Using E-TRF to recognize deuterated compounds – Milja’s master’s thesis
Our Application Scientist Milja Virtanen finished her master’s thesis for Aqsens Health this autumn. Her MSc thesis in Chemistry focused on finding out if E-TRF can be used to differentiate between deuterated and non-deuterated compounds. In this blog post Milja answers a few questions about her thesis and the research process.
What does “deuterated” mean?
A substance, compound or organism is deuterated if one or more of its hydrogen atoms have been changed to deuterium atoms.
Why are some compounds deuterated, for example in the pharmaceutical industry?
Deuteration usually stabilizes the structure of a compound. This can come useful, for example, when a drug-molecule breaks down too quickly in metabolism or when the breakdown causes adverse effects. By slowing down the breakdown time with deuteration, it is possible to make sure that the drug has time to reach its destination in our bodies. It can also decrease the unwanted side-effects.
What was your research hypothesis?
In my thesis I set out to see whether E-TRF can be used to detect and differentiate between deuterated and non-deuterated compounds. I wanted to know if we can form individual fingerprints for the different compounds. I also wanted to find out whether we could see the effects of deuterium oxide on the enzyme-color reaction in the sample liquid, and more specifically, whether the speed of the color reaction was affected. Our Chief Scientific Officer Janne was also interested in evaluating E-TRF’s ability to detect molecular level changes in samples.
Why was it interesting to see whether E-TRF can differentiate between deuterated and non-deuterated chemical compounds?
The differences between deuterated and non-deuterated compounds are extremely small. So, seeing whether E-TRF could detect these small changes could tell us a lot about the method’s performance. It was also interesting to see the differences in detection performance. Would E-TRF be able to differentiate only fully deuterated compounds from non-deuterated compounds, or could it possibly detect the structural change caused by only a single deuterium atom?
How did you set out to see if your hypothesis was correct?
I chose five model compounds with different sizes and complexities. The model compounds that were fully deuterated were deuterium oxide and acetophenone-d8. More complex and less deuterated were caffeine-(trimethyl-d3) and L-dopa-(phenyl-d3). The non-deuterated equivalents for the four model compounds were MQ-water, acetophenone, caffeine and L-dopa. Also, D-glucose and its two deuterated versions were tested (D-glucose-1-d1 and D-glucose-1,2,3,4,5,6,6-d7).
I began the experimentation by looking for potential modulators that could help differentiate the compounds from one another. When I had found several modulators that enhanced the separation between the compounds, I performed parallel evaluations and gathered each compound’s individual fingerprint. By comparing these individual fingerprints, I could make determinations about E-TRF’s capability in detecting deuterated and non-deuterated compounds.
Every thesis writer usually comes across a few problems. Were there any bumps on the road for you?
The thing I feel like I struggled with the most was with finding suitable sources that could help me in assessing my results. Because E-TRF is a completely new method, it is difficult to compare it to the older methods.
What did your MSc thesis prove in the end?
It was possible to form a clearly different fingerprint for simple and fully deuterated compounds and accurately differentiate between these compounds. I also concluded that the changes caused by deuterium oxide in the enzyme color reaction could be observed more clearly with E-TRF than with absorbance alone.
Do you think E-TRF could be a useful tool, for example, in the pharmaceutical industry?
I think that E-TRF shows real potential in the area of disease detection, and it also might become a useful tool in the pharmaceutical industry. With further research and more analyses, I think we could determine the strengths of E-TRF in the detection of small molecular changes, such as deuteration. I think my thesis created a good starting point for more pharmaceutical research using E-TRF.
Do you think you will have the chance to continue the research into E-TRF and deuterated compounds at Aqsens?
I think that the research will continue in some parts of the area that my thesis covered. For example, what effects does deuterium oxide have on the different enzyme reactions, and is it possible to follow them with E-TRF. I think it would also be interesting to see how deuterium oxide affects our biosensors and whether it changes their behavior. So, there are many interesting opportunities ahead.
Milja Virtanen
Application Scientist
Aqsens Health Oy
