Global (Untargeted) Metabolomics by Chemical Isotope Labeling LC-MS

To increase metabolome coverage and achieve accurate quantification for all detectable metabolites, we use a Chemical Isotope Labeling (CIL) Metabolomics Platform (Zhao et al., 2019, Zhao et al., 2020) for metabolome analysis. 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. However, a user can choose the labelling method(s) to analyze one or more chemical-group-based submetabolomes, depending on budget and desired coverage. Using one submetabolome analysis, we detect and perform accurate relative quantification of up to 2,500 metabolites (not features) in many different types of samples, including urine, blood, plant extract and cell extracts. Using a combination of four submetabolome analyses, we can cover over 7,000 to 10,000 metabolites (sample dependent) per sample. For the analysis, we will do sample preparation, metabolite extraction, chemical labelling, LC-MS analysis, data analysis, metabolite ID, and statistical analysis (PCA, PLS-DA, Volcano plots). We will provide a summary of our analysis and statistical results. The peak pairs (metabolites) detected, positive or putative IDs, and their relative quantification data will be summarized in an Excel file, which can be used for further analysis such as uploading to MetaboAnalyst for more statistical analysis or pathway analysis. References: Shuang Zhao, Hao Li, Wei Han, Wan Chan and Liang Li, “Metabolomic Coverage of Chemical-Group-Submetabolome Analysis: Group Classification and 4-Channel Chemical Isotope Labeling LC-MS”, Anal. Chem. 2019, 91, 12108-12115. Shuang Zhao and Liang Li, “Chemical Derivatization in LC-MS-Based Metabolomics Study”, Trends in Analytical Chemistry (TrAC), 2020, 131, 115988. For more information about our different options and levels of metabolome coverage:

• Basic Global Metabolomics Analysis by Chemical Isotope Labeling LC-MS
• Elevated Global Metabolomics Analysis by Chemical Isotope Labeling LC-MS
• Comprehensive Global Metabolomics Analysis by Chemical Isotope Labeling LC-MS

Service Workflow

TMIC’s Li Node uses a flexible and staged analysis strategy to maximize results and fit all budgets. With global metabolomics profiling, clients may be uncertain about the outcome of the analysis and feel that there is a high investment risk to generate extensive datasets during early stage discovery that may or may not support study goals. We therefore offer a one-channel global metabolomics analysis which allows identification of 2000-3000 metabolites, not features, for $120 CAD per sample. This basic analysis includes results (metabolite identification, accurate relative concentration), and basic statistics, and can be very useful in designing the next stage of global analysis. The basic one-channel analysis is cost effective, and provides useful metabolomics information as is or as the first step towards more comprehensive metabolomics studies. A typical workflow for applying the flexible and staged analysis strategy includes:

1. Receiving raw samples from a client (e.g., unprocessed urine, plasma, serum, cell pellet, tissue, etc.).
2. Aliquoting each sample into multiple aliquots, depending on the number of submetabolomes (channels) to be performed and whether global lipidome analysis is also needed.
3. Using one aliquot for Basic Global Metabolomics Analysis (one channel) (@$120 per sample).
4. Sending the one-channel analysis report to the client and followed by consultation to discuss results.
5. Client evaluates the results from the one-channel analysis and makes a decision on:
   1. If the client wishes to continue the analysis of other channels for more comprehensive coverage (e.g., Elevated or Comprehensive Metabolomics Analysis) and/or lipidome analysis, we will continue in this manner. The final cost of the service will depend on the total number of channels analyzed and whether lipidome analysis is performed (refer to price list).
   2. If the client does not wish to proceed with further analysis, only the cost of the one-channel analysis will be invoiced.

Platform Strengths

High-coverage accurate relative quantification of metabolites and simultaneous profiling of multiple pathways for discovery or targeted metabolomics.  

Metabolite Identification (Tier 1, Tier 2, Tier 3)

Metabolite identification is based on three-tiered ID approach: Tier 1: CIL Library based on accurate mass, RT and peak pairs. Tier 2: Linked Identity Library, high confidence putative identifications (90% accurate) based on accurate mass and predicted RTs. Tier 3: MyCompoundID based on masses.  

Example Data (Human serum; other types of samples should give similar results)

Sample Metabolite Identifications for BASIC Global Metabolomics by CIL LC-MS

For the lists of metabolite identifications from the elevated or comprehensive analysis, please contact Professor Liang Li (email: liang.li at ualberta.ca).

Statistical Analysis

Basic statistics included.  

Prices

BASIC Global Metabolomics by CIL LC-MS:

• Academic: $120/sample
• Industry: $156/sample

ELEVATED Global Metabolomics by CIL LC-MS:

• Academic: $213/sample
• Industry: $277/sample

COMPREHENSIVE Global Metabolomics by CIL LC-MS:

• Academic: $399/sample
• Industry: $519/sample

Flexible Global Metabolomics by CIL LC-MS:

• Academic: $120 CAD/sample for the first channel
• $93 CAD for any additional channel. For example, choose 3 channels: $120+2x$93=$306 per sample.

Industry: $156 CAD/sample for the first channel

• $121 CAD for any additional channel. For example, choose 3 channels: $156+2x$121=$398 per sample.

Notes: The above prices apply to a sample size of >20. If less than 20 samples, add $400 extra per channel. A flat fee of $200 for sample handling per project is added to each project.