by - PURPOSE:
To lay down a procedure for General chromatographic practices, procedures to be followed also describe the requirement of records to be maintained.
2.0 SCOPE:
This SOP is applicable in Quality Control.
RESPONSIBILITY:
QC chemist / Officer : To follow the procedure
- DEFINITION:
Head Quality Control : For SOP compliance
- ENVIRONMENT, HEALTH AND SAFETY:
NA
- Procedure:
- Precaution:
- After completion of analysis, solvent-carrying lines shall be purged with HPLC water/ Ultra Q water.
- In Ideal Condition Keep the HPLC System in water or mixture of water / methanol or mixture of water Acetonitrile. If any instruments fault to meet the specified tolerance limit, during calibration immediately inform to Manager QC.
- Do not keep HPLC System in buffer when NOT IN USE.
- Do not SWITCH OFF the instrument directly when it is controlled through software.
- Column shall be wash immediately after completion of analysis.
- Before starting analysis allow at least 15 minutes for lamp to warm up & system to equilibrate for analysis.
- Chromatographic Procedure:
- Precaution:
Standard Practices:
Mobile Phase:
- Water
for preparation of mobile phase or diluents shall be collected freshly from
Ultra-Q water.
- Chromatographic conditions shall be set as per the product specific standard test procedure.
- During Preparation of mobile Phase if pH adjustment is required then the pH Meter shall be ensured for the daily calibration of the instrument.
- Prepare mobile phase, as per standard test procedure or pharmacopoeia and finally filter with 0.45 µ filter and degas before use.
- HPLC System Hot water flushing and online filters / reservoir filters used on HPLC to be cleaned on fortnight basis with 1% nitric acid solution.
- Mobile Phase Bottles and all solvents bottles in use for mobile phase shall be properly labeled.
- Mobile Phase shall be prepared freshly for analysis and shall be used within 2 days. If mobile phase appears to be hazy or turbid, same shall be discarded.
- HPLC water / Ultra-Q on HPLC System shall be change on daily basis. Solvent and solvent mixture shall be use on weekly basis. If the appears to be hazy or turbid same shall be discarded.
- Mobile Phase Preparation for HPLC analysis shall be recorded in raw data sheet / test data sheet.
- If more than one batch are analyzed together than reference of general test data sheet for mobile phase preparation recording shall be given in data sheet of next batches.
- For HPLC Analysis use of column is maintained in the column usage logbook.
- For HPLC Analysis uses of instrument is referenced in the instrument usage logbook.
- For HPLC Analyst must in his/her log in, and should be logout after completion of the analysis.
- For any new analysis (first time), New instrument method shall be created in application software by the authorized person, as per parameter specified in respective STP.
- After Creation method shall be verified by the second analyst/ reviewer for correctness and same shall be attached with test data sheet.
- Adjustment of chromatographic conditions:
Adjustment of chromatographic conditions shall be done on the basis of analytical method validation with prior approval of Head Quality Control.
- Thin-Layer chromatography and paper chromatographyComposition of mobile phase:
The amount of the minor solvent component may be adjusted by ±30% relative or ±2% absolute, whichever is the larger. No other component is altered by more than 10% absolute.
- pH of the aqueous component of the mobile phase:
±0.2 pH, unless otherwise stated in the monograph, or ±1.0 pH when neutral substances are being examined.
- Concentration of salts:
In the buffer component of a mobile phase±10%.
- Application Volume:
Decreased to 20% of the prescribed volume if using fine particle size plates (2-10µm).
- Migration distance of the solvent front:
Not Less than 50mm
- Liquid chromatography:Composition of the Mobile Phase:
The amount of the minor solvent component may be adjusted by ±30% relative or ±2% absolute, whichever is the larger. No other component is altered by more than 10% absolute.
- pH of the aqueous component of the mobile phase:
±0.2 pH, unless otherwise stated in the monograph.
- Concentration of salts:
In the Buffer component of a mobile phase ±10%.
- Detector wavelength:
No adjustment permitted.
- Column Parameter:
Column length up to ±70%
Column internal diameter up to ±25%
Particle size maximal reduction of 50% , No increase permitted.
- Flow rate:
Not more than±10%.
- Temperature:
Not More than±10% a maximum of 60º.
- Injection Volume:
May be decreased, provided detection and repeatability of the peak (s) to be determined are satisfactory.
- System Suitability and Documentation:
- Analyst may run standard or resolution solution to confirm the retention time and system suitability parameter before start the actual sequence of the analysis.
- The qualification method for the 1st injection of system suitability or retention time check shall be separate with name “system suitability check”. In the actual sequence specific product qualification shall be used Unsuccessful system suitability check shall be invalidated with appropriate justification.
- In case of gradient program method, the gradient saturation may inject prior to the sequence run.System suitability shall be established as per the procedure and determined by the first chromatogram of replicate injections of standard solution except resolution.
- All the standard and sample solution shall be filtered through 0.45µ membrane filter or as defined in the individual procedure.Run time of the chromatogram shall be as defined in the standard test procedure (STP).For the analysis of related substances the run time shall be at least 2.5 times of the retention time of principle peak or as defined in the individual procedure.In case if the sequence summary not defined in standard testing procedure then the same shall be prepare as follows.
- For assay, sequence of injections shall be as follow
- Blank (As per requirement for base line stable and avoid the carry over).
- 5 or 6 replicate injections of standard solution or as specified in the respective test procedure.
- Duplicate injection of sample solution.
- Bracketing standard (standard solution) in single after every 10 injections of sample and at the end of the sequence or inject one standard after every 8 hours to check the system suitability whichever is earlier. The 8 Hours start from completion of last standard used for calculation purpose (Initial standard).
- For related substance, sequence of injections shall be as follow
- Blank (As per requirement for base line stable and avoid the carry Over).
- Placebo solution (if defined in the procedure)
- Resolution Solution (if specified)
- 6 replicate injections of standard solution or as specified in the respective test procedure.
- Single injection of sample solution.
- Bracketing standard (standard solution) in single after every 10 injections of sample and at the end of the sequence or inject one standard after every 8 hours to check the system suitability.
- Generating and Processing of Chromatogram:
- After Completion of analysis analyst shall confirm for the compliance of system suitability Parameters.
- Analyst shall take the print of all chromatograms of the sequence.
- Each printed Chromatograms shall have the information like instrument ID, Analyst name, sample detail, method name, data file name, date and time acquired, date and time of processing.
- The whole sequence shall be processed /Integrated by one method except system suitability check. Manual integration shall not allow.
- In case of RS, AMV and on other specific requirement then the multiple qualification method can be used for processing of chromatographic data.
- The scaling parameters shall be uniform for all chromatograms of a sequence.
- Modification to processing parameters or the use of multiple processing methods within a chromatographic run is not permitted without documented scientific justification. “Modification of integration parameters’ and laboratory management approval to ensure that the integrity of data is not compromised. The chromatographic shall be locked by the reviewer.The chromatogram shall be locked by locking the sequence.
- If a chromatogram is required to be reprocessing after release of batch or after review for any reason, ‘Reprocessing request shall be raised. Such reprocessing of the chromatogram shall be done only after approval from Head QC and Head QA or their designee. The reason for reprocessing shall be recorded on initially processed chromatogram and singed by chromatograph processer, Reviewer, Head QC & Head QA or designed. If due to any reason (like system suitability failure or system failure) chromatographic analysis needs to be repeated than original chromatograms shall be reteamed and marked as invalid with proper justification and authorization .Both the reprocessed chromatogram as well as original chromatogram, shall be attached with the record of analysis, original chromatograms shall be invalidated.
- Along with chromatograms analyst shall take the prints of sequence and method parameters for analysis.
- Analyst shall also take the print of summary report for % RSD of initial standard injection s and with all bracketing standard injection separately and attached with the chromatograms.
- If the more than one sample/batches are analyzed under single sequence than chromatograph of initial standard injection and methods –print shall be attached with chromatogram of first sample/batch.
- Sequence print shall be attached with chromatogram of each sample/batch.
- Analyst shall attach the Sample information format with General test datasheet of subsequences sample and reference of product and batch no. /AR no. shall mention on the same.
- Checking the precision of duplicate sample injection.
- In the test of assay, RSD should be not more than 2.0%.
- For reporting the results. If the difference between two injections is within the acceptance criteria average of two injections should be considered for Assay and maximum value should be considered for related substances.
- If the difference found more than specified limit. Immediately inform, to Supervisor, repetition of duplicate injection from the same aliquot shall be carried out after preliminary investigation.
- If any of the value of duplicated injection id out of specification then shall be investigated As per Reporting of laboratory incident.
- The analyst with date shall sign each page of chromatograms.
- Processing of chromatograms shall be done preferably within next working day, after completion of sample set analysis If the concerned Analyst is not available in the office or allocated some other job than the Lab-in charge/ Manager/Designee shall allocate the task to another Analyst for processing of the chromatogram in case of reason other than mentioned above. Dept. Head or designee approval shall be required.
- Processing method used for processing of all the chromatograms shall be attached along with record of analysis.
- For assay, sequence of injections shall be as follow
- Instruments Shutdown:
- After completion of analysis switch off the detector and flush the column with water or methanol or with prescribed solvent for about half an hour at the flow rate 1ml. It the Mobile phase contains any buffer solution then flush the column with Mehoanol. Water (60:40) mixture or prescribed solvent at the flow rate 1ml per min for about one hour. Preserve the column with Acetonitrile/Methanol. Remove the column and close the close the column and with end closures.Anayst shall restart the HPLC/GC and respective client on weekly basis.
Solvents: Used for HPLC should be HPLC grade.
Chemicals: used for HPLC should ne AR grade
- following are the precaution for using the instrument when it is used after a long time
- Run the HPLC system for at least four hours with water and solvents prior to starting of analysis
- Check the value and reservoir filters should be sonicate Purge the injector.
- Syringe should be free from air bubble.
- Prevention of carry over.
- In the last of residual solvents always inject one blank after the standard to prevent the
Carryover of standard in the sample analyst can increase the number of such blank according to the requirement. In the test of related substance always inject one blank before bracketing standard. To prevent the carryover sample. Analyst can increase the number of such blanks according to the requirement.
- If still the carry over is observed use the solution of Acetonile.isopropyl alcohol and water in the ratio 5:1:1 and select the needle wash time as ‘Extended’.
- Precautions.
- Suitability covers the HPLC waste reservoir to avoid the evaporation of waste solvents.
- Always label the HPLC waste reservoir as ‘WASTE’.
- The HPLC vials should not be filled more than 75% of total volume of HPLC vials.
- After using of filler (Paper, Nylon, PVDF and etc.)Put the cut mark with glass marker on used filter and immediately put the waste bin.
- Before start of analysis, analyst shall create the sequence of injection as motioned in STP & analysis plan System generated printout of injection sequence from the software shall be taken, The analyst shall confirm from the printout of the sample set for the entire sequence and sign off with date, The same shall be verified & signed by the second analyst/reviewer/ designee for correctness and also verify/check the standard and test dilution as per standard test procedure on working bench and HPLC vial position as per sequence in HPLC injector port. The sequence shall be reviewed as per annexure-V.
- Reference(S):
- In house
- ABBREVIATIONS:
- SOP :Standard Operating Procedure
- QC :Quality Control
- HPLC :High Performance Liquid Chromatography
- RSD :Relative Standard Deviation
- OOS :Out of Specification
- CU :Content Uniformly
- STP :Standard Test Procedure
- RS :Related Substance
- AMV :Analytical Method Validation
- Revision History
| Rev.No. | Details of changes | Reason for Change |
| 00 | Nil | New SOP |
https://pharm85.wp9.hostingraja.info/good-laboratory-practices-2/
Reprocessing request for Liquid Chromatography
A. Request Details
Product:_______________________ Batch No.___________________________
Specs/STP/Pharma copoeial Method ______________________________________________
Test/Parameter/Station ______________________________________________
______________________________________________
Reason for Re-processing/unlocking _______________________________________________
Of chromatogram _______________________________________________
| Request initiated by | Name | Sign\Date | ||
| Reviewed by Head-QC/designee | Name | Sign\Date | ||
| Authorized By Head QA/designee | Name | Sign\Date |
B. Corrective Action
Data is available in the system (Month of Analysis:______________________________________________
Restoration Required:______________________________________________________________________
Impact on reporting results:_________________________________________________________________
Certificate of analysis:_____________________________________________________________________
Record of analysis:________________________________________________________________________
Batch approved or rejected:_________________________________________________________________
- REVISION HISTORY:
| S. No. | Document No. with Version No. | Superseded Document No. with Version No. | Reference Change Control No. | Page no. | Point No./Section | Description of Change |
| 1.0 | 01 | NA |
