Metabolomics for toxicity analysis using the chlorophyte Scenedesmus vacuolatus:
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| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Leipzig
Helmholtz Centre for Environmental Research - UFZ
2009
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| Series: | PhD dissertation
2009,2 |
| Subjects: | |
| Online Access: | Volltext Inhaltsverzeichnis |
| Item Description: | Auch als Internetausgabe |
| Physical Description: | 116, VII S. graph. Darst. |
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Record in the Search Index
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| adam_text | Titel: Metabolomics for toxicity analysis using the chlorophyte Scenedesmus vacuolatus
Autor: Klünder, Christina
Jahr: 2009
Contents
Danksagung...................................................................................................I
Contents.......................................................................................................H
Abbreviations..............................................................................................IV
Summary.......................................................................................................6
1 Introduction........................................................................................11
Ecotoxicological Bioassays.......................................................................................11
Aquatic Toxicology...................................................................................................11
Phytotoxicity Assessment with Scenedesmus vacuolatus.........................................12
i-Triazines.................................................................................................................12
Ecotoxicological Parameters of Effect......................................................................13
Metabolites, Metabolome and Metabolomics............................................................14
Environmental Metabolomics...................................................................................16
Linking the Metabolome with the Phenotype............................................................17
Aims of this Work.....................................................................................................19
2 Material and Methods........................................................................21
2.1 Solvents and Reagents...................................................................21
2.2 Test Organism, Cultural Maintenance and Sampling................21
2.2.1 Test Organism and Cultural Maintenance...................................................21
2.2.2 Determination of Cell Density and Algal Growth.......................................22
2.2.3 Harvest........................................................................................................22
2.2.4 5. vacuolatus Sample Storage and Conservation........................................23
2.3 Bioassay to Study Effects of Toxicity...........................................24
2.3.1 Used Chemical to Study Effects of Toxicity...............................................24
2.3.2 Treatment of S. vacuolatus..........................................................................24
2.3.3 Phenotype Investigation..............................................................................25
2.4 Algal Metabolite Extraction and Separation of Hydrophilic and
Lipophilic Metabolite Fractions...................................................26
2.4.1 Metabolite Extraction..................................................................................26
2.4.2 Separation of Hydrophilic and Lipophilic Metabolites...............................27
2.5 Analysis of Hydrophilic and Lipophilic Metabolites with a GC-
MS system.......................................................................................28
2.5.1 Derivatisation of Algal Metabolites............................................................28
2.5.2 Retention Index Calibration Standard.........................................................29
2.5.3 Gas Chromatography-Mass Spectrometry (GC-MS) to Analyse Hydrophilic
and Lipophilic Metabolites.........................................................................30
2.6 Qualitative and Quantitative Data Analysis................................31
2.6.1 Qualitative Data Analysis...........................................................................31
2.6.2 Quantitative Comparison of Chromatographic Results...............................32
2.6.3 Analysis of Variability of Analytical GC-MS Data....................................33
.. 2.6.4 Multivariate Statistical Analysis (PCA)....................................... ..........11.34
3 Results.................................................................................................37
3.1 Qualitative and Quantitative Analysis of Metabolomics Results
for Hydrophilic and Lipophilic Algal Extracts...........................37
3.1.1 Qualitative Analysis of TIC s Using the System s ChemStation Software 37
3.1.2 Qualitative Analysis of TIC s Using the AMDIS Software....................37
3.1.3 Identification of S. vacuolatus Metabolites Using Custom Libraries..........
(Qualitative Analysis).................................................... 38
II
Contents
3.1.4 Selection of Quantitative Data from S. vacuolalus and Reproducibility of
the Analytical Results.................................................................................40
3.2 Application of Metabolomics for the Qualitative and
Quantitative Analysis of S. vacuolatus Metabolites in Response to
Time and Prometryn Toxicity.......................................................41
3.2.1 Concentration - Responses of the Algal Metabolites..................................42
3.2.2 Time - Responses during Algal Growth.....................................................44
3.2.3 Characterisation of the Algal Metabolome in Response to Time and
Prometryn Toxicity.....................................................................................49
3.2.4 Characterisation of the Algal Growth in Response to Time and Prometryn
Toxicity.......................................................................................................57
3.3 Summary of Results.......................................................................59
4 Discussion...........................................................................................62
4.1 Metabolomics Approach - Workflow...........................................62
4.1.1 Experimental Design and Harvest...............................................................63
4.1.2 Sample Preparation.....................................................................................66
4.1.3 Metabolite Analysis....................................................................................68
4.1.4 Data-Processing..........................................................................................72
4.1.5 Data-Analysis..............................................................................................74
4.2 Compound Identification..............................................................81
4.2.1 Method of Compound Identification...........................................................81
4.3 Metabolite Composition in S. vacuolatus Extracts......................82
4.3.1 S. vacuolatus Metabolites in Comparison with those of Chlorella- and
Chlamydomonas-species.............................................................................82
4.3.2 Algal Growth during Reference Conditions................................................84
4.4 Application of Metabolomics for the Analysis of S. vacuolatus
Metabolites in Response to Time and Prometryn Toxicity........ 88
4.4.1 General Effects of 5-Triazines observed on the Chlorophyte Scenedesmus
vacuolatus...................................................................................................88
4.4.2 Observed Effects of Prometryn on the Metabolome and the Cell Volume of
Scenedesmus vacuolatus.............................................................................91
4.4.3 Primary and Secondary Responses to Toxicity over Time..........................96
4.4.4 Combining the Information of the Metabolome with that of the Phenotype
from S. vacuolatus.......................................................................................98
5 Conclusions Perspectives.............................................................101
References.................................................................................................106
Appendix.......................................................................................................ii
Liste der Publikationen.................................................................................v
Curriculum Vitae.........................................................................................vi
III
|
| any_adam_object | 1 |
| author | Klünder, Christina 1979- |
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| dewey-full | 571.9 571.95929833 572.4 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 571 - Physiology & related subjects 572 - Biochemistry |
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| dewey-search | 571.9 571.95929833 572.4 |
| dewey-sort | 3571.9 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
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| spelling | Klünder, Christina 1979- Verfasser (DE-588)137347316 aut Metabolomics for toxicity analysis using the chlorophyte Scenedesmus vacuolatus vorgelegt von Christina Klünder Leipzig Helmholtz Centre for Environmental Research - UFZ 2009 116, VII S. graph. Darst. txt rdacontent n rdamedia nc rdacarrier PhD dissertation 2009,2 Auch als Internetausgabe Zugl.: Freiberg, Techn. Univ. Bergakad., Diss., 2008 Metabolom (DE-588)4807318-0 gnd rswk-swf Metabolit (DE-588)4138579-2 gnd rswk-swf Toxikologie (DE-588)4060538-3 gnd rswk-swf Grünalgen (DE-588)4072124-3 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Metabolit (DE-588)4138579-2 s Metabolom (DE-588)4807318-0 s Toxikologie (DE-588)4060538-3 s Grünalgen (DE-588)4072124-3 s DE-604 PhD dissertation 2009,2 (DE-604)BV035421074 2009,2 http://www.ufz.de/data/ufzdiss_2_2009_9947.pdf Volltext HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=017112774&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Klünder, Christina 1979- Metabolomics for toxicity analysis using the chlorophyte Scenedesmus vacuolatus PhD dissertation Metabolom (DE-588)4807318-0 gnd Metabolit (DE-588)4138579-2 gnd Toxikologie (DE-588)4060538-3 gnd Grünalgen (DE-588)4072124-3 gnd |
| subject_GND | (DE-588)4807318-0 (DE-588)4138579-2 (DE-588)4060538-3 (DE-588)4072124-3 (DE-588)4113937-9 |
| title | Metabolomics for toxicity analysis using the chlorophyte Scenedesmus vacuolatus |
| title_auth | Metabolomics for toxicity analysis using the chlorophyte Scenedesmus vacuolatus |
| title_exact_search | Metabolomics for toxicity analysis using the chlorophyte Scenedesmus vacuolatus |
| title_full | Metabolomics for toxicity analysis using the chlorophyte Scenedesmus vacuolatus vorgelegt von Christina Klünder |
| title_fullStr | Metabolomics for toxicity analysis using the chlorophyte Scenedesmus vacuolatus vorgelegt von Christina Klünder |
| title_full_unstemmed | Metabolomics for toxicity analysis using the chlorophyte Scenedesmus vacuolatus vorgelegt von Christina Klünder |
| title_short | Metabolomics for toxicity analysis using the chlorophyte Scenedesmus vacuolatus |
| title_sort | metabolomics for toxicity analysis using the chlorophyte scenedesmus vacuolatus |
| topic | Metabolom (DE-588)4807318-0 gnd Metabolit (DE-588)4138579-2 gnd Toxikologie (DE-588)4060538-3 gnd Grünalgen (DE-588)4072124-3 gnd |
| topic_facet | Metabolom Metabolit Toxikologie Grünalgen Hochschulschrift |
| url | http://www.ufz.de/data/ufzdiss_2_2009_9947.pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=017112774&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV035421074 |
| work_keys_str_mv | AT klunderchristina metabolomicsfortoxicityanalysisusingthechlorophytescenedesmusvacuolatus |