tag:blogger.com,1999:blog-9209375845415652006.post1369218500516913198..comments2024-01-24T09:56:18.495-05:00Comments on HPLC CHROMATOGRAPHY HINTS and TIPS FOR CHROMATOGRAPHERS [HPLC TRAINING ARTICLES] : HPLC Peak Splitting. Common Reasons For ItHPLC EXPERThttp://www.blogger.com/profile/04810178046245936465noreply@blogger.comBlogger3125tag:blogger.com,1999:blog-9209375845415652006.post-29816605658521999862016-09-26T11:51:35.358-04:002016-09-26T11:51:35.358-04:00BTW: Sample degradation could also explain the &qu...BTW: Sample degradation could also explain the "extra" peak seen.HPLC EXPERThttps://www.blogger.com/profile/04810178046245936465noreply@blogger.comtag:blogger.com,1999:blog-9209375845415652006.post-49634406738924392222016-09-24T17:34:14.032-04:002016-09-24T17:34:14.032-04:00By increasing the sample concentration (and volume...By increasing the sample concentration (and volume) by ten times, it is likely you have overloaded the HPLC column (see #1 above). To avoid overload, try and keep the injection volume below 3% of the column dead volume. You may want to perform a loading study to find out what the limits of the specific column and method are.<br /><br />The reason you experienced sensitivity problems when transferring the method to a second HPLC system is most likely that the two systems are not configured the same. Many differences could exist which can result in different response outputs. <br />For example:<br />(1) Difference in each detectors flow cell path length can have a dramatic effect on the observed signal (i.e. 6 mm vs 10 mm). For more information, please refer to the article: http://hplctips.blogspot.com/2011/03/flow-cell-volume-path-length.html<br /><br />(2) For UV/VIS detectors, the wavelength bandwidth value also can have a large effect on the observed signal too. For more information, please refer to the article, http://hplctips.blogspot.com/2011/09/uv-vis-hplc-detector-signal-bandwidth.html<br /><br /> *You need to carefully compare each system to first. HPLC EXPERThttps://www.blogger.com/profile/04810178046245936465noreply@blogger.comtag:blogger.com,1999:blog-9209375845415652006.post-57588844033349918532016-09-24T17:04:22.906-04:002016-09-24T17:04:22.906-04:00We recently started to see our API as two connecte...We recently started to see our API as two connected peaks instead of one with a dip in between the two peaks. We moved the method to a different HPLC system and could not see the sample anymore so we increased the injection from 10 ul to 100 ul and now see it, but as a split peak. What can we do to fix this?Anonymousnoreply@blogger.com