In this video, we’ll show you how to detect and analyze dopamine and serotonin levels in brain microdialysis samples in 5 minutes. For this analysis, we use HPLC-ECD because it is a cost-effective way to get fast and ultra-sensitive data down to the femtomole range.
While HPLC-ECD is a common analysis method, careful consideration must be taken regarding analytical conditions, such as mobile phase preparation, column selection, and other factors. Ideally, we want:
- Prompt elution to save time
- Clear separation of the signal peaks from the solvent-front peaks
- Fewer peaks around the signal peaks
- A low noise level and a stable baseline
Optimizing these conditions can take time and resources. That’s why many labs require a dedicated person to handle HPLC. Thus, we’ve created an application to take out all the guesswork thereby maximizing the separation conditions for dopamine and serotonin.
For this application, we’re going to use the HTEC, which includes everything you need, integrated into one single unit. Let’s take a look inside:
- Dual-piston pump with a unique algorithm to reduce noise without any pulse damper.
- Degasser to remove small air bubbles for better pump performance.
- Temperature control for consistent results.
- Separation column uniquely selected for separation of DA and 5-HT.
- Electrochemical detector cell with a 3 electrode DC potentiostat and an amperometric electrode.
The electrically active analyte is detected as it flows over the surface of a smooth electrode with an applied voltage.
We’ve optimized conditions to make it as easy as possible. Simply inject the sample using a manual injector or autosampler if you’re using laboratory automation. You can monitor signal response in real-time using the dedicated chromatography software. In 5 minutes you will have your results and you’re ready for the next sample. It’s highly sensitive down to 0.16 femtomoles.
Here’s an example of a chromatogram showing dopamine and serotonin.
For more information on dopamine and serotonin analysis, contact us today!