目的 建立基于高效液相色谱-四级杆飞行时间质谱(HPLC-Q/TOF-MS)的分析方法,对硫酸多黏菌素E中的组分进行结构解析,结合光化学反应鉴定双键化组分。方法 通过HPLC-Q/TOF-MS/MS对主要组分多黏菌素E1和E2质谱数据的分析,总结了多黏菌素类化合物结构解析的规律,采用精确分子质量、特征碎片离子确定组分结构,并选择氨基酸残基离子来辅助判断结构;多黏菌素E样品在254 nm紫外光灯下经照射发生光化学反应(帕特诺-比希反应),通过MS/MS验证多黏菌素E1双键化组分的环氧化产物。结果 本实验首次总结了一条新的裂解途径。根据以上方法共解析了21种组分的结构,包括2个多黏菌素E1和E2的双键化组分,结合光化学产物的二级质谱特征离子推断双键位于酰基脂肪链末端,将该组分命名为7'-烯基多黏菌素E1和6'-烯基多黏菌素E2。结论 7'-烯基多黏菌素E1及其光反应环氧化产物、6'-烯基多黏菌素E2均为首次报道,本方法为多肽类抗生素组分包括含双键的未知杂质的结构分析提供了一种解决思路。
Abstract
OBJECTIVE To establish a high performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (HPLC-Q/TOF-MS) method for the structural analysis of the components in polymyxin E sulfate, and identify the vinyl components in aid of photochemical reaction. METHODS Based on the mass data of polymyxin E1 and E2, the strategy for structure analysis of polypeptide antibiotics was summarized. The structure of polymyxins was determined by accurate molecular weight and characteristic fragment ions. The amino acids contained in the components were judged by amino acid residue ions. The photochemical (Paterno-Büchi reaction) was performed under a low-pressure mercury lamp (emission band at 254 nm), and the epoxidation product of the vinyl polymyxin E1 was verified by MS/MS. RESULTS A new fragmentation route was proposed for the first time in this paper. According to the above-mentioned methods, the structures of 21 components were analyzed. Two components of vinyl polymyxin E1 and E2 were found and it was the first time to identify the structure of vinyl polymyxin E1 by Paterno-Büchi reaction combined with characteristic ions analysis. These two novel components were named as 7'-vinyl polymyxin E1 and 6'-vinyl polymyxin E2, respectively. CONCLUSION 7'-Vinyl polymyxin E1 and its photochemical product, 6'-vinyl polymyxin E2, are reported for the first time. The study provides an effective solution for the identification of the components including vinyl ones in polymyxins.
关键词
多黏菌素E组分 /
液质联用 /
光化学反应 /
结构分析 /
7'-烯基多黏菌素E1
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Key words
polymyxin E sulfate /
HPLC-MS/MS /
Paterno-Büchi reaction /
structure analysis /
7'-vinyl polymyxin E1
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中图分类号：
R917
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脚注
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基金
上海市2020年度“科技创新行动计划”科学仪器领域资助项目(20142202200)
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