Tips for Troubleshooting in Chromatography

Description:   Peak of Lanreotide not detected on HPLC of Thermo make. Investigation:  Investigation was done to find the root cause for above issue. Mobile phase changed multiple times. All HPLC tubings were checked for any kind of leakage but no issue was observed. Root Cause : To identify the root cause service engineer of Thermo was contacted. He took multiple trails but not succeeded.  After that he identifies that during method preparation calibration curve value taken was 5. As per his opinion it should be 6 for Thermo HPLC otherwise principle will not be detected. The same method was previously run on waters make HPLC where calibration curve was 6 and the same wss latter included in method, but when run on Thermo make HPLC,  peak was not detected. Conclusion: It is to be noted that calibration curve value play significant role during run on different HPLC systems. It is to be modified during development studies so that final method should include the correct value.  If anybody

  Description of Incident:  During analysis of one of the Active content during GC analysis, peak interference of Toluene observed.  Investigation:  During Investigation it was found that in every injection on GC, peak of Toluene was eluting. It was checked and found that Toluene was not used in that particular analysis then also Toluene peak was eluting. To rule out the possibility of chemical contamination, new bottle of DMSO was opened. After using new bottle reagent also, Toluene peak was eluting  and no significant drop in area was observed. Root cause:  During further Investigation  it was observed that during previous analysis Toluene was used as an standard.  Currently the oven temperature  was kept as 120°C (as boiling point of Toluene is 110°C) to remove the traces of Toluene but then also Toluene peak was observed.  Remedy :  As a corrective action, oven temperature was increased to about 200°C and column condition done for about half an hour. After that blank was infected a

Trouble description : During HPLC analysis, sometimes it happens that there is significantly less area observed as compared to previous trend od peak area. Investigation :  During investigation everything was checked i.e.: Checke Solution dilutions prepared for sample and standard and found Ok. Checked Column leakage and found no leakage. On changing the HPLC system, same solutions were used and arwawere checked  Mobile phase filteration checked and it was found that analyst did not filterd the mobile phase As unfiltered mobile phase was used,so solvent inlet filter was checked On inevstigating it was found that they were completely choked. Action taken : Solvent inlet filter was changed and analysis performed. Area observed in trend. CAPA: Solvent filter should be sonicated for 30 mins if any variation is observed during the analyis.