Standard Operating Procedure (SOP) for UV-Visible Spectrophotometer (Shimadzu UV-1800)

Purpose

This Standard Operating Procedure (SOP) outlines the steps required for the safe and accurate operation of the Shimadzu UV-1800 UV-Visible Spectrophotometer. The document is designed to ensure optimal performance, reproducibility, and compliance with laboratory standards.

Scope

This SOP applies to all laboratory personnel trained to use the Shimadzu UV-1800 UV-Visible Spectrophotometer for the analysis of liquid samples in the UV-Visible spectrum.

Safety Precautions

1. Always wear appropriate personal protective equipment (PPE), including gloves, lab coat, and safety goggles.
2. Handle chemicals and samples in compliance with their respective Material Safety Data Sheets (MSDS).
3. Ensure the spectrophotometer is placed on a stable, clean surface, away from direct sunlight and vibrations.
4. Avoid touching the optical components, such as the sample holder or cuvettes, with bare hands.
5. Dispose of samples and solvents according to laboratory waste disposal guidelines.

Materials and Equipment

1. Shimadzu UV-1800 UV-Visible Spectrophotometer
2. Quartz or glass cuvettes (depending on the wavelength range)
3. Sample solutions
4. Blank solution (e.g., solvent used for sample preparation)
5. Kimwipes or lint-free tissue
6. Calibration standards (if required)
7. Computer with UV Probe software installed

Procedure

1. System Preparation

1.1. Turn on the spectrophotometer and allow it to warm up for at least 30 minutes to stabilize the light source. 1.2. Ensure the instrument is connected to the computer and the UV Probe software is launched.

2. Initial Setup

2.1. Verify that the instrument is in good working condition (e.g., no error messages). 2.2. Clean the sample compartment and cuvettes using appropriate cleaning methods. 2.3. Select the mode of operation (e.g., spectrum mode, photometric mode, or kinetics mode) in the UV Probe software based on the analysis requirements.

3. Baseline Correction

3.1. Fill a clean cuvette with the blank solution.

3.2. Insert the blank solution cuvette into the sample holder, ensuring proper alignment.

3.3. Close the lid of the sample compartment.

3.4. Perform a baseline correction using the software’s “Baseline” function to ensure accurate measurements.

4. Sample Analysis

4.1. Prepare the sample solution and ensure it is free from bubbles or particulate matter.

4.2. Transfer the sample solution into a clean cuvette, ensuring it is filled at least two-thirds full.

4.3. Wipe the exterior of the cuvette with a lint-free tissue to remove any smudges or droplets.

4.4. Place the cuvette in the sample holder, aligning it properly.

4.5. Close the sample compartment lid.

4.6. Select the wavelength range or specific wavelength for measurement.

4.7. Initiate the measurement by clicking the “Start” or equivalent button in the software.

4.8. Record the absorbance or transmittance values displayed.

5. Data Management

5.1. Save the recorded data in the designated folder on the computer.

5.2. Export the data in the desired format (e.g., CSV, PDF) for further analysis or reporting.

5.3. Document the results in the laboratory notebook or electronic record-keeping system.

6. Shutdown Procedure

6.1. Remove the cuvettes from the sample compartment and clean them immediately.

6.2. Turn off the spectrophotometer and the computer.

6.3. Cover the instrument with a dust cover to protect it from contamination.

Maintenance

1. Perform routine maintenance as specified in the Shimadzu UV-1800 user manual.
2. Replace the light source (deuterium or tungsten lamp) as needed, following manufacturer guidelines.
3. Clean the sample compartment regularly to prevent residue buildup.
4. Calibrate the instrument periodically using certified standards.

Troubleshooting

Issue	Possible Cause	Solution
No power	Loose power cable	Check and secure the power connection.
Unstable baseline	Dirty optical components	Clean cuvettes and sample compartment.
Low sensitivity	Aging light source	Replace the light source.
Incorrect readings	Improper baseline correction	Re-run the baseline correction procedure.

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References

• Shimadzu UV-1800 User Manual
• Laboratory Safety Guidelines
• ISO/IEC 17025: General Requirements for the Competence of Testing and Calibration Laboratories

Revision History

Revision	Date	Description	Author
1.0	January 21, 2025	Initial draft	[Your Name]