Pesticide Analytical Manual Vol. I Regulatory Operations eneral Analytical Operations and Information Chapter 3 Chapter 4 Multiclass Selective MRMS MRMS Chapter 5 GLC Chapter 6 HPLC Table of Contents 501: General Information 501A: Principles 501-11/94 01 B: Equipment for glC Gas Chromatographic Components 501-1 Other A. 501-21/94 R 501-31/94 501 C: Residue Methodology for GLC Determination 501-9 1/ 1/94 Reagent Blanks 501-5 Choice of soly 501 D: Injection Techniques 501-6 Manual injection 5016 1/94 0171/94 01 E: Reference standards 501-81/94 Refe 501-8 l/94 m60 500-1
Transmittal No. 96-E1 (9/96) Form FDA 2905a (6/92) Pesticide Analytical Manual Vol. I SECTION 500 500–1 Table of Contents page date 501: General Information 501 A: Principles 501-1 1/94 501 B: Equipment for GLC 501-1 1/94 Gas Chromatographic Components 501-1 1/94 Other Apparatus 501-2 1/94 Reagents and Gases 501-3 1/94 501 C: Residue Methodology for GLC Determination 501-3 1/94 Cleanup 501-4 1/94 Reagent Blanks 501-5 1/94 Choice of Solvent 501-6 1/94 501 D: Injection Techniques 501-6 1/94 Manual Injection 501-6 1/94 Autoinjectors 501-7 1/94 501 E: Reference Standards 501-8 1/94 References 501-8 1/94 Chapter 1 Regulatory Operations Chapter 2 General Analytical Operations and Information Chapter 3 Multiclass MRMs Chapter 5 GLC Chapter 4 Selective MRMs Chapter 6 HPLC
Pesticide Analytical Manual Vol I 502: Columns 50211/9 Column Specifications 502-1 1/94 Column parameters 50221/94 502 B: Packed Columns 502-6 /94 Components of Packed Columns 502-6 1/94 Preparation of Packed Columns 5028 1/94 Installation of Packed column 502-9 1/94 Conditioning of Packed columns 502101/94 Rejuvenation of Packed Columns 502-101/94 Criteria for Acceptable Packed Columns 502-111/94 for Packed Colume ating Procedures 502-121/94 502 C: Open Tubular Capillary Columns 502-131/9 Column Description 502141/94 502-161/94 Capillary column Systems 502-161/94 Installation and Conditioning of 502201/94 Capillary columns Rejuvenation of capillary columns 502211/94 Recommended Operating Procedure for 502-22 1/94 Wide bore Columns(Isothermal) Apparatus and Reagents 502221/94 System Startup and Inspection 502251/94 References 502251/94 Detectors 503 A: Introduction 50311/94 Definitions of Detector Characteristics 503-11/94 503 B: Electron Capture Detector 50321/94 Principles 50331/94 Design 50331/94 pparatus and reagents 50331/94 Detector Characteristics 50341/94 Other Influences on Detector Performance 503-7 1/94 d Operating Procedures /9 503 C: Flame Photometric Detector 50391/94 Principle 50391/94 Design 503-91/94 Apparatus and Reagents 503101/94 Detector Characteristics 503101/94 Other Influences on Detector Performance 508-11 1/94 Procedu 503121/9 Troubleshooting 503131/94 503 D: Electrolytic Conductivity Detector 503-141/94 Principles 503141/94 5002 Form FDA 2905a(6/92)
Transmittal No. 96-E1 (9/96) 500–2 Form FDA 2905a (6/92) SECTION 500 Pesticide Analytical Manual Vol. I page date 502: Columns 502 A: Introduction 502-1 1/94 Column Specifications 502-1 1/94 Column Parameters 502-2 1/94 502 B: Packed Columns 502-6 1/94 Components of Packed Columns 502-6 1/94 Preparation of Packed Columns 502-8 1/94 Installation of Packed Columns 502-9 1/94 Conditioning of Packed Columns 502-10 1/94 Rejuvenation of Packed Columns 502-10 1/94 Criteria for Acceptable Packed Columns 502-11 1/94 Recommended Operating Procedures 502-12 1/94 for Packed Columns 502 C: Open Tubular Capillary Columns 502-13 1/94 Column Description 502-14 1/94 Injection onto Capillary Columns 502-16 1/94 Capillary Column Systems 502-16 1/94 Installation and Conditioning of 502-20 1/94 Capillary Columns Rejuvenation of Capillary Columns 502-21 1/94 Recommended Operating Procedure for 502-22 1/94 Wide Bore Columns (Isothermal) Apparatus and Reagents 502-22 1/94 System Startup and Inspection 502-25 1/94 References 502-25 1/94 503: Detectors 503 A: Introduction 503-1 1/94 Definitions of Detector Characteristics 503-1 1/94 503 B: Electron Capture Detector 503-2 1/94 Principles 503-3 1/94 Design 503-3 1/94 Apparatus and Reagents 503-3 1/94 Detector Characteristics 503-4 1/94 Other Influences on Detector Performance 503-7 1/94 Recommended Operating Procedures 503-8 1/94 503 C: Flame Photometric Detector 503-9 1/94 Principles 503-9 1/94 Design 503-9 1/94 Apparatus and Reagents 503-10 1/94 Detector Characteristics 503-10 1/94 Other Influences on Detector Performance 503-11 1/94 Recommended Operating Procedures 503-12 1/94 Troubleshooting 503-13 1/94 503 D: Electrolytic Conductivity Detector 503-14 1/94 Principles 503-14 1/94
Pesticide Analytical Manual Vol. I Design 503-141/94 EICD-X 503-161/94 Principles 503-161/94 Apparatus and ro 503-161/94 Detector characteristics 503-171/94 Other Influences on Detector Performance 503-19 1/94 Recommended Operating Procedures 503-211/94 System Suitability Test 503-211/94 Troubleshooting 503221/94 EICD-N 503-231/94 Apparatus and Reagents 503-231/94 etector Characteristics 503-231/94 Other Influences on Detector Performance 503-24 1/94 Recommended Operating Procedures 503-251/94 System Suitability 503251/94 ableshooting 503-251/94 General Precautions for eIcds 503261/94 503 E: Nitrogen/Phosphorus Detector 503-271/94 Principles 503-271/94 503-271/94 Apparatus and Reagents 503-281/94 Detector Characteristics 503-281/94 Other Influences on Detector Performance 508-30 1/94 Recommended operating Procedures 503-311/94 References 503-321/94 504: Quantitation 504 A: Introduction 504-1 1/94 504 B: Manual Quantitation 1/94 Measurement of peak height Measurement of Area by Triangulation 504-21/94 504 C: Electronic Integration 504-31/94 504 D: Special Considerations for Complex 504-6 1/94 Chromatograms BHC 50461/94 Chlordane 5048 1/94 PCBs 504-101/94 504-141/94 504-151/94 505: Bibliography General Texts 05-11/94 Inlets 05-1 1/94 Columns 05-1 1/94 Detectors 05-1 1/94 m60 500-3
Transmittal No. 96-E1 (9/96) Form FDA 2905a (6/92) Pesticide Analytical Manual Vol. I SECTION 500 500–3 page date Design 503-14 1/94 ElCD-X 503-16 1/94 Principles 503-16 1/94 Apparatus and Reagents 503-16 1/94 Detector Characteristics 503-17 1/94 Other Influences on Detector Performance 503-19 1/94 Recommended Operating Procedures 503-21 1/94 System Suitability Test 503-21 1/94 Troubleshooting 503-22 1/94 ElCD-N 503-23 1/94 Principles 503-23 1/94 Apparatus and Reagents 503-23 1/94 Detector Characteristics 503-23 1/94 Other Influences on Detector Performance 503-24 1/94 Recommended Operating Procedures 503-25 1/94 System Suitability Test 503-25 1/94 Troubleshooting 503-25 1/94 General Precautions for ElCDs 503-26 1/94 503 E: Nitrogen/Phosphorus Detector 503-27 1/94 Principles 503-27 1/94 Design 503-27 1/94 Apparatus and Reagents 503-28 1/94 Detector Characteristics 503-28 1/94 Other Influences on Detector Performance 503-30 1/94 Recommended Operating Procedures 503-31 1/94 References 503-32 1/94 504: Quantitation 504 A: Introduction 504-1 1/94 504 B: Manual Quantitation 504-2 1/94 Measurement of Peak Height 504-2 1/94 Measurement of Area by Triangulation 504-2 1/94 504 C: Electronic Integration 504-3 1/94 504 D: Special Considerations for Complex 504-6 1/94 Chromatograms BHC 504-6 1/94 Chlordane 504-8 1/94 PCBs 504-10 1/94 Toxaphene 504-14 1/94 References 504-15 1/94 505: Bibliography General Texts 505-1 1/94 Inlets 505-1 1/94 Columns 505-1 1/94 Detectors 505-1 1/94
Pesticide Analytical Manual Vol I Fi igures Polysiloxane Stationary Phases 50221/9 502-b GLC Column parameters 50241/94 02-c Ferrules for Connecting Glass and Metal 5029 1/94 502-d Capillary column Cross-Section 502151/94 02-e Inlet Adapters for Capillary Columns 502-171/94 02-f Van Deemter Curves 502191/94 502-g Capillary Column Inlet System 502-2 1/94 503-a Two EC Detector Designs 503-3 1/94 603-b Reproducibility 503-5 1/94 03C Dynamic Response Ranges 50361/9 503-d Linear Response Range 50361/94 508-e Effect of Detector Temperature 50371/94 503-f Effect of Carrier Gas Flow Rate 50371/94 503-g Single- Flame FPD 50391/94 0%-h Dual-Flame fpd 503101/94 508-i Block diagram of the elcd 503-151/94 3-j ElCD Reactor and Conductivity Cell 503-151/94 508-k N/P Detector Components 503281/94 03-1 N/P Detector Configurations 5032 1/94 504-a Manual Peak measurement 504-21/94 504-b Triangulation of Peak on Sloping Baseline 1/94 504-c Technical Chlordane 50481/94 504-d Chlordane, Heptachlor, Heptachlor Epoxide 504-9 1/94 504-e PCBs in chinook salmon 504-131/94 504-f Toxaphene 504141/94 Tables 02-a: Common GlC Liquid Phases Used in Pesticide 502-3 1/94 Residue determination 502-b: Operating Conditions for Packed Columns 502-18 1/94 504-a: Effects of Changing Electronic Integrator 5046 1/94 Setting 504-b: Response of Two Detectors to Four 50471/94 BHC Isomers 504-c: Weight Percent Factors for Individual Gas 504121/94 Chromatographic Peaks in Aroclor Reference Standard 50o4 Form FDA 2905a(6/92)
Transmittal No. 96-E1 (9/96) 500–4 Form FDA 2905a (6/92) SECTION 500 Pesticide Analytical Manual Vol. I page date Figures 502-a Polysiloxane Stationary Phases 502-2 1/94 502-b GLC Column Parameters 502-4 1/94 502-c Ferrules for Connecting Glass and Metal 502-9 1/94 502-d Capillary Column Cross-Section 502-15 1/94 502-e Inlet Adapters for Capillary Columns 502-17 1/94 502-f Van Deemter Curves 502-19 1/94 502-g Capillary Column Inlet System 502-23 1/94 503-a Two EC Detector Designs 503-3 1/94 503-b Reproducibility 503-5 1/94 503-c Dynamic Response Ranges 503-6 1/94 503-d Linear Response Range 503-6 1/94 503-e Effect of Detector Temperature 503-7 1/94 503-f Effect of Carrier Gas Flow Rate 503-7 1/94 503-g Single-Flame FPD 503-9 1/94 503-h Dual-Flame FPD 503-10 1/94 503-i Block Diagram of the ElCD 503-15 1/94 503-j ElCD Reactor and Conductivity Cell 503-15 1/94 503-k N/P Detector Components 503-28 1/94 503-l N/P Detector Configurations 503-29 1/94 504-a Manual Peak Measurement 504-2 1/94 504-b Triangulation of Peak on Sloping Baseline 504-3 1/94 504-c Technical Chlordane 504-8 1/94 504-d Chlordane, Heptachlor, Heptachlor Epoxide 504-9 1/94 504-e PCBs in Chinook Salmon 504-13 1/94 504-f Toxaphene 504-14 1/94 Tables 502-a: Common GLC Liquid Phases Used in Pesticide 502-3 1/94 Residue Determination 502-b: Operating Conditions for Packed Columns 502-13 1/94 504-a: Effects of Changing Electronic Integrator 504-6 1/94 Settings 504-b: Response of Two Detectors to Four 504-7 1/94 BHC Isomers 504-c: Weight Percent Factors for Individual Gas 504-12 1/94 Chromatographic Peaks in Aroclor Reference Standards
Pesticide Analytical Manual Vol. I SECTION 501 501: GENERAL INFORMATION Multiresidue methodology by definition requires determinative steps capable of separating analytes from one another so each can be detected and measured individually. Both gas-liquid chromatography (GLC) and high performance liquid chromatography(HPLC) provide these capabilities, and both are used in modern laboratories GLC has been the predominant determinative step in pesticide multiresidue methodology for over 30 years. Because GlC involves interaction between a vapor phase and liquid phase, its application is restricted to analytes that can be vapor ized without degradation. For heat-labile chemicals, HPLC offers a variety of alter native schemes for separating analytes according to chemical or physical charac minative step of choice for residues to which it is applicable t remain the deter- teristics, but GLC's relative simplicity and ruggedness cause it 501 A: PRINCIPLES paration in glc is achieved by differences in distribution of analytes between mobile and stationary phases, causing them to move through the column at dif ferent rates and from it at different times [1]. A measured aliquot of solution is injected into a gas chromatographic column through an inlet heated to a suffi- gas that forms the mobile phase sweeps analytes through the cole flow of inert ciently high temperature that analytes are vaporized. In this state, th this movement is the analyte's solubilization in the liquid phase. e n: retarding passag through the column, analytes that were injected in the same solution separate from one another because of their different vapor pressures and selective interac- tions with the liquid phase [2]. When analytes elute from the column and enter a detector, the detector responds to the presence of a specific element or func- tional group within the molecule. The detector's response causes a change in electronic signal, which is proportional to the amount of residue; the signal is amplified and recorded as a chromatogram Analytes are identified by the time it takes them to pass through a column of specific liquid phase (retention time), at a specified temperature and gas flow Quantities are calculated from the detector response. Both retention time and response are compared to values obtained for a reference standard solution in- jected into the same system 501 B: EQUIPMENT FOR GLC Gas Chromatographic Components The basic gas chromatograph consists of an inlet system, column, detector, elec- tronic equipment to amplify the detector signal, and a recorder or other data- handling device. Carrier gas(es), with appropriate pneumatic system(s), are also integral to the glc system. The inlet system, column, and detector are maintained in temperature-controlled environment The following are desirable features in GLC hardware: 1)Inlet, column oven, and detector should be individually heated and temperature-controlled. Temperature should be maintained to #O1C 501-1
Pesticide Analytical Manual Vol. I SECTION 501 501–1 Transmittal No. 94-1 (1/94) Form FDA 2905a (6/92) 501: GENERAL INFORMATION Multiresidue methodology by definition requires determinative steps capable of separating analytes from one another so each can be detected and measured individually. Both gas-liquid chromatography (GLC) and high performance liquid chromatography (HPLC) provide these capabilities, and both are used in modern laboratories. GLC has been the predominant determinative step in pesticide multiresidue methodology for over 30 years. Because GLC involves interaction between a vapor phase and liquid phase, its application is restricted to analytes that can be vaporized without degradation. For heat-labile chemicals, HPLC offers a variety of alternative schemes for separating analytes according to chemical or physical characteristics, but GLC’s relative simplicity and ruggedness cause it to remain the determinative step of choice for residues to which it is applicable. 501 A: PRINCIPLES Separation in GLC is achieved by differences in distribution of analytes between mobile and stationary phases, causing them to move through the column at different rates and from it at different times [1]. A measured aliquot of solution is injected into a gas chromatographic column through an inlet heated to a sufficiently high temperature that analytes are vaporized. In this state, the flow of inert gas that forms the mobile phase sweeps analytes through the column; retarding this movement is the analyte’s solubilization in the liquid phase. During passage through the column, analytes that were injected in the same solution separate from one another because of their different vapor pressures and selective interactions with the liquid phase [2]. When analytes elute from the column and enter a detector, the detector responds to the presence of a specific element or functional group within the molecule. The detector’s response causes a change in electronic signal, which is proportional to the amount of residue; the signal is amplified and recorded as a chromatogram. Analytes are identified by the time it takes them to pass through a column of specific liquid phase (retention time), at a specified temperature and gas flow. Quantities are calculated from the detector response. Both retention time and response are compared to values obtained for a reference standard solution injected into the same system. 501 B: EQUIPMENT FOR GLC Gas Chromatographic Components The basic gas chromatograph consists of an inlet system, column, detector, electronic equipment to amplify the detector signal, and a recorder or other datahandling device. Carrier gas(es), with appropriate pneumatic system(s), are also integral to the GLC system. The inlet system, column, and detector are maintained in temperature-controlled environments. The following are desirable features in GLC hardware: 1) Inlet, column oven, and detector should be individually heated and temperature-controlled. Temperature should be maintained to ±0.1° C