Simultaneous Analysis of Glutamate and GABA for Neuroscience Applications

Simultaneous Analysis of Glutamate and GABA for Neuroscience Applications

In this video, we’ll show you how to detect and analyze Glutamate and GABA in brain tissue or microdialysis samples in just 12 minutes. For this analysis, we’ll be derivatizing our analytes with OPA and beta-mercaptoethanol and analyze with high-performance liquid chromatography or HPLC, coupled with an electrochemical detector. But don’t let any of that scare you. We’ve simplified and automated the process to make it easier than ever.

The generation of good data and consistent results can take time and resources. That’s why many labs require highly-trained specialists in HPLC. However, we’ve created an application, which includes everything you need to run analyses right away. Everything from the mobile phase, reagent, and column selection, we have everything spelled out for you so all you have to do is take your sample and start running analysis. We test everything in house to make sure it works, ensuring there are no surprises.

For this application, we recommend use with our autosampler, called the AS-700. This enables proper mixing and incubation times, which provide consistent results sample after sample. Even with the precolumn derivatization process, It’s fast and highly sensitive, down to 1 picomole for glutamate, and 100 femtomoles for GABA.

At Amuza, we always have the end-user in mind. That’s why when you purchase one of our machines, you also have access to our self-help support center as well as our support specialists that can assist you with troubleshooting or replacement of parts as needed.

For more information on glutamate and GABA analysis, contact us today!

Questions?

Fast analysis of acetylcholine for neuroscience applications

Fast analysis of acetylcholine for neuroscience applications

We show you how to detect and analyze acetylcholine in brain microdialysis samples in 18 minutes, with minimal or no need to add an acetylcholine esterase inhibitor or AEI.

For this analysis, we use high-performance liquid chromatography or HPLC, coupled with an immobilized enzymatic reactor column and an electrochemical detector. It’s fast and highly sensitive, down to the femtomole range.

Acetylcholine is separated by a polymer-based reverse phase column, which has been selected specifically for this application. Not only is the column we’ve selected great at separating acetylcholine, but it can also handle the higher pH required for optimal enzyme activity. After exiting the separation column, acetylcholine and other compounds enter an immobilized-enzyme reactor column. Inside the reactor column, compounds are broken down to produce hydrogen peroxide, which is then selectively oxidized by an applied voltage across the flow path and changes to current are detected with a platinum electrode.

Optimizing the right conditions can take time and resources. That’s why many labs require highly-trained specialists in HPLC. However, we’ve created an application, which includes everything you need to run the acetylcholine analysis right away.

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 acetylcholine
-Enzymatic reactor column
-Electrochemical detector cell with a platinum working electrode for selective detection of the enzymatic reaction products.

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 18 minutes you will have your results and you’re ready for the next sample. It’s highly sensitive down to the femtomole range. Here’s an example of a chromatogram showing acetylcholine.

For more information on acetylcholine analysis, contact us today!

Questions?

Benefits of using HPLC-ECD for Neurotransmitter Detection

Benefits of using HPLC-ECD for Neurotransmitter Detection

Here’s why we recommend HPLC-ECD for neurotransmitter detection.

While there are a few ways to analyze neurotransmitters, like liquid chromatography-mass spectrometry, or LC-MS, ELISA or Radioimmunoassays. However, for qualitative and quantitative analysis of neurotransmitters, we recommend high-performance liquid chromatography detected with an electrochemical detector, known as HPLC-ECD.

The first reason is sensitivity. HPLC-ECD is highly sensitive, down to the femtomolar range. This is suitable for most neuroscience applications, including quantitative analysis of in-vivo microdialysis samples.

The second reason is time. Applications using HPLC-ECD can run anywhere from 5 to 30 minutes, where you can detect multiple analytes from a single injection. For example, we have an application for the simultaneous detection of dopamine and serotonin in just 5 minutes. That means you can run over a hundred samples overnight for high throughput using laboratory automation like an autosampler. We’ve also developed applications, saving you the economy of time by already optimizing conditions. All the applications on our site we know work, because we have tested them in-house.

The third reason is price. HPLC-ECD is priced with intended use in your own lab, thus, no need to outsource analysis in another lab or core facility. This gives you, the user, more confidence in your data with the knowledge that you are in control. Pricing for the equipment typically ranges from $20,000 to $60,000. If you’re not quite ready to commit to a new machine, ask us about our leasing options.

Finally, usability. While HPLC can seem overwhelming and complicated, we’ve simplified it with the HTEC. The HTEC system is an all-in-one device that integrates everything you need into one instrument. It’s straightforward and easy to use. Should you decide to acquire one of our instruments, on-site installation and training at your lab will be provided. Once installed, you can immediately start running samples. If you have questions or need assistance, there’s no need to worry. Our self-help support site and knowledgeable support staff are available to guide you through the process, including how to use as well as help with troubleshooting. At Amuza, it’s important for us that our instruments work just as well for you in your hands as they do for us.

Therefore, we recommend HPLC-ECD for the detection of neurotransmitters. For more information contact us today.

Questions?

5-minute Analysis of Dopamine and Serotonin in Brain Microdialysate

5-minute Analysis of Dopamine and Serotonin in Brain Microdialysate

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!

Questions?

New Loaner Program for Neurotransmitter Detection Systems

New Loaner Program for Neurotransmitter Detection Systems

Are you interested in analyzing neurotransmitters in brain microdialysate or brain tissue?

Perhaps you have a number of samples you’d like to run, but don’t want to fully commit to a new machine. Today, we’re happy to announce a new loaner program for an HTEC system fully ready to use.

The Eicom HTEC is a fully integrated HPLC-ECD for the detection of neurotransmitters from brain microdialysis and/or tissue samples. It is a fast and highly sensitive system that can be placed on a lab benchtop. Sample analysis can range from 5 to 30 minutes per sample, depending on the application. Sample runs can include quantitative analysis neurotransmitters like dopamine, serotonin, norepinephrine, acetylcholine, amino acids like glutamate and GABA. The HTEC is simple and straightforward to use. You don’t have to be an expert on HPLC to run analysis, anyone can be an expert!

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 specific for the application
  • 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.

How the loaner program works:

Contact us to discuss the scope of your research or questions you may be asking. Our support specialists will be happy to discuss what kind of analytes you’re interested in as well as the brain region. We’ll get you set up with an application specifically designed for your experiments. We’ll come to your lab and do the installation and training on-site over a 2 day period. From there, you can start running samples. Should you have any questions or need assistance, our support specialists will be available to help along with a self-help support site. At Amuza, it’s important that our instruments work just as well for you as they do in our facility.

We’ll provide shipping materials once your loan of the equipment is completed.

Contact us today for more questions including pricing options.

Price Comparison of 3 Neurotransmitter Detection Methods

Price Comparison of 3 Neurotransmitter Detection Methods

When it comes to neurotransmitter detection such as Dopamine, Serotonin, Norepinephrine, Glutamate, GABA, Acetylcholine, etc., in microdialysis samples, tissue homogenates, blood plasma, and others, there are three (3) main methods; LC-Mass Spectrometry (LC-MS), HPLC-ECD (Electrochemical Detection) & ELISA/Radioimmunoassays.

The first option is LCMS, it provides the highest sensitivity and specificity to the detecting neurotransmitters and also the most expensive option. The price range is between US$200,000 to $600,000.

The second method is HPLC-ECD which is much more cost affordable while offering great sensitivity. The price range is between US$20,000 to $60,000.

The third method is ELISA or Radioimmunoassays which offers less sensitivity. There is also a concern of selectivity due to cross-talk among antibodies.

Therefore, when choosing which method would work best for you, we would recommend HPLC-ECD.