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CC1 LECTURE DEAN > SPECTROPHOTOMETRY PART 2 > Flashcards

SPECTROPHOTOMETRY PART 2 Flashcards

1
Q
  • not routinely done
A

SPECIAL

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2
Q

excitation: unstable

A

FLAME PHOTOMETRY

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3
Q

ground state: emit (emission) radiant energy

A

FLAME PHOTOMETRY

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4
Q

heating: dissociation of atoms

A

FLAME PHOTOMETRY
ATOMIC ABSORPTION SPECTROPHOTOMETRY

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5
Q

Detection of trace elements

A

ATOMIC ABSORPTION SPECTROPHOTOMETRY

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6
Q
  • in combination with proteins
A

mercury

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7
Q

: absorb (absorption) radiant energy

A

hollow cathode lamp

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8
Q
  • allowed to pass through; converted to electrical signal
A

transmitted light

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9
Q

Light Source: FEP

A

Flame

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10
Q

Light Source: AAS

A

Hollow Cathode Lamp

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11
Q

State of atoms: FEP

A

Excited

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12
Q

State of atoms: AAS

A

Ground State

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13
Q

Energy measured: FEP

A

Thermal

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14
Q

Energy measured: AAS

A

Radiant

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15
Q

Basis of measurement: FEP

A

Light emission

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16
Q

Basis of measurement: AAS

A

Light absorbance

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17
Q

Sensitivity: FEP

A

Lesser

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18
Q

Sensitivity: AAS

A

Greater

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19
Q
  • failure of the flame to dissociate the sample
A
  1. CHEMICAL INTERFERENCE
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20
Q

*unique trace elements in CHEMICAL INTERFERENCE

A
  1. Lanthanum
  2. Strontium
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21
Q

*calcium phosphate remedy:

A

displacement principle

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22
Q
  • excitation instead of dissociation
A
  1. IONIZATION INTERFERENCE
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23
Q

*absorbs radiant energy at ground state = EXCITATION instead

A
  1. IONIZATION INTERFERENCE
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24
Q

*cause: HIGH TEMPERATURE

A
  1. IONIZATION INTERFERENCE
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25
Q
  • interference on the absorption by evaporation of solvents or formation of solids
A
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26
Q
  1. MATRIX INTERFERENCE

*examples:

A
  2. Use of organic compounds
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27
Q

USE OF FLAMELESS AAS

A

carbon rod or graphite furnace

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28
Q

heated until dried/charred

A

FLAMELESS AAS

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29
Q

FLAMELESS AAS: dissociation of atoms

A

atomized

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30
Q

more sensitive

A

FLAMELESS AAS

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31
Q

permits determination of trace metals in small samples

A

FLAMELESS AAS

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32
Q

Atoms absorb light of a particular wavelength and emit light of a longer wavelength (lower energy)

A

FLOURESCENCE SPECTROPHOTOMETRY

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33
Q

USE OF FLOURESCENCE SPECTROPHOTOMETRY

A

Porphyrins, hormones, amino acids, vitamins, cathecolamines

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34
Q

FLOURESCENCE SPECTROPHOTOMETRY COMPONENTS

A

Energy: mercury or xenon arc lamp, slit 1 degree monochromator, quartz cuvettes, 2 degree monochromator, detector, readout device

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35
Q
  1. Xenon
  2. Entrance slit
  3. Primary monochromator
  4. Secondary filter
A

FLOURESCENCE SPECTROPHOTOMETRY

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36
Q

*absorption causes excitation
*light emission
*filtered by 2nd filter
*PHOTOMULTIPLIER TUBE

A

FLOURESCENCE SPECTROPHOTOMETRY

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37
Q
  • has the ability to magnify the radiant energy converted to electical energy
A

*PHOTOMULTIPLIER TUBE

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38
Q
  • excitation filter (intends to isolate the specific wavelength of light: short wavelength, high intensity/energy)
A

Primary monochromator

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39
Q
  • an energy emission that occurs when certain compounds absorbs electromagnetic radiation become excited and then return to an energy levelnthat is usually slightly hugher than their original level
A

FLOURESENCE

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40
Q
  • emitted energy is equal to or lower than the absorbed energy
A

PHOSPHORESCENCE

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41
Q

absorbs short wavelength, high energy, great intensity
excitation (unstable)
ground state
emit radiant energy

A

XENON LAMP

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42
Q

measurement of light blocked by a suspension of particulate matter

A

TURBIDIMETRY

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43
Q

measurement of the reduction in light transmission caused by particle formation

A

TURBIDIMETRY

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44
Q

protein determination

A

TURBIDIMETRY

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45
Q

TURBIDIMETRY PRECIPITATION AGENTS

(turbid solution)

A

Sulfosalicylic acid and Trichloroacetic acid

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46
Q

Factors affecting turbidimetry and nephelometry:

A

Size and number of particles
The depth of the tube
Cross-sectional area of each particle

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47
Q
  • measure turbidity of samples to detect bacterial growth in broth cultures
A

MICROBIOLOGY ANALYZERS

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48
Q
  • measure the antibiotic sensitivity from such cultures
A

MICROBIOLOGY ANALYZERS

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49
Q
  • detect clot formation in sample cuvets
A

COAGULATION ANALYZERS

50
Q
  • quantify protein concentration in biological fluids such as urine and CSF
A

CLINICAL CHEMISTRY ANALYZERS

51
Q

measurement of light scattered by small particles at an angle to the beam incident on the cuvet

A

NEPHELOMETRY

52
Q

more specific than turbidimetry

A

NEPHELOMETRY

53
Q

immunoglobulins, complement, immune complexes

A

NEPHELOMETRY

54
Q

When light passes from one medium into another, the light beam changes its direction at the boundary surface if its speed in the second medium is different from the first

A

REFRACTOMETRY

55
Q

Refractivity depends:

A

wavelength of the incident light
temperature
nature of the liquid medium
concentration of the solute dissolved in the medium

56
Q

Determines osmolality based on freezing point depression

A

Component parts

57
Q

Component parts: OSMOMETRY

A

Cooling bath, thermistor probe, stirring wire, galvanometer

58
Q

OSMOMETRY USE:

A

Serum and urine ormolality

59
Q

compounds are separated based on differences in affinity/attraction between a mobile phase and a stationary phase

A

CHROMATOGRAPHY

60
Q

– solid support (coated or uncoated)

A

Stationary phase

61
Q

not treated with any substance

A

Stationary phase

62
Q

CHROMATOGRAPHY

General types:

A

Adsorption Chromatography
Partition Chromatography

63
Q

CHROMATOGRAPHY

Kinds of Chromatography

A
  1. Paper chromatography
  2. Thin Layer Chromatography
  3. Ion-Exchange Chromatography
  4. Gel Filtration / Molecular Sieve / Gel Permeation / Size Exclusion / Molecular Exclusion
  5. Gas Chromatography
  6. High Performance Liquid Chromatography
64
Q
  • molecules adhere on the surface of the uncoated solid support (non-mobile)
A

Adsorption Chromatography

65
Q
  • solid support is coated with a film of water or non-volatile organic liquid
A

Partition Chromatography

66
Q

Partition Chromatography Examples:

A

TLC, GLC

67
Q

– flowing gas or liquid (eluent or carrier fluid)

A

Mobile phase

68
Q

coated/moistened with either water or organic solvent

A

Mobile phase

69
Q

Rf

A

retardation factor

70
Q

ratio of the distance of movement by a compound to the distance of the solvent front

A

Rf (retardation factor)

71
Q

Rf =

A

a/B

72
Q

= distance travelled by compound from origin to front of spot

A

a

73
Q

= distance travelled by solvent

A

B

74
Q
  • coated stationary phase
A

PARTITION

75
Q
  • compact spots easier to detect
A

Thin Layer Chromatography

76
Q
  • used for screening of drugs of abuse
A

Thin Layer Chromatography

77
Q
  • Separation is based on electrical charge
A

Ion-Exchange Chromatography

78
Q

– capture anions

A
  • Anion exchangers
79
Q

– capture cations

A
  • Cation exchangers
80
Q
  • Separation is based on differences in molecular size
A

Gel Filtration

81
Q
  • confirmation and quantitation of drugs
A

Gas Chromatography

82
Q
  • Some substances dissolve more in the immobile film of wax or oil-like material on a solid support medium while others in the surrounding steam of flowing gas
A

Gas Chromatography

83
Q
  • selective adsorption with the application of pressure
A

High Performance Liquid Chromatography

84
Q
  • for high MW compounds
A

High Performance Liquid Chromatography

85
Q

immobile phase:

  1. Paper chromatography
A

cellulose paper (coated with H2O)

86
Q

immobile phase:

  1. Thin Layer Chromatography
A

H2O (bound to silica)

87
Q

immobile phase:

  1. Ion-Exchange Chromatography
A

H2O (bound to silica)

88
Q

immobile phase:

  1. Gel Filtration / Molecular Sieve / Gel Permeation / Size Exclusion / Molecular Exclusion
A

a. Polyacrylamide (plastic)
b. Sephadex (cross-linked polysaccharide)
c. Porous beads

89
Q

immobile phase:

  1. Gas Chromatography
A
  • silica-coated (organic liquid) or uncoated
  • diatomaceous earth (silica) coated with a non-volatile organic liquid
90
Q

mobile phase:

  1. Paper chromatography
A

organic solvent

91
Q

mobile phase:

  1. Thin Layer Chromatography
A

organic solvent

92
Q

mobile phase:

  1. Ion-Exchange Chromatography
A

water

93
Q

mobile phase:

  1. Gel Filtration / Molecular Sieve / Gel Permeation / Size Exclusion / Molecular Exclusion
A

flowing water

94
Q

mobile phase:

  1. Gas Chromatography
A

Inert carrier gas (Helium or Nitrogen)
a. Gas – Liquid Chromatography (GLC) – based on partition
b. Gas – Solid Chromatography (GSL) – based on adsorption

95
Q

– base on partition
– based on adsorption

A

a. Gas – Liquid Chromatography (GLC)
b. Gas – Solid Chromatography (GSL)

96
Q

– polystyrene beads

A

Synthetic resins

97
Q
  • The stationary phase contains fixed charged group (constant) and a mobile charge with an opposite charge (exchangeable)
A

Ion-Exchange Chromatography

98
Q

separation of a substance in a pure form and then determining its dry weight

A

GRAVIMETRIC METHOD

99
Q

Example: Total Lipid determination

A

GRAVIMETRIC METHOD

100
Q

detection of light energy scattered or reflected toward a detector that is not in the direct path of the transmitted light (90° to the incident beam)

A

NEPHELOMETRY

101
Q

The factors affecting turbidimetric measurements are the same factors affecting

A

NEPHELOMETRY

102
Q

It is more specific than turbidimetry

A

NEPHELOMETRY

103
Q

measurement of the amount of light blocked by a particulate matter suspended in solution (180° to the incident beam)

A

TURBIDIMETRY

104
Q

Factors affecting turbidimetry:

A

Size and number of particles
The depth of the tube
Cross-sectional area of each particle

105
Q

Types of Radiation in Scintillation Counters:

A

Alpha
Beta
Gamma

106
Q

Exists in two forms Scintillation Counters:

A

soft and hard gamma

107
Q

Types of Scintillation Counters:

A

Solid Scintillation Counter
Liquid Scintillation Counter

108
Q

– measures gamma radiation using thallium activated NaI crystal as scintillator and PM tube as detector with preamplifier circuit

A

Solid Scintillation Counter

109
Q

– measures beta radiation using liquid flour as scintillator

A

Liquid Scintillation Counter

110
Q

– positively charged particles; resemble the nucleus of helium atom with a mass of 4
Have very litenergytle

A

Alpha

111
Q

– resembles an electron with both negative (β-) and positive (β+) charges but essentially no mass
Exists in two forms: soft and hard beta

A

Beta

112
Q

– a form of electromagnetic energy with no mass, only energy

A

Gamma

113
Q

SEPARATION OF SUBSTANCES

A

I. PRECIPITATION
II. ULTRAFILTRATION and DIALYSIS
III. CENTRIFUGATION

114
Q

• Separation is based on solubility

A

PRECIPITATION

115
Q

• The precipitate is studied by:

A

a. Turbidimetric method
b. Chemical reaction (after being dissolved)
c. Gravimetric method

116
Q
  • separates dissolved materials
A

ULTRAFILTRATION

117
Q
  • removes particulate matter
A

ULTRAFILTRATION

118
Q
  • movement through a semi-permeable membrane driven by a force or pulled through a vacuum
A

ULTRAFILTRATION

119
Q

Cellulose esters
Cellulose acetate
Polyamide
Polyvinyl chloride
Sheets, disks or hair thin fibers, conical

A

ULTRAFILTRATION

120
Q
  • For desalting, fractionation of protein solutions and the preparation of pff
A

ULTRAFILTRATION

121
Q

180° to the incident beam

A

TURBIDIMETRY

122
Q

90° to the incident beam

A

NEPHELOMETRY

CC1 LECTURE DEAN (14 decks)

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