Metabolomic Stuff Flashcards
How do Primary & Intermediary Metabolites differ?
Primary: Normal physiological function (ie. Cell Growth)
Intermediary: Maintains homeostasis (ie. Energy Intermediates).
What is the “metabolome”?
Complete set of small molecules (ie. Carbs, Lipids) in a biological sample… Excludes enzymes, genetic materials, structural molecules.
What techniques are used in metabolomics?
LC-MS (espec. HPLC)
GC-MS
NMR
Capillary Electrophoresis
Rs formula…?
Rs = [(V2 - V1) / (W1 + W2)] x [2]
What effect does resin particle size have on barometric pressure & resolution in columnar liquid chromatography?
Pressure: Will be higher.
Resolution: Will be better.
If flow rate is too high in liquid chromatography, what happens to resolution?
Reduced
Typical pressure & flow rate in HPLC?
Pressure: 200-400 bar
Flow Rate: 1mL / min
Particulate Retention in Normal Phase HPLC is determined by what interaction?
Polar portion of the Stationary Phase with particles flowing through the column.
The Stationary Phase in Normal Phase HPLC is _____, whereas the Mobile Phase is _____.
Polar; Non-Polar
What are some examples of compounds contained within the Stationary Phase of Normal Phase HPLC?
Silica, CN, NH2
What are some examples of compounds contained within the Mobile Phase of Normal Phase HPLC?
Hexane
Reverse Phase HPLC differs from Normal Phase HPLC how?
Normal: Stationary Phase = Polar; Mobile = Non-Polar.
Reverse: Stationary Phase = Non-Polar; Mobile = Polar.
Common ligands used in the Stationary Phase of Reverse Phase HPLC?
C8, C18
Common ligands used in the Mobile Phase of Reverse Phase HPLC?
Methanol, Acetonitrile
Average particle size & columnar pressure in UPLC?
Particle Size: 1.7 - 1.8um
Pressure: 1030 bar
-Makes this extremely sensitive, provides highest resolution, & increases processing speeds.
Primary usage of UPLC in comparison to standard LC or HPLC?
Reduces band spreading (narrows down the population of analyte molecules).
Primary gasses used as Mobile Phases in GC?
Helium (most common & $$$), Hydrogen, Nitrogen.
Compounds used in the Stationary Phase of GC Capillary Columns?
PEG, HCs, Esters, Amides, Polysiloxanes
According to the Human Metabolome Database (HMDB), what molecular size would classify a metabolite as either ‘abundant’ or ‘relatively rare’?
Abundant: > 1uM
Rare: < 1nM
A lipid profile is an example of a Metabolite ______ ______, as it provides a quantification of a specific group of metabolites.
Target Analysis
Is Metabolite Profiling or Metabolite Fingerprinting more targeted?
Profiling
Utilization of urine metabolomic data in the characterization of biomarkers associated with elderly diabetic patients is an example of what?
Metabolite Profiling
Cancer staging based upon H-NMR samples of metabolites from studied patients would be an example of what Metabolomics study?
Metabolite Fingerprinting
