It has been a long while, almost two months, since I wrote about pharmaceutical drug analysis on this blog in post, titled, Charged Aerosol Detection Speeds Analysis of Pharma Drug Impurities (link to post), and I found the UHPLC application featured here while looking for something else on our website. I am pleased to write about this application because it features our latest UHPLC system (Thermo Scientific Vanquish UHPLC system) and can be used with our method transfer tool to speed and streamline the process of adapting HPLC parameters for UHPLC analysis.
Application Brief 170, Fast Nevirapine Impurity Profiling Using UHPLC-DAD, (downloadable PDF), describes the development of a fast ballistic gradient UHPLC method optimized for simultaneous analysis of an Active Pharmaceutical Ingredient (API) and impurities in a nevirapine tablet. This medication is a non-nucleoside reverse transcriptase inhibitor used in treatment of HIV-1 infected adults. The method was developed using the aforementioned UHPLC system plus one of our fast, high-resolution HPLC columns (Thermo Scientific Syncronis C18 HPLC column) and the separation is completed in 2.8 min as compared to the 80 min of the isocratic USP method.
When transferring methods from HPLC to UHPLC, it is usually sufficient to maintain the resolution of the original method. Therefore, a popular strategy is to use smaller particles in shorter columns–this approach maintains resolution and provides faster separations. Rather complex calculations are required to adapt parameters, such as flow rate, injection volume, or gradient profile to the new column characteristics and you can use our HPLC to UHPLC Method Transfer Tool (downloadable tool) to automate these calculations.The tool is a calculation sheet supporting 12 selectable operating languages that does the calculations for you.
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To understand the theoretical background and the equations for an application’s transfer to UHPLC, do check out Technical Note 75, A Universal Tool for Method Transfer From HPLC to UHPLC, (downloadable PDF). The technical note also describes the tool, explains how to enter application details, and familiarizes you with the calculated results. Beyond the basic parameters, it also provides valuable features on how to deal with changing gradient delay volume, the adaption of data collection rates and recommended column reconditioning times, making it to a valuable tool for any HPLC or UHPLC user.
- A large library of chromatography applications for pharmaceutical analysis on this blog
- Our recently released Chromatography Solution Online Center features many useful and complimentary chromatography tools which can help speed up your analysis. The site is updated on a monthly basis; therefore, do check out the Archives section to see what was previously featured.
Let us know if you have any questions on the article featured in this post using the Comments box below. We look forward to hearing from you.