GUIDELINE FOR PREPARATION OF MOBILE PHASE REQUIRED FOR HPLC

       Important Points:

•  Use HPLC grade/ AR Grade / GR grade chemicals.
• Weigh / Measure each component of the mobile phase separately.
• Degas each solvent and buffer by sonication and then filter each component of the mobile phase separately through 0.45µ filter (Use the filter paper as per suitability with solvent)
• Mix the individual solvent and buffer as pr the specified proportion.
• Adjust the pH of the mobile phase wherever specified (i.e. before or after mixing of individual components) between ± 0.05 unless otherwise specified. 
• Sonicate the mobile phase for 10 minutes or more as required
• Ensure that the mobile phase is free from particles. Do not use the mobile phase if particles are observed.
• Enter the details of mobile phase preparation in the laboratory notebook.

Procedure for mobile phase validation

1.  Prepare the mobile phase as per the test procedure and keep on the benchtop in a well-closed condition.

2.  Evaluate system suitability parameter(s) as per the test procedure at initial, after one day and after two days.

3.  The mobile phase is considered stable if all the system suitability parameter are within the specified limits.

Stability:

The mobile phase is to be prepared daily if the mobile phase validation is not done.

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