Applications of LC/MS in structure identifications of small molecules and proteins in drug discovery.

With advancements in ionization methods and instrumentation, liquid chromatography/mass spectrometry (LC/MS) has become a powerful technology for the characterization of small molecules and proteins. This article will illustrate the role of LC/MS analysis in drug discovery process. Examples will be given on high-throughput analysis, structural analysis of trace level impurities in drug substances, identification of metabolites, and characterization of therapeutic protein products for process improvement. Some unique MS techniques will also be discussed to demonstrate their effectiveness in facilitating structural identifications.

[1]  B. Pramanik,et al.  Self‐ionization of nitrocompounds under negative chemical ionization conditions , 1987 .

[2]  Neil L. Kelleher,et al.  Peer Reviewed: Top-Down Proteomics , 2004 .

[3]  R. Subramanian,et al.  Detecting and characterizing reactive metabolites by liquid chromatography/tandem mass spectrometry. , 2006, Journal of mass spectrometry : JMS.

[4]  R. Cody,et al.  Versatile new ion source for the analysis of materials in open air under ambient conditions. , 2005, Analytical chemistry.

[5]  Jonathan P. Williams,et al.  The use of recently described ionisation techniques for the rapid analysis of some common drugs and samples of biological origin. , 2006, Rapid communications in mass spectrometry : RCM.

[6]  A. Makarov,et al.  Orbitrap Mass Analyzer – Overview and Applications in Proteomics , 2006, Proteomics.

[7]  M. Ford,et al.  Thin-layer chromatography and mass spectrometry coupled using desorption electrospray ionization. , 2005, Analytical chemistry.

[8]  Alexander Makarov,et al.  Dynamic range of mass accuracy in LTQ orbitrap hybrid mass spectrometer , 2006, Journal of the American Society for Mass Spectrometry.

[9]  J. R. Engen,et al.  Ultra performance liquid chromatography (UPLC) further improves hydrogen/deuterium exchange mass spectrometry , 2006, Journal of the American Society for Mass Spectrometry.

[10]  Li-Kang Zhang,et al.  LC/MS: Theory, Instrumentation, and Applications to Small Molecules , 2006 .

[11]  M. Karas,et al.  Matrix-assisted ultraviolet laser desorption of non-volatile compounds , 1987 .

[12]  M. Mann,et al.  Parts per Million Mass Accuracy on an Orbitrap Mass Spectrometer via Lock Mass Injection into a C-trap*S , 2005, Molecular & Cellular Proteomics.

[13]  M. Mann,et al.  Electrospray ionization for mass spectrometry of large biomolecules. , 1989, Science.

[14]  M. Thevis,et al.  Determination of N-Desmethyl- and N-Bisdesmethyl Metabolites of Sibutramine in Doping Control Analysis Using Liquid Chromatography-Tandem Mass Spectrometry , 2006, European journal of mass spectrometry.

[15]  Michael S. Bereman,et al.  Direct high-resolution peptide and protein analysis by desorption electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. , 2006, Rapid communications in mass spectrometry : RCM.

[16]  Chowdhury,et al.  Liquid chromatography/mass spectrometry methods for distinguishing N-oxides from hydroxylated compounds , 2000, Analytical chemistry.

[17]  Jonathan P. Williams,et al.  Rapid accurate mass desorption electrospray ionisation tandem mass spectrometry of pharmaceutical samples. , 2005, Rapid communications in mass spectrometry : RCM.

[18]  M. Fathi,et al.  Desorption electrospray ionization mass spectrometry: direct toxicological screening and analysis of illicit Ecstasy tablets. , 2006, Rapid communications in mass spectrometry : RCM.

[19]  B. Chait Mass Spectrometry: Bottom-Up or Top-Down? , 2006, Science.

[20]  Ronald C. Beavis,et al.  Matrix-assisted laser desorption/ionization mass spectrometry of biopolymers. , 1991, Analytical chemistry.

[21]  Richard M. Caprioli,et al.  Imaging Mass Spectrometry: Principles and Potentials , 2005, Toxicologic pathology.

[22]  B. Pramanik,et al.  Special techniques of fast atom bombardment mass spectrometry for the study of oligosaccharide containing macrotetronolide antibiotic, kijanimicin. , 1984, The Journal of antibiotics.

[23]  A. Makarov,et al.  Performance evaluation of a hybrid linear ion trap/orbitrap mass spectrometer. , 2006, Analytical chemistry.

[24]  R. Cooks,et al.  Ambient Mass Spectrometry , 2006, Science.

[25]  A. Kalgutkar,et al.  Application of a linear ion trap/orbitrap mass spectrometer in metabolite characterization studies: Examination of the human liver microsomal metabolism of the non-tricyclic anti-depressant nefazodone using data-dependent accurate mass measurements , 2006, Journal of the American Society for Mass Spectrometry.

[26]  R. Aebersold,et al.  Mass Spectrometry and Protein Analysis , 2006, Science.

[27]  R. Cooks,et al.  Desorption electrospray ionization using an Orbitrap mass spectrometer: Exact mass measurements on drugs and peptides. , 2006, Rapid communications in mass spectrometry : RCM.

[28]  M. Mann,et al.  Electrospray Mass Spectrometry , 1992 .

[29]  Giles,et al.  High throughput liquid chromatography/mass spectrometric analyses using a novel multiplexed electrospray interface. , 1999, Rapid communications in mass spectrometry : RCM.

[30]  J. Gebler,et al.  Integration of multidimensional chromatographic protein separations with a combined "top-down" and "bottom-up" proteomic strategy. , 2006, Journal of Proteome Research.

[31]  J. Jorgenson,et al.  Ultrahigh-pressure reversed-phase liquid chromatography in packed capillary columns. , 1997, Analytical chemistry.

[32]  P. Newton,et al.  Characterization of Solid Counterfeit Drug Samples by Desorption Electrospray Ionization and Direct‐analysis‐in‐real‐time Coupled to Time‐of‐flight Mass Spectrometry , 2006, ChemMedChem.

[33]  L. F. Waanders,et al.  Top-down Protein Sequencing and MS3 on a Hybrid Linear Quadrupole Ion Trap-Orbitrap Mass Spectrometer*S , 2006, Molecular & Cellular Proteomics.

[34]  Richard D. Smith,et al.  Proteomics by FTICR mass spectrometry: top down and bottom up. , 2005, Mass spectrometry reviews.

[35]  Neil L Kelleher,et al.  Shotgun annotation of histone modifications: a new approach for streamlined characterization of proteins by top down mass spectrometry. , 2004, Journal of the American Chemical Society.

[36]  Huanwen Chen,et al.  Desorption electrospray ionization mass spectrometry for high-throughput analysis of pharmaceutical samples in the ambient environment. , 2005, Analytical chemistry.

[37]  J. Yates,et al.  Performance of a linear ion trap-Orbitrap hybrid for peptide analysis. , 2006, Analytical chemistry.

[38]  Iain Beattie,et al.  Ultra-performance liquid chromatography coupled to quadrupole-orthogonal time-of-flight mass spectrometry. , 2004, Rapid communications in mass spectrometry : RCM.

[39]  R. Cooks,et al.  Desorption electrospray ionization mass spectrometry for the analysis of pharmaceuticals and metabolites. , 2006, Rapid communications in mass spectrometry : RCM.

[40]  A. Makarov,et al.  The Orbitrap: a new mass spectrometer. , 2005, Journal of mass spectrometry : JMS.

[41]  A. Makarov,et al.  Interfacing the orbitrap mass analyzer to an electrospray ion source. , 2003, Analytical chemistry.

[42]  F. McLafferty,et al.  Extending Top-Down Mass Spectrometry to Proteins with Masses Greater Than 200 Kilodaltons , 2006, Science.

[43]  S. E. Rodriguez-Cruz Rapid analysis of controlled substances using desorption electrospray ionization mass spectrometry. , 2006, Rapid communications in mass spectrometry : RCM.

[44]  M. Mann,et al.  The human urinary proteome contains more than 1500 proteins, including a large proportion of membrane proteins , 2006, Genome Biology.

[45]  R. Kostiainen,et al.  Liquid chromatography/atmospheric pressure ionization-mass spectrometry in drug metabolism studies. , 2003, Journal of mass spectrometry : JMS.

[46]  J. Jorgenson,et al.  Ultrahigh-pressure reversed-phase capillary liquid chromatography: isocratic and gradient elution using columns packed with 1.0-micron particles. , 1999, Analytical chemistry.

[47]  Robert S Plumb,et al.  Statistical search space reduction and two-dimensional data display approaches for UPLC-MS in biomarker discovery and pathway analysis. , 2006, Analytical chemistry.

[48]  J. Henion,et al.  Structural characterization of protein tryptic peptides via liquid chromatography/mass spectrometry and collision-induced dissociation of their doubly charged molecular ions. , 1991, Analytical chemistry.

[49]  Huanwen Chen,et al.  Combining desorption electrospray ionization mass spectrometry and nuclear magnetic resonance for differential metabolomics without sample preparation. , 2006, Rapid communications in mass spectrometry : RCM.

[50]  B. Pramanik,et al.  The role of mass spectrometry in the drug discovery process. , 1999, Current opinion in drug discovery & development.

[51]  R. Plumb,et al.  Investigating the human metabolism of acetaminophen using UPLC and exact mass oa-TOF MS. , 2005, Journal of pharmaceutical and biomedical analysis.

[52]  R. Cooks,et al.  Ambient mass spectrometry using desorption electrospray ionization (DESI): instrumentation, mechanisms and applications in forensics, chemistry, and biology. , 2005, Journal of mass spectrometry : JMS.

[53]  Beatrix Ueberheide,et al.  Protein identification using sequential ion/ion reactions and tandem mass spectrometry. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[54]  Andrew B. Feldman,et al.  Top-down proteomics for rapid identification of intact microorganisms. , 2005, Analytical chemistry.

[55]  B. Pramanik,et al.  Structural analysis of biologically active peptides and recombinant proteins and their modified counterparts by mass spectrometry. , 1991, Journal of chromatography.

[56]  Koichi Tanaka,et al.  Protein and polymer analyses up to m/z 100 000 by laser ionization time-of-flight mass spectrometry , 1988 .

[57]  B. Green,et al.  Collision-induced fragmentation pathways including odd-electron ion formation from desorption electrospray ionisation generated protonated and deprotonated drugs derived from tandem accurate mass spectrometry. , 2006, Journal of mass spectrometry : JMS.

[58]  G. Guiochon,et al.  Heterogeneous adsorption of 1-indanol on cellulose tribenzoate and adsorption energy distribution of the two enantiomers. , 2004, Analytical chemistry.

[59]  C. Creaser,et al.  Direct analysis of pharmaceutical drug formulations using ion mobility spectrometry/quadrupole-time-of-flight mass spectrometry combined with desorption electrospray ionization. , 2005, Analytical chemistry.

[60]  J E Patrick,et al.  Fragmentation of N-oxides (deoxygenation) in atmospheric pressure ionization: investigation of the activation process. , 2001, Rapid communications in mass spectrometry : RCM.

[61]  F. McLafferty,et al.  Top down characterization of larger proteins (45 kDa) by electron capture dissociation mass spectrometry. , 2002, Journal of the American Chemical Society.

[62]  G. Siuzdak,et al.  Nonlinear data alignment for UPLC-MS and HPLC-MS based metabolomics: quantitative analysis of endogenous and exogenous metabolites in human serum. , 2006, Analytical chemistry.

[63]  Jennifer H Granger,et al.  A rapid screening approach to metabonomics using UPLC and oa-TOF mass spectrometry: application to age, gender and diurnal variation in normal/Zucker obese rats and black, white and nude mice. , 2005, The Analyst.

[64]  N. Kelleher,et al.  Top-down approaches for measuring expression ratios of intact yeast proteins using Fourier transform mass spectrometry. , 2006, Analytical chemistry.

[65]  R. Cooks,et al.  Mass Spectrometry Sampling Under Ambient Conditions with Desorption Electrospray Ionization , 2004, Science.

[66]  B. Pramanik,et al.  Isolation and characterization of a resistant core peptide of recombinant human granulocyte‐macrophage colony‐stimulating factor (gm‐csf); confirmation of the gm‐csf amino acid sequence by mass spectrometry , 1993, Protein science : a publication of the Protein Society.