摘要
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)的纽莫康定B_(0)(PB_(0))及其组分的结构分析方法。方法纽莫康定B_(0)及其组分通过HPLC分离,流动相A为0.1%甲酸水溶液,流动相B为乙腈,梯度洗脱后经在线Q/TOF-MS得到PB_(0)及其组分的一级及二级质谱信息。对PB_(0)的精确分子量、特征碎片离子及低端氨基酸残基离子进行解析,系统归纳了3条碎片离子裂解途径,可用于其他组分的结构分析。而针对单羟基鸟氨酸取代的同源组分,建立了区别于PB_(0)的裂解规律。结果本文首次发现了在裂解过程中,脂肪酰基链可以从双羟基鸟氨酸上转移至其他氨基酸上,单羟基鸟氨酸取代的组分可以形成亚胺离子。依据裂解规律共解析了11个组分的结构,首次发现了含有不饱和脂肪酰基链的PB组分(PB_(0)-1、PB_(0)-2和PB_(0)-5)、含有羟基化脂肪酰基链的PB组分(PB_(0)-4)、羰基化鸟氨酸的组分(PB_(0)-8)及环肽开环(5、6位氨基酸处开环)且1位氨基酸转变为5-氨基脯氨酸的组分(PB_(0)-9、PB_(0)-10和PB_(0)-11)。结论本研究拓宽了对PB_(0)组分结构多样性的认识,归纳了快速、合理解析PB_(0)及其组分的结构分析方法,适用于纽莫康定类药物的质量研究。
Objective To develop a structural analysis method for pneumocandin B_(0)(PB_(0))and its components using HPLC-Q/TOF-MS.Methods PB_(0)and its components were separated through HPLC with gradient elution,employing 0.1%formic acid in water as mobile phase A(mpA)and acetonitrile as mobile phase B(mpB).After gradient elution,the primary and secondary mass spectrometry information of PB_(0)and its components were obtained by online Q/TOF-MS.Precise molecular weights,characteristic fragmentation ions,and amino acid residue ions of PB_(0)were identified using Q/TOF-MS.From these data,three fragmentation pathways were systematically outlined to aid in the structural elucidation of PB_(0)and its related components.A distinct fragmentation pattern was also established for homologous components substituted with monohydroxyornithine.Results This study firstly identified the transfer of the fatty acyl chain from dihydroxyornithine to other amino acids,and the formation of imine ions in monohydroxyornithine-substituted components during fragmentation.The structures of eleven components were elucidated using this analysis strategy.Notably,new pneumocandin components were reported for the first time,including those with unsaturated fatty acyl chains(PB_(0)-1,PB_(0)-2,and PB_(0)-5),hydroxylated fatty acyl chains(PB_(0)-4),carbonylated ornithine(PB_(0)-8),and components of ring-opening peptides at positions 5 and 6(PB_(0)-9,PB_(0)-10,and PB_(0)-11)with ornithine replaced by 5-amino proline at position 1.Conclusion The research expanded the understanding of the structural diversity of PB_(0)components and offered a method for rapid and precise structural analysis of pneumocandins,which could be applicable for quality control in pneumocandin drug production.
作者
张含智
王夏昆
田振华
Zhang Hanzhi;Wang Xiakun;and Tian Zhenhua(Abiochem Biotechnology(Group)Co.,LTD,Shanghai 200240)
出处
《中国抗生素杂志》
北大核心
2025年第11期1299-1307,共9页
Chinese Journal of Antibiotics
基金
科技部国家重点研发计划资助项目(No.2022YFC2303100)
国家自然科学基金(No.22193070)。
