UV Cabinet Operation and Calibration

1.0 OBJECTIVE :

To describe a procedure for Operation and Calibration of UV Cabinet.

2.0 SCOPE :

This procedure is applicable to Expo-Hi Tech make UV cabinet.

3.0 RESPONSIBILITY :

Quality control Chemist and above

4.0 ACCOUNTABILITY:

Quality Control Head

5.0 PROCEDURE :

5.1 SHORT GENERAL INFORMATION

An instrument consist of a mercury vapour lamp, which is used as the source of ultraviolet light and a filter which gives an emission band with maximum intensity at about 254 nm or 365 nm.

UV cabinets are mainly used for Chromatographic analysis by Ultraviolet fluorescence. These Cabinets are used for viewing Chromatogram combine both long wave and short wave Ultraviolet lights and also White light in a compact table-top unit. This self-contained unit is designed for easy viewing of Chromatograms.

5.2 PRECAUTION

Ultraviolet radiation is harmful to eyes and skin, protect your skin and your eyes against the direct contact with UV rays.

Never look at the lighted UV lamp directly.

5.3 OPERATION

Connect the plug to main switch and switch on the mains.

Switch ON the ON/OFF switch.

Red coloured light will glow, indicating power supply is connected.

Switch ON DAY LIGHT marked switch.

Red indicator light will be ON.

Open the front door of cabinet and place the pre-coated (coated with silica gel G) TLC plate inside UV cabinet and close the door.

Three switches are provided on the UV cabinet.

Switch 1 – For General DAY LIGHT.

Switch 2 – For UV Light at short wavelength (254 nm).

Switch 3 – For UV light at long wavelength (365 nm).

Switch off DAY LIGHT.

Switch on the required UV light, i.e., short wave (254 nm), long wave (365 nm).

Visualize the sample from outside the chamber through glass provided to visualize the sample inside the chamber.

After visualizing the sample note down the observation, switch off the UV light and remove plate from chamber.

Switch ‘OFF’ the instrument when not required.

5.4 CALIBRATION

Solution Preparation:

Ethanol (95 %): Add 12.5 ml of Water in 250 ml of Ethanol.

Test solution for 254 nm: Dissolve 40 mg of Sodium Salicylate in 100 ml of Ethanol (95 %).

Test solution for 365 nm: Dissolve 200 mg of Sodium Salicylate in 100 ml of Ethanol (95 %).

Stationary phase: Thin layer chromatographic plate coated with a 0.25 mm layer of chromatographic silica gel G mixture.

Procedure:

Do not touch the TLC plate on the side with the white surface.

In order to obtain an imaginary start line, draw a thin line with pencil which is 0.5 to 1.5 cm from the bottom of the plate.

Apply separately 5 µL of both test solutions to the starting line of a TLC plate (Keep the distance between the spots to about 3 cm) with the help of capillary,

Make a spot about diameter of 5 mm, don’t allow spot to become too large, if necessary, touch the capillary, lift and touch again.

Allow it to dry in air.

Place the plate in UV cabinet and observe under ultraviolet light at 254 nm and 365 nm.

Acceptance criteria:

The spot of Test solution for 254 nm should be visible at about 254 nm.

The spot of Test solution for 365 nm should be visible at about 365 nm.

FREQUENCY : Quarterly

6.0 ABBREVIATIONS:

Abbreviation Expanded form

w/v Weight per volume

% Percentage

nm Nanometer