Metabolomic Stuff

Question 1

How do Primary & Intermediary Metabolites differ?

Answer 1

Primary: Normal physiological function (ie. Cell Growth)

Intermediary: Maintains homeostasis (ie. Energy Intermediates).

Question 2

What is the “metabolome”?

Answer 2

Complete set of small molecules (ie. Carbs, Lipids) in a biological sample… Excludes enzymes, genetic materials, structural molecules.

Question 3

What techniques are used in metabolomics?

Answer 3

LC-MS (espec. HPLC)
GC-MS
NMR
Capillary Electrophoresis

Question 4

Rs formula…?

Answer 4

Rs = [(V2 - V1) / (W1 + W2)] x [2]

Question 5

What effect does resin particle size have on barometric pressure & resolution in columnar liquid chromatography?

Answer 5

Pressure: Will be higher.
Resolution: Will be better.

Question 6

If flow rate is too high in liquid chromatography, what happens to resolution?

Answer 6

Reduced

Question 7

Typical pressure & flow rate in HPLC?

Answer 7

Pressure: 200-400 bar
Flow Rate: 1mL / min

Question 8

Particulate Retention in Normal Phase HPLC is determined by what interaction?

Answer 8

Polar portion of the Stationary Phase with particles flowing through the column.

Question 9

The Stationary Phase in Normal Phase HPLC is _____, whereas the Mobile Phase is _____.

Answer 9

Polar; Non-Polar

Question 10

What are some examples of compounds contained within the Stationary Phase of Normal Phase HPLC?

Answer 10

Silica, CN, NH2

Question 11

What are some examples of compounds contained within the Mobile Phase of Normal Phase HPLC?

Answer 11

Hexane

Question 12

Reverse Phase HPLC differs from Normal Phase HPLC how?

Answer 12

Normal: Stationary Phase = Polar; Mobile = Non-Polar.

Reverse: Stationary Phase = Non-Polar; Mobile = Polar.

Question 13

Common ligands used in the Stationary Phase of Reverse Phase HPLC?

Answer 13

C8, C18

Question 14

Common ligands used in the Mobile Phase of Reverse Phase HPLC?

Answer 14

Methanol, Acetonitrile

Question 15

Average particle size & columnar pressure in UPLC?

Answer 15

Particle Size: 1.7 - 1.8um
Pressure: 1030 bar

-Makes this extremely sensitive, provides highest resolution, & increases processing speeds.

Question 16

Primary usage of UPLC in comparison to standard LC or HPLC?

Answer 16

Reduces band spreading (narrows down the population of analyte molecules).

Question 17

Primary gasses used as Mobile Phases in GC?

Answer 17

Helium (most common & $$$), Hydrogen, Nitrogen.

Question 18

Compounds used in the Stationary Phase of GC Capillary Columns?

Answer 18

PEG, HCs, Esters, Amides, Polysiloxanes

Question 19

According to the Human Metabolome Database (HMDB), what molecular size would classify a metabolite as either ‘abundant’ or ‘relatively rare’?

Answer 19

Abundant: > 1uM
Rare: < 1nM

Question 20

A lipid profile is an example of a Metabolite ______ ______, as it provides a quantification of a specific group of metabolites.

Answer 20

Target Analysis

Question 21

Is Metabolite Profiling or Metabolite Fingerprinting more targeted?

Answer 21

Profiling

Question 22

Utilization of urine metabolomic data in the characterization of biomarkers associated with elderly diabetic patients is an example of what?

Answer 22

Metabolite Profiling

Question 23

Cancer staging based upon H-NMR samples of metabolites from studied patients would be an example of what Metabolomics study?

Answer 23

Metabolite Fingerprinting