FSCI 5355 Flashcards

1
Q

Capillary Electrophoresis and Chromatography (CE) is one of the most powerful liquid phase separation techniques because of:

A

High speed
High efficiency
Different selectivity
Ability to sample small volumes

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

What is CE?

A

Movement of ionic species in conductive media under applied electric field.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

Basic premise of electrophoretic separation:

A

Different charged species move at different velocity and can be separated.

Complex interplay between analyte, ions, capillary surfaces and additives in electrolyte.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

Electroomosis and electromootic flow:

A

Observed when electric field is applied to conductive solution in capillary with fixed charges on its interior wall.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

Potential difference close to wall called:

A

Zeta potential

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

Innermost layer close to capillary surface is static and called:

A

Helmholtz (stern) Layer

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

Second layer more diffuse layer and migrates in direction of cathode called:

A

Outer Helmholtz Plane (OHP)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

During CE migration, associated solvent is drags molecules, which creates:

A

Electromootic flow

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

During CE migration, force propelling liquid originates at charged surface, causes flow to move in ____like fashion as opposed to when pressure pumps liquid.

This difference in flow profile main reason for CE efficiency.

A

Plug

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

Modes of separation (ZE)

Zone electrophoresis is most widely used mode and simplest form of CE:

A
  • Capillary filled w/buffer
  • Separation occurs because solutes migrate to discrete zones at different velocities
  • Neutral compounds cannot be separated
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

Modes of separation

Electrophoretic Chromatography (EC):

A

Allows separation of neutral compounds w/addition of additive.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

Mode of Separation

Sieving Electrophoresis (SE)

A
  • Separation based on differences in size and shape of charged analytes
  • Basis for DNA Generation Sequencing
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

Modes of separation

Electrochromatography:

A
  • Combination of CE and HPLC

- Heterogeneous phase inside capillary

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Modes of separation

Isoelectric Focusing:

A
  • High resolution technique
  • Used to separate proteins
  • Acids and based
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

Modes of separation

Isotachophoresis is when injection of the sample is between leading electrolyte and terminating electrolyte:

A

When voltage is applied, formation of zones is proportional to concentration of analyte.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

Injection/ Main advantage of CE:

A
  • injection of extremely small volumes
  • electric voltage applied to end of capillary, while immersed in DNA sample
  • Negative charge draws sample into capillary
17
Q

Interpretations of electropherograms

Two thresholds:

A

Analytical - average fluorescent signal converted to peak height

Stochastic - signal levels that cannot be used

18
Q

Chromatography:

A

Powerful and versatile technique that is routinely used for single chemical species.

19
Q

Classification of Chromatographic Techniques

Physical set/up of stationary phase:

A

Planar (thin layer)

Column based

20
Q

Chromatographic technique mobile phase:

A

Gas, liquid, supercritical fluid

21
Q

Chromatography further subclassified by separation mechanism:

A
Adsorption
Partition
Ion exchange 
Size exclusion 
Affinity
22
Q

Liquid chromatography based on:

A

Adsorption

23
Q

Gas chromatography based on:

A

Partition mechanism

24
Q

Ion exchange chromatography separation based on:

25
Ion exchange chromatography: Cation retains cations with
Negative charge
26
Anion exchange:
Retains anions with positive charge
27
Column types:
Packed column - Fill with small particles
28
Open Tubular Capillary Column:
- Wall coated - Support Coates - Adsorption (solid particles attach to wall)
29
Chromatography Distribution Equilibria is:
Chromatography separation depend on distribution of sample components between stationary phase and mobile phase.
30
Retention time is:
Time taken for each analyte to move from injection into column until detection.
31
Un-retained compounds elute first:
Void time
32
Tr= tr - tm
Subtract void time from retention time.
33
Spectroscopy is:
Study of interaction between matter and electromagnetic radiation.
34
Mass Spectroscopic Principles:
Ionizer, Mass analyzer and detector
35
What does a mass spectrum do?
Give info on chemical structures.
36
General schematic of mass spectrum:
- Creation of ions - Separation of ions - Detection of ions