4) Procedure

The following steps outline the procedure for performing hematological analysis in preclinical studies:

1. Step 1: Blood Collection
   1. Collect blood samples from animals according to the study protocol. Ensure that the collection method (e.g., tail vein, jugular vein, cardiac puncture) is species-appropriate and follows ethical guidelines for animal welfare.
   2. Use appropriate anticoagulants (e.g., EDTA, heparin) to prevent coagulation and ensure sample integrity.
   3. Label each blood sample with the animal identification number, time of collection, and any other relevant information.
2. Step 2: Blood Sample Processing
   1. Process the blood samples immediately after collection to prevent degradation of blood components.
   2. If necessary, centrifuge the blood samples to separate plasma, serum, and cellular components such as red blood cells, white blood cells, and platelets.
   3. Store processed samples at appropriate temperatures (e.g., plasma at -20°C, whole blood at 4°C) until analysis.
3. Step 3: Hematological Testing
   1. Perform hematological analysis using automated hematology analyzers or manual methods, such as hemocytometer counts, to measure key blood parameters, including red blood cells (RBCs), white blood cells (WBCs), hemoglobin (Hb), hematocrit (Hct), and platelets (PLT).
   2. Ensure that the hematology analyzer is calibrated and that reagents are properly prepared and stored.
   3. For manual methods, ensure proper pipetting techniques and accurate counts when using a hemocytometer or other manual instruments.
4. Step 4: Data Analysis
   1. Record the results of the hematological tests, ensuring that all data is accurately transcribed into laboratory logs or electronic databases.
   2. Analyze the data to evaluate the effects of the drug candidate on hematological parameters. Compare treated animals with control groups to assess potential toxicity or changes induced by the drug.
   3. Review and interpret the results based on the study objectives, considering normal ranges for each blood parameter.
5. Step 5: Reporting and Documentation
   1. Prepare a comprehensive report summarizing the hematological analysis, including methodology, results, and interpretations.
   2. Document any deviations from the protocol, including reasons for adjustments or any issues encountered during the blood collection or analysis.
   3. Ensure that all data, including raw data, analysis, and reports, are securely stored and accessible for future reference or audits.
6. Step 6: Sample Disposal
   1. Dispose of all used blood samples and consumables (e.g., needles, pipettes) in accordance with biosafety and waste disposal regulations.
   2. Ensure that all biological waste is handled in designated biohazard disposal containers.

5) Documents

The following documents should be maintained for hematological analysis in preclinical studies:

1. Blood Collection Logs
2. Hematology Analysis Records
3. Test Result Reports
4. Data Analysis and Interpretation Logs
5. Quality Control and Calibration Records
6. Waste Disposal Records

6) Abbreviations

• GLP: Good Laboratory Practices
• IACUC: Institutional Animal Care and Use Committee
• RBC: Red Blood Cell
• WBC: White Blood Cell
• PLT: Platelets
• Hb: Hemoglobin
• Hct: Hematocrit
• QA: Quality Assurance
• FDA: Food and Drug Administration

7) References

References to regulatory guidelines and scientific literature that support this SOP:

• OECD Principles of Good Laboratory Practice (GLP)
• FDA Guidelines for Hematological Testing in Toxicology Studies
• ICH Guidelines for Nonclinical Safety Testing

8) Version

Version 1.0: Initial version of the SOP.

9) Annexure

Hematological Analysis Report Template