌酶相对分子质量与理论值一致；肽图结果显示各样品的氨基酸序列均与理论一致；二硫键连接方式均为Cys6-Cys127、Cys30-Cys115、Cys64-Cys80、Cys76-Cys94。结论:UPLC-Q TOF MS法可用于溶菌酶一级结构的分析。更多还原

【Abstract】 Objective:To analyze the primary structure of the lysozyme from three different domestic and foreign manufacturers using UPLC-Q TOF MS. Methods: The UPLC-Q TOF MS analysis was performed on a BEH300 C₄（50 mm×2.1 mm, 1.7 μm） column and a BEH300 C₁₈（150 mm×2.1 mm, 1.7 μm） column with 0.1% formic acid aqueous solution as mobile phase A and 0.1% formic acid-acetonitrile solution as mobile phase B, with gradient elution at a flow rate of 0.2 mL·min^-1. Q TOF MS electrospray positive ion mode was used, and for relative molecular mass detection, MS scan mode was applied. For amino acid sequence and disulfide bond connection, MS^E scan mode was applied. Results: The measured relative molecular mass of the three lysozyme samples was consistent with theory; the results of peptide map showed that the amino acid sequences of all sampleswere consistent with the theory; the connection modes of disulfide bonds of all lysozymes were Cys6-Cys127, Cys30-Cys115,Cys64-Cys80 and Cys76-Cys94. Conclusion: UPLC-Q TOF MS method can be used to analyze the primary structure of the lysozyme.更多还原