to all UV-Visible spectrophotometers used in the Analytical Method Development (AMD) department for both qualitative and quantitative analysis of pharmaceutical substances.

3. Responsibilities

• Analytical Chemist: Performs calibration activities, records data, and ensures instruments are within specifications.
• QA Personnel: Reviews calibration results and ensures compliance with regulatory requirements.
• AMD Head: Approves calibration reports and ensures timely execution of schedule.

4. Accountability

The AMD Section Head is accountable for ensuring all UV-Visible spectrophotometers are calibrated as per schedule and documentation is reviewed and approved as per GMP guidelines.

5. Procedure

5.1 Calibration Frequency

1. Perform calibration monthly and after major maintenance or relocation of the instrument.
2. Record all calibration activities in the UV Calibration Logbook (Annexure-1).

5.2 Wavelength Accuracy Check

1. Use Holmium Perchlorate solution or a certified UV wavelength standard.
2. Scan spectrum between 200 nm and 600 nm using a 1 cm quartz cell.
3. Verify the presence of characteristic peaks (e.g., 241.15 nm, 279.4 nm, 287.15 nm).
4. Acceptance Criteria: ±1.0 nm from certified reference values.

5.3 Photometric Accuracy Check

1. Prepare potassium dichromate solution in 0.005M H₂SO₄ at 60, 40, 20, and 10 ppm.
2. Measure absorbance at 235, 257, 313, and 350 nm against the solvent blank.
3. Acceptance Criteria: Absorbance values must be within ±0.01 AU of expected values for each concentration.

5.4 Stray Light Test

1. Use a 1.2% KCl solution and scan at 200 nm.
2. Acceptance Criteria: Absorbance should be greater than 2.0 AU.

5.5 Resolution Test

1. Use Toluene in Hexane and scan from 260–280 nm.
2. Calculate resolution using absorbance at 269 nm and 266 nm.
3. Acceptance Criteria: Ratio A₂₆₉/A₂₆₆ ≥ 1.5.

5.6 Documentation

1. Record all observations, spectra, and absorbance values in Annexure-2.
2. Fill calibration checklist (Annexure-3) and attach with report.
3. Obtain QA review and approval before using instrument for analysis.

5.7 Handling Out of Calibration (OOC)

1. If any parameter is out of acceptance criteria, stop use of the instrument immediately.
2. Log deviation and initiate investigation as per deviation SOP.
3. Perform maintenance and recalibrate before resuming use.

6. Abbreviations

• UV: Ultraviolet
• AU: Absorbance Unit
• QA: Quality Assurance
• OOC: Out of Calibration
• AMD: Analytical Method Development

7. Documents

1. UV Calibration Logbook – Annexure-1
2. Calibration Raw Data Sheet – Annexure-2
3. Calibration Checklist – Annexure-3

8. References

• ICH Q2(R1) – Validation of Analytical Procedures
• USP General Chapter <857> – Ultraviolet-Visible Spectroscopy
• 21 CFR Part 211 – GMP for Finished Pharmaceuticals

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name	Neha Mehta	Priya Nair	Dr. Harshita Goyal
Designation	AMD Analyst	QA Reviewer	Head – Analytical Method Development
Department	Analytical Method Development	Quality Assurance	Analytical Method Development

11. Annexures

Annexure-1: UV Calibration Logbook

Includes calibration dates, performed by, instrument ID, and remarks for traceability.

Annexure-2: Calibration Raw Data Sheet

Scanned spectra, absorbance readings, calculation tables, and peak listings for each parameter tested.

Annexure-3: Calibration Checklist

Parameter	Test Performed	Within Limits	Remarks
Wavelength Accuracy	Yes	Yes	Complies
Photometric Accuracy	Yes	Yes	Complies
Stray Light	Yes	Yes	Complies
Resolution	Yes	Yes	Complies

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
01/06/2025	2.0	Added resolution check and updated annexures	Periodic SOP review	Dr. Harshita Goyal
10/08/2022	1.0	Initial release	New SOP	QA Head