Laboratory Procedure Manual Analyte: Specific Organophosphorous Pesticides, Synthetic Pyrethroids, Selected Herbicides and DEET Matrix: Urine Method High-Performance Liquid Chromatography- Atmospheric Pressure Chemical Ionization & Heated Electrospray Ionization Mass Spectrometry Method No: 6103.01 Revised: as performed by: Organic Analytical Toxicology Branch Division of Laboratory Sciences National Center for Environmental Health contact: Dr. Dana Barr Phone: 770-488-7886 Fax: 770-488-7609 Email: [email protected] Dr. Eric J. Sampson, Director Division of Laboratory Sciences Important Information for Users CDC periodically refines these laboratory methods. It is the responsibility of the user to contact the person listed on the title page of each write-up before using the analytical method to find out whether any changes have been made and what revisions, if any, have been incorporated Universal Pesticides OAT-DLS Table Of Contents 1. Clinical Relevance and Summary of Test Principle ..............................................................................1 2. Safety Precautions................................................................................................................................2 a. Reagent Toxicity or Carcinogenicity..................................................................................................2 b. Radioactive Hazards.........................................................................................................................2 c. Microbiological Hazards....................................................................................................................2 d. Mechanical Hazards .........................................................................................................................3 e. Protective Equipment........................................................................................................................3 f. Training .............................................................................................................................................3 g. Personal Hygiene..............................................................................................................................3 h. Disposal of Wastes ...........................................................................................................................3 3. Computerization: Data-System Management .......................................................................................3 a. Software and Knowledge Requirements...........................................................................................3 b. Sample Information...........................................................................................................................3 c. Data Maintanance.............................................................................................................................4 4. Specimen Collection, Storage and Handling Procedures; Criteria for Specimen Rejection .................4 a. Sample Handling...............................................................................................................................4 b. Sample Rejection..............................................................................................................................4 5. Procedures for Microscopic Examinations; Criteria for Rejecting Inadequately Prepared Slides.........4 6. Preparation of Reagents, Calibrators (Standards), Controls, and All Other Materials; Equipment and Instrumentation .............................................................................................................................................4 a. Reagents and Sources .....................................................................................................................4 b. Reagent Preparation.........................................................................................................................6 c. Standard Preparation........................................................................................................................6 1) Stock Solutions of Analytes (200 ng/mL) ......................................................................................6 2) Stock Solutions of Labeled Isotopes (200 ng/mL).........................................................................6 3) Working Labeled Isotopes Solution (ISTD) ...................................................................................6 4) Working Standard Solutions..........................................................................................................6 5) Calibration-Verification Materials ..................................................................................................7 6) Proficiency-Testing Standards (PT 1-3) ........................................................................................7 d. Equipment/Supplies ..........................................................................................................................7 e. Instrumentation .................................................................................................................................7 1) Mass-Spectrometer TSQ Quantum...............................................................................................7 2) Agilent 1100 Series LC System ....................................................................................................9 7. Calibration and Calibration-Verification Procedures ................................................................................9 a. Calibration Plot......................................................................................................................................9 b. Verification of Calibration....................................................................................................................10 c. Proficiency Testing..............................................................................................................................10 Universal Pesticides OAT-DLS Table Of Contents 8. Operating Procedures; Calculations; Interpretation of Results...............................................................11 a. Analytical Runs ...............................................................................................................................11 b. Sample Preparation ........................................................................................................................11 c. Liquid Chromatography Conditions.................................................................................................11 d. Mass Spectrometry Conditions .......................................................................................................12 e. Processing data ..............................................................................................................................13 f. Quantification ..................................................................................................................................13 g. Rearrangement of Data Files..........................................................................................................14 h. Transfer of Data ..............................................................................................................................14 i. Importation of Data into Database ..................................................................................................14 j. Statistical analysis and interpretation of data..................................................................................14 k. Routine and Periodic Maintenance of Key Components.................................................................14 1) Routine Maintenance......................................................................................................................14 2) Periodic Maintenance .....................................................................................................................15 l. Calibration.......................................................................................................................................15 9. Reportable Range of Results..................................................................................................................18 a. Linear Limits....................................................................................................................................18 b. Analytical Sensitivity .......................................................................................................................18 c. Accuracy .........................................................................................................................................19 d. Precision .........................................................................................................................................20 e. Analytical Specificity .......................................................................................................................20 10. Quality Control (QC) Procedures..........................................................................................................21 11. Remedial Action If Calibration or QC Systems Fail to Meet Acceptable Criteria ..................................21 12. Limitations of Method; Interfering Substances and Conditions.............................................................22 13. Critical-Call Results (“Panic Values”)....................................................................................................22