title: A Strategy Combining Higher Energy C-Trap Dissociation with Neutral Loss- and Product Ion-Based MSnAcquisition for Global Profiling and Structure Annotation of Fatty Acids Conjugates.
abstract: Fatty acids conjugates (FACs) are ubiquitous but found in trace amounts in the natural world. They are composed of multiple unknown substructures and side chains. Thus, FACs are difficult to be analyzed by traditional mass spectrometric methods. In this study, an integrated strategy was developed to global profiling and targeted structure annotation of FACs in complex matrix by LTQ Orbitrap. Dicarboxylic acid conjugated bufotoxins (DACBs) in Venenum bufonis (VB) were used as model compounds. The new strategy (abbreviated as HPNA) combined higher-energy C-trap dissociation (HCD) with product ion- (PI), neutral loss- (NL) based MSn(n ≥ 3) acquisition in both positive-ion mode and negative-ion mode. Several advantages are presented. First, various side chains were found under HCD in negative-ion mode, which included both known and unknown side chains. Second, DACBs with multiple side chains were simultaneously detected in one run. Compared with traditional quadrupole-based mass method, it greatly increased analysis throughput. Third, the fragment ions of side chain and steroids substructure could be obtained by PI- and NL-based MSnacquisition, respectively, which greatly increased the accuracy of the structure annotation of DACBs. In all, 78 DACBs have been discovered, of which 68 were new compounds; 25 types of substructure formulas and seven dicarboxylic acid side chains were found, especially five new side chains, including two saturated dicarboxylic acids [(azelaic acid (C9) and sebacic acid (C10)] and three unsaturated dicarboxylic acids (u-C8, u-C9, and u-C10). All these results greatly enriched the structures of DACBs in VB. Graphical Abstract ᅟ.
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China.|The College of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing, 210009, China.
authors.forename: Qi-Rui
authors.initials: QR
authors.lastname: Bi
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China.
authors.forename: Jin-Jun
authors.initials: JJ
authors.lastname: Hou
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China.
authors.forename: Min
authors.initials: M
authors.lastname: Yang
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Qi
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China.
authors.forename: Rui-Hong
authors.initials: RH
authors.lastname: Feng
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Dai
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China.|The College of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing, 210009, China.
authors.forename: Bing-Peng
authors.initials: BP
authors.lastname: Yan
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China.|The College of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing, 210009, China.
authors.forename: Jian-Wei
authors.initials: JW
authors.lastname: Wang
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China.|The College of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing, 210009, China.
authors.forename: Xiao-Jian
authors.initials: XJ
authors.lastname: Shi
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China. wanyingwu@simm.ac.cn.
authors.forename: Wan-Ying
authors.initials: WY
authors.lastname: Wu
authors.affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai, 201203, China. daguo@simm.ac.cn.|The College of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing, 210009, China. daguo@simm.ac.cn.
authors.forename: De-An
authors.initials: DA
authors.lastname: Guo
mesh_terms: D002018:Bufanolides / Q000737:chemistry; D003652:Decanoic Acids / Q000737:chemistry; D003998:Dicarboxylic Acids / Q000737:chemistry; D005227:Fatty Acids / Q000032:analysis* / Q000737:chemistry*; D015394:Molecular Structure; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013256:Steroids / Q000737:chemistry; D057188:Workflow
chemical_list: D002018:Bufanolides; D003652:Decanoic Acids; D003998:Dicarboxylic Acids; D005227:Fatty Acids; D013256:Steroids; C095591:chan su; C011107:sebacic acid; C010038:azelaic acid; C026738:bufotoxin
keywords: Fatty acid conjugates; Higher-energy C-trap dissociation; Linear ion-trap quadrupole Orbitrap mass spectrometry; Neutral loss based Ms3 acquisition; Product ion based Ms4 acquisition; Venenum bufonis