4) Procedure

The following steps outline the procedure for conducting biochemical analysis in preclinical studies:

1. Step 1: Sample Collection
   1. Collect the required biological samples (e.g., blood, urine, tissue) from animals according to the study protocol, ensuring that collection methods are species-appropriate and follow ethical guidelines.
   2. Label each sample clearly with the animal’s identification number, time of collection, and any other relevant information.
   3. If necessary, separate plasma, serum, or other fractions from whole blood using appropriate centrifugation techniques.
2. Step 2: Sample Preparation
   1. Prepare samples for biochemical analysis by homogenizing tissue samples or diluting biological fluids, as required by the assay protocol.
   2. If necessary, store samples at appropriate temperatures (e.g., -80°C for long-term storage) until analysis, ensuring they are kept stable and free from degradation.
3. Step 3: Biochemical Assay Selection
   1. Select the appropriate biochemical assays based on the study objectives. This may include enzyme assays (e.g., ALT, AST), metabolic assays (e.g., glucose, lipids), or molecular markers (e.g., cytokines, DNA/RNA quantification).
   2. Ensure that assay kits are properly validated for the intended application and that reagents are prepared according to the manufacturer’s instructions.
4. Step 4: Biochemical Analysis
   1. Perform biochemical assays using automated equipment (e.g., spectrophotometers, microplate readers) or manual methods as specified in the study protocol.
   2. Ensure that all equipment is calibrated and properly maintained to ensure accurate results.
   3. Follow the appropriate procedures for sample incubation, reagent addition, and detection to ensure consistent and reliable results.
5. Step 5: Data Analysis and Interpretation
   1. Record all raw data, including the results of the biochemical assays, and ensure proper documentation of any deviations from the protocol.
   2. Analyze the data to assess the biochemical effects of the drug candidates, comparing treated groups to control groups as needed.
   3. Interpret the results in the context of the study objectives, considering any significant changes in biochemical markers that could indicate therapeutic efficacy or toxicity.
6. Step 6: Reporting and Documentation
   1. Prepare a report summarizing the methodology, results, and interpretations of the biochemical analysis. Include any relevant graphs, tables, and statistical analysis of the data.
   2. Ensure that the report includes a discussion of any observed trends or unexpected findings, and document any protocol deviations or issues encountered during the analysis.
   3. Store all data, including raw results, analysis logs, and final reports, in a secure manner to ensure availability for future reference or audits.
7. Step 7: Sample Disposal
   1. Dispose of all used reagents, consumables, and biological samples in accordance with biosafety and waste disposal regulations.
   2. Ensure that all waste is categorized and handled properly, with biological samples being disposed of in designated biohazard containers.

5) Documents

The following documents should be maintained during the biochemical analysis process:

1. Sample Collection and Handling Logs
2. Biochemical Assay Protocols
3. Raw Data and Analysis Logs
4. Test Result Reports
5. Quality Control and Calibration Records
6. Waste Disposal Records

6) Abbreviations

• GLP: Good Laboratory Practices
• ALT: Alanine Aminotransferase
• AST: Aspartate Aminotransferase
• 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 Biochemical Testing in Toxicology Studies
• ICH Guidelines for Nonclinical Safety Testing

8) Version

Version 1.0: Initial version of the SOP.

9) Annexure

Biochemical Analysis Report Template