METABOLOMICS is the systematic study of the small molecular metabolites in a cell, tissue, biofluid, or cell culture media that are the tangible result of cellular processes or responses to an environmental stress.

1. METABOLOMICS
Presented by S.M GHARIB NAWAZ JAN

2. CONTENTS:
 Introduction
 History
 Advancements
 Techniques
 Applications
 Significances
 Challenges & Problems

3. What is METABOLOMICS?

4. METABOLOMICS:
 Metabolomics is the systematic study of the small molecular metabolites in a
cell, tissue, biofluid, or cell culture media that are the tangible result of cellular
processes or responses to an environmental stress.

5. METABOLOMICS:
 The metabolome is the total complement of metabolites present in a biological
sample under given genetic, nutritional or environmental conditions.

6. METABOLOMICS:
 Metabolomics technologies yield many insights into basic biological research in
areas such as systems biology and metabolic modelling, pharmaceutical research,
nutrition and toxicology.

7. METABOLITES:
 Intermediates and products of metabolism
 MW < 1500 Da
 Examples:
• antibiotics,
• pigments,
• carbohydrates,
• fatty acids and amino acids
 Primary and secondary metabolites

8. METABOLITES:

10. INTER RELATIONSHIP:

11. HISTORICAL BACKGROUND:
 The first paper was titled, “Quantitative Analysis of Urine Vapor and Breath by
Gas-Liquid Partition Chromatography”, by Robinson and Pauling in 1971.
 The name metabolomics was coined in the late 1990s (the first paper using the
word metabolome is Oliver, S. G., Winson, M. K., Kell, D. B. & Baganz, F.
(1998).
 Many of the bioanalytical methods used for metabolomics have been adapted (or
in some cases simply adopted) from existing biochemical techniques.

12.  Metabolomics is expanding to catch up with other multiparallel
analytical techniques (transcriptomics, proteomics) but remains far
less developed and less accessible.
 Human Metabolome project – first draft of human metabolome:
23rd January 2007

13. APPLIED IN OTHER FIELDS:
 Drug assessment
 Clinical toxicology
 Nutrigenomics
 Functional genomics

14. ADVANCEMENT:
 Metabolomics depicts the functional end-point of genetic and environment
 Targeted metabolomics data are analytically reproducible and allow immediate
biochemical interpretation
 Proof-of-concept has been achieved in routine diagnostics of inborn errors of
metabolism

15. ADVANCEMENT:
 Many metabolic biomarkers are valid across species and enable translational
research
 Comprehensive targeted metabolomics bridges the gap to open profiling
approaches

16. ANALYSIS OF METABOLOMICS DATA
TECHNIQUES:
 Separation Techniques
Gas Chromatography (GC)
Capillary Electrophoresis (CE)
High Performance Liquid Chromatography (HPLC)
Ultra Performance Liquid Chromatography (UPLC)
 Combination of Techniques
GC-MS
HPLC-MS
 Detection Techniques
Nuclear Magnetic Resonance Spectroscopy (NMR)
Mass Spectrometry (MS)

17. STEPS IN METABOLOMIC TECHNIQUES:

18. HOW METABOLOMICS WORKS ?

19. GAS CHROMATOGRAPHY:
 Involves a sample being vaporized to a gas and injected into a
column
 Sample is transported through the column by an inert gas mobile
phase
 Column has a liquid or polymer stationary phase that is adsorbed
to the surface of a metal tube
 Columns are 1.5-10 m in length and 2-4 mm in internal diameter
 Samples are usually derivatized with TMS to make them volatile

20. GAS CHROMATOGRAPHY:

24. High Pressure (Performance) Liquid Chromatography -
HPLC
 Developed in 1970’s
 Uses high pressures (6000 psi) and smaller (5 mm), pressure-stable
particles
 Allows compounds to be detected at ppt (parts per trillion) level
 Allows separation of many types of polar and nonpolar compounds

27. HPLC MODALITIES
 Reversed phase – for separation of non-polar molecules (nonpolar
stationary phase, polar mobile phase).
 Normal phase – for separation of non-polar molecules (polar
stationary phase, non-polar/organic mobile phase).

30. CAPILLARY ELECTROPHORESIS:
 Higher theoretical separation efficiency than HPLC (although
requiring much more time per separation)
 Suitable for use with a wider range of metabolite classes than is
GC.
 As for all electrophoretic techniques, it is most appropriate for
charged analytes.

31. MASS SPECTROMETRY
 Mass spectrometry is a technique to measure the
mass of ions (m/z)
 All mass spectrometers perform three main tasks:
1. Ionize molecules.
2. Acceleration: Use electric and magnetic fields to
accelerate ions and manipulate their flight.
3. Detect ions (convert to electronic signal)

33. DIFFERENT TYPES OF MS:
 GC-MS - Gas Chromatography MS
separates volatile compounds in gas column and ID’s by mass
 LC-MS - Liquid Chromatography MS
separates delicate compounds in HPLC column and ID’s by mass
 MS-MS - Tandem Mass Spectrometry
separates compound fragments by magnetic or electric fields and
ID’s by mass fragment patterns

34. TARGETED VS UNTARGETED METABOLOMICS:
TARGETED METABOLOMICS:
 Pre-defined set of metabolites to
quantify
 Typically carried out in diagnostics
 Pros: Technically simple
 Cons: Limited scope, missing
information
UNTARGETED METABOLOMICS:
 Global analysis of metabolic changes in
response to disease, environmental or genetic
perturbations.
 Typically carried out for hypothesis generation,
followed by targeted profiling for more
confident quantification of relevant metabolites.
 Pros: Unbiased (no selection of metabolites)
 Cons: Technically challenging (both the analysis
and the bioinformatics), risk of getting too many
unknowns

35. TARGETED & UNTARGETED METABOLOMICS:

36. EXOMETABOLOMICS OR METABOLIC
FOOTPRINTING:
 Study of extracellular metabolites.
 Uses many techniques from other subfields of metabolomics.
 Has applications in:
- biofuel development,
- bioprocessing,
- determining drugs' mechanism of action,
- studying intercellular interactions.

37. NUCLEAR MAGNETIC RESONANCE
SPECTROSCOPY

38. NMR SPECTROSCOPY
METABOLOMIC FINGER PRINTING
 Detection technique which does not rely on separation of the analytes,
and the sample can thus be recovered for further analyses.
 All kinds of small molecule metabolites can be measured simultaneously
- in this sense, NMR is close to being a universal detector.
 The main advantages of NMR are high analytical reproducibility and
simplicity of sample preparation.
 Most metabolites have unique chemical shift fingerprints that helps
reduce redundancy.
 Such analysis is known as finger printing.

40. LIST OF SOFTWARES FOR METABOLOMIC ANALYSIS:

41. APPLICATIONS:
 Basic research:
- Functional genomics in biochemistry, physiology, cell biology, microbiology, ecology, …
 Agricultural & nutrition industry:
- Plant intermediary metabolism
- Health effects of functional food products
 Biotechnology:
- Optimization and monitoring of fermentation processes
 Pharmaceutical R&D:
- Pathobiochemistry / characterization of disease models
- Safety / toxicology
- Efficacy / pharmaco-dyanamics and mode-of-action
 Clinical diagnostics & theranostics:
- Early diagnosis and accurate staging
- Specific monitoring of therapeutic effects

42. SIGNIFICANCES:
 Genetic Disease Tests
 Nutritional Analysis
 Clinical Blood Analysis
 Clinical Urinalysis
 Cholesterol Testing
 Drug Compliance
 Transplant Monitoring
 MRS and CS imaging

43. CHALLENGES:
 Metabolomics is not only concerned with the identification and
quantification of metabolites.
 It is also concerned with relating metabolite data to biology and
metabolism.
 Metabolomics requires chemical information which is linked to
both biochemical causes and physiological consequences. This
means that metabolomics must combine the two very different
fields of informatics: bioinformatics and chemoinformatics.

44. ANY QUESTIONS ???
FEEL FREE TO ASK.