HPLC LC/MS Troubleshooting
What is Chromatography?
A. Introduction Chromatography is an important biophysical technique that enables separation, identification, and purification of components of a mixture for qualitative analysis and quantitative determinations. Three components are central to any chromatography system. Figure 1 demonstrates chromatography principles using a filter paper as a stationary phase for separation of mixture components. This type of chromatography …
Drifting HPLC Peak Retention Times
A drift in analyte retention time over time is sometimes observed in HPLC chromatograms. This drift takes analytes outside their ‘retention window’ within the data analysis software leading to analyte peaks being missed (Figure 1). Peak drifting is often seen during long analysis sequence, analyzing “dirty” samples, or when installing a new HPLC column. A …
HPLC Column Cleaning
As HPLC chemists we know that columns do have a finite lifetime. You column life time (total number of samples you can run) is dependent on factors such as: the mode of separation the type of packing on the column the type of stationary phase Column care Increase Your Column’s Lifetime While most of these …
Poor HPLC Peak Shapes: Troubleshooting
Figure 1 shows various peak shape issues and their root causes observed while running an HPLC. These peak shape issues include peak tailing, peak splitting, and peak fronting. In this post, we discuss the root causes for these and their troubleshooting. Poor peak shapes result from factors related to sample, stationary phase, mobile phase, and …