Refractory Organic Substancesin the Environment

Edited byF. H. Frimmel, G. Abbt-Braun, K. G. Heumann, B. Hock,H.-D. Lüdemann and M. Spiteller

Innodata9783527614448.jpg

Refractory Organic Substancesin the Environment

Edited by

F. H. Frimmel, G. Abbt-Braun,

K. G. Heumann, B. Hock,

H.-D. Lüdemann and M. Spiteller

Related JournalActa hydrochimicaet hydrobiologica

ISSN 0323-4320www.wiley-vch.de/home/actahydro

Refractory Organic Substancesin the Environment

Edited byF. H. Frimmel, G. Abbt-Braun, K. G. Heumann, B. Hock,H.-D. Lüdemann and M. Spiteller

Editors

Prof. Dr. Fritz H. FrimmelEngler-Bunte-InstitutBereich WasserchemieUniversität KarlsruheEngler-Bunte-Ring 1D-76131 KarlsruheGermany

Dr. Gudrun Abbt-BraunEngler-Bunte-InstitutBereich WasserchemieUniversität KarlsruheEngler-Bunte-Ring 1D-76131 KarlsruheGermany

Prof. Dr. Klaus G. HeumannInstitut für Anorganischeund Analytische ChemieJohannes-Gutenberg-Universität MainzDuesbergweg 10�14D-55099 MainzGermany

Prof. Dr. Berthold HockLehrstuhl für BotanikTechnische Universität MünchenAlte Akademie 12D-85350 FreisingGermany

Prof. Dr. Hans-Dietrich LüdemannInstitut für Biophysikund Physikalische BiochemieUniversität RegensburgD-93040 RegensburgGermany

Prof. Dr. Michael SpitellerInstitut für UmweltforschungUniversität DortmundD-44221 DortmundGermany

This book was carefully produced. Never-theless, editors, authors and publisher donot warrant the information containedtherein to be free of errors. Readers areadvised to keep in mind that statements,data, illustrations, procedural details orother items may inadvertently be inaccurate.

Library of Congress Card No.: applied for

A catalogue record for this book is availablefrom the British Library.

Die Deutsche Bibliothek –CIP-Cataloguing-in-Publication-DataA catalogue record for this publication isavailable from Die Deutsche Bibliothek.

� Wiley-VCH Verlag GmbH,69469 Weinheim (Federal Republic ofGermany). 2002

All rights reserved (including those oftranslation in other languages). No part ofthis book may be reproduced in any form –by photoprinting, microfilm, or any othermeans – nor transmitted or translatedinto machine language without writtenpermission from the publishers. Registerednames, trademarks, etc. used in this book,even when not specifically marked as such,are not to be considered unprotected by law.

Printed in the Federal Republic of Germany.

Printed on acid-free paper.

Typesetting Hagedorn Kommunikation,ViernheimPrinting betz-druck gmbh, DarmstadtBookbinding J. Schäffer GmbH + Co. KG,Grünstadt

ISBN 3-527-30173-9

Dedication

This volume is dedicated to the memory of Dr. Ulrich de Haar, former Secretary ofthe Senate Commission for Water Research of the Deutsche Forschungsge-meinschaft (DFG), whose brilliance, enthusiasm and creativity permeated thiswork. Dr. de Haar’s extensive geochemical background and experience in water re-search was instrumental in bridging the often difficult gap between research ideasand funding mechanisms. We are also indebted to him for the project title ROSIG(Refraktäre Organische Substanzen in Gewässern).

V

Contents

VII

Preface XIX

List of Authors XXI

List of Abbreviations XXXI

1 Setting the Scene 11.1 The Relevance of Reference Materials � Isolation and General

Characterization 11.1.1 Introduction 11.1.2 Concept of Standard and Reference Material 31.1.3 Reference Materials in the ROSIG Priority Program 41.1.4 Experimental Details 51.1.4.1 Origin of the Samples 51.1.4.2 Isolation 71.1.4.3 Analytical Methods 101.1.5 Discussion 111.1.5.1 Mass Balance of the Isolates 131.1.5.2 Spectroscopic Characterization in the UV and Visible Range 161.1.5.3 Elemental Analysis 191.1.5.4 Proton and Complexation Capacities 251.1.5.5 Size-exclusion Chromatography 281.1.6 Conclusions 33

References 36Additional References for ROS Literature 38

1.2 Element Determination and its Quality Control in Fractions of RefractoryOrganic Substances and the Corresponding Original Water Samples 39

1.2.1 Introduction 391.2.2 Description of Analytical Methods 411.2.2.1 Sample Treatment 411.2.2.2 Flame and Graphite Atomic Absorption Spectrometry

(FAAS, GF-AAS) 421.2.2.3 Inductively Coupled Plasma Mass Spectrometry (ICP-MS) 42

VIII Contents

1.2.2.4 Inductively Coupled Plasma Isotope Dilution Mass Spectrometry(ICP-IDMS) 42

1.2.2.5 Instrumental Neutron Activation Analysis (INAA) 431.2.2.6 Total Reflection X-Ray Fluorescence Spectrometry (TXRF) 441.2.3 Results of the Interlaboratory Study 451.2.3.1 Element Concentrations in ROS Fractions and their Original

Samples 451.2.3.2 Relative Enrichment Factors of Elements in Separated FA and

HA Fractions 491.2.3.3 Quality Control of Analytical Data 49

References 53

2 Structural Investigations 552.1 Heavy Metal and Halogen Interactions with Fractions of Refractory

Organic Substances Separated by Size-exclusion Chromatography 552.1.1 Introduction 552.1.2 SEC/ICP-MS System and the Isotope Dilution Technique 562.1.3 Determination of Heavy Metal�ROS Complexes in Fractions Separated

by SEC 592.1.4 Correlation of Metal�ROS Complexes with other Analytical Data 622.1.5 Kinetically Stable Cr(III) and Co(III) Complexes of ROS 642.1.6 Determination of Halogen�ROS Compounds in Fractions separated

by SEC 682.1.7 Conclusions 71

References 72

2.2 Characterization of Refractory Organic Substances and their MetalSpecies by Combined Analytical Procedures 73

2.2.1 Introduction 732.2.2 Size Distribution and Stability of ROS�Metal Species Assessed by

Ultrafiltration, EDTA Exchange, and Atomic Spectrometry 742.2.3 On-site Characterization of Aquatic Refractory Organic Substances and

their Metal Species by Mobile Fractionation and ExchangeTechniques 79

2.2.4 Conclusions 86References 87

2.3 Application of Nuclear Magnetic Resonance Spectroscopy to StructuralInvestigations of Refractory Organic Substances � Principles andDefinitions 89

2.3.1 Potential of Nuclear Magnetic Resonance (NMR) Spectroscopy for ROSStructure Elucidation 89

2.3.2 Quantification of Solution-state and Solid-state NMR Spectra of ROS 912.3.3 Assignment of ROS NMR Signals 92

References 94

2.4 Structural Characterization of Refractory Organic Substances bySolid-state High-resolution 13C and 15N Nuclear Magnetic Resonance 96

2.4.1 Introduction 962.4.2 Pulse Sequences Used 972.4.2.1 Cross Polarization Magic Angle Spinning (CPMAS) 972.4.2.2 Magic Angle Spinning (MAS) or Bloch-decay Spectra 972.4.2.3 Dipolar Dephasing Techniques (DD Techniques) 972.4.3 Lake Hohloh Samples 982.4.3.1 13C Solid-state Spectra 982.4.3.2 Proton Solution Spectra 1032.4.3.3 15N CPMAS Spectra 1042.4.4 Comparison of Samples from Different Locations 1072.4.4.1 13C Solid-state Spectra 1072.4.4.2 15N CPMAS Spectra 1102.4.5 Conclusions 1122.4.5.1 Methods 1122.4.5.2 Structural Results 113

References 114

2.5 Quantification of Substructures of Refractory Organic Substances byMeans of Nuclear Magnetic Resonance 115

2.5.1 Introduction 1152.5.2 Experimental Details 1162.5.2.1 Nuclear Magnetic Resonance 1162.5.2.2 Multistage Ultrafiltration 1162.5.3 Results from Solution-state NMR investigations 1172.5.3.1 General Considerations 1172.5.3.2 Aliphatic Structural Moieties 1182.3.3.3 Aromatic Structural Moieties 1182.5.4 Combination of Multistage Ultrafiltration and Two-dimensional

HMQC NMR 1252.5.5 Conclusions 127

References 128

2.6 Nuclear Magnetic Resonance Spectroscopy Investigations of SilylatedRefractory Organic Substances 129

2.6.1 Introduction 1292.6.1.1 Functional Group Analysis in Refractory Organic Substances 1292.6.1.2 Silylation of Refractory Organic Substances 1302.6.2 Materials and Methods 1312.6.3 Two-dimensional NMR Spectroscopy of Silylated Natural Organic

Matter 1322.6.3.1 Comparative Analysis of 2D NMR Spectra; Visualization of Conformity

and Variation by Calculation of Minimum and Difference Spectra 136

IXContents

2.6.4 NMR Functional Group Analysis of FA and HA Fractions Isolated fromSamples of Different Origin 137

2.6.4.1 Comparison of BS1 FA, HO10 FA, FG1 FA, HO13 FAand SUW FA 137

2.6.4.2 Comparison of BS1 HA, HO13 HA, and SUW HA 1402.6.4.3 Comparison of Results from NMR Functional Group Analysis with Data

Obtained from Complementary Methods 1412.6.5 Conclusions 143

References 144

2.7 Isotopic Evidence for the Origin and Formation of Refractory OrganicSubstances 146

2.7.1 Variations of Stable Isotope Abundances in the Course of the Synthesisand Diagenesis of Biomass 146

2.7.2 Bulk Isotope Analysis 1492.7.3 Pyrolysis-Gas Chromatography/Mass Spectrometry-Isotope-ratio

Mass Spectrometry (Py-GC/MS-IRMS) 1502.7.4 Bulk Isotopic Analyses of the Reference Samples 1512.7.4.1 Differences Between the Isotope Content of the Reference Samples 1512.7.5 Formation of Refractory Organic Substances in a Peat Profile of the

Hohlohsee 1542.7.5.1 Bulk Isotope Analyses on Hohlohsee Samples 1552.7.5.2 �13C Values of Pyrolysis Products of Sphagnum Peat and DOM from

Bog Water 156References 160

2.8 Analytical Pyrolysis of Humic Substances and Dissolved Organic Matterin Water 163

2.8.1 Introduction 1632.8.2 Methods 1642.8.3 Characterization and Differentiation of Structural Properties 1652.8.4 Pyrolysis-Field Ionization Mass Spectrometry 1722.8.5 Structural Variations as a Result of Sampling Season and Year 1752.8.6 Quality Control and Changes During Sample Storage 1762.8.7 Correlations between 13C NMR and Py-FIMS 1792.8.8 Developments of Quantitative GC/MS 1802.8.8.1 Determination of Hexose Subunits of Cellulose by Py-GC/MS 1802.8.8.2 Determination of Solvent-extracted Fatty Acids by GC/MS 1832.8.9 Conclusions 184

References 186

2.9 Characterization of Refractory Organic Substances by HPLC/MS 1882.9.1 Introduction 1882.9.2 Experimental Details 1892.9.2.1 Mass Spectrometry 189

X Contents

XIContents

2.9.2.2 Size-exclusion Chromatography (SEC) 1912.9.3 Results and Discussion 1912.9.3.1 Mass Spectra of Refractory Organic Substances 1912.9.3.2 Size-exclusion Chromatography of Refractory Organic Substances 197

References 199

2.10 UV-Visible Spectroscopy and the Potential of Fluorescent Probes 2002.10.1 UV-Visible Absorption Spectroscopy 2002.10.2 Colorimetric Analysis 2052.10.3 Fluorescence of Refractory Organic Substances 2082.10.4 Aging of Refractory Organic Substances 2092.10.5 Complexing of Heavy Metals 211

References 214

2.11 Stationary and Time-resolved Fluorescence for Refractory OrganicSubstances Characterization 215

2.11.1 Introduction 2152.11.2 Experimental Details 2172.11.3 Results and Discussion 2182.11.3.1 Steady-state Fluorescence of Humic Substances 2182.11.3.2 Time-resolved Fluorescence of Humic Substances 2212.11.3.3 Fluorescence of Chemically Altered Humic Substances 2252.11.4 Conclusions 227

References 229

2.12 Structural Characterization of Refractory Organic Substancesby Pyrolysis-GC/FTIR 232

2.12.1 Introduction 2322.12.2 Experimental Details 2332.12.2.1 Sample Preparation 2332.12.2.2 Curie-point Pyrolysis 2332.12.2.3 Flash Pyrolysis 2342.12.2.4 GC/FTIR Spectroscopy 2342.12.3 Results and Discussion 236

References 238

2.13 X-ray Microscopy Studies of Refractory Organic Substances 2392.13.1 Introduction 2392.13.2 X-ray Microscopy 2402.13.3 Results 2432.13.3.1 Refractory Organic Substances in Soils 2432.13.3.2 Flocculation of Refractory Organic Substances by Cations 2442.13.3.3 Spectromicroscopy of Refractory Organic Substances 2442.13.4 Conclusions 246

References 247

2.14 Fractionation of Refractory Organic Substances by Electrophoresis 2492.14.1 Introduction 2492.14.2 Experimental Details 2502.14.3 Results and Discussion 2512.14.4 pH-dependence 2542.14.5 Electrophoresis with Organic Modifiers 2552.14.6 Micellar Electrokinetic Chromatography 2552.14.7 Electrophoresis in Sieving Media 2562.14.8 Isoelectric Focusing 2572.14.9 Characterization of Degradation Products of Refractory Organic

Substances 2582.14.10 Hydrothermolysis 2602.14.11 Pyrolysis 2612.14.12 Conclusions 262

References 263

2.15 Occurrence of Amino Acids, Carbohydrates, and Low-molecular-weightOrganic Acids in Refractory Organic Substances 264

2.15.1 Introduction 2652.15.2 Experimental Details 2672.15.2.1 Samples 2672.15.2.2 Determination of Amino Acids 2682.15.2.3 Determination of Carbohydrates 2682.15.2.4 Determination of Low-molecular-weight Organic Acids 2692.15.3 Results and Discussion 2702.15.3.1 Acid-hydrolyzable Amino Acids and Carbohydrates 2702.15.3.2 Enzymatically Hydrolyzable Amino Acids and Carbohydrates 2772.15.3.3 Alkaline-hydrolyzable Low-molecular-weight Organic Acids 2782.15.4 Conclusions 279

References 280

2.16 Serological Characterization of Refractory Organic Substancesby Serotyping 282

2.16.1 Introduction 2822.16.2 Experimental Details 2832.16.2.1 Material 2832.16.2.2 Antibody Production 2832.16.2.3 Precipitation Test 2842.16.2.4 Dot Blot Test 2842.16.2.5 Enzyme Immunoassay 2842.16.2.6 Fractionation of Refractory Organic Substances 2862.16.2.7 Serotyping 2872.16.3 Results 2872.16.3.1 Immunochemical Reactions of Refractory Organic Substances 2872.16.3.2 Fractionation of Refractory Organic Substances 293

XII Contents

2.16.3.3 Serotyping 2942.16.4 Discussion 297

References 301

2.17 Chemical and Spectroscopic Data of the Reference Samples �Comparison and Evaluation 302

2.17.1 Evaluation of the Isolation Procedure by Combining 13C NMR Spectrawith DOC Concentration Results 302

2.17.2 Comparison of NMR Results with Data from Other Methods ofStructural Characterization 304

2.17.2.1 NMR and UV Spectroscopy 3042.17.2.2 NMR and Analysis of the Content of Carbohydrates and Amino

Acids 3052.17.3 Complementary Results from Data Comparisons 3062.17.3.1 Carboxylic Groups Bound in Esters, Amides and Free Carboxylic Acid

Groups 3062.17.3.2 Phenolic Groups 3072.17.3.3 Exchangeable Hydrogens � Comparison of 1H and 13C NMR Data 3082.17.4 Conclusions 309

3 Biochemical and Biological Characterization 3113.1 Formation, Utilization, and Transformation of Some Refractory Organic

Substances by Aquatic Microorganisms 3113.1.1 Introduction 3113.1.2 Experimental Details 3123.1.2.1 Materials and Methods 3123.1.3 Results and Discussion 313

References 320

3.2 Effect of Microorganisms on the Formation and Transformationof Iodine Species of Refractory Organic Substances 321

3.2.1 Introduction 3213.2.2 Formation of Iodine�ROS Species from Inorganic Iodine 3223.2.3 Microbiological Influence on the Iodine�ROS Formation 3243.2.4 Transformation of Iodine�ROS Species by Aging 3263.2.5 Conclusion 328

References 329

3.3 The Influence of Refractory Organic Substances on BacterialColonization and Diversity Patterns 330

3.3.1 Introduction 3303.3.2 Materials and Methods 3313.3.2.1 Experimental Design 3313.3.2.2 Physicochemical Investigations 332

XIIIContents

XIV Contents

3.3.2.3 Microbiological Monitoring 3323.3.2.4 Molecular Methods 3333.3.2.5 Cluster Analysis 3333.3.2.6 Scanning Electron Microscopy 3343.3.3 Results 3343.3.3.1 Physicochemical Investigations 3343.3.3.2 Microbiological Monitoring 3363.3.3.3 Molecular Methods 3393.3.3.4 Cluster Analysis 3393.3.3.5 Scanning Electron Microscopy 3413.3.4 Discussion 342

References 344

3.4 Influence of Refractory Organic Substances on Enzyme Activity in-vivoand DNA Damage of Aquatic Microorganisms 346

3.4.1 Introduction 3463.4.2 Methods 3483.4.2.1 Detection and Evaluation of Microbial Enzyme Activity in Batch

Experiments 3483.4.2.2 Flow Injection Analysis of Microbial Enzyme Activity (FAME) 3483.4.2.3 Detection of Genotoxicity by Use of the umu Test 3493.4.2.4 Detection of Genotoxicity by Use of the Comet Assay 3503.4.2.5 Origin of Water Samples and Microorganisms 3503.4.2.6 Refractory Organic Substances 3503.4.2.7 Spiking of Refractory Organic Substances and Pollutants 3513.4.2.8 Influence of pH and Conductivity on Refractory Organic Substances 3513.4.3 Results 3513.4.3.1 Effects of Different Concentrations of Refractory Organic Substances

on Enzyme Activities in Batch Experiments 3513.4.3.2 Interaction of Refractory Organic Substances with Pollutants in Batch

Experiments 3533.4.3.3 Application of the FAME System Looking for Effects of Different

Refractory Organic Substances on Enzyme Activity 3543.4.3.4 Addition of HgCl2 and its Influence on Peroxidase Activity Using the

FAME System 3543.4.3.5 Detection of Genotoxic Effects by Use of the umu Test 3563.4.3.6 Detection of Genotoxic Effects by Use of the Comet Assay 3573.4.4 Discussion 3583.4.4.1 ROS as Nutrients or Toxicants or as a Trap for Nutrients and

Toxicants 3583.4.4.2 Interactions of ROS with Added Toxicants 3593.4.4.3 Do pH and Electrical Conductivity Influence the Effects of ROS and

Toxicants? 3593.4.4.4 Are ROS Genotoxic or Can They Mask Genotoxins? 359

References 360

3.5 Effects of Dissolved Organic Matter on the Bioconcentration of OrganicContaminants and on Reproduction in Aquatic Invertebrates 361

3.5.1 General Introduction 3613.5.2 Influence of Dissolved Organic Matter on the Bioconcentration of

Hydrophobic Organic Contaminants � Quantitative Aspects 3623.5.2.1 Introduction 3623.5.2.2 Influence of the Concentration of Dissolved Organic Matter 3633.5.2.3 Influence of the Origin of Dissolved Organic Matter 3653.5.2.4 Influence of the Type of Contaminant 3693.5.2.5 A General Model for Describing the Effect of Dissolved Organic Matter

on the Bioconcentration of Hydrophobic Organic Contaminants 3703.5.3 Effect of Refractory Organic Substances on the Reproduction of a

Nematode (Caenorhabditis elegans) 3723.5.3.1 Introduction 3723.5.3.2 Experimental Details 3733.5.3.3 Results 3733.5.3.4 Discussion 3763.5.3.5 Ecological Implications 377

References 378

4 Molecular Interactions 3834.1 Sorption of Dissolved Organic Matter on Soil Particles and its

Dependence on their Surface-charge Properties 3834.1.1 Introduction 3834.1.2 Materials and Methods 3844.1.2.1 Samples 3844.1.2.2 Methods 3854.1.3 Results and Discussion 3864.1.3.1 Charge of Dissolved Organic Matter 3864.1.3.2 Charge of the Soil Samples 3874.1.3.3 Sorption of Dissolved Organic Matter 3884.1.4 Conclusions 392

References 393

4.2 Dissolved Organic Carbon in Seepage Water �Production and Transformation During Soil Passage 394

4.2.1 Introduction 3954.2.2 Materials and Methods 3964.2.2.1 Experiment (1) � Laboratory Experiments with Forest Soils � DOC

Production as a Function of Litter Quality and Ash Addition 3964.2.2.2 Experiment (2) � DOC Production in Agricultural Soils � Continuous

Rye and Silage Maize Fields 3984.2.2.3 Experiment (3) � Field and Laboratory Studies with Forest Soils �

Transformation of DOC in Seepage Water During Passage ThroughSoil 399

XVContents

4.2.3 Results and Discussion 4004.2.3.1 Experiment (1) � Laboratory Experiments with Forest Soils � DOC

Production as a Function of Litter Quality and Ash Addition 4004.2.3.2 Experiment (2) � DOC Production in Agricultural Soils – Continuous

Rye and Silage Maize Fields 4034.2.3.3 Experiment (3) � Field and Laboratory Studies with Forest Soils �

Transformation of DOC in Seepage During Soil Passage 406References 409

4.3 Refractory Organic Substances in Aggregated Forest Soils � Retentionversus Translocation 411

4.3.1 Introduction 4114.3.2 Materials and Methods 4134.3.2.1 Sorption Experiments with Homogenized Soil Samples and Intact

Soil Cores 4134.3.2.2 Field Experiment on Dissolved Organic Matter in Slightly Developed,

Shallow Forest Soils 4164.3.3 Sorption of Dissolved Organic Matter in Aggregated Forest Soils 4174.3.3.1 Sorption of Dissolved Organic Matter to Homogenized Soil Samples

and Intact Cores 4174.3.3.2 Ionic Strength and Competitive Inorganic Anions Effects on the

Sorption of Dissolved Organic Matter 4214.3.3.3 Competition of Dissolved Organic Matter Constituents During

Sorption 4224.3.3.4 Dissolved Organic Matter Adsorption to Surface and Interior Material

of Soil Aggregates 4234.3.3.5 Sorption of Dissolved Organic Matter in Aggregated Soils �

Synopsis 4244.3.4 Dissolved Organic Matter in Slightly Developed,

Shallow Forest Soils 4244.3.4.1 Release of Dissolved Organic Matter from the Forest Floor 4244.3.4.2 Dissolved Organic Matter in the Mineral Soil 4264.3.4.3 Fluxes of Organic Matter and Nutrients � Rainstorm Events 4284.3.4.4 Chemical Characteristics of Leached Organic Matter 4294.3.4.5 Dissolved Organic Matter in Slightly Developed Shallow Soils �

Synopsis 430References 432

4.4 Refractory Organic Substances Derived from Organic Amendmentsin Soil � Formation, Translocation, and Interaction withXenobiotics 435

4.4.1 Introduction 4354.4.2 Material and Methods 4364.4.2.1 Crop Residues 4364.4.2.2 Xenobiotics 436

XVI Contents

4.4.2.3 Soil-column Experiments 4374.4.2.4 Sampling and Analysis 4384.4.2.5 Extraction of the Soil Samples 4394.4.2.6 Calorimetric Studies of the Interaction of Refractory Organic Substances

and Methabenzthiazuron 4404.4.2.7 Biosurfactant Properties of Refractory Organic Substances from Crop

Residues 4404.4.3 Results and Discussion 440

References 445

4.5 Analysis of the Binding of Amitrole and Anilazine to Aquatic andTerrestrial Refractory Organic Substances 446

4.5.1 Introduction 4464.5.2 Materials and Methods 4484.5.2.1 Chemicals 4484.5.2.2 Synthesis 4484.5.2.3 Refractory Organic Substances and Dissolved Organic Matter 4504.5.2.4 Chromatographic Methods 4514.5.2.5 NMR Spectroscopy 4524.5.2.6 Selective Reaction Monitoring (SRM) of Bound Residues 4524.5.2.7 Formation and Clean-up of Dissolved Organic Matter�Amitrole

Complexes 4524.5.2.8 Formation of Bound Residues 4544.5.2.9 Interaction of Anilazine with Dissolved Organic Matter 4544.5.2.10 Determination of kf Values According to the Freundlich Equation for

Amitrole and Dissolved Organic Matter 4554.5.2.11 Incubation of Amitrole with Five Different Soils 4554.5.2.12 Water � Sediment Systems with Anilazine and Dihydroxyanilazine

456

4.5.2.13 Water � Sediment Systems with Amitrole 4574.5.2.14 Quantification of Mineralization 4574.5.2.15 Release of Amitrole from DOM�Amitrole Complexes 4574.5.2.16 Extraction Methods for Purified Bound Residues 4584.5.2.17 Extraction of Soil Samples 4584.5.2.18 Silylation of Soil Samples 4584.5.2.19 Matrix Effects 4584.5.2.20 Biotests 4594.5.3 Results and Discussion 4594.5.3.1 Pesticides in Soil 4594.5.3.2 Pesticides in Water � Sediment Systems 4604.5.3.3 Pesticides and Dissolved Organic Matter 462

References 472

XVIIContents

4.6 Sorption and Chemical Reactions of Polycyclic Aromatic Hydrocarbonswith Dissolved Refractory Organic Substances and Related ModelPolymers 475

4.6.1 Introduction to the Problems 4754.6.2 The Solid-phase Microextraction Technique 4774.6.3 Kinetics of the Interaction of Dissolved Organic Matter and PAH 4844.6.4 Reversibility of Sorptive Interactions in the Aqueous Domain 4894.6.5 Relationships between Structural Properties of HOC and their Sorption

Affinity 4914.6.6 The Modified Flory-Huggins Concept 4924.6.7 Relationships between Properties of Dissolved Organic Matter and their

Sorption Potential 4974.6.8 Poly(Acrylic Acid) Esters as Surrogates of Dissolved Organic Matter

for Sorption Studies 4994.6.9 Long-term Interactions between Dissolved Organic Matter and PAH

under Strictly Abiotic Conditions 5064.6.10 Summary 510

References 512

4.7 Investigation of the Interactions between Polycyclic AromaticCompounds and Refractory Organic Substances with Stationary andTime-resolved Fluorescence and Absorption Spectroscopy 516

4.7.1 Introduction 5164.7.2 Experimental Details 5184.7.2.1 Samples 5184.7.2.2 UV-Visible and Fluorescence Spectroscopy 5194.7.3 Results and Discussion 5214.7.3.1 Absorption and Fluorescence Spectroscopic Characterization of

Refractory Organic Substances 5214.7.3.2 Interactions between Refractory Organic Substances and Pyrene in

Electronic Singlet States 5244.7.3.3 Interactions between Refractory Organic Substances and Polynuclear

Aromatic Components in Electronic Triplet States 5274.7.3.4 Strong Interactions of Refractory Organic Substances and Polycyclic

Aromatic Compounds � Triplet Quenching of Methylene Blue 5294.7.4 Conclusions 531

References 532

Index 535

Author index 545

XVIII Contents

Preface

The research described herein is the result of six years of cross-disciplinary scien-tific study of the properties of a uniform set of samples of refractory organic sub-stances (ROS). Sources sampled included a brown water lake, soil seepage water,ground water, a secondary effluent from a waste water treatment plant, and awaste water lake from a brown coal processing plant. Each source was sampled sea-sonally, and the ROS from each sample were isolated by use of a common tech-nique before being made available to the cross-disciplinary research teams.Thus, this work comprises a unique data base in addition to providing new insightsinto the structures, transformations, and interactions of ROS in the environment.

Most of the results were obtained in the course of a priority research programfunded by the Deutsche Forschungsgemeinschaft (DFG), to whom we are deeplyindebted. Two members of the DFG were particularly influential in obtaining sup-port for this program, Dr. Ulrich de Haar (see Dedication) and Professor Fritz Führ,a member of the Senate of the DFG, both of whom effectively advocated support ofthis research. Such advocacy was vital to this research, because economic condi-tions at the time placed constraints upon funding and competition among manydisciplines was intense.

Cross-disciplinary interest and support for this program was extraordinaryamong German scientists and without the generous and thoughtful time commit-ments of many individuals this work would not have been possible. Notable amongthese are the project referees who assisted in numerous proposal reviews andoffered valuable suggestions at annual workshops. This group was chaired withgentle authority by J. Schwoerbel, and included K.-H. Ballschmiter, U. Förstner,K. Haider, G. Matthess, K. Cammann, H. J. Knackmuss, J. Reichert, A. Schäfer,and C. J. Soeder. The principal contributors to the work of this project were, ofcourse, the individual scientists who wrote the proposals and wrote the chapterscontained herein.

Steady contributions were made by Axel Heidt and Dr. Gudrun Abbt-Braun inrunning the sampling campaigns and organizing two international symposia. Inaddition, Dr. Abbt-Braun handled the day-to-day tasks of project managementwith extraordinary effectiveness and sensitivity. Dagmar Schirner supervised the fi-nancial management of the project, and Dr. Claudia Boukis, Markus Delay, SabrinaLehringer, and Carola Neidhardt assisted in the organization of symposia and in

XIX

the onerous tasks associated with preparation of this book. I am grateful to otherpersonnel of the DFG for skillful administration of this project, notably Dr. UteWeber, Dr. Susanne Faulhaber, and Frau Michels. Special thanks are due to Dr.Pauly of Wiley-VCH, for expertise with this publication and for understandingthe tight schedules of research scientists, and to Professor Russ Christman foreditorial assistance.

As a part of our project, two international symposia were held at the Universityof Karlsruhe under the name of ROSE (Refractory Organic Substances in the En-vironment). In these settings, project scientists were able to discuss their findingswith each other and with invited experts from around the world. Two books ofextended abstracts of symposia presentations are available. In addition, a selectionof peer-reviewed papers appeared in two special issues of Acta hydrochimica ethydrobiologica.

On behalf of the project editorial group, I am happy to present this unique col-lection of scientific reports which represents the most important findings of ourproject. It is my hope that this book will find many interested readers and thatit will provide an encouraging platform for young scientists who wish to devotetheir careers to addressing the many fascinating and unanswered questionsabout the properties and roles of refractory organic substances in the environment.

Karlsruhe, Fritz FrimmelJanuary 2002

XX Preface

List of Authors

Dr. Gudrun Abbt-BraunEngler-Bunte-InstitutDivision of Water ChemistryUniversität KarlsruheEngler-Bunte-Ring 1D-76131 KarlsruheGermanyTel.: �49-(0)721-6084309Fax: �49-(0)721-6087051e-mail: gudrun.abbt-braun@ciw.uni-karlsruhe.de

Professor Fritz BeeseInstitute of Soil Scienceand Forest NutritionUniversität GöttingenBüsgenweg 2D-37077 GöttingenGermanyTel.: �49-(0)551-399765Fax: �49-(0)551-393310e-mail: fbeese@gwdg.de

Dipl.-Chem. Kai BehrensInstitut für Anorganischeund Angewandte ChemieUniversität HamburgMartin-Luther-King-Platz 6D-20146 HamburgGermany

Dipl.-Chem. Stefan BrandtIm Kleinfeld 9D-67227 Frankenthal

Dipl.-Chem. Thomas BrinkmannEngler-Bunte-InstitutDivision of Water ChemistryUniversität KarlsruheEngler-Bunte-Ring 1D-76131 KarlsruheGermany

Professor Joachim BuddrusInstitut für Spektrochemieund Angewandte Spektroskopie (ISAS)Universität DortmundBunsen-Kirchhoff-Str. 11Postfach 10 13 52D-44139 DortmundGermanyTel.: �49-(0)231-1392214Fax: �49-(0)231-1392120e-mail: buddrus@isas-dortmund.de

XXI

XXII List of Authors

Dr. Peter BurbaInstitut für Spektrochemieund Angewandte Spektroskopie (ISAS)Universität DortmundBunsen-Kirchhoff-Str. 11Postfach 10 13 52D-44139 DortmundGermanyTel.: �49-(0)231-1392181Fax: �49-(0)231-1392120e-mail: burba@isas-dortmund.de

Dr. B. Kent BurnisonCanada Center of Inland Waters867 Lakeshore RoadP. O. Box 5050Burlington Ontario L7R 4A6Canada

Dr. Frank ButtchereitInstitut für BodenkundeUniversität HannoverHerrenhäuser Str. 2D-30419 HannoverGermany

Dr. Harald ClausUmweltbundesamtDienstgebäude LangenPaul-Ehrlich-Str. 29D-63225 LangenGermany

Dr. Andrea DankwardtLehrstuhl für BotanikTechnische Universität MünchenAlte Akademie 12D-85350 FreisingGermany

Dr. Antony N. DaviesInstitut für Spektrochemieund Angewandte Spektroskopie (ISAS)Universität DortmundBunsen-Kirchhoff-Str. 11D-44139 DortmundGermanyTel.: �49-(0)231-1392218Fax: �49-(0)231-1392120e-mail: davies@isas-dortmund.de

Dr. Ralf DunkelStraße der Jugend 6AD-06179 LangenbogenGermany

Dipl.-Biol. Monika ErbesMeixlerstraße 4D-55120 MainzGermany

Professor Zdenek FilipUmweltbundesamtInstitut für Wasser-, Boden-und LufthygieneDienstgebäude LangenPaul-Ehrlich-Str. 29D-63225 LangenGermanyTel.: �49-(0)6103-704160Fax: �49-(0)6103-704147e-mail: zdenek.filip@uba.de

Professor Walter R. FischerInstitut für BodenkundeUniversität HannoverHerrenhäuser Str. 2D-30419 HannoverGermanyTel.: �49-(0)511-7622623Fax: �49-(0)511-7625559e-mail: fischer@ifbk.uni-hannover.de

XXIIIList of Authors

Dr. Heiner FlessaInstitute of Soil Scienceand Forest NutritionUniversität GöttingenBüsgenweg 2D-37077 GöttingenGermany

Dr. Dieter FreitagGSF-Forschungszentrum für Umweltund GesundheitInstitut für Ökologische ChemieSchulstraße 10D-85356 Freising/AttachingGermany

Professor Fritz H. FrimmelEngler-Bunte-InstitutDivision of Water ChemistryUniversität KarlsruheEngler-Bunte-Ring 1D-76131 KarlsruheGermanyTel.: �49-(0)721-6082580Fax: �49-(0)721-699154e-mail: fritz.frimmel@ciw.uni-karlsruhe.de

Professor Fritz FührForschungszentrum Jülich GmbHInstitut für RadioagronomiePostfach 19 13D-52425 JülichGermany

Dr. Anett GeorgiUFZ � Center for EnvironmentalResearch Leipzig-HallePermoserstr. 15D-04318 LeipzigGermany

Dr. Gerd GleixnerMax-Planck-Institut für BiogeochemiePostfach 100 164D-07701 JenaGermanyTel.: �49-(0)3641-643707Fax: �49-(0)3641-643710e-mail: gerd.gleixner@bgc-jena.mpg.de

Dr. Annette GünzlLeopoldstr. 8D-85399 HallbergmoosGermany

PD Dr. Georg GuggenbergerLehrstuhl für Bodenkundeund BodengeographieUniversität BayreuthD-95440 BayreuthGermany

Dr. Stephan HaiberInstitut für Spektrochemieund Angewandte Spektroskopie (ISAS)Universität DortmundBunsen-Kirchhoff-Str. 11Postfach 10 13 52D-44139 DortmundGermany

Dr. Markus Haitzer(since 2000) US Geological Survey3215 Marine StreetBoulderColorado 80303USA

Dipl.-Biol. Ines HeiberNaheweinstr. 37D-55452 LaubenheimGermany

XXIV List of Authors

Dr. Bálint HeilInstitute of Soil Scienceand Forest NutritionBüsgenweg 2D-37077 GöttingenGermany

Dr. Norbert HertkornGSF-Forschungszentrumfür Umwelt und GesundheitInstitut für Ökologische ChemieIngolstädter Landstr. 1D-85764 NeuherbergGermanyTel.: �49-(0)89-31874248Fax: �49-(0)89-31873371e-mail: hertkorn@gsf.de

Dipl.-Ing. Helmut HerzogInstitut für Spektrochemieund Angewandte Spektroskopie (ISAS)Universität DortmundBunsen-Kirchhoff-Str. 11Postfach 10 13 52D-44139 DortmundGermany

Professor Klaus Gustav HeumannInstitut für Anorganischeund Analytische ChemieJohannes-Gutenberg-Universität MainzDuesbergweg 10�14D-55099 MainzGermanyTel.: �49-(0)6131-3925882Fax: �49-(0)6131-3923369e-mail: heumann@mail.uni-mainz.de

Dr. Wieland HillInstitut für Spektrochemieund Angewandte Spektroskopie (ISAS)Universität DortmundBunsen-Kirchhoff-Str. 11D-44139 DortmundGermany

Professor Bertold HockLehrstuhl für BotanikTechnische Universität MünchenAlte Akademie 12D-85350 FreisingGermanyTel.: �49-(0)8161-713396Fax: �49-(0)8161-713044e-mail: hock@weihenstephan.de

Dr. Sebastian Höss(since 2000) Boden und SedimentBewertung(BSB)Thierschstr. 43D-80538 MünchenGermany

Dipl.-Biol. Svenja HonschoppParkstr.D-28209 BremenGermany

Dr. Jutta JahnelEngler-Bunte-InstitutDivision of Water ChemistryUniversität KarlsruheEngler-Bunte-Ring 1D-76131 KarlsruheGermany

Dr. Brit Jakubowski (née Aster)Institut für Spektrochemieund Angewandte Spektroskopie (ISAS)Universität DortmundBunsen-Kirchhoff-Str. 11Postfach 10 13 52D-44139 DortmundGermany

XXVList of Authors

Dr. Gerald JandlInstitut für BodenkundeUniversität RostockJustus-von-Liebig-Weg 6D-18051 RostockGermany

Oliver Janssen-WeetsPappelstr. 88D-28199 BremenGermany

Dr. Klaus KaiserLehrstuhl für Bodenkundeund BodengeographieUniversität BayreuthD-95440 BayreuthGermanyTel.: �49-(0)921-552178Fax: �49-(0)921-552246e-mail: klaus.kaiser@uni-bayreuth.de

Dipl.-Ing. agr. Matthias KaniaLehrstuhl für BotanikTechnische Universität MünchenAlte Akademie 12D-85350 FreisingGermany

Professor Martin KaupenjohannInstitut für Bodenkundeund Standortslehre (310)Fachgebiet BodenchemieUniversität HohenheimD-70599 StuttgartGermany

Professor Antonius KettrupGSE-Forschungszentrum für Umweltund GesundheitInstitut für Ökologische ChemieIngolstädter Landstr. 1D-85764 NeuherbergGermanyTel.: �49-(0)89-31874047Fax: �49-(0)89-31873371

Dr. Uwe KlausInstitut für UmweltforschungUniversität DortmundD-44221 DortmundGermany

Professor Arndt KnöchelInstitut für Anorganischeund Angewandte ChemieUniversität HamburgMartin-Luther-King-Platz 6D-20146 HamburgGermanyTel.: �49-(0)40-428383982Fax: �49-(0)40-428382893e-mail: knoechel@chemie.uni-hamburg.de

Dr. Thomas KochKurfürstenallee 29D-28211 BremenGermany

Professor Frank-Dieter KopinkeUFZ � Center for EnvironmentalResearch Leipzig-HallePermoserstr. 15D-04318 LeipzigGermanyTel.: �49-(0)341-2353264Fax: �49-(0)341-2352492e-mail: kopinke@san.ufz.de

XXVI List of Authors

Dr. Oliver KrachtMax-Planck-Institut für BiogeochemiePostfach 100 164D-07701 JenaGermany

Dipl.-Ing. Rüdiger KuckukInstitut für Spektrochemieund Angewandte Spektroskopie (ISAS)Universität DortmundBunsen-Kirchhoff-Str. 11D-44139 Dortmund

Professor Jussi KukkonenUniversität JoensuuDepartment of BiologyP. O. Box 11180101 JoensuuFinnland

Dr. Michael Kumke(since 2000) Institut für Physikalischeund Theoretische ChemieUniversität PotsdamKarl-Liebknecht-Str. 24�25D-14476 GolmGermanyTel.: �49-(0)331-9775209Fax: �49-(0)331-9775088e-mail: kumke@chem.uni-potsdam.de

Dr. Jörg LambertInstitut für Spektrochemieund Angewandte Spektroskopie (ISAS)Universität DortmundPostfach 10 13 52D-44139 DortmundGermany

Professor Heinz LanghalsInstitut für Organische ChemieUniversität MünchenButenandtstr. 5�13 (Haus F)D-81377 MünchenGermanyTel.: �49-(0)89-21807699Fax: �49-(0)89-21807640e-mail: langhals@lrz.uni-muenchen.de

Dr. Ulrich Lankes(since 2000) Engler-Bunte-InstitutDivision of Water ChemistryUniversität KarlsruheEngler-Bunte-Ring 1D-76131 KarlsruheGermany

Professor Peter LeinweberInstitut für BodenkundeUniversität RostockJustus-von-Liebig-Weg 6D-18051 RostockGermanyTel.: �49-(0)381-4982088Fax: �49-(0)381-4982159e-mail: peter.leinweber@agrarfak.uni-rostock.de

Professor Hans-Gerd LöhmannsröbenInstitut für Physikalischeund Theoretische ChemieUniversität PotsdamKarl-Liebknecht-Str. 24�25Haus 25D-14476 GolmGermanyTel.: �49-(0)331-9775222Fax: �49-(0)331-9775088e-mail: loeh@chem.uni-potsdam.de

XXVIIList of Authors

PD Dr. Bernard LudwigInstitute of Soil Scienceand Forest NutritionBüsgenweg 2D-37077 GöttingenGermanyTel.: �49-(0)551-393506e-mail: bludwig@gwdg.de

Professor Hans-Dietrich LüdemannInstitut für Biophysikund Physikalische BiochemieUniversität RegensburgD-93040 RegensburgGermanyTel.: �49-(0)941-9432492Fax: �49-(0)0941-9432479e-mail: hans-dietrich.luedemann@biologie.uni-regensburg.de

Dr. Katrin MackenzieUFZ � Center for EnvironmentalResearch Leipzig-HalleSektion SanierungsforschungPermoserstr. 15D-04318 LeipzigGermany

Dr. Gieselher MarxInstitut für Anorganischeund Analytische ChemieJohannes-Gutenberg-Universität MainzDuesbergweg 10–14D-55099 MainzGermany

Dr. Jörg MielckeEngler-Bunte-InstitutBereich WasserchemieUniversität KarlsruheEngler-Bunte-Ring 1D-76131 KarlsruheGermany

Dr. Rommy Müller-StarckLehrstuhl für BotanikTechnische Universität MünchenAlte Akademie 12D-85350 FreisingGermany

Professor Alexander NehrkornHeinrichsweg 1D-26209 HattenGermany

PD Dr. Ursula ObstForschungszentrum Karlsruhe GmbHITC-WGTPostfach 3640D-76021 KarlsruheGermanyTel.: �49-(0)7247-826806Fax: �49-(0)7247-826858e-mail: Ursula.Obst@itc-wgt.fzk.de

Dr. Thomas PfeiferInstitut für UmweltforschungUniversität DortmundD-44221 DortmundGermany

Dr. Thomas PützForschungszentrum Jülich GmbHInstitut für RadioagronomiePostfach 1913D-52425 JülichGermanyTel.: �49-(0)2461-616182Fax: �49-(0)2461-612518e-mail: t.puetz@fz-juelich.de

XXVIII List of Authors

Dr. Gunther RädlingerInstitut für Anorganischeund Analytische ChemieJohannes Gutenberg UniversitätMainzDuesbergweg 10�14D-55099 MainzGermany

Dr. Hans-Hermann RüttingerInstitut für Pharmazeutische ChemieFachbereich PharmazieMartin-Luther-Universität HalleWolfgang-Langenbeck-Str. 4D-06120 HalleGermanyTel.: �49-(0)345-5525182Fax: �49-(0)345-5527028e-mail: ruettinger@pharmazie.uni-halle.de

Dr. Claudia SchmidtInstitut für RöntgenphysikGeorg-August-Universität GöttingenGeiststr. 11D-37073 GöttingenGermany

Professor Hanns-Ludwig SchmidtPrielhofweg 2D-84036 LandshutGermanyTel.: �49-(0)871-44497Fax: �49-(0)871-44497

Professor Hans-Rolf SchultenInstitut für BodenkundeUniversität RostockJustus-von-Liebig-Weg 6D-18051 RostockGermanyTel./Fax: �49-(0)381-4982137e-mail: hans-rolf.schulten@agrarfak.uni-rostock.de

Dr. Ute Schultze (née Zimmermann)Institut für Physikalischeund Theoretische ChemieUniversität Erlangen-NürnbergEgerlandstr. 3D-91058 ErlangenGermany

Professor Ernst-Detlef SchulzeMax-Planck-Institut für BiogeochemiePostfach 100 164D-07701 JenaGermanyTel.: �49-(0)3641-643702Fax: �49-(0)3641-643710e-mail: eschulze@bgc-jena.mpg.de

Dr. Thomas SkrivanekInstitut für Physikalischeund Theoretische ChemieUniversität Erlangen-NürnbergEgerlandstr. 3D-91058 ErlangenGermany

Dipl.-Ing. Christian SpechtEngler-Bunte-InstitutDivision of Water ChemistryUniversität KarlsruheEngler-Bunte-Ring 1D-76131 KarlsruheGermany

Professor Michael SpitellerInstitut für UmweltforschungUniversität DortmundD-44221 DortmundGermanyTel.: �49-(0)231-7554080Fax.: �49-(0)231-7554085e-mail: spiteller@infu.uni-dortmund.de