Automated HPLC techniques

Automated HPLC can be used to support a number of technologies including:

• fraction collection
• LC-MS

The use of a robot system linked into either system can load racks of samples, or remove racks of fractions.

Metabolite profiling from radiolabelled pre-clinical and Phase I ADME samples can be a time consuming process. The radiolabelled drug-related material in plasma and excreta from these studies is commonly quantified using high-performance liquid chromatography (HPLC) followed by radiochemical detection.

Microplate scintillation counting e.g. TopCount has allowed lower limits of detection, enabling more samples containing lower levels of radioactivity to be analysed. Samples are fraction-collected following HPLC separation into 96-well plates. However metabolite profiling using conventional HPLC can require long run times, which equates to fractionation into up to 6 plates. Using most commercially available fraction-collectors, samples have to be injected individually so that the scientist can change plates between each analysis. Only 18-20 plates (which equates to 3-10 samples depending on the HPLC method) can generally be run per day with no overnight or weekend runs possible.

By automating the Aglient 1100 HPLC instrument in the process, the samples can be processed overnight.

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Poster: Automating HPLC Fractionation for Profiling of Low Level Radioactivity

Analysis by LC-MS is a common technique, however, the whole process can be made very effective by automating the sample preparation step of the process and loading the samples directly onto the LC-MS.

This process has been fully automated with a Mitsubishi robot on a 5m track with 3 individual Hamilton 4200/STAR processing pods and a range of sample preparation equipment:

• Thermo Cytomat 189 plate storage
• SIAS centrifuge
• Thermo (Matrix) 2D barcode reader
• BMG LABTECH nephelometer
• Thermo ALPS plate sealer
• LC-MS with CTC autosampler
• Ultrasonic bath