SOP FOR ANALYSIS OF HPLC SAMPLES

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         OBJECTIVE

         To lay down a standard procedure for analysis of HPLC samples.

     SCOPE

This SOP is applicable for analysis of HPLC samples in Quality Control Department

       RESPONSIBILITY

    • Officers / Executive – Quality Control shall be responsible for complies of this SOP                                                                                      ACCOUNTABILITY
      • Manager- Quality Control and Head QA Shall be accountable for compliance of this SOP.
  • PROCEDURE
    • Initialization of the Instrument.
    • Instrument make it on by starting with pumping system, an injector, a chromatographic column (a column temperature controller may be used), a detector and a data acquisition system.
    • Used the specified username and password to login software.
    • Place the mobile phase with the respective channel. Connect the required column which specified as per STP.
    • HPLC Column Saturation:
      • Equilibrate the column with the prescribed mobile phase and flow rate at room temperature or at the temperature specified in STP until a stable baseline is achieved.
    • Creating the Sequence:
      • Fill the information of vial No. /inj. No., sample name, method set, no. of injection, injection volume, run time and column information in respective place.
      • For Assay / Dissolution / Content Uniformity sample sequence like to be:

Diluent     —  Resolution (if any)  —  System Suitability Standard —  Standard — Sample — Standard Bracketing Where injection for Resolution – Single; System suitability replicate injection; Standard – two; Assay – two; Dissolution, Content Uniformity – one & Standard Bracketing – two.

  • For Related Substance sequenece like to be:

Diluent— placebo (if any) —  Resolution (if any)  —  System Suitability Standard /Standard — Sample

Note: where before sample injection purge the injector with specified diluents.

  • In sample sequence, after 10 sample / 6 hour inject Bracketing standard or at the end of analysis.
  • The RSD of two standards and bracketing standard should not be more than 2%.
  • Run the Assay sample 5 minutes more of retention time of principal peak or prescribed in the STP.
  • Run the Related Substance sample 3 times more of retention time of principal peak or prescribed in the STP.
  • Saving the Sequence:
    • All sequence save by the following way:
    • For In-Process, Finished sample – “Name of the Product” “Date, Month, year” e.g. CARNIVIT 240508 where “CARNIVIT” Name of the Product & “240508” Date, Month, Last digit of current year.
    • For Raw material – “Name of the sample” “Date, Month, Year” e.g. Azithromycin 240508 where “Azithromycin” Name of the sample” & “240508” Date, Month, Last digit of current year.
  • Processing the Chromatogram:
    • Select the Chromatogram, open required processing method; integrate it; identified principal peak; process it with required peak width and threshold and other integration parameters.
    • For Assay integration parameters for standard and sample should be same.
  • Recording the Chromatogram:
    • Take out the print of all chromatogram in respective Reporting method.
    • Analyst shall sign all the chromatogram with date.

   REFERENCES

In-House

  • ABBREVIATIONS

         SOP: Standard Operating Procedure

QC: Quality Control

STP: Standard Testing Procedure

HPLC: High Performance Liquid Chromatography

RSD: Relative Standard Deviation

No.: Number

 DISTRIBUTION LIST

Quality Assurance Department

Quality Control Department

  HISTORY OF REVISION

Version No. Date of Revision Reason for Revision

 

 

SOPs