Global, untargeted metabolic profiling requires probing as much chemical space as possible, which includes everything from small, polar nucleotides to long chain fatty acids and free lipids. This vast diversity of metabolites and chemical space poses a problem for global metabolomics: no individual platform can sufficiently measure the breath of metabolites that exist in biological samples. Therefore, specific platforms are required to capture the variety of molecules that are of interest to researchers. At HMT, we utilize two approaches to cover the wide range of chemical classes: Capillary Electrophoresis – Mass Spectrometry (CE-MS) and Liquid Chromatography – Mass Spectrometry (LC-MS).
HMT’s unique CE-MS technology provides accurate and robust profiling of polar metabolites, including:
- phosphorylated sugars
- amino acids
- short and medium chain fatty acids
- bacterial metabolites
- and much more!
Capillary electrophoresis provides extremely high levels of separation and resolution, which allows us to provide polar metabolite profiling with the highest specificity and widest dynamic range.
Basic Scan / Dual Scan – Multipurpose Profiling for Global Metabolism
Global metabolic profiling targets more than 1,200 metabolites involved in primary and secondary metabolism pathways shared by many organisms.
Our unique CE-MS analysis provides accurate and robust profiling including the metabolism of sugars, amino acids, nucleotides, lipids, and stress response biomarkers. Together with CE-MS, LC-MS based lipidomics provides changes in lipid turnover engaged in cellular energy and signaling pathways.
This package is useful for researchers in a variety of areas and sample species.
- Global measurement of > 1,200 metabolites involved in primary and secondary metabolism Basic Scan: CE-MS analysis of sugars, amino acids, nucleotides and other ionic metabolites Dual Scan: LC-MS analysis of fatty acids, steroids and other lipids in addition to Basic Scan
- Report including pathway mapping, PCA, HCA and statistical analysis
- Suitable for a variety of research areas including medical, agriculture, bioprocessing, etc.
OMEGA Scan – Ultrasensitive Global Profiling (CE + LC)
OMEGA Scan makes use of CE-MS, but utilizes our more sensitive FTMS mass spectrometer, which more than doubles the number of metabolites reported in Basic Scan and boasts improved sensitivity and dynamic range. OMEGA Scan can also be used in conjunction with any lipidomics method (OMEGA LC, Basic LC or MSCAN).
OMEGA-LC is the right companion to OMEGA-Scan providing LC-MS untargeted coverage of hydrophobic (lipid) metabolites. Using the same ultrasensitive FTMS as in OMEGA Scan, OMEGA-LC uses lipid extraction and affinity selection to provide hundreds of lipid-like metabolites, phytochemicals and xenobiotics. Affinity selection removes large diacyl and triacyl glycerides to facilitate a deeper look into the lipidome.
Aside from Basic Scan and OMEGA Scan having more than one lipid option, one may also choose one of two quantitative platforms: Q110 or Q353. The Q110 is a list of 110 metabolites for which quantitative measurements will be provided along with the untargeted list of metabolites from Basic Scan or OMEGA Scan. This list contains amino acids, nucleic acids, antioxidants, glycolytic intermediates, and other polar metabolites. The Q110 is provided with each Basic and OMEGA Scan data set. The Q353 is an extended list of 353 metabolites, that includes middle chain fatty acids, neurotransmitters and more. The Q110 list is provided with each CE-MS data set while the Q353 is an additional fee. These quantitative panels can be customized to the client’s needs and requests.
Novel Metabolite Discovery
Another option for Global profiling is the provision of raw data; data that includes unannotated features and metabolites with unvalidated identifications. For the Basic Scan this data is referred to as PRTGa data, while for OMEGA Scan this data is referred to as the Advanced Scan option. The PRTGa and Advanced options include comparisons to our unique peptide library. Our OMEGA-SEARCH option leads clients through a process of novel metabolite identification and validation.
Combination provides ultimate coverage
Our optimum platform for Global Profiling is OMEGA2-353, a combination of OMEGA Scan with Q353 and OMEGA-LC. This service, especially with the advanced option, delivers up to 1000 or more metabolites for biomarker discovery and broad metabolite profiling of both polar and lipid metabolomic space.
These packages are useful for researchers in a variety of areas and sample types by providing high resolution data and comprehensive solutions:
- OMEGA2-353 global untargeted measurements uses our polar and lipid libraries, and can include unannotated and tentatively assigned metabolites.
- Multiple lipid options include OMEGA-LC, Basic-LC and MSCAN.
- Quantitative data using either Q110 or Q353 preselected panels that can be customized with clients’ metabolites of interest.
- Data reports include pathway mapping, PCA, HCA, statistical analysis, and biological interpretation.
- Interpretations can include RNA sequencing, 16s sequencing, metagenomic sequencing, cytokine/chemokine measurements, clinical phenotypes and/or drug regime – comorbidities and more.
- OMEGA Search provides pipeline of activities to identify and validate new metabolite discovery.
- These platforms are suitable for a variety of research areas including medical, agriculture, bioprocessing, microbiome, cancer, skincare, cellular agriculture and more.
Deep Dive Advanced Scan – Ultimate Metabolomics Platform for Biomarker Discovery
Our Advanced profiling mode covers the complete metabolome including unknown metabolites observed in CE-TOFMS data analysis. In addition to cell derived metabolites, peptides, exogenous compounds, and disease specific novel compounds will be detected and screened as biomarkers for disease or bioprocessing.
All projects are performed under QC conditions based on established SOPs and experience of over 10 years of operation. Together with multivariate and statistical analysis, this package is the ultimate tool for biomarker screening and metabolism based profiling.
- Profiling of whole compounds measured by CE-MS including unknown metabolites
- Unknown compounds accompany precise m/z assisting the prediction of chemical structure
- Report including pathway mapping, PCA, HCA, statistical analysis and follow up discussion
- Suitable for biomarker screening and metabolome based profiling
HMT Deep Dive relies on three fundamental properties unique to HMT
- High resolution and sensitivity of CE
- Unique chemical space observed by CE-MS
- Metabolite identifications based on common physical properties and accurate migration times to predict metabolite structures within specific chemical classes.
Such advantages of CE-MS allows HMT to delve deeply into the metabolomic space, adding to the understanding of molecular mechanisms, biological processes, diseases, and the identification of potential new biomarkers.
Mediator Scan – Targeted for Signaling and the Inflammasome
In contrast to the LC-MS method for Dual Scan, M-SCAN represents a novel targeted method for measuring lipid mediators, critical signaling modulators involved in many pathways such as inflammation, allergic response, oxidative stress, and the various metabolic diseases.
The sample protocol for M-SCAN also minimizes large triglycerides in favor of 400 lipid mediators. While the current protocol is designed for relative expression, quantitative profiling for selected lipids can be requested by our clients.
M-Scan Targets Include:
- Fatty acid-derived – prostaglandins, thromboxanes, leukotrienes, lipoxins etc.
- Phosphate structures – platelet activating factors, endogenous cannabinoids (anandamide, 2-acylglycerol), lysophosphatidic acids, lysophotidylcholines, lysophosphatidylserines, sphingosine-1-phosphates etc.
- Sterol structures – glucocorticoids (glucose corticoid), aldosterones (mineral corticoid), sex steroids (estrogen, androgen etc.), bile acids, vitamin D, etc.
Lipid mediators have a wide variety of effects on physiological functions, including immunity, biological defense, blood-pressure regulation, pain or fever, gastrointestinal tract activity and growth, division, and differential regulation of cells. Many diseases are associated with failure in functional balance of such effects. A large portion of lipid mediators act by binding with G-protein coupled receptors found on the cell membrane. Meanwhile, steroid hormones, vitamin A, vitamin D, and others bind with nuclear receptors and induce gene expression changes.
- Absolute quantitation of 110 target metabolites for CE-MS analysis
- LC-MS analysis for lipidomics
- CE-MS analysis for short peptides
- Additional data analysis (PCA, HCA, pathway mapping etc.)
- Feasibility experimental test for sample assessment