Method Development

The development of mass spectrometry and NMR methods to aid in metabolic profiling and unknown metabolite identification is a major focus of the NW-MRC.

NMR and MS methods are powerful methods and their combination promises new routes in the metabolomics field. However, the inability to correlate the data between the two platforms is a major bottleneck. To alleviate this problem and exploit the combined strength of the two powerful methods, major research efforts in our laboratory are focused on the following:

  1. Development of isotope labeled approaches for the detection of enhanced pool of quantifiable metabolites using NMR spectroscopy.
  2. Development of smart isotope tags for efficient detection of the same metabolites by both NMR and MS, which enable bringing the NMR and MS data on the same platform.
  3. Development of ratio analysis spectroscopy for unknown metabolites identification.
  4. Simplification of complex spectra following selective detection or selective suppression approaches.

Recent Developments

Targeted Metabolites

We are continually adding compounds to our targeted LC-MS assay. Of note, we have recently added nucleotides, vitamins, and several sugars to our current list of 300+ molecules. To further optimize our offerings, we may be shifting certain compounds to other instruments. Please contact us for more information.

Oxylipin Assay

In the past, analysis of oxylipins were commonly conducted by immunoassay techniques or GC-MS, which have their limitations due to cross-reactivity, molecular interference, and a lack of sensitivity due to low concentrations or require a time consuming derivation step. As potential biomarkers for their regulatory roles in inflammation, cardiovascular protection, blood clotting and apoptosis, it became necessary to implement a powerful method for oxylipin quantitation. We have recently added a targeted 98 oxylipin assay to our Sciex 5550 instrument, which quantitates by using isotopic standard compounds.

Bile Acids

Bile acids (BAs) are signaling molecules that play critical roles in lipid absorption, metabolism, cholesterol homeostasis, vitamin circulation, and can reflect internal conditions for intestinal and liver abnormalities and pathology. However, due to its poly-ring structure and various hydroxylation patterns, their complex structure makes it difficult to differentiate between compounds. We have developed a targeted 55 compound assay for conjugated and unconjugated BAs on our Waters Micro MS instrument.

Short Chain Fatty Acids

Short Chain Fatty Acids (SCFAs) have become an important biomarker for identifying the condition of the microbiota, immunological regulation, obesity, and other metabolic and disease states. They were once commonly analyzed by GC-MS, but we have created an isotope-labeled derivatization and quantification protocol for 10 SCFAs on our Waters Micro MS instrument.

Tryptophan

As an essential amino acid and a precursor for many biologically active substances, tryptophan is an important identifier of physiopathological states such as cardiovascular disease, inflammation, metabolism, and cancer. We are currently able to target 22 tryptophan related metabolites.

Tissue Preparation

Although we run biofluids such as serum/plasma, we are able to develop new methods for appropriate metabolite extraction. Some other sample types we offer include tissue (brain, liver, etc), drosophila (whole fly, hemolymph, and head), cells, urine, fecal, vaginal fluids, etc.

Please note:

Our assays target the number of compounds stated here and on our Assays & Fees page, but the actual number of compounds reported for analysis will depend on type of sample, concentration, type of assay, batch variance, and other factors.

We reserve the right to refuse certain preparation styles if we are not equipped to do so or if it is more favorable for your samples to come to us prepared (eg. cells already counted, hemolymph already extracted, etc). There is an additional cost for sample and/or tissue prep.

If you do not see your target or sample type of interest here or in our Assays & Fees page, please contact us to discuss possibilities for specialized assay or sample prep development.