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Comparison Of Three ISD Models > FSCI 5355 > Flashcards

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

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2
Q

What is CE?

A

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

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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.

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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.

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5
Q

Potential difference close to wall called:

A

Zeta potential

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6
Q

Innermost layer close to capillary surface is static and called:

A

Helmholtz (stern) Layer

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7
Q

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

A

Outer Helmholtz Plane (OHP)

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8
Q

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

A

Electromootic flow

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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

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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
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11
Q

Modes of separation

Electrophoretic Chromatography (EC):

A

Allows separation of neutral compounds w/addition of additive.

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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
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13
Q

Modes of separation

Electrochromatography:

A
  • Combination of CE and HPLC

- Heterogeneous phase inside capillary

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14
Q

Modes of separation

Isoelectric Focusing:

A
  • High resolution technique
  • Used to separate proteins
  • Acids and based
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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.

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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:

A

Charge

25
Q

Ion exchange chromatography:

Cation retains cations with

A

Negative charge

26
Q

Anion exchange:

A

Retains anions with positive charge

27
Q

Column types:

A

Packed column

  • Fill with small particles
28
Q

Open Tubular Capillary Column:

A
  • Wall coated
  • Support Coates
  • Adsorption (solid particles attach to wall)
29
Q

Chromatography Distribution Equilibria is:

A

Chromatography separation depend on distribution of sample components between stationary phase and mobile phase.

30
Q

Retention time is:

A

Time taken for each analyte to move from injection into column until detection.

31
Q

Un-retained compounds elute first:

A

Void time

32
Q

Tr= tr - tm

A

Subtract void time from retention time.

33
Q

Spectroscopy is:

A

Study of interaction between matter and electromagnetic radiation.

34
Q

Mass Spectroscopic Principles:

A

Ionizer, Mass analyzer and detector

35
Q

What does a mass spectrum do?

A

Give info on chemical structures.

36
Q

General schematic of mass spectrum:

A
  • Creation of ions
  • Separation of ions
  • Detection of ions

Comparison Of Three ISD Models (3 decks)

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