SOP for Calibration of High Performance Thin Layer Chromatography (HPTLC)

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

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)

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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

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

ABBREVIATIONS

%= Percentage

RSD= Relative standard deviation

N.M.T = Not more than

N.L.T = Not less than

μl = Microlitter

HPTLC = High-performance thin layer chromatography

API= Active pharmaceuticals ingredient

ANNEXURE

REVISION HISTORY

Nil

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