SOP for Performance Check of High-Performance Thin Layer Chromatography

Objective

To check the performance of High-performance thin layer chromatography.

Scope

This procedure is applicable to calibrate high-performance thin layer chromatography.

Responsibility

Doing: Technical Assistant

Checking: Executive /Manager

Accountability

Head of the Department

Frequency

once in a 3 month

Procedure

• Check the performance of the instrument in terms of

1. Linearity of spotting ( Annexure-I)
2. Reproducibility of spotting ( Annexure-II)

• Prepare the mobile phase as per the method given in Annexure-I.
• Prepare the solution as mentioned in Annexure-I
• Use 10 x 10 cm HPTLC plate (Kieselgel 60F254 or Identical)
• For Linearity of spotting

1. Apply 2 µl, 4µl, 6µl, 8µl and 10 µl of solution on the HPTLC plate with a spotter.
2. Allow the plate to run in the mobile phase.
3. Dry the plate with a drier.
4. Scan the plate with the scanner.
5. Check the linearity and correlation coefficient.
6. Fill in the Data as mentioned in Annexure-I

• Reproducibility of spotting

1. Apply 10µl solution on the HPTLC plate for five times in sequence
2. Allow the plate to run in the mobile phase.
3. Dry the plate with a drier.
4. Scan the plate with the scanner.
5. Calculate the RSD for five tracks ( RSD Limit: N.M.T 3.0%)
6. Fill in the data mentioned in Annexure-II

Abbreviations

RSD= Relative standard deviation

N.M.T = Not more than

N.L.T = Not less than

HPTLC = High-performance thin layer chromatography

API= Active pharmaceuticals ingredient

Annexure

Revision History

Nil

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