2. Responsibility: It is the responsibility of the Technical Assistant, Technical Manager, Quality Manager, and all those individuals who are working on the instrument.
3. Instrument Details:
Make-......................., Model- HPTLC
4. Procedure:
· Enter in the log book (In and out time)
· Connect the power supply. Switch “ON” the main power supply and instrument mains like the Sample applicator, Visualizer, Scanner, and the connected computer.
4.1 Preparation of sample
· Obtain HPTLC plate silica gel 60 F 254 (20X10 cm).
· Record the batch number. Inspect the plate no under UV254 nm for any damage to the layer. if damaged discards the plate with a soft pencil label the plate in the upper right corner with your initials, date (dd/mm/yy) -consecutive numbed of the day.
· On the right side of the plate mark a developing distance of 80 mm from the lower edge of the plate.
4.2. Preparation of chamber (Manual development only)
· Obtain a twin trough chamber for 20X10 cm plates.
· Fit the rear trough of the chamber with a filter paper into the rear trough ensuring complete wetting. Pour a sufficient amount of developing solvent into the front trough to have a level of 5 mm.
· Close the lid of the chamber and allow 20 min for saturation.
4.3.Sample Application
4.3.1. Automatic application (ADC-2)
· Select the following application parameters on the application device:
· Band length 8mm
· Number of tracks 15
· First application position X:20 mm
· Application Position Y:8mm
· Distance between tracks: automatic(Minimum 11 mm)
· Sample solvent Methanol
· Disable any unused track
· Apply the application volume as according to the standardized procedure for selected herbal drugs.
· Dry the plate in the air drier.
4.3.2. Manual Application (Linomat-2)
· Designed the application parameter and noted it down.
· Draw the band length up to 8 mm
· Decide the no of the track according to the no of the sample.
· Mix the sample with methanol and fill in the syringe.
· Ally on the y axis of the plate which contains an 8 mm distance.
· Dry the place using an air drier.
4.2.1.1.Plate conditioning (Manual development only)
After sample application places the plate for 45 minutes in a suitable desiccator containing a saturated solution of MgCl2.
4.2.1.2.Manual development
· Slowly open the lid of the saturated chamber and insert the conditioned plate into the front trough so that the back of the plate rests against the front wall of the chamber and the layer faces inside the chamber. Close the lid.
· Let the mobile phase ascend until it reaches the mark.
· Open the lid and remove the plate. Place it upright in a rack under the fume hood.
· Dry the plate with cold air from the hair drier for 5 minutes.
4.2.1.3. Automatic development
Use the following set of the automatic chamber:
· Enable pre-drying
· Saturation with filter paper 2m min.
· Humidity control 10 min with MgCl2
· Migration distance 70 or 80 mm
· Drying time 5 minutes
· 10 ml of developing solvent
· 25 ml of saturation solvent
· Set humidity control up to 70%.
4.2.2. Derivitization:
Ø Use the reagent specified in the individual monograph.
Ø Derivatization is either performed by using an automatic dipping device (typical immersion time 5cm/s and ni dwell time) or an automatic spraying device (typically 2-4 ml of reagent).
Ø Use the reagent specified in the individual monograph.
Ø Derivatization is either performed by using an automatic dipping device (typical immersion time 5cm/s and ni dwell time) or an automatic spraying device (typically 2-4 ml of reagent).
4.2.1. Documentation
30 min after the second derivatization step, take an image of the derivatized plate under UV
366NM.
4.2.2. Reporting
Create a copy of the software base report or use your own reporting documents.
END OF THE DOCUMENT
You may like to read these links:
1. List of All SOPs and Documents for Laboratory Instruments and Equipment
2. List of All SOPs and Documents for Chemistry Laboratory
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