MS-1-supplemental reading

MS-1-supplemental reading - An Introduction to Mass...

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An Introduction to Mass Spectrometry CONTENTS 1. What is mass spectrometry (MS)? What Information does mass spectrometry provide? 2. Where are mass spectrometers used? 3. How can mass spectrometry help biochemists? 4. How does a mass spectrometer work? 1. Introduction 2. Sample introduction 3. Methods of sample ionisation 4. Analysis and separation of sample ions 5. Detection and recording of sample ions 5. Electrospray ionisation 1. Electrospray ionisation 2. Nanospray ionisation 3. Data processing 6. Matrix assisted laser desorption ionisation 7. Positive or negative ionisation? 8. Tandem mass spectrometry (MS-MS): Structural and sequence information from mass spectrometry 1. Tandem mass spectrometry 2. Tandem mass spectrometry analyses 1. What is mass spectrometry (MS)? What information does mass spectrometry provide? Mass spectrometry is an analytical tool used for measuring the molecular mass of a sample. For large samples such as biomolecules, molecular masses can be measured to within an accuracy of 0.01% of the total molecular mass of the sample i.e. within a 4 Daltons (Da) or atomic mass units (amu) error for a sample of 40,000 Da. This is sufficient to allow minor mass changes to be detected, e.g. the substitution of one amino acid for another, or a post-translational modification. For small organic molecules the molecular mass can be measured to within an accuracy of 5 ppm or less, which is often sufficient to confirm the molecular formula of a compound, and is also a standard requirement for publication in a chemical journal. Structural information can be generated using certain types of mass spectrometers, usually those with multiple analysers which are known as tandem mass spectrometers. This is achieved 1
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by fragmenting the sample inside the instrument and analysing the products generated. This procedure is useful for the structural elucidation of organic compounds and for peptide or oligonucleotide sequencing. 2. Where are mass spectrometers used? Mass spectrometers are used in industry and academia for both routine and research purposes. The following list is just a brief summary of the major mass spectrometric applications: Biotechnology: the analysis of proteins, peptides, oligonucleotides Pharmaceutical: drug discovery, combinatorial chemistry, pharmacokinetics, drug metabolism Clinical: neonatal screening, haemoglobin analysis, drug testing Environmental: PAHs, PCBs, water quality, food contamination Geological: oil composition 3. How can mass spectrometry help biochemists? Accurate molecular weight measurements: sample confirmation, to determine the purity of a sample, to verify amino acid substitutions, to detect post-translational modifications, to calculate the number of disulphide bridges Reaction monitoring: to monitor enzyme reactions, chemical modification, protein digestion Amino acid sequencing: sequence confirmation, de novo characterisation of peptides, identification of proteins by
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MS-1-supplemental reading - An Introduction to Mass...

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