Hi, I have a question on HPLC (high performance liquid chromatography).
The procedure says to use a
4.6 x 150mm x 3um column.
The relative retention time of Impurity B with reference to the main peak is about 0.5min.
If I use a 4.6 x 100mm x 2.7um column, will the relative retention time of Impurity B with reference to the main peak still be about 0.5min?
See your post above and the response by Bob Pursley.
To determine whether the relative retention time of Impurity B with reference to the main peak will still be about 0.5 minutes when using a different column, we need to consider the factors that affect retention time in high-performance liquid chromatography (HPLC).
The retention time in HPLC depends on several factors, including the column dimensions (length, inner diameter), particle size, stationary phase, and mobile phase composition. Changes in any of these factors can influence the relative retention time.
In this case, you are comparing two columns with different dimensions and particle sizes. The first column specified is 4.6 x 150mm x 3um, while the second column you want to use is 4.6 x 100mm x 2.7um.
When comparing two columns, the important factor to consider is the length and the particle size. The length of the column affects the overall separation efficiency, while the particle size affects the resolution between different peaks. Smaller particle sizes generally provide higher resolution.
In your case, the length of the second column is shorter (100mm) compared to the first column (150mm), which means that the separation will be faster in the second column. However, the particle size of the second column (2.7um) is slightly smaller than the first column (3um), which can help in improving resolution.
Considering these factors, it is difficult to predict the exact relative retention time of Impurity B on the second column without experimental confirmation. However, it is likely that the relative retention time will be different from 0.5 minutes, as the changes in column length and particle size can affect retention times.
To determine the new relative retention time on the second column, you will need to conduct experiments with the specific conditions (mobile phase composition, flow rate, etc.) and evaluate the relative retention time of Impurity B with respect to the main peak. It may be necessary to develop a new method optimization based on the characteristics of the new column.