Chromatography troubleshooting is a key part of a chromatographer’s daily work. Here, we present articles and protocols to help a chromatorgher achieve this efficiently.
1. What is Void Volume? The void volume (column void volume or dead volume) is the volume that the mobile phase occupies in a column. For instance, if 80% of the entire column’s capacity is filled up with stationary phase material, the void volume would be 20% of the total column volume. 2. How to Calculate …
Ensuring optimum HPLC column performace is important for saving time and cost. The HPLC column peformance and retention characteristics should be monitored at regular intervals using standards. MoniFollowing recommendations are provided for routine HPLC column care and maintainance. Montoring for following signs of potential breakdown in HPLC column performance. A. Signs of a Damaged HPLC …
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 …
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 …
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 …
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 …
