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    Validation Plan Format

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    Nora Buan
    This document outlines a validation plan for a qualitative LC-QTOF mass spectrometry method to identify cocaine in seized samples from the Philippines. The plan details the equipment, materials, reagents, reference standards, and supplies used. It describes the instrument parameters, acquisition method, and calibration procedures. The validation parameters to be tested are outlined as peak symmetry, precision, robustness, stability, and carryover. Acceptance criteria and validation procedures are proposed for each parameter. Appendices will include sample data and a proposed validation report format.

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    Validation Plan Format

    Uploaded by

    Nora Buan
    43 views8 pages
    This document outlines a validation plan for a qualitative LC-QTOF mass spectrometry method to identify cocaine in seized samples from the Philippines. The plan details the equipment, materials, reagents, reference standards, and supplies used. It describes the instrument parameters, acquisition method, and calibration procedures. The validation parameters to be tested are outlined as peak symmetry, precision, robustness, stability, and carryover. Acceptance criteria and validation procedures are proposed for each parameter. Appendices will include sample data and a proposed validation report format.

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    Analytical Chemistry

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    validation plan format

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    This document outlines a validation plan for a qualitative LC-QTOF mass spectrometry method to identify cocaine in seized samples from the Philippines. The plan details the equipment, materials, reagents, reference standards, and supplies used. It describes the instrument parameters, acquisition method, and calibration procedures. The validation parameters to be tested are outlined as peak symmetry, precision, robustness, stability, and carryover. Acceptance criteria and validation procedures are proposed for each parameter. Appendices will include sample data and a proposed validation report format.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    43 views8 pages

    Validation Plan Format

    Uploaded by

    Nora Buan
    This document outlines a validation plan for a qualitative LC-QTOF mass spectrometry method to identify cocaine in seized samples from the Philippines. The plan details the equipment, materials, reagents, reference standards, and supplies used. It describes the instrument parameters, acquisition method, and calibration procedures. The validation parameters to be tested are outlined as peak symmetry, precision, robustness, stability, and carryover. Acceptance criteria and validation procedures are proposed for each parameter. Appendices will include sample data and a proposed validation report format.

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    © All Rights Reserved
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    Download as docx, pdf, or txt
    of 8

    VALIDATION PLAN INFORMATION

    Organization Name
    Division Performing Validation
    Method Validation for
    Standard Operating
    Procedure (SOP) Title

    Equipment
    Validation Number MVAL-01-2019-001
    Number of Revision None

    Authors

    Reviewed by: Signature Date Signed

    Approved by: Signature Date Signed

    (TITLE) Method Validation Plan for

    1
    1. Scope and Purpose of Validation

    To establish a qualitative Liquid Chromatograph-Quadrupole Time of Flight (LC-


    QTOF) Mass Spectrometer method for positive identification of Cocaine in seized
    samples in the Philippines.

    2. Types of materials for which the method will be applied

     Standard Cocaine (Solid)


     Unknown powders and other solid materials which are suspected to contain
    Cocaine.

    3. Equipment, Chemicals, and Supplies

    3.1 Equipment

    Agilent 6530 QTOF LC/MS equipped with Agilent 1260 Infinity II high speed
    pump, multi-sampler, and multi-column thermostat with Mass Hunter Software.

    3.2 Chemicals and Sample Materials

    3.2.1 Reagents

     Distilled water, LC-MS grade (Honeywell)


     Acetonitrile, LC-MS grade (Sigma-Aldrich)
     Methanol, LC-MS grade (Sigma-Aldrich)
     Isopropanol, LC-MS grade (Sigma-Aldrich)
     Formic Acid, Reagent grade (Agilent)
     ES-L low concentration tuning mix (Agilent)
     ES-TOF biopolymer analysis kit (Agilent)
     API-TOF reference mass solution kit (Agilent)
     HPLC flushing solvent

    3.2.2 Reference Standard

     Cocaine, solid, 50mg (Australian Government National Measurement Institute)

    3.2.3 Actual Sample

     Cocaine powder samples without on-going case.

    4. Supplies

    2
     LC vials (amber) with septum and cap (1.5ml)
     Volumetric flasks-glass (2ml, 5ml, 10ml, 100ml, 500ml, 1000ml)
     Volumetric flasks-plastic (2ml, 5ml, 10ml, 100ml, 500ml, 1000ml)
     Autopipettors (0.1-2µl, 1-10µl, 10-100µl, 100-1000µl)
     Pipette tips (0.1-2µl, 1-10µl, 10-100µl, 100-1000µl)
     Disposable plastic syringe (10 ml)
     Filter membrane units (0.22 µm)
     Nalgene bottles (100ml, 500ml, 1000ml)
     Amber bottle with plastic hose and plastic cap for LC
     Ultra high purity Nitrogen gas
     Vortex mixer
     Sonicator

    5. Procedure

    5.1 Preparation of standard solutions

    Certified reference material of Cocaine dissolved in acetonitrile will be used for


    standard stock solution preparation. Standard stock solutions will must contain a
    concentration in parts per billion (ppb) and will be placed in a closed flask in a
    refrigerator if not needed. Pipette accurately appropriate amounts of the stock standard
    solution into appropriate volumetric flasks. Dilute solutions in appropriate amount of
    acetonitrile up to mark and calculate the concentration of each subsequent standard
    solution in ppb. Then transfer prepared standard solutions into a LC vial and inject 1-2
    μL into the LC-QTOF instrument. On the other hand, all Cocaine standard and sample
    solutions will be analyzed with a minimum of 3 trials in every validation parameter
    conducted, and all values will be reported up 2 to 4 decimal places.

    5.2 Instrument Parameters

    5.2.1 LC Conditions

    Column Agilent ZORBAX Eclipse Plus C18


    2.1mm x 50mm, 1.8µm

    Column Temperature 60oC

    Mobile Phase A: 0.1% Formic acid B: 0.1% Formic acid in


    in water acetonitrile

    Flow Rate 0.5ml/min

    Isocratic Program 85% A and 15% B

    Analysis Time 4 minutes

    Post Time 2 minutes

    3
    Overall Cycle Time 8 minutes

    Gas Ultra High Purity Nitrogen

    Injection Volume 1µl (with 5 seconds needle wash in flushport)

    5.2.2 QTOF Conditions

    Electrospray AP-ESI
    Source (Using Agilent Jet Stream Technology)

    Ionization Polarity Positive

    Sheath Gas Temperature and 380oC at 12L/min


    Flow

    Nozzle Voltage 500 V

    Drying Gas Temperature and 320oC at 8L/min


    Flow

    Nebulizer Gas Pressure 27psi

    Capillary Voltage 3.750 V

    Fragmentor Voltage 150 V

    Collision Energy 10 V

    Personal Compound
    Database and Library Forensics (ForTox PCDL)
    (PCDL)

    5.2.2 MS Acquisition Method Parameters

    Reference Ion Masses 121.050873 and 922.009798


    Enabled

    Acquisition Mode MS1

    Minimum Mass Value 50 m/z

    Maximum Mass Value 1000 m/z

    Scan Rate 3 Hz

    4
    5.2.3 Mass Calibration, Tuning and Corresponding Accepted Values

    Corrected Residual Must be less than 0.5ppm


    Mass Error

    Resolution 2GHz: 118 ion > 4,500


    1521 ion > 13,000

    Abundance 200,000 to 500,000

    Note: Check tune and mass calibration must be performed before the start of each
    analysis for the day. Check tune results must be accepted in order to proceed with the
    validation analysis.

    6. Validation Parameters

     Peak Symmetry
     Precision (Repeatability/Reproducibility)
     Robustness
     Stability
     Carryover

    5
    7. Validation Procedure and Acceptance Criteria

    Acceptance Proposed
    Parameter Validation Procedure Criteria Duration

    1. Peak Symmetry

    -a measure of
    chromatographic
    peak’s symmetry.

    2. Precision

    - Measure of the
    closeness of the
    analytical results
    obtained from a
    series of replicate
    measurements of the
    same measure under
    the conditions of the
    method.

    3. Robustness
    - Performance of an
    analytical method in
    the presence of
    minor variation of
    some experimental
    conditions of the
    method within the
    laboratory.

    4. Stability
    - a method that
    demonstrates the
    extent to which the
    analytes are stable
    6
    during the whole
    analytical
    procedure, including
    storage before and
    after analysis.

    5. Carryover
    - a type of test to
    check possible
    significant peaks in
    the blank run after
    testing a sample of
    typically higher
    concentration.

    8. References

    8.1 United Nations Office on Drugs and Crime, Validation of Analytical Methodology
    and Calibration of Equipment used for Testing Illicit Drugs in Seized Materials and
    Biological Specimens, ST/NAR/41 2009, pp. 5-50.

    8.2 United Nations Office on Drugs and Crime, Recommended Methods for the
    Identification and Analysis of Cocaine in Seized Materials, ST/NAR/7/REV.1 2012,
    pp. 17-34.

    8.3 Health Sciences Authority, Singapore, Training Course on Qualitative Method


    Validation, April 05, 2017, Training Manual, pp. 1-15.

    8.4 University of the Philippines- Natural Sciences Research Institute, Training Course
    on Quality Assurance in Chemical Analysis for the Philippine Drug Enforcement
    Agency (PDEA) Laboratory Service, February 17-19, 2016, Training Manual, pp.
    155-178.

    8.5 Agilent Technologies, The Secret of Good Peak Shape in HPLC, Available:
    https://www.agilent.com/cs/library/eseminars/Public/secrets%20of%20good
    %20peak%20shape%20in%20hplc.pdf.

    8.6 SWGDRUG Supplemental Document SD-2, Quality Assurance/Validation of


    Analytical Methods, SWGDRUG 2006-02-09, pp. 6-8, 18-20, 47-51.

    7
    9. Appendices

    9.1 Appendix A:

    9.1.1 Peak symmetry


    9.1.2 Precursor ion and two (2) candidate ions and their corresponding % abundances
    9.1.3 Mass spectrum and library search score
    9.1.4 Mean, standard deviation and relative standard deviation

    9.2 Appendix B:

    9.2.1 Proposed Validation Report Format

    Note: Proposed submission period of validation report which includes calculation,


    interpretation and reporting will be on ___________.

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