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Clinical Chemistry > Module 3d: Chromatography > Flashcards

Module 3d: Chromatography Flashcards

1
Q

Refers to the group of techniques used to separate complex mixtures on the basis of different physical and chemical interactions between the compounds and the stationary phase.

A

Chromatography

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2
Q

Four basic components in any chromatographic technique:

A
  1. stationary phase
  2. mobile phase
  3. column
  4. eluate
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3
Q

Component that carries the complex mixture (sample)

A

mobile phase

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4
Q

component which can either be solid or liquid through which the mobile phase flows

A

stationary phase

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5
Q

component that holds the stationary phase

A

column

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6
Q

The separated components

A

eluent

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7
Q

5 separation mechanisms of chromatography

A
  1. ion-exchange
  2. partition
  3. adsorption
  4. size exclusion
  5. affinity mechanisms
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8
Q

Separation mechanism that is based on the differences or magnitude of their ionic charge.

A

ion-exchange chromatography

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9
Q

An exchangable ion found in close proximity with the fixed charge to maintain electrochemical neutrality.

A

counterion

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10
Q

Ion exchange chromatography used primarily for strongly acidic groups

A

Cation-exchange particles

acids have high H+ that can exchange with negatively charged stationary phase

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11
Q

Ion exchange chromatography used primarily for strongly basic quarternary amines with positive charges.

A

anion exchange packings

basic solutions have less H+ and adhere to negatively charged stationary phase

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12
Q

Chromatography involving separation of solutes based on their differential distribution of between two immiscible liquids

A

partition chromatography

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13
Q

partition chromatography separates solutes based on

A

the difference in solubility of the solute molecules between the stationary and mobile phase

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14
Q

Partition chromatography is classified into

A

gas liquid chromatography

liquid liquid chromatography

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15
Q

Liquid liquid chromatography is further classified into:

A

normal phase

reverse phase

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16
Q

Normal phase LLC has

A

stationary - polar

mobile - non- polar

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17
Q

Reverse phase LLC has

A

stationary - non polar

mobile - polar

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18
Q

Version of partition chromatography wherein analytes elute in order of increasing polarity

A

hydrophilic interaction chromatography

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19
Q

Separation technique based on the differences between the adsorption and desorption of solutes at the surface of a solid.

A

Adsorptiom chromatography

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20
Q

adsorption chromatography is also known as

A

liquid- solid chromatography

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21
Q

In adsorption chromatography, the molecules that are most soluble in the mobile phase move the ___________

A

fastest

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22
Q

Separation method based on the molecular sizes and shape

A

Size exclusion/ steric exclusion chromatography

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23
Q

In size exclusion chromatography, the molecules that remain entirely in the mobile phase are large or small??

A

large

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24
Q

Process where hydrophilic gel isused in separation of enzymes, antibodies and proteins.

A

Gel filtration

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25
Q

example of hydrophilic gels

A

dextran

agarose

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26
Q

Process where hydrophobic gel is used to separate triglycerides and fatty acids

A

Gel permeation

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27
Q

example of hydrophobic gels

A

sephadex

28
Q

separation method that uses the the unique and specific biological interaction of the analyte and ligand

A

affinity chromatography

29
Q

Two forms of chromatography

A

planar

column

30
Q

two types of planar chromatography

A
  1. paper chromatography (PC)

2. thin layer chromatography (TLC)

31
Q

type of paper commonly used in paper chromatography

A

Whatman paper

32
Q

separation mechanism in paper chromatography

A

normal phase partition

non polar mobile phase, polar stationary phase

33
Q

Applications of papaer chromatography

A

fractionation of sugar and amino acid

34
Q

Thinl layer chromatography setup is consist of

A

thin layer of silica gel coated on glass plate

35
Q

separation method of thin layer chromatography

A

Any mode depending on the sorbent or thin layer and solvent applied

36
Q

Migration of solutes in PC and TLC is expressed as

A

Retention Factor (Rf) value

solute migration/ solvent migration

37
Q

Two categories of column chromatography

A

gas chromatography

liquid chromatography

38
Q

Column chromatography which usually provides higher resolution due to much longer columns and higher linear flow rate

A

Gas chromatography

39
Q

Column chromatography of choice for volatile components, including analytes of lower molecular weight.

A

gas chromtography

40
Q

column chromatography usually used for larger molecules like peptides, proteins, nucleic acids, and large polymers.

A

liquid chromatography

41
Q

Setup of Gas chromatography

A

mobile phase: gas

statiomary phase: nonvolatile liquid bonded to the surface of capillary

42
Q

examples of carrier gases (typically inert)

A

N, He, Ar

43
Q

In gas chromatography, molecules that are ______ spend more time in the gas phase

A

more volatile with higher pressure

44
Q

two types of gas chromatography

A

Gas solid chromatography

gas liquid chromatography

45
Q

Basic components of a gas chromatograph

A
  1. carrier gas with flow controller
  2. injector or inlet
  3. chromatographic column
  4. column oven
  5. detector
  6. computer
46
Q

Two types of chromatographic column in gas chromatograph

A

packed column

capillary column

47
Q

These columns can be filled with uncoated support particles.

A

packed column

48
Q

carrier gas most usually used for packed columns

A

nitrogen

49
Q

Chromatographic column which whose fused silica tubular columns are wall-coated with a thin film of liquid phase. They are very efficient but have low sample capacities.

A

Capillary column

50
Q

carrier gas usually used for capillary column

A

high purity hydrogen or helium

51
Q

Most commonly used detector for gas chromatography clinical analysis because of its simplicity, reliability, versatility, sensitivity and ease of operation.

A

flame ionization detector

52
Q

Column chromatography that is based on the distribution of solutes between a liquid mobile phase and a stationary phase

A

liquid chromatography

53
Q

most widely used form of LC where particles of small diameter are employed as the stationary phase support

A

High performance liquid chromatography

54
Q

Basic components of a liquid chromatograph

A
  1. solvent reservoir
  2. pumps
  3. injector
  4. chromatographic column
  5. column oven
  6. online detector
  7. computer
55
Q

Identify type of chromatography:

Important tool for monitoring drugs and their metabolites.

A

LC

56
Q

Identify type of chromatography:

Identify and quantify such drugs as alcohol, used in forensics to compare fibers found on a victim

A

GC

57
Q

Identify type of chromatography:
Detecting pesticide or insecticide residues in food, also used in forensics to analyze the
dye composition of fibers

A

TLC

58
Q

Identify type of chromatography:

Separating amino acids and anions, RNA, fingerprinting, separating and testing histamines, antibiotics

A

PC

59
Q

Used to analyze metal ions and organic compounds in solutions using liquids which
may incorporate hydrophilic, insoluble molecules.

A

LC

60
Q

Used to analyze volatile gases where Helium is used to move the gaseous mixture through a
column of absorbent material

A

GC

61
Q

Simple and rapid method to check the purity of the organic compound using an absorbent material on flat glass plates

A

TLC

62
Q

Most common type of chromatography which uses capillary action to pull the solutes up

A

PC

63
Q

Considered as the gold standard for drug testing

A

GC- MS

64
Q

Used for xenobiotics, anabolic steroids and pesticides.

A

GC-MS

65
Q

Drugs are detected through the presence of

A

decomposition fragments

66
Q

Has great potential for measuring low-level and mixed-polarity analytes such as vitamin D, testosterone, and immunosuppressant drugs

A

LC- MS

Clinical Chemistry (14 decks)

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