Interview with TMIC Young Scientists

We are pleased to have an interview with TMIC young scientists who were awarded at the 4th Annual Metabolomics Association of North America (MANA) Conference in Edmonton. Let’s get to know who they are and learn more about their research:

  • Metabolomics Service Cores Best Presentation Award – Oral Presentation: Shuang Zhao
  • Early Career Member (ECM) Best Poster Award– Graduate Students: Michal Lazarek
  • Early Career Member (ECM) Best Poster Award– Undergraduate Students: Ewenet Mesfin and Abby Kropielnicki

Metabolomics Service Cores Best Presentation Award

This award is available to the MANA members who hold a position in a service core and will present (poster or oral) at MANA 2022 under one of the scientific themes of the conference.

Shuang Zhao

This award was given to Dr. Shuang Zhao for his oral presentation entitled “Development of a High-Coverage and Quantitative Metabolomics Assay for Targeted Analysis of Multiple Pathways”. Shuang received his Ph.D. degree in Chemistry from the University of Alberta and currently holds his position as a laboratory coordinator at Dr. Liang Li Node

What is the key point or highlight that you want the audience to take away from your presentation?

The highlight is the assay itself. We use our unique chemical isotope labeling LC-MS and also a dual processing approach. We develop this assay based on both technologies and the performance is outstanding as it can analyze multiple pathways.

Can you summarize your presentation in a 30-second elevator pitch?

In the current targeted metabolomic approaches, there are several analytical challenges related to conventional targeted analysis, such as the limited availability of internal standards, the limited number of metabolites, etc. In our lab, we developed the assay by using the two technologies to improve analytical performance by detecting more metabolites and making the quantification more accurate, thus, we can generate high-performance results targeting multiple pathways.

Where do you think the future of this technology lies?

One of the main features of the assay is expandable/ flexible, our future direction is to generate similar assays e.g. microbiome metabolite assay and plant metabolome assay, expanding those assays to different areas such as cellulosome metabolomics or energy metabolomics. We also plan to render this assay as a service to provide high-quality results to the users in the community.

The difference between conventional and multiple pathways analysis

Early Career Member (ECM) Best Poster Award

The Best Poster award is a competitive award for an Early Career Member who clearly and enthusiastically describes the scientific merit of their research. Awards are based on effective visual and oral presentation, quality of research, and depth of understanding. Total of awards for 5 postdocs, 5 graduate students, and 2 undergraduate students. TMIC researchers received one award for graduate students and both for the undergraduate category.

Michal Lazarek

The ECM Best poster award for graduate students was awarded to Michal Lazarek with the poster presentation entitled “Metabolomic profiling of acyl-CoA metabolites using chemical isotope labeling LC-MS/ Liquid Chromatography-Mass Spectrometry”. Michal is presently a Ph.D. student under Dr. Liang Li.

What is the key point or highlight that you want the audience to take away from your

Acyl-CoA metabolites can be derivatized with base-activated dansylation for hydroxyl metabolites, subsequently separated on a C18 column and detected using mass spectrometry.

Can you summarize your presentation in a 30-second elevator pitch?

Our group developed a four-channel chemical isotope labeling method for global metabolomics that
uses a divide and conquer approach to increase the number of metabolites we can detect. By
separating the metabolites according to the moiety present in their molecule: hydroxyl, amine and phenyl, carbonyl, and carboxyl, each metabolite is tagged with a functional group that increases its hydrophobicity and the ionizability in the ion source. Until now, our group was not able to detect acyl- CoA metabolites that are both very polar and present in low concentrations in biological samples. By confirming that we can label these analytes using one of the four reactions optimizing the sample diluent and elution gradient conditions, I was able to significantly improve the peak shapes and confidently detect short-chain acyl-CoAs using the LC-MS.

Where do you think the future of this technology lies?

Even though the coverage we achieve with our approach is great, there are still many more metabolites we cannot currently detect. It leaves space for future work in our research. As for my next plan, I am excited to try the optimized conditions for detecting the acyl-CoAs on real biological samples and see the results.

Ewenet Mesfin

The ECM Best poster award for undergraduate students was both awarded to our TMIC young scientists. Congratulations to Ewenet Mesfin with the poster presentation entitled “Analysis of Plant Volatiles Using TD-GCxGC-TOFMS (Thermal Desorption-comprehensive Two Dimensional Gas Chromatography-time of Flight Mass Spectrometry)”. Ewenet is a student in the Chemistry department and working as a research assistant at Dr. Harynuk’s laboratory throughout the summer.

What is the key point or highlight that you want the audience to take away from your presentation?

  • Sampling plant Volatile Organic Compounds (VOCs) using Solid-phase Micro-extraction (SPME) vs TDU shows a significant difference in the number of total peaks detected by GC×GC-TOFMS.
  • For untargeted plant VOCs the results suggest that the Thermal Desorption Unit (TDU) sampling method shows more reliable results.
  • This methodology could be applied to hundreds of crops for further metabolite discovery and validation.

Can you summarize your presentation in a 30-second elevator pitch?
My poster presentation was about the Analysis of plant volatiles using TD-GCxGC-TOFMS. In this study, we have compared two different techniques of collecting in-vivo plant VOCs. SPME is one method where it shows a lower number of compounds being trapped compared to Thermal desorption (TD) tubes. As a conclusion for the untargeted study of plant VOC, using TD coupled to GC×GC-TOFMS essentially helps to observe more compounds with greater peak areas.

Where do you think the future of this technology lies?

The techniques of collecting VOCs using TD tubes will be essentially used for a wide range of applications for hundreds of plant crops metabolite studies.

Abby Kropielnicki

Another ECM best poster award for the undergraduate category was honored to Abby Kropielnicki with the poster entitled “Colorimetric Analysis of L-carnitine: a Biomarker for Sheep Pregnancy Status and Human Disease”. Abby is a student at the University of Alberta with a specialization in Immunology & Infection. She has been working at Dr. Wishart’s laboratory as research assistant since middle of 2021.

What is the key point or highlight that you want the audience to take away from your presentation?

 The aim of my research is to create a simple, innovative detection method for the pregnancy and litter size of sheep, and other livestock, for farmers in the agricultural industry. In addition, this new, L-carnitine-based assay has the potential to detect and manage human health status for various diseases associated with this important metabolomic biomarker. Overall, this detection method could be taken on-farm or in-clinic as a pen-side instrument, allowing for it to be a well-rounded test for the benefit of both the farmer and the general public. 

Can you summarize your presentation in a 30-second elevator pitch?

The core concept of my research is to translate complex, expensive lab diagnostic tools into a pen-side instrument for the public. An alternative option for evaluating animal pregnancy (the current gold standard being ultrasonography) as well as human disease (blood tests and trained personnel required) is a critical area of research with increasing demand. The objective of my research was to create an accurate, affordable, and user-friendly detection method for disease prediction which also has the potential to be used towards various applications related to the biomarker which this assay is centralized around. L-carnitine is the most prominent metabolomic biomarker among a panel of 8 biomarkers for detecting sheep pregnancy. These panels of biomarkers were identified after an analysis of 400 sheep serum samples by Nuclear Magnetic Resonance in the Wishart node of The Metabolomic Innovation Centre. Downstream of the workflow, a colorimetric assay for L-carnitine was developed under the supervision of Dr. David Wishart. With increased yellow colour pigmentation of the assay, increased L-carnitine concentrations are detected. We can extend our studies to other animals such as cows and porcines. This point-of-care detection method was developed for those who are looking to bypass the costs associated with accessing qualified technicians, saving both time and money when evaluating animal and human health.

Where do you think the future of this technology lies?

In the field of veterinary and physician diagnostics, point-of-care tests are highly advantageous over current bench-side laboratory tests (as can be seen with point-of-care Covid-19 tests). These cost-effective, fast, innovative technologies are more easily accessible to remote and/or disadvantageous populations. This new detection method is in the process of being transitioned from a successful single-step stable, dry colorimetric assay using serum into an automated, pen-side instrument with colour detection sensors and fluidics. Once the device is complete, the final goal is to conduct a large population study, validating the pen-side tool. After successfully launching the new point-of-care test, similar technologies can be developed for other metabolites and targeted species such as in the cattle industry and in human disease diagnosis.

There were a few assays mentioned in the presentation that offered by different nodes of TMIC, check them out further:

Global (Untargeted) Metabolomics by Chemical Isotope Labeling LC-MS increases metabolome coverage and achieves accurate quantification for all detectable metabolites. The whole metabolome is analyzed by combining the analysis of four submetabolomes: amine/phenol, carboxyl, carbonyl and hydroxyl submetabolome. The combined results from four channels are able to cover 85% to 95% of the entire chemical space of the metabolome. (Zhao S. et al., Anal. Chem. 2019, 91, 12108−12115

NMR Analysis supports absolute quantification of up to 340 metabolites. This targeted analysis can be used for the identification and quantitation of water-soluble metabolite classes including amino acids, sugars, alcohols, organic acids, amines, TCA cycle intermediates, and short chain fatty acids. We recently discussed the benefits and importance of NMR in the future of Metabolomics (which is so-called “sleeping giant”, which has to be awakened).

Untargeted Metabolomics by GCxGC TOF MS is a semi-quantitative method for known and unknown metabolite profiling. It is an ideal discovery platform for analysis of volatile and semi-volatile compounds, as well as compounds that can be derivatized with standard chemistries, allowing us to also detect simple sugars, sterols and larger molecules. Aside from simple liquid injections of extracts, a wide range of injection methods are available including solid-phase microextraction, static headspace, dynamic headspace, and thermal desorption. This service typically allows for the detection of 2000-9000 features (sample dependent). Relative quantification is provided based on peak areas, normalized to appropriate internal standards.

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