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本書詳細介紹了一系列測定茶葉中農(nóng)藥殘留的快速高通量分析方法，具有高精度、高可靠性、高靈敏度的特點，適用廣泛。全書共7章：茶葉農(nóng)藥殘留分析基礎(chǔ)研究和檢測方法建立；不同樣品制備技術(shù)提取凈化效能對比研究；茶葉水化對農(nóng)藥多殘留方法效率的影響；茶葉農(nóng)藥殘留測定基質(zhì)效應及其補償作用研究；方法耐用性系統(tǒng)評價，誤差原因分析，關(guān)鍵控制點建立；茶葉陳化樣品和污染樣品農(nóng)藥降解規(guī)律研究；經(jīng)11個國家和地區(qū)的30個實驗室國際協(xié)同研究，建立茶葉中653種農(nóng)藥化學污染物高通量分析方法AOAC標準。

本書可作為科研單位、高等院校、質(zhì)檢機構(gòu)等各類專業(yè)技術(shù)人員從事食品安全、環(huán)境保護、農(nóng)業(yè)科技及農(nóng)藥開發(fā)等技術(shù)研究與應用的參考書，也可作為大學教學參考書。

In China, tea has a long history. It is a unique Chinese health drink which is popular with people all over the world. It is also an important cash crop and a traditional bulk export agricultural product in China. As tea plants like to grow in hot and humid environment, so their growth process is very susceptible to diseases and insect pests and weeds, and chemical pesticide spraying is the most effective solution so far. However, chemical pesticides leave undesirable residues in tea in accompany with the increment of yield and these residues will affect human health through the food chain. In addition, the developed countries and regions, such as the European Union, the United States, and Japan, are important importers of Chinese tea. The increasingly strict pesticide residue limit standards (MRLs) have become an important bottleneck in Chinas tea export. Therefore, from the protection of human health, to eliminate trade barriers, and to safeguard the perspective of international trade of tea, the thorough study of the detection technique of multipesticide residues in tea and the establishment of a highsensitivity, accuracy, selectivity, and throughput detection method have extremely important practical significance.

This book is the systematic summarization of the research of our team on the detection technology theory and application of the multipesticide residues in tea products in last 10 years. The book describes in detail the high-throughput analytical techniques for 653 multiclasses and multikinds of pesticide residues and chemical pollutants in tea, while the mainstream and advanced detection means such as gas chromatographymass spectrometry (GCMS), gas chromatographytandem mass spectrometry (GC-MS/MS), liquid chromatographytandem mass spectrometry (LCMS/MS), gas chromatography quadrupole tandem time-of-flight mass spectrometry (GC-Q-TOF), and liquid chromatography quadrupole tandem timeof- flight mass spectrometry (LC-Q-TOF) are concerned.

Compared with other plant-derived agricultural products such as vegetables and fruits, the composition of tea is extremely complex for it is rich in physiologically active compounds such as polyphenols, alkaloids, pigments, aromatic substances, amino acids, vitamins, and so on, and the composition varies greatly with processing procedure, so it is considerable difficult for the detection of multiclasses and multikinds of trace pesticide residues in tea.

Considering multiclasses and multikinds of pesticides may exist in different tea matrix, deep research on the comparison of the efficiency of various sample preparation technology and sample purification technology is introduced in this book, and the Cleanert TPT tea purification column, which has their own independent intellectual property rights, was developed. The simultaneous extraction and purification pretreatment method was established, while the matrix effect and its compensation of 28 different areas and species tea was deeply studied and the solution, using protecting agent for the compensation of matrix effect, was put forward.

In view of peoples drinking habits, the comparison of two extraction methods, hydration method and nonhydration method, on the extraction efficiency, purification effect of residual pesticides in tea, and the applicability of different pesticides was conducted in-depth. At the same time, each component of the uncertainty is discussed in detail, and the measurement uncertainty is regarded as evaluation criteria of the method.

In addition, in order to further evaluate the repeatability, reproducibility, and durability of the method, the authors also carried out the evaluation of the ruggedness of the method for up to 3 months. At the same time, concrete analysis has been made to error types from different analytical stages and their causes, while elaborated discussions have been conducted on the key control points in different stages of the method to ensure the accurate and reliable of the method.

Furthermore, the dynamics of pesticide degradation of aged oolong tea samples and contaminated green tea samples at different field test and room temperature storage conditions using the high-throughput detection method established were carried by the authors work team. The stability of the contaminated tea samples within 3 months was also investigated, so as the degradation rule of pesticides in tea can be obtained. So, the prediction of the content of pesticide residues using degradation dynamic equation can be accomplished and the results are basically consistent with the measured values. This has a very important significance in offering guidance to tea growers to the applications of pesticide and to the import and export trade of tea.

Based on the aforementioned series of important research, the international collaborative research, participated by a total of 30 laboratories from 11 countries and regions, was organized and implemented by our team as the Study Director of AOAC international and the purpose of the research is the evaluation of the reproducibility of the analysis method for the 653 kinds of pesticide residues in tea and investigation whether the method can meet the requirements of AOAC international official method. This is overall comprehensive collaborative research of AOAC international on the detection technical standards of pesticide residue in tea, meanwhile the detailed report of the study is also included in this book.

In short, a series of important studies about the detection technology of multipesticide residues in tea are involved in this book and they are the deepest and most comprehensive summary of high-throughput detection of multipesticide residues in tea till now. The method, high-throughput detection of 653 multiclasses and multikinds of pesticide residues and chemical pollutants using GCMS, GCMS/MS, and LCMS/MS, was selected as AOAC international priority research projects in 2010, and the method passed the AOAC international collaborative study and was approved as AOAC international official method for the detection of pesticide residues in tea (AOAC Official Method 2014.09) in 2014. Nevertheless, due to the limitations of the level, there may be unavoidable errors. We would kindly ask the users of this publication to provide feedback to the authors so that subsequent editions may be improved upon.

Guo-Fang Pang

October 10, 2017

Preface xi

Introduction xiii

1 Fundamental Research: Analytical

Methods for Multiresidues in Tea

1.1 Simultaneous Determination of 653

Pesticide Residues in Teas by Solid

Phase Extraction (SPE) with Gas

ChromatographyMass Spectrometry

(GCMS) and Liquid Chromatography

Tandem Mass Spectrometry

(LCMS/MS) 3

1.1.1 Introduction 3

1.1.2 Experimental 4

1.1.2.1 Reagents and Materials 4

1.1.2.2 Apparatus 5

1.1.2.3 Extraction 5

1.1.2.4 Cleanup 5

1.1.2.5 Determination 5

1.1.3 Results and Discussion 6

1.1.3.1 Optimization of Gas

ChromatographyMass Spectrometry

Conditions and Selection of Pesticide

Varieties Suitable for Analysis 6

1.1.3.2 Optimization of Liquid

ChromatographyTandem Mass

Spectrometry Conditions and

Selection of Pesticide Varieties

Suitable for Analysis 6

1.1.3.3 Optimization of Sample Extraction

Conditions 10

1.1.3.4 Optimization of Sample Cleanup

Conditions 12

1.1.3.5 Evaluation of Method Effi ciency

LOD, LOQ, Recovery and Relative

Standard Deviations of Both Gas

ChromatographyMass Spectrometry

and Liquid ChromatographyTandem

Mass Spectrometry 48

References 90

1.2 Research on the Evaluation of the

Effectiveness of the Nontargeting,

High-Throughput Method for the

Detection of 494 Pesticides Residue

Using GC-Q-TOF/MS Technique

in Tea 91

1.2.1 Introduction 91

1.2.2 Experimental 92

1.2.2.1 Reagents, Standard, and Materials 92

1.2.2.2 Equipment 93

1.2.2.3 Condition of Gas Chromatography and Mass Spectrometry 93

1.2.2.4 Pretreatment Methods for the Screening Analysis of Tea Samples 93

1.2.3 Establishment of Database 93

1.2.4 Evaluation of the Effi ciency of the Method 93

1.2.4.1 Qualitative Analysis 94

1.2.4.2 Screening Limit 95

1.2.4.3 Recovery (Rec. 60%120%&RSD < 20%) 111

1.2.4.4 Comprehensive Analysis

(Comparative Analysis the Infl uence

of Matrix on the Screening Limit

and Recovery of Pesticides) 113

1.2.5 Conclusion 114

1.3 A Study of Effi ciency Evaluation for

Nontarget and High-Throughput

Screening of 556 Pesticides Residues

by LC-Q-TOF/MS 115

1.3.1 Introduction 115

1.3.2 Experimental 116

1.3.2.1 Reagents and Materials 116

1.3.2.2 HPLC Conditions 116

1.3.2.3 Mass Spectrometry Conditions 116

1.3.2.4 Sample Preparation 117

1.3.3 Creation of the Pesticide Database 117

1.3.4 Method Evaluation 117

1.3.4.1 Qualitative Analysis 117

1.3.4.2 Screening Limits 117

1.3.4.3 Accuracy and Precision 119

1.3.4.4 Comprehensive Analysis of Screening Limits and Recoveries 124

1.3.5 Conclusions 124

1.4 A Study of Effi ciency Evaluation for

Nontarget Screening of 1050 Pesticide

By TOFMS and

LCQTOF/MS 143

1.4.1 Introduction 143

1.4.2 Experimental 143

1.4.2.1 Reagents and Materials 143

1.4.2.2 Standard Solution Preparation 144

1.4.2.3 Sample Collection and Pretreatment 144

1.4.2.4 Instrumental Analysis 144

1.4.2.5 Creation of the Pesticides Database 144

1.4.2.6 Method Evaluation 145

1.4.3 Results and Discussion 145

1.4.3.1 Screening Limits Analysis 145

1.4.3.2 Accuracy and Precision 147

1.4.3.3 Comparison of the Common-Detected Pesticides 150

1.4.4 Conclusion 152

2 Comparative Study of Extraction

and Cleanup Efficiencies of Residue

Pesticides in Tea

2.1 Review of Sample Preparation

Techniques for Residue Pesticides

in Tea 223

2.1.1 Review of Solid Phase Extraction Technique 223

2.1.2 Review of QuEChERS Method 224

References 225

2.2 Comparative Study of Extraction

Effi ciencies of the Three Sample

Preparation Techniques 227

2.2.1 Introduction 227

2.2.2 Experimental 228

2.2.2.1 Reagents and Materials 228

2.2.2.2 Apparatus 228

2.2.2.3 Experimental Method 228

2.2.3 Preparing Pesticides Incurred Tea

Samples and Deciding the Precipitated

Content of the Target Pesticide 229

2.2.4 Optimization of Experimental Conditions for Method-3 229

2.2.5 Optimization of the Evaporation

Temperatures and Degrees of the

Sample Solutions for Three Methods 245

2.2.5.1 Optimization Selection

of Temperatures of Rotary

Evaporation 245

2.2.5.2 Optimum Selection of the Degree of Rotary Evaporation 245

2.2.6 Stage I: Evaluation on the Extraction

Effi ciencies of the Multiresidue

Pesticides in Green Tea Samples

by Method-1, Method-2, and

Method-3 246

2.2.6.1 Test Data Analysis 246

2.2.6.2 Analysis of Experimental Phenomenon 246

2.2.7 Stage II: Evaluation on the Extraction

Effi ciencies of the Multiresidue

Pesticides in Green Tea and Oolong

Tea Samples by Method-1 and

Method-2 256

2.2.8 Stage III: Evaluation on the Extraction

Effi ciencies of Green Tea and Oolong

Tea Sample the Incurred 201 Pesticides

After 165 days by Method-1 and

Method-2 266

2.2.8.1 Test Data Analysis 266

2.2.8.2 Test Phenomenon Analysis 266

2.2.9 Conclusions 277

2.3 The Evaluation of the Cleanup

Effi ciency of SPE Cartridge Newly

Developed for Multiresidues

in Tea 279

2.3.1 Introduction 279

2.3.2 Reagents and Materials 280

2.3.3 Apparatus 280

2.3.4 Experimental 280

2.3.5 Stage I: The Comparative Test of the

Cleanup Effi ciencies of 12 SPE

Combined Cleanup Cartridges

Against the Spiked 84 Pesticides

in Tea 280

2.3.6 Stage II: The Comparative Test on

Cleanup Effi ciencies of Spiked 201

Pesticides in Tea with 4# Envi-Carb PSA

Tandem Cartridge and Cleanert TPT 281

2.3.7 Stage III: The Comparative Test on the

Cleanup Efficiencies of Green Tea and

Oolong Tea Youden Pair Samples

Incurred by 201 Pesticides with

Cleanert TPT Cartridge and 4# Envi-

Carb PSA Combined Cartridge 287

2.3.7.1 Comparison of the Determined

Values of Target Pesticide

Content 287

2.3.7.2 Comparison of the Reproducibility

of the Target Pesticide

Determination 295

2.3.8 Conclusions 329

References 464

3 Study on the Influences of Tea

Hydration for the Method Efficiency

and Uncertainty Evaluation of

the Determination of Pesticide

Multiresidues in Tea Using Three

Sample Preparation Methods/

GCMS/MS

3.1 A Comparative Study on the Influences

of Tea Hydration for the Method

Efficiency of Pesticide Multiresidues

Using Three Sample Preparation

Methods/GCMS/MS 467

3.1.1 Introduction 467

3.1.2 Experimental Method 469

3.1.2.1 Three Different Sample Preparation Methods 469

3.1.2.2 Aged Sample Preparation 469

3.1.2.3 Incurred Sample Preparation 469

3.1.3 Experimental Results and Discussion 470

3.1.3.1 Comparison of Accuracy and

Precision for Fortification

Recovery Experiments by the

Three Methods 470

3.1.3.2 Correlation Comparison of the

Three Methods Extraction Efficiency

with Pesticide log Kow Values 471

3.1.3.3 General Analysis of Method Applicability 474

3.1.3.4 Comparison of the Cleanup Efficiency for the Three Methods 478

3.1.4 Conclusions 478

References 482

3.2 Uncertainty Evaluation of the

Determination of Multipesticide

Residues in Tea by Gas

ChromatographyTandem Mass

Spectrometry Coupled with Three

Different Pretreatment Methods 483

3.2.1 Introduction 483

3.2.2 Reagents and Materials 484

3.2.3 Apparatus 485

3.2.4 Experimental Method 485

3.2.4.1 Method 1 (M1) 485

3.2.4.2 Method 2 (M2) 486

3.2.4.3 Method 3 (M3) 486

3.2.5 Estimation of Uncertainty 487

3.2.5.1 Identification of Uncertainty Sources 487

3.2.5.2 Calculation of Standard Uncertainty 489

3.2.6 Combined Uncertainty 497

3.2.7 Expanded Uncertainty 497

3.2.8 Result and Discussion 497

3.2.8.1 Uncertainty of M1 497

3.2.8.2 Uncertainty of M2 499

3.2.8.3 Uncertainty of M3 499

3.2.8.4 Comparison of the Three Methods for Uncertainty Analysis 500

3.2.9 Conclusion 503

References 504

4 Matrix Effect for Determination of Pesticide Residues in Tea

4.1 Review 505

4.1.1 Current Situation of Matrix Effect 505

4.1.2 Cluster Analysis and Its Application in Chemical Analysis 509

4.2 Study on the Matrix Effects of Different

Tea Varieties from Different

Producing Areas 510

4.2.1 Introduction 510

4.2.2 Experimental Materials 510

4.2.3 Preparation of a Test Sample 511

4.2.4 Instrumental Analysis Condition 512

4.2.5 Qualitative Analysis of the Sample and Data Processing 513

4.2.6 Matrix Effect Evaluation of GCMS 513

4.2.7 Evaluation of the Matrix Effect in Determination of GCMS/MS 517

4.2.8 Evaluation of the Matrix Effect in Determination of LCMS/MS 520

4.3 Compensation for the Matrix

Effects in the Gas Chromatography

Mass Spectrometry Analysis of 186

Pesticides in Tea Matrices Using

Analyte Protectants 523

4.3.1 Introduction 523

4.3.2 Experiment and Material 523

4.3.3 Experimental Method 524

4.3.4 Effect of Different Analyte Protectants on Solvent Standards 524

4.3.5 Effects of Different Analyte Protectant Combinations on Tea 527

4.3.6 Effectiveness Evaluation of Analyte

Protectant Compensation Matrix

Effect Method 530

4.4 Compensation for the Matrix

Effects in the Gas Chromatography

Mass Spectrometry Analysis of 205

Pesticides in Tea Matrices 533

4.4.1 Introduction 533

4.4.2 Experimental Materials 533

4.4.3 Experimental Method 533

4.4.4 Effect of Different Analyte Protectants on Solvent Standards 534

4.4.5 Effects of Different Analyte Protectant Combinations on Tea 534

4.4.6 Effectiveness Evaluation of

Analyte Protectant Compensation

Matrix Effect Method 537

References 541

5 The Evaluation of the Ruggedness

of the Method, Error Analysis, and

the Key Control Points of

the Method

5.1 Introduction 567

5.2 Experiment 568

5.2.1 Reagents and Materials 568

5.2.2 Apparatus 569

5.2.3 Preparing Pesticides-Aged Tea Samples

and the Evaluation of Uniformity for

Preparation 569

5.2.4 Experimental Method 569

5.3 Evaluation of the Ruggedness of the Method 570

5.3.1 Reproducibility Comparison

of the First Determination

RSD of Parallel Samples from

Pesticide-Aged Youden Pair

Samples with 18 Circulative

Determination Values 570

5.3.2 Evaluation of the Ruggedness of the Method

Based on the RSD of the Youden Pair Ratios

from the 3-Month Circulative

Experiment 571

5.4 Error Analysis and Key Control Points 573

5.4.1 Error Analysis of Sample Pretreatment and Key Control Points 573

5.4.2 The Effects of Equipment Status

on Test Results (Error Analysis)

and Critical Control Point 574

5.4.3 Chromatographic Resolution (Error Analysis)and Key Control Points 578

5.5 Conclusions 593

References 783

6 Study on the Degradation of Pesticide Residues in Tea

6.1 Study on the Degradation of 271

Pesticide Residues in Aged Oolong Tea

by Gas Chromatography-Tandem Mass

Spectrometry and Its Application in

Predicting the Residue Concentrations of

Target Pesticides 787

6.1.1 Introduction 787

6.1.2 Reagents and Material 788

6.1.2.1 Reagents 788

6.1.2.2 Material 788

6.1.3 Apparatus and Conditions 788

6.1.4 Sample Pretreatment 789

6.1.4.1 The Preparation Procedures for Aged Tea Samples 789

6.1.4.2 Extraction 789

6.1.4.3 Cleanup 789

6.1.5 The Degradation of 271 Pesticides in Aged Oolong Tea 789

6.1.5.1 Degradation Trend A 790

6.1.5.2 Degradation Trend B 791

6.1.5.3 Degradation Trend C 791

6.1.5.4 Degradation Trend D 792

6.1.5.5 Degradation Trend E 792

6.1.5.6 Degradation Trend F 792

6.1.6 Pesticides in Different Classes 793

6.1.7 The Practical Application of Degradation Regularity 794

6.1.7.1 The Degradation Regularity of 20 Representative Pesticides 794

6.1.7.2 The Prediction of Pesticide Residues in Aged Oolong Tea 795

6.1.8 Conclusions 797

References 799

6.2 A GCMS, GCMS/MS and LCMS/MS

Study of the Degradation Profiles of

Pesticide Residues in Green Tea 801

6.2.1 Introduction 801

6.2.2 Reagents and Materials 802

6.2.3 Design of Field Trials 802

6.2.3.1 Selection of Pesticide Varieties 802

6.2.3.2 Selection and Planning of Tea Plantations 802

6.2.3.3 Pesticide Application and Incurred Tea Sample Preparations 803

6.2.4 Analytical method 804

6.2.4.1 Extraction and Cleanup 804

6.2.4.2 Apparatus Conditions 804

6.2.5 The Pesticide Degradation Profiles within 30 days for the Field Trials 805

6.2.6 Study on Pesticide Degradation by Three Techniques 805

6.2.7 Comparison of the Pesticides Degradation

to MRL Actual Values in Field Trials with

Power Function Equations Predicted Values

Detected by Three Techniques 807

6.2.8 Stability Study on the Pesticides in Incurred

Tea Samples at Room Temperature Storage

Conditions (1825C) 809

6.2.9 Conclusions 810

6.2.10 Acknowledgments 810

References 810

7 High-Throughput Analytical

Techniques for Determination of

Residues of 653 Multiclass Pesticides

and Chemical Pollutants in Tea by

GCMS, GCMS/MS and LCMS/MS:

Collaborative Study

7.1 The Pre-Collaborative Study of AOAC Method Efficiency Evaluation 813

7.1.1 Introduction 813

7.1.2 Design of Collaborative Study 814

7.1.2.1 Selection of Representative Pesticides 814

7.1.2.2 Preparation and Homogeneity Evaluation of Aged Samples 815

7.1.2.3 Types and Quantity of Collaborative Study Samples 815

7.1.2.4 Organizing and Implementing of Collaborative Study 815

7.1.2.5 System Suitability Check 816

7.1.2.6 Requirement for Participating in Collaborative Laboratories 816

7.1.3 Experiment 816

7.1.3.1 Instruments and Reagents 816

7.1.3.2 Preparation of Standard Solutions 816

7.1.3.3 Sample Preparation 817

7.1.3.4 Test Condition 818

7.1.4 Method Efficiency Acceptance Criteria 819

7.1.4.1 General Principle 819

7.1.4.2 Standard Curve Linear Correlation Coefficient Acceptance Criteria 820

7.1.4.3 Target Pesticide Ion Abundance Ratio Criteria 820

7.1.4.4 Recovery, RSDr, and RSDR Acceptance Criteria 821

7.1.4.5 Outliers Elimination Via Grubbs and Dixon Double Checking 821

7.1.5 Result and Discussion 821

7.1.5.1 The Labs That Deviated

from the Operational Method

Have Been Eliminated 821

7.1.5.2 Eliminations of Outliers Via

Double-Checking of Grubbs

and Dixon 821

7.1.5.3 Method Efficiency 821

7.1.5.4 Qualification and Quantification 825

7.1.5.5 Error Analysis and Tracing 829

7.1.6 Conclusions 832

References 832

7.2 Collaborative Study 835

7.2.1 Introduction 835

7.2.2 Collaborative Study Protocol 837

7.2.2.1 Need/Purpose 837

7.2.2.2 Scope/Applicability 837

7.2.2.3 Materials/Matrices 837

7.2.2.4 Concentrations/Ranges of Analytes 837

7.2.2.5 Spiked Samples of Blind Duplication 837

7.2.2.6 Naturally Incurred Residues Matrices 837

7.2.2.7 Sample Preparation and Handing Homogeneity 837

7.2.2.8 Quality Assurance 839

7.2.3 Method Performance Parameters for Single Laboratory 840

7.2.3.1 Limit of Detection and Limit of Quantification 840

7.2.3.2 Accuracy and Precision for a Single Laboratory 841

7.2.3.3 Linearity of 653 Pesticides for Single Laboratory 841

7.2.3.4 Ruggedness of Multiresidue Method 841

7.2.4 Collaborators 841

7.2.4.1 Instrument and Materials Used by Collaborators 841

7.2.5 AOAC Official Method: High-Throughput

Analytical Techniques for Determination of

Residues of 653 Multiclass Pesticides and

Chemical Pollutants in Tea by GCMS, GC

MS/MS, and LCMS/MS 844

7.2.5.1 Principle 844

7.2.5.2 Apparatus and Conditions 844

7.2.5.3 Reagents and Materials 847

7.2.5.4 Preparation of Standard Solutions 849

7.2.5.5 Extraction and Cleanup Procedure 849

7.2.5.6 Qualitative and Quantitative 850

7.2.6 Results and Discussion 851

7.2.6.1 Evaluation of Collaborative Study Results 851

7.2.6.2 The Method Efficiency of the

Fortification Samples by GCMS,

GCMS/MS, and LCMS/MS 853

7.2.6.3 The Method Efficiency of the Aged

Samples by GCMS, GCMS/MS,

and LCMS/MS 859

7.2.6.4 The Method Efficiency of the

Incurred Samples by GCMS,

GCMS/MS, and LCMS/MS 864

7.2.7 Qualification and Quantification 865

7.2.7.1 Qualification of Target Pesticides 865

7.2.7.2 Quantification of Target Pesticides 865

7.2.8 Error Analysis and Traceability 871

7.2.8.1 GCMS Data Error Analysis and Traceability 871

7.2.8.2 GCMS/MS Data Error Analysis and Traceability 871

7.2.8.3 LCMS/MS Data Error Analysis and Traceability 875

7.2.9 Collaborators Comments on Method 877

7.2.9.1 Collaborators Comments on GCMS 877

7.2.9.2 Collaborators Comments on GCMS/MS 877

7.2.9.3 Collaborators Comments on LCMS/MS 878

7.2.10 Conclusions 879

References 880

Index 883

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