Standard Operating Procedure for Operation of UV Spectrophotometer in API Manufacturing
| Department | API Manufacturing |
|---|---|
| SOP No. | SOP/API/136/2025 |
| Supersedes | SOP/API/136/2022 |
| Page No. | Page 1 of 9 |
| Issue Date | 13/04/2025 |
| Effective Date | 15/04/2025 |
| Review Date | 13/04/2026 |
1. Purpose
To provide a standardized procedure for the safe and effective operation of the UV-Visible Spectrophotometer, including calibration, sample analysis, maintenance, and documentation for use in testing of APIs and related substances.
2. Scope
This SOP applies to all Quality Control personnel responsible for using UV-Visible spectrophotometers in the API manufacturing unit for qualitative and quantitative analysis of raw materials, intermediates, and finished APIs.
3. Responsibilities
- QC Analyst: Operate, calibrate, and maintain the UV spectrophotometer as per the procedure.
- QC Supervisor: Review and verify instrument performance, calibration records, and usage logs.
- QA Officer: Ensure compliance with this SOP during audits and routine checks.
4. Accountability
The QC Head is accountable for ensuring proper functioning, documentation, and calibration compliance of UV-Visible spectrophotometers in the Quality Control laboratory.
5. Procedure
5.1 Instrument Description
UV-Visible Spectrophotometer is used for the measurement of absorbance or transmittance of samples at specified wavelengths in the ultraviolet (190–400 nm) and visible (400–800 nm) regions.
5.2 Instrument Start-Up
- Ensure the instrument is connected to a stable power source.
- Switch on the instrument and allow it to warm up for at least 30 minutes before calibration or use.
- Open the instrument software on the connected system (e.g., UV-Probe, LabSolutions, UVWin).
5.3 Calibration Procedure
- Perform wavelength accuracy check using Holmium oxide or certified filter standards.
- Perform absorbance accuracy using potassium dichromate solution in 0.005 M sulfuric acid at 235 nm, 257 nm, 313 nm, and 350 nm.
- Check stray light using 1.2% KCl solution at 200 nm (absorbance ≥ 2.0).
- Check baseline flatness and noise using air blank scans from 200–800 nm.
- Record all calibration results in the Calibration Log (Annexure-1).
5.4 Sample Analysis
- Prepare sample and blank as per the Standard Test Procedure (STP).
- Use clean, matched quartz cuvettes with appropriate path length (e.g., 1 cm).
- Rinse cuvettes with sample and blank before measurement.
- Place blank in reference holder and sample in the sample holder.
- Select required wavelength and record absorbance or full spectrum as per STP.
- Print results and attach to the analytical worksheet with analyst signature and date.
5.5 Post-Analysis and Shutdown
- Remove and rinse cuvettes with distilled water, then dry.
- Close instrument software and turn off the UV spectrophotometer.
- Cover the instrument to protect from dust and light exposure.
5.6 Maintenance and Precautions
- Do not touch the optical surfaces (cuvettes, lenses) with bare hands.
- Ensure the instrument chamber is free from any contamination or spills.
- Record any unusual observations or performance issues in the Instrument Usage Log (Annexure-2).
- Schedule preventive maintenance and service calibration every 6 months.
6. Abbreviations
- UV: Ultraviolet
- QC: Quality Control
- STP: Standard Test Procedure
- nm: Nanometer
- SOP: Standard Operating Procedure
7. Documents
- Calibration Log (Annexure-1)
- Instrument Usage Log (Annexure-2)
- Analytical Worksheet
- Preventive Maintenance Report
8. References
- Pharmacopoeia (USP <857>, IP, BP)
- ICH Q2(R1) – Validation of Analytical Procedures
- Instrument Operating Manual
- 21 CFR Part 11
9. SOP Version
Version: 2.0
10. Approval Section
| Prepared By | Checked By | Approved By | |
|---|---|---|---|
| Signature | |||
| Date | |||
| Name | |||
| Designation | |||
| Department |
11. Annexures
Annexure-1: Calibration Log
| Date | Parameter | Standard Used | Observed Value | Limit | Remarks |
|---|---|---|---|---|---|
| 13/04/2025 | Wavelength Accuracy | Holmium Oxide | 279.8 nm | 279–281 nm | Pass |
Annexure-2: Instrument Usage Log
| Date | Analyst | Batch No. | Test | Instrument ID | Remarks |
|---|---|---|---|---|---|
| 13/04/2025 | Ravi Kumar | API-20250413 | Assay | UV-01 | Normal operation |
Revision History:
| Revision Date | Revision No. | Details | Reason | Approved By |
|---|---|---|---|---|
| 01/01/2022 | 1.0 | Initial SOP | New Equipment SOP | QA Head |
| 13/04/2025 | 2.0 | Updated software, added annexures | Audit Observation | QA Head |