碰撞诱导解离和高能碰撞解离方式在褐藻胶寡糖结构分析中的比较和应用

摘要

利用组合式高分辨质谱仪(LTQ Orbitrap XL)对均一褐藻胶寡糖,包括甘露糖醛酸寡糖和古洛糖醛酸寡糖进行二级质谱分析,比较了常规的碰撞诱导解离(CID)和高能碰撞解离(HCD)在寡糖结构分析中的差异,并应用HCD技术分析甘露糖醛酸寡糖和古洛糖醛酸寡糖的结构差异.研究发现,HCD可避免CID中的1/3效应,而且可以提供更多的糖环断裂碎片信息,如2.55A2 (m/z 291)、2.44A(m/z 235、411)、2,5A脱水(m/z 101、277、453)以及Z2脱羧碎片(m/z 307、483、659)等,进一步验证了Z2脱羧碎片是区别甘露糖醛酸与古洛糖醛酸的特征碎片.同时,HCD能够提供丰富的裂解碎片信息,这为褐藻胶寡糖的结构解析提供了依据.%Alginate homooligosaccharides including oligomannuronate (M) and oligoguluronate (G) with degree of polymerization from 2 to 7 were analyzed by linear ion trapOrbitrap mass spectrometer (LTQ Orbitrap XL) using collision induced dissociation (CID) and high energy collision induced dissociation (HCD).First of all,comparison of CID MS/MS and HCD MS/MS of either M or G was conducted.Taking the trimannuronate (M3) as an example,CID MS/MS showed the m/z 369,193 and 175 ions were assigned to C2/Y2,C1/Y1 and B1/Z1,respectively,which were formed due to single glycosidic bond cleavage,while HCD MS/MS caused cleavage of all glycosidic bonds and showed weak A-type ions,such as 0,2A (m/z 309,485),2,5A (m/z 291,467) and 0,4A3 ions (m/z 471).Interestingly,an internal M produced a Zint-CO2 ions at m/z 307,which was unique and found only in HCD MS/MS.Furthermore,CID MS/MS and HCD MS/MS of M3 and triguluronate (G3) were executed.For CID,M3 and G3 did not show significant difference,and they gave identical glycosidic bond cleavage ions C1 (m/z 193),B1 (m/z 175) together with the dehydrated and decarboxylated ions (m/z 157 and m/z 131).These results suggest that it is impossible to differentiate the two residues at a nonreducing terminal of M3 and G3 by CID.However,a significant difference was observed in HCD spectra for M3 and G3.It was found that besides decarboxylation fragment m/z 307,debris 2,4A3 (m/z 411),2,5A3 (m/z 467) and 0,4A3 (m/z 471)were found in M3,whereas these ions were not found in HCD MS/MS of G3,thus demonstrating that HCD MS/MS can be used to differentiate M3 and G3.In the CID MS/MS of pentamannuronate (M5),fragment ions 0,2A3 (m/z 837),0,4A3 (m/z 823) and 2,5A3(m/z 819) was found apparently,however,HCD displayed rich fragment ions in the low mass region,which should be ascribed to the characteristics of high-energy dissociation of HCD.For heptamannuronate (M7) and heptaguluronate (G7),doubly charged protonated species were collected as parent ions for CID and HCD.HCD MS/MS provided more fragment ions than CID MS/MS,especially,Zint-CO2 ion (m/z 307,483,659,835 and 1 011) signal was apparently showed in M7 HCD spectra.Taken together,this work demonstrates that HCD MS/MS provides rich product ions and has no low mass cut off.The fragment ions in HCD spectra have high mass accuracy and resolution.These characteristics of HCD spectra complement the power of CID and allow easy spectrametric interpretation and high confidence in structural elucidation for alginate oligosaccharides.