| SOP for Good Chromatographic (HPLC) Practices |
1.0 OBJECTIVE 1.1 To lay down the procedure for good chromatographic (HPLC) Practices. 2.0 SCOPE 2.1 This SOP is applicable for good chromatographic (HPLC) Practices in quality control department. 3.0 RESPONSIBILITY 3.1 QC – Chemist / Officer / Executive 4.0 ACCOUNTABILITY 4.1 Head-QC 5.0 PROCEDURE 5.1 Analysis: 5.1.1 For operation of HPLC, refer SOP number XXX. 5.1.2 Perform the analysis of samples as per respective specification/STP. 5.1.3 Injection sequence to be followed should be as per the respective STP. The following injections sequence may be used as given below: Sr. no. No of injections Procedure of sequence 1 1 System suit 2 1 Blank 3 5 Standard 4 2 Sample 5 1 Standard bracketing OR > [System suitability (or system suit. > Blank) > Placebo (if required> Impurity standard (if applicable) > Standards>Samples>Bracketing standard or bracketing system suitability. 5.2 Primary System Suitability: 5.2.1 Firstly sequence is to be prepared by the analyst and should be named as “DDMMYYYY_ sample name_ version no”. 5.2.2 Before running the system the batch sequence and the method should verified by the data reviewer. 5.2.3 Now the system suit parameters like theoretical plates, tailing factor, resolution are checked in primary system suit as per STP. 5.2.4 If primary system suitability parameter does not meet the acceptance criteria then the system is to be altered/paused and following the check list is to be checked. Sr.no. Check list HPLC System- Pressure, Leakage, (Delta psi for Waters system). Mobile Phase- Buffer Solution mixed Properly, pH, Filtered, Sonication Time Column Used as per STP- Properly Washed 5.2.5 Now the print out is taken of processed chromatogram of system suit and sequence is stamped as “invalid”and proper justification will be written on the chromatogram. Now the Fresh sequence with new Version No. is prepared and system is run. 5.3 Final system suitability: 5.3.1 If the primary system suitability parameteris achieved then the system should be proceeded for final check of “% Area RSD”(NMT 2.0 for 5 replicate injections of standard Solution and NMT 3.0 for 6 replicate injections of Standard solution or individual monograph). 5.3.2 If final system suitability parameter gives abnormal results, then the system should be altered/paused andagain the check list should be followed. 5.3.3 Now again theprint out is to be taken of processed chromatogram of system suit and sequence is stamped as “invalid”and proper justification will be written on the chromatogram. The final batch sequence with new Version No is prepared and system is run. For preparation of standard and sample, refer individual specification/STP. 5.3.4 If sequence is modified for the addition or reduction of injections, the batch sequence shall be reprinted and checked by supervisor. 5.4 Bracketing: 5.4.1 The system suitability should be demonstrated throughout the run by intermittent evaluation. The intermittent system suitability should be performed by following method. 5.4.2 Refer below table for injecting bracketing standard: Sr. No. Test Bracketing Assay After every 5 batches Dissolution After every 1 batches or six injections Content uniformity After every 1 batches Related substances After every 5 batches or at the end of sequence 5.4.3 For example, if the batch sequence continues for 24 hours and sequence stops after 8 hour due to any reason (e.g. electricity breakdown, column or system pressure, missing vial etc.), in this case, inject the standard or system suitability solution, process it and check the system suitability criteria. If the system suitability passes, then continue the sequence. If the system suitability does not passing the criteria, Print out is taken of processed chromatogram of system suit and sequence is stamped as “invalid”and proper justification is to be written on the chromatogram. Now the Fresh sequence with update Version No is prepared and system is run. 5.5 Processing: 5.5.1 Process the batch sequence by auto integration. No manual integration shall be done while processing for assay, dissolution and CU. In case of related substances, if auto integration is not achieved due to the low response of peaks, then manual integration can be done by giving the justification for the same on chromatogram. 5.5.2 If due to shift of retention time or any other reason, processing parameters are not giving the same results for standard and test, and then Print out is taken of processed chromatogram of system suit and sequence is stamped as “invalid”and proper justification is to be written on the chromatogram. Now the Fresh sequence with updated Version No is prepared and system is run. 5.5.3 After correcting the processing method, process the whole data. 5.6 Recording Chromatograms: 5.6.1 Standard or system suitability chromatogram shall have following parameters: Theoretical plates Tailing factor Resolution (if specified) Relative retention time (if specified). 5.6.2 Each chromatogram should be duly signed off by the chromatographer and reviewed and signed off by the superior/section in-charge or data reviewer. 5.6.3 The invalid chromatogram should be duly signed and checked by the chromatographer and section in-charge or data reviewer respectively. 6.0 ABBREVIATIONS 6.1 SOP : Standard operating procedure 6.2 HPLC : High performance liquid chromatography 6.3 RSD : Relative standard deviation 6.4 STP : Standard test procedure 6.5 NMT : Not more than 6.6 % : Percentage 6.7 QC : Quality Control 6.8 NLT : Not less than |