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MT6317 CLIN CHEM 1 > U3 LAB: INSTRUMENTATION > Flashcards

U3 LAB: INSTRUMENTATION Flashcards

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1
Q

Four Major Disciplines of Analytic Techniques

A
  • Spectrometry
  • Luminescence
  • Electroanalytic methods
  • Chromatography
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2
Q

under Spectrophotometry

A
  • Spectrophotometry
  • Flame Emission Spectrophotometry
  • Atomic Absorption
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3
Q

Measures intensity of light or light transmitted

A

Spectrophotometry

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4
Q

Spectrophotometry

directly proportional to concentration

A

Absorbance

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5
Q

Spectrophotometry

inversely proportional to concentration

A

Transmittance

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6
Q

Measures intensity of light after an ion is burned (light of a single atom)

A

Flame Emission Spectrophotometry

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7
Q

Principle of Flame Emission Spectrophotometry

A

excitation of electrons from lower to higher energy

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8
Q

Measure of light absorbed after ions are dissociated by heat

A

Atomic Absorption

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9
Q

Atomic Absorption is used to measure?

A

trace elements (Ca, Mg)

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10
Q

Measures the light produced when ions are excited to unexcited

A

Fluorometer

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11
Q

Luminescence

Material absorbs at high energy but short wavelength, emits light at lower energy (visible light)

A

Fluorescence

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12
Q

Luminescence

Measures analyte produced in the reaction vessels

A

Chemiluminescence

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13
Q

Chemiluminescence produces?

A

electromagnetic radiation of UV, visible, infrared

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14
Q

Luminescence

Chemical yields electronically excited intermediate/product responsible for the emission

A

Chemiluminescence

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15
Q

Luminescence

for proteinous analytes

A

Nephelometry

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16
Q

Luminescence

measures the light scattered by a particulate matter suspended in a solution

A

Nephelometry

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17
Q

Luminescence

Most common example of Nephelometry

A

measurement of antigen-antibody complexes (turbidimetry)

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18
Q

Separation of molecules by their molecular weight

A

Electrophoresis

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19
Q

This refers to separation techniques for soluble components in a solution by specific differences in physical or chemical characteristics.

A

Chromatography

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20
Q

Separation of compounds that are naturally volatile or chemically converted to a volatile form

A

Gas Chromatography

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21
Q

Beer Lambert’s Law

A

concentration of analyte is directly proportional to absorbance

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22
Q

The darker the sample?

A

the higher concentration, intensity, absorbance

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23
Q

<400nm

A

Ultraviolet (UV)

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24
Q

400-700nm

A

Visible Light

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25
Q

> 700nm

A

infrared

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26
Q

750-1600nm

A

mid IR / fingerprint region

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27
Q

longer wavelength, _____ energy and frequency

A

lower

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28
Q

shorter wavelength, _________ energy and frequency

A

higher

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29
Q

Planck’s Formula

A

E = hv

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30
Q

Planck’s Formula

E

A

energy of photon in Joules

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31
Q

Planck’s Formula

h (constant)

A

6.626 x 10-34

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32
Q

Planck’s Formula

v

A

frequency of electromagnetic radiation

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33
Q

Beer Lambert’s Law

What is inversely proportional?

A

Transmittance and absorbance

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34
Q

T/F: A solution transmits light corresponding in wavelength to its color, and usually absorbs light of wavelengths complementary to its color.

A

True

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35
Q

This minimizes unwanted stray light.

A

Entrance slit

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36
Q

This eliminates unwanted wavelengths.

A

Monochromater

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37
Q

Holds the solution

A

Cuvette

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38
Q

Two types of Light Source

A
  • Continuum Source
  • Line Source
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39
Q

Continuum Source

A
  • Tungsten
  • Deuterium
  • Xenon
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40
Q

Continuum Source

Most common for visible infrared light sources

A

Tunsgten

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41
Q

Continuum Source

Routinely used for UV based light sources

A

Deuterium

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42
Q

Continuum Source

Used in most spectrofluorometers

A

Xenon

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43
Q

under Line source

A

Mercury-vapor lamps

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44
Q

Light under infrared region

A

Silicon Carbide

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45
Q

Stray light is considered as?

A
  • any wavelength outside band
  • causes absorbance error
  • limits the maximum absorbance
  • most common cause of lost linearity
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46
Q

Stray light can come from?

A

deteriorating light source

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47
Q

Kinds of Monochromator

A
  • Prisms
  • Diffraction Gratings
  • Filters
  • Holographic Gratings
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48
Q

Kinds of Monochromator

made up of glass

A

Prisms

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49
Q

Kinds of Monochromator

has grooves

A

Diffraction Gratings

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50
Q

This controls the width of the light beam.

A

Exit slit

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51
Q

Described as the total range of wavelengths transmitted

A

bandpass

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52
Q

Cuvette is also known as?

A

Analytical / Absorption / Sample Cell

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53
Q

Kinds of Cuvette

A
  • Alumina Silica Glass
  • Quartz Plastic
  • Borosilicate glass
  • Soft glass
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54
Q

Kinds of Cuvette

most common, wide range absorbance

A

Alumina Silica Glass

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55
Q

Kinds of Cuvette

Alumina Silica Glass has an absorbance range of?

A

350 - 2000nm

56
Q

Kinds of Cuvette

measurement of solution requiring a UV and visible spectra

A

Quartz Plastic

57
Q

Notes to remember for cuvette

A

1.) discard cuvettes with scratches (cause interferences)

2.) silica cuvettes transmit light effectively at wavelengths above 220nm

3.) prolonged alkaline solution may cause cuvette to corrode or dissolve

58
Q

This detects and converts transmitted light into photoelectric energy.

A

Photodetector

59
Q

Kinds of Photodetector

A
  • Photocell
  • Phototube
  • Photomultiplier tube
  • Photodiode
60
Q

Kinds of Photodetector

most simple

A

Photocell

61
Q

Kinds of Photodetector

most sensitive as it amplifies light first before converting to electricity

A

Photomultiplier tube

62
Q

This splits monochromatic light into 2

A

Beam splitters

63
Q

Double beam in space

A

2 photodetectors

64
Q

Double beam in time

A

1 photodetector

65
Q

FEP

Principle

A

excitation of electron from low energy to high energy

66
Q

FEP

Photodetector

A

photocell

67
Q

FEP

Light source

A

flame (also serves as cuvette)

68
Q

FEP

Sodium

A

yellow-orange

69
Q

FEP

Potassium

A

pink

70
Q

FEP

Calcium

A

orange

71
Q

FEP

Magnesium

A

bright white

72
Q

FEP

Copper I

A

blue

73
Q

FEP

Copper II

A

green

74
Q

FEP

Lithium

A

red

75
Q

FEP

Cesium

A

purple

76
Q

FEP

known as internal standards and collects variation in flames

A

Lithium and Cesium

77
Q

Most specific and most sensitive spectrophotometry

A

Atomic Absorption

78
Q

AAS

Principle

A

element is not excited but merely dissociates from its chemical bond and placed in an unionized gram stain

79
Q

AAS

Photodetector

A

Photomultiplier tube (measures trace elements)

80
Q

AAS

Light source

A

Hollow cathode lamp

81
Q

Volumetric (Titrimetric)

Principle

A

unknown sample is made to react with a known solution in the presence of an indicator

82
Q

Volumetric (Titrimetric)

Examples

A
  • Schales and Schales
  • EDTA Titration method
83
Q

Volumetric (Titrimetric)

Schales and Schales

A

for chloride

84
Q

Volumetric (Titrimetric)

EDTA Titration

A

for calcium

85
Q

Nephelometry

Principle

A

determines amount of scattered light by a particulate matter suspended in a turbid solution

86
Q

Nephelometry

Light scattering depends on:

A
  • particle size
  • wavelength
87
Q

Nephelometry

Angle

A

15 to 90 degrees

88
Q

Nature of antigen

A

provokes immune response

89
Q

Detection methods of the antibody molecule

A
  • Direct neutralization
  • Opsonization
  • Complement activation
  • Somatization
  • Control of inflammatory response
90
Q

Phenomenon of forward scatter

A

Mie scatter

91
Q

Mie scatter occurs because most complexes have a diameter of around?

A

250-1500nm

92
Q

Wavelengths in Mie scatter

A

320 to 650nm

93
Q

Light source of nephelometry

A
  • laser (most common)
  • light amplification (stimulated emission of radiation)
  • Tungsten Iodide lamp
94
Q

Turbidimetry

Principle

A

measures reduction in light transmission by one particle formation

95
Q

measure of blocked light or light reduced in large particles by a particulate matter in a solution

A

Turbidimetry

96
Q

Clinical applications of Turbidimetry

A
  • CSF
  • urine
97
Q

Phenomenon in Turbidimetry

A

Raleigh scatter

98
Q

This refers to scattered light in any directions, particles are smaller than wavelengths of light.

A

Raleigh scatter

99
Q

T/F: Raleigh scatter will scatted light in many directions, but not equally forward and backward.

A

False

equal lang

100
Q

separate DNA and RNA based on size and electrical charge

A

Electrophoresis

101
Q

separating charged constitutents of a sample by means of an electrical current

A

Electrophoresis

102
Q

Migration of charged macromolecules in presence of electrical power through porous support

A

Zone electrophoresis

103
Q

Porous support for electrophoresis

A
  • Paper
  • Cellulose acetate (densitometry)
  • Agarose gel (DNA, RNA, proteins)
104
Q

has a net charge that can be either positive or negative depending on pH conditions

A

Amphoteric

105
Q

movement of buffer ions and solvent relatives to the fixed support

A

Electroendosmosis / Endosmosis

106
Q

migration of small charged ions

A

Iontrophoresis

107
Q

Five components of Electrophoresis

A
  1. driving force (electrical power)
  2. support medium (gel, cellulose paper)
  3. buffer
  4. sample
  5. detecting system
108
Q

Detecting system for DNA

A

UV Transilluminator

109
Q

Detecting system for Proteins

A

Densitometry

110
Q

Support media for Electrophoresis

A
  • Cellulose acetate
  • Agarose gel
  • Polyacrylamide gel
111
Q

This separates serum into 5 bands.

A

Cellulose acetate

112
Q

Support media for DNA, separates 10-15 bands, predominant component of agar

A

Agarose gel

113
Q

For protein, separates based on charge and molecular size, 30 fractions, neurotoxic, used to study isoenzymes

A

Polyacrylamide gel

114
Q

This is lighter than the molecule of interest.

A

Tracking dye

115
Q

T/F: The brighter the band, the higher the protein is in electrophoresis

A

True

116
Q

Procedure for Electrophoresis

A
  1. Loading of sample
  2. Electrophoretic migration
  3. Wash and fix
  4. Staining
  5. Visualization
  6. Quantification
117
Q

5 Bands of Separated Proteins

A
  • Albumin
  • Alpha 1
  • Alpha 2
  • Beta
  • Gamma
118
Q

Stains for Proteins

A
  • Amido Black
  • Ponceau S
  • Oil Red O
119
Q

Stains for Lipids

A
  • Sudan Black
  • Fat Red 7B
120
Q

Other stains

A
  • Coomassie Blue
  • Gold/Silver Stain
121
Q

Stain that is very sensitive to nanograms of proteins

A

Gold/Silver Stain

122
Q

Reagent to measure protein in CSF

A

Coomassie brilliant blue

123
Q

Elution of volatile compounds based on boiling point, used to separate steroids, lipids, alcohols

A

Gas Chromatography

124
Q

Fragmentation and ionization of molecules

A

Mass Spectroscopy

125
Q

GC-MS is gold standard for?

A

drug testing

126
Q

MS/MS is gold standard for?

A

newborn screening

127
Q

used to determine structure of organic compound (e.g. MRI)

A

Nuclear Magnetic Resonance Spectroscopy

128
Q

Migration based on electrical charge

A

Electrophoresis

129
Q

Migration based on physical/chemical properties

A

Chromatography

130
Q

Migration is through a pH gradient

A

Isoelectric focusing

131
Q

Migration is through electro-osmosis flow

A

Capillary Electrophoresis

132
Q

Measures light from excited to unexcited

A

Fluorometry

133
Q

Fluorometry

Primary filter

A

UV light

134
Q

Fluorometry

Secondary filter

A

Visible light

135
Q

Detection systems in PCR is based on?

A

fluorescence

136
Q

Chemiluminescence

Principle

A

measurement of luminescence produced by chemical reaction producing light emission

MT6317 CLIN CHEM 1 (8 decks)

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