Novel mass spectrometric methodologies for the structural and compositional analysis of labile sulfated glycosaminoglycans:
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| Main Author: | |
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2013
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | IX S., S. 11 - 144 S. Ill., graph. Darst. 30 cm |
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| adam_text | TABLE OF CONTENTS
PREFACE I
ACKNOWLEDGEMENTS ILL
TABLE OF CONTENTS V
LIST OF ABBREVIATIONS VIII
1 INTRODUCTION 11
1.1 GLYCOSAMINOGLYCANS 11
1.2 STRUCTURE 12
1.2.1 HEPARIN AND HEPARAN SULFATE 13
1.2.2 CHONDROITIN SULFATE 15
1.2.3 BIOSYNTHESIS 16
1.3 FUNCTIONS 20
1.4 GAG ANALYSIS 22
1.5 INSTRUMENTATION 29
1.5.1 MASS SPECTROMETRY 29
1.5.2 HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHY 35
1.5.3 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY 36
1.6 MOTIVATION 38
2 MATERIALS AND METHODS 40
2.1 REAGENTS 40
2.2 SAMPLES 40
2.3 DERIVATIZATION OF SAMPLES 41
2.3.1 ANILINE 42
2.3.2 DANSYLHYDRAZINE 43
2.3.3 2-AMINOACRIDONE 43
2.3.4 TRIMETHYLSILYLDIAZOMETHANE 44
2.4 PURIFICATION OF SAMPLES 44
2.4.1 IN-SOLUTION DIGEST WITH CHONDROITINASE ABC AND HEPARINASES
1,11,111 44
2.4.2 SEPARATION OF HEPARIN AND CHONDROITIN SULFATE DIGESTION PRODUCTS
45
2.4.3 DESALTING OF DIGESTS AND DERIVATIZATION PRODUCTS 46
HTTP://D-NB.INFO/1047345927
TABLE OF CONTENTS
2.5 CHROMATOGRAPHY 47
2.5.1 HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHY 47
2.5.2 REVERSED PHASE ION-PAIRING HPLC 49
2.5.3 NANO-HPLC COUPLED TO ESI-MS 51
2.6 NANOESI MASS SPECTROMETRY 55
2.7 MALDI MASS SPECTROMETRY 55
2.7.1 ULTRAVIOLET MALDI-O-TOF-MS 55
2.7.2 INFRARED MALDI-O-TOF-MS 57
3 RESULTS 59
3.1 DERIVATIZATION OF SULFATED DISACCHARIDES WITH
TRIMETHYLSILYL-DIAZOMETHANE 59
3.2 ANALYSIS OF SULFATED DISACCHARIDES BY REVERSED PHASE ION-PAIRING
HPLC 62
3.2.1 DERIVATIZATION OF DISACCHARIDES WITH ANILINE 62
3.2.2 SEPARATION OF DERIVATIZED SPECIES 64
3.3 ZIC-HILIC-BASED NANO-HPLC-MS OF GLYCOSAMINOGLYCANS 67
3.3.1 SEPARATION CHARACTERISTICS OF SULFATED SPECIES 67
3.3.2 DISACCHARIDE ANALYSIS 69
3.3.3 OLIGOMER ANALYSIS 72
3.3.4 APPLICATION TO ENZYMATICALLY DEGRADED GAG POLYMERS 77
3.4 GLYCOSAMINOGLYCAN ANALYSIS BY HPTLC AND IR-MALDI MASS SPECTROMETRY
80
3.4.1 SEPARATION OF SULFATED DISACCHARIDES ON SILICA GEL 81
3.5 IMPROVING THE IONIZATION CONDITIONS OF GAGS IN MALDI MASS
SPECTROMETRY 88
3.5.1 TEST OF DIFFERENT MATRIX COMPOUNDS 88
3.5.2 UV-MALDI 89
3.5.3 IR-MALDI USING GLYCEROL AS A MATRIX 92
3.5.4 WATER ICE AS SOFT MATRIX FOR THE STRUCTURAL CHARACTERIZATION OF
HEPARAN SULFATE
BY IR-MALDI MASS SPECTROMETRY 95
4 DISCUSSION 110
4.1 IMPROVING GLYCOSAMINOGLYCAN ANALYSIS BY DERIVATIZATION 110
4.1.1 PERMETHYLATION 110
4.1.2 INTRODUCTION OF CHROMOPHORES 112
4.2 HPLC-BASED APPROACHES IN THE ANALYSIS OF GLYCOSAMINOGLYCANS 114
4.2.1 RP IP HPLC 114
4.2.2 ZIC-HILIC-BASED NANO-HPLC 115
LUKASWITT VII
NOVEL MASS SPECTROMETRIC METHODOLOGIES FOR THE STRUCTURAL AND
COMPOSITIONAL ANALYSIS OF LABILE SULFATED GLYCOSAMINOGLYCANS
4.3 HYPHENATION OF TLC AND IR-MALDI 117
4.4 WATER ICE AS A SOFT MATRIX FOR IMPROVED IONIZATION OF
GLYCOSAMINOGLYCANS IN IR-
MALDI 118
5 CONCLUSIONS 122
6 REFERENCES 125
7 APPENDIX 137
7.1 ZUSAMMENFASSUNG (GERMAN) 137
7.2 INDEX OF TABLES 140
7.3 INDEX OF FIGURES..... 141
7.4 CURRICULUM VITAE 144
|
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| dewey-ones | 610 - Medicine and health |
| dewey-raw | 610 |
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| dewey-sort | 3610 |
| dewey-tens | 610 - Medicine and health |
| discipline | Medizin |
| format | Thesis Book |
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| institution | BVB |
| language | English |
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| physical | IX S., S. 11 - 144 S. Ill., graph. Darst. 30 cm |
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| spelling | Witt, Lukas 1983- Verfasser (DE-588)1046419862 aut Novel mass spectrometric methodologies for the structural and compositional analysis of labile sulfated glycosaminoglycans vorgelegt von Lukas Alexander Witt 2013 IX S., S. 11 - 144 S. Ill., graph. Darst. 30 cm txt rdacontent n rdamedia nc rdacarrier Münster (Westfalen), Univ., Diss., 2013 (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027337054&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Witt, Lukas 1983- Novel mass spectrometric methodologies for the structural and compositional analysis of labile sulfated glycosaminoglycans |
| subject_GND | (DE-588)4113937-9 |
| title | Novel mass spectrometric methodologies for the structural and compositional analysis of labile sulfated glycosaminoglycans |
| title_auth | Novel mass spectrometric methodologies for the structural and compositional analysis of labile sulfated glycosaminoglycans |
| title_exact_search | Novel mass spectrometric methodologies for the structural and compositional analysis of labile sulfated glycosaminoglycans |
| title_full | Novel mass spectrometric methodologies for the structural and compositional analysis of labile sulfated glycosaminoglycans vorgelegt von Lukas Alexander Witt |
| title_fullStr | Novel mass spectrometric methodologies for the structural and compositional analysis of labile sulfated glycosaminoglycans vorgelegt von Lukas Alexander Witt |
| title_full_unstemmed | Novel mass spectrometric methodologies for the structural and compositional analysis of labile sulfated glycosaminoglycans vorgelegt von Lukas Alexander Witt |
| title_short | Novel mass spectrometric methodologies for the structural and compositional analysis of labile sulfated glycosaminoglycans |
| title_sort | novel mass spectrometric methodologies for the structural and compositional analysis of labile sulfated glycosaminoglycans |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027337054&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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