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Clinical Chemistry 1 > Analytical Methods and Instrumentation > Flashcards

Analytical Methods and Instrumentation Flashcards

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1
Q

____ provide the foundation for all measurements made in a modern clinical chemistry laboratory.

A

Analytic Techniques and Instrumentation

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2
Q

Four Basic Disciplines

A
  • Spectrometry
  • Luminescence
  • Electroanalytic Methods
  • Chromatography
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3
Q

Methods under Spectrometry

A
  • Spectrophotometry
  • Atomic Absoption Spectrometry (AAS)
  • Mass Spectrometry (MS)
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4
Q

Methods under Luminescence

A
  • Fluorescence
  • Chemiluminescence
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5
Q

Methods under Electroanalytic Methods

A
  • Electrophoresis
  • Potentiometry/Amperometry
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6
Q

Methods under Chromatography

A
  • Gas
  • Liquid
  • Thin-layer
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7
Q

Other Analytical Methods

A
  • Colorimetry
  • Volumetric (Titrimetric)
  • Turbidimetry
  • Nephelometry
  • Fluorometry
  • Osmometry
  • Electrochemistry Techniques
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8
Q

The primary goal of ____ is the isolation of discreet portions of the sprectrum.

A

Photocolorimetry

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9
Q

Photocolorimetry is used for measuring ____ in the laboratory.

A

analytes

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10
Q

Measurements under Photocolorimetry

A
  • Spectrophotometric measurement
  • Photometric measurement
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11
Q

____ is the measurement of light intensity in a narrower wavelength.

A

Spectrophotometric measurement

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12
Q

____ is the measurement of light intensity without consideration of the wavelength.

A

Photometric measurement

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13
Q

____ involves the measurement of the light transmitted by a solution to determine the concentration of the light-absorbing substance.

A

Spectrophotometry

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14
Q

Two Types of Spectrophotometer

A
  • Single Beam
  • Double Beam
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15
Q

____ is the simplest type of spectrophotometer.

A

Single Beam Spectrophotometer

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16
Q

____ is designed to make one measurement at a time at one specified wavelength.

A

Single Beam Spectrophotometer

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17
Q

In ____, the absorpition maximum of the analyte must be known in advanced.

A

Single Beam Photometer

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18
Q

In single beam spectrophotometer, the ____ must be known in advanced.

A

maximum absorption

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19
Q

____ splits the monochromatic light into two components.

A

Double Beam Spectrophotometer

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20
Q

In ____, one beam passes throughout the sample and the other beam passes through reference solution or reagent blank.

A

Double Beam Spectrophotomer

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21
Q

What is the purpose of the additional beam in Double Beam Spectrophotometer?

A

To correct for variation of light source intensity

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22
Q

In double beam spectrophotometry, absorbance of the sample can be recorded directly as the ____ of the sample beam.

A

electrical output

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23
Q

2 Types of Double Beam Spectrophotometer

A
  1. Double-beam in space
  2. Double-beam in time
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24
Q

____ uses two photodetectors for the sample and reference beam.

A

Double-beam in space spectrophotometer

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25
Q

Using ____, the beam is split into two paths and measured by matched detectors.

A

double-beam in space spectrophotometer

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26
Q

____ uses one photodetector, and alternately passes the monochromatic light using a chopper or rotating sector minor.

A

Double-beam in time spectrophotometer

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27
Q

Double-beam in time spectrophotometer uses a ____ to alternately pass the monochromatic light through the sample cuvette and reference cuvette.

A

chopper or rotating sector minor

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28
Q

Using ____, the beam is split into two but is measured by the same detector.

A

double-beam in time spectrophotometer

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29
Q

6 Basic Components of Single or Double-Beam Configuration Spectrophotometer

A
  1. Stable source of radiant energy
  2. Filter
  3. Sample holder
  4. Radiation detector
  5. Signal processor
  6. Readout device
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30
Q

____ isolates the specific region of the EFS.

A

Filter

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31
Q

____ is also called “photomultiplier”.

A

Radiation detector

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32
Q

Visible light travels at a speed of ____.

A

300,000 km/s

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33
Q

Visible light can be broken down into ____ colors.

A

7

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34
Q

Colors of the visible spectrum

A
  • Red
  • Orange
  • Yellow
  • Green
  • Blue
  • Indigo
  • Violet
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35
Q

Visible spectrum range

A

350 nm - 700 nm

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36
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 350-430
Color Absorbed:
Complementary Color: Yellow - Blue

A

Violet

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37
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 350-430
Color Absorbed: Violet
Complementary Color:

A

Yellow - Blue

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38
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength:
Color Absorbed: Violet
Complementary Color: Yellow - Blue

A

350 - 430

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39
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 431-475
Color Absorbed:
Complementary Color: Yellow

A

Blue

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40
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 431-475
Color Absorbed: Blue
Complementary Color:

A

Yellow

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41
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength:
Color Absorbed: Blue
Complementary Color: Yellow

A

431 - 475

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42
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 476-495
Color Absorbed:
Complementary Color: Orange

A

Green - blue

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43
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 476-495
Color Absorbed: Green-blue
Complementary Color:

A

Orange

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44
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength:
Color Absorbed: Green-blue
Complementary Color: Orange

A

476 - 495

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45
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 496-505
Color Absorbed:
Complementary Color: Red

A

Blue - green

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46
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 496-505
Color Absorbed: Blue-green
Complementary Color:

A

Red

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47
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength:
Color Absorbed: Blue-green
Complementary Color: Red

A

496 - 505

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48
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 506-555
Color Absorbed:
Complementary Color: Purple

A

Green

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49
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 506-555
Color Absorbed: Green
Complementary Color:

A

Purple

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50
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength:
Color Absorbed: Green
Complementary Color: Purple

A

506 - 555

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51
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 556-575
Color Absorbed:
Complementary Color: Violet

A

Yellow - green

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52
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 556-575
Color Absorbed: Yellow-green
Complementary Color:

A

Violet

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53
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength:
Color Absorbed: Yellow-green
Complementary Color: Violet

A

556 - 575

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54
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 576-600
Color Absorbed:
Complementary Color: Blue

A

Yellow

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55
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 576-600
Color Absorbed: Yellow
Complementary Color:

A

Blue

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56
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength:
Color Absorbed: Yellow
Complementary Color: Blue

A

576 - 600

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57
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 601-650
Color Absorbed:
Complementary Color: Green-blue

A

Orange

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58
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 601-650
Color Absorbed: Orange
Complementary Color:

A

Green - blue

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59
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength:
Color Absorbed: Orange
Complementary Color: Green-blue

A

601 - 650

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60
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 651-700
Color Absorbed:
Complementary Color: Blue-green

A

Red

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61
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength: 651-700
Color Absorbed: Red
Complementary Color:

A

Blue - green

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62
Q

Colors and the Complementary Colors of the Visible Spectrum

Wavelength:
Color Absorbed: Red
Complementary Color: Blue-green

A

651 - 700

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63
Q

____ provides polychromatic light and must generate sufficient energy or power to measure the analyte of interest.

A

Light source or Radiant source

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64
Q

To give accurate absorbance measurements throughout its absorbance range, the response to change of light source in light intensity must be ____.

A

linear

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65
Q

Types of Light or Radiant Source

A
  • Continuum
  • Line
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66
Q

____ emits radiation that changes in intensity.

A

Continuum source

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67
Q

Which type of light or radiant source is widely used in the laboratory?

A

Continuum source

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68
Q

Examples of Continuum Source

A
  • Tungsten
  • Deuterium
  • Xenon
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69
Q

____ emits limited radiation and wavelength.

A

Line Source

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70
Q

Examples of Line Source

A
  • Mercury and Sodium vapor lamps
  • Hallow cathode tube
  • Laser
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71
Q

____ minimizes unwated or stray light and prevents the entrance of scattered light into the monochromator system.

A

Entrance slit

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72
Q

Entrance slit minimizes ____ and prevents the entrance of ____.

A

unwatanted/stray light; scattered light

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73
Q

____ is any wavelength outside the band transmitted by the monochromator.

A

Stray light

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74
Q

____ limits the maximum absorbance that a spectrophotometer can achieve.

A

Stray light

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75
Q

____ is the most common cause of linearity at high analyte concentration.

A

Stray light

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76
Q

Stray light is the most common cause of ____ at high analyte concentration.

A

linearity

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77
Q

____ isolates specific or individual wavelength of light.

A

Monochromator

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78
Q

Kinds of Monochromator

A
  • Prisms
  • Diffraction gratings
  • Filters
  • Holographic gratings
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79
Q

____ are wedge-shaped pieces of glass, quartz, or sodium chloride.

A

Prisms

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80
Q

____ can be rotated, allowing only the desired wavelength to pass through an exit slit.

A

Prisms

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81
Q

A narrow light focused on a prism is ____ as it enters a more dense glass.

A

refracted

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82
Q

____ are most commonly used for it has a better resolution than prisms.

A

Diffraction gratings

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83
Q

____ are made by cutting grooves or slits into an illuminated surface of a flat piece of crown glass-wavelengths.

A

Diffraction gratings

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84
Q

Grooves of diffraction gratings

A

15,000 to 30,000 per inch

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85
Q

____ is simple and less expensive, but is not precise.

A

Filter

86
Q

____ are made by placing semi-transparent silver films on both sides.

A

Filters

87
Q

Filters are made by placing ____ on both sides.

A

semi-transparent silver films

88
Q

____ produces monochromatic light based on the principle of constructive interference of waves.

A

Filter

89
Q

Filters produce ____ based on the principle of constructive interference of waves.

A

monochromatic light

90
Q

Filters produce monochromatic light based on the ____.

A

principle of constructive interference of waves

91
Q

____ controls the width of the light beam.

A

Exit slit

92
Q

Width is also called as ____.

A

bandpass

93
Q

____ is the total range of wavelength transmitted.

A

Bandpass

94
Q

____ allows only a narrow fraction of the spectrum to reach the cuvette.

A

Exit slit

95
Q

The narrower the bandpass, the ____ the resolution.

A

greater

96
Q

The ____ the bandpass, the greater the resolution.

A

narrower

97
Q

The ____ holds the solution whose concentration is to be measured.

A

cuvette

98
Q

Other names for cuvette

A
  • Absorption cell
  • Analytical cell
  • Sample cell
99
Q

Kinds of Cuvette

A
  • Alumina silica glass
  • Quartz/Plastic
  • Borosilicate glass
  • Soft glass
100
Q

____ is the most commonly used cuvette.

A

Alumina silica glass

101
Q

____ is used for the measurement of solution requiring visible and ultraviolet spectra.

A

Quart/Plastic Cuvette

102
Q

____ transmit light effectively at a wavelength greater or equal to 220.

A

Silica Cuvette

103
Q

The path length of a cuvette is ____.

A

1 cm

104
Q

To increase sensitivity, some cuvettes are designed to have a path length of ____.

A

10 cm

105
Q

____ detects and converts transmitted light into photoelectric energy.

A

Photodetector

106
Q

Photodetector detects and converts transmitted light into ____.

A

photoelectric energy

107
Q

Kinds of Photodetector

A
  • Barrier Layer Cells/Photocell/Photovultaic Cell
  • Phototube
  • Photomultiplier tube
  • Photodiode
108
Q

____ is used in filter photometers with a wide bandpass.

A

Barrier Layer Cells/Photocell/Photovultaic Cell

109
Q

____ is a basic photo transducer that is used for detecting and measuring radiation in the visible region.

A

Barrier Layer Cells/Photocell/Photovultaic Cell

110
Q

____ is the simplest detector, least expensive, temperature sensitive, and non-linear.

A

Barrier Layer Cells/Photocell/Photovultaic Cell

111
Q

____ has a photosensitive material that gives off electron when light energy strikes it.

A

Phototube

112
Q

____ contain cathodes and anodes enclosed in a glass case.

A

Phototube

113
Q

____ requires external voltage.

A

Phototube

114
Q

Phototube requires ____.

A

external voltage

115
Q

____ has an excellent sensitivity and has a rapid response.

A

Photomultiplier Tube

116
Q

____ detects very low level of light.

A

Photomultiplier Tube

117
Q

____ is the most commonly used detector for visible and UV region.

A

Photomultiplier Tube

118
Q

____ can detect and amplify radiant energies.

A

Photomultiplier Tube

119
Q

____ contains dynodes.

A

Photomultiplier Tube

120
Q

____ are a series of anodes with high positive voltage.

A

Dynodes

121
Q

____ is 200x more sensitive than phototube.

A

Photomultipler Tube

122
Q

____ is not as sensitive as PMT but with excellent linearity.

A

Photodiode

123
Q

____ measures light at a multitude of wavelength.

A

Photodiode

124
Q

____ detects less amount of light.

A

Photodiode

125
Q

____ is most useful as simultaneous multichannel detector.

A

Photodiode

126
Q

____ displays output of the detection system.

A

Meter or read-out device

127
Q

____ states that the concentration of the uknown substance is directly proportional to the absorbed light and inveresely related to the transmitted light.

A

Beer’s Law

128
Q

According to Beer’s Law, the concentration of the unknown substance is ____ to the absorbed light.

A

directly proportional

129
Q

According to Beer’s Law, the concentration of the unknown substance is ____ to the transmitted light.

A

inversely proportional

130
Q

____ is the amount of light absorbed.

A

Absorbance

131
Q

Absorbance cannot be measured directly by ____.

A

Spectrophotometer

132
Q

____ states how much of the light the sample absorb.

A

Absorbance

133
Q

Absorbance is also known as ____.

A

optical density

134
Q

____ is the fraction of the incident light which is transmitted.

A

Transmittance

135
Q

____ is the amount of light that successfully passes through the substance and comes out the other side.

A

Transmittance

136
Q

____ contains serum but without the reagent to complete the assay.

A

Blank

137
Q

____ corrects absorbance caused by the color of the reagent.

A

Reagent blank

138
Q

Measures not effective in some cases of turbidity, ____ may be necessary.

A

ultracentrifugation

139
Q

To correct for artefactual absorbance reading, ____ may be used.

A

blanking procedures or dual wavelength method

140
Q

____ means that the wavelength indicated on the control dial is the actual wavelength of light passed by the monochromator.

A

Wavelength accuracy

141
Q

____ is demonstrated when a change in concentration results in a straight line calibration curve.

A

Linearity

142
Q

____ measures the light transmitted by a single atom burned in a flame.

A

Flame Emission Photometry (FEP)

143
Q

Principle of Flame Emission Photometry (FEP)

A

Excitation of electrons from lower to higher state of energy

144
Q

Principle: Excitation of electrons from lower to higher state of energy.

A

Flame Emission Photometry (FEP)

145
Q

Flame Emission Photometry (FEP)

Light source

A

Flame

146
Q

Flame Emission Photometry (FEP)

Method

A

Indirect Internal Standard Method

147
Q

Flame Emission Photometry (FEP)

Internal standard

A

Lithium or Cesium

148
Q

____ is used for the measurement of excited atoms such as Na and K.

A

Flame Emission Photometry (FEP)

149
Q

Flame Emission Photometry (FEP)

____ indicates changes in the fuel reading of the instrument.

A

Flickering of light

150
Q

____ measures the light absorbed by atoms dissociated by heat.

A

Atomic Absorption Spectrometry (AAS)

151
Q

Principle of Atomic Absorption Spectrophotometry (AAS)

A

Element is not excited but merely dissociated from its chemical bond and placed in an unionized, unexcited, ground state.

152
Q

Principle: Element is not excited but merely dissociated from its chemical bond and placed in an unionized, unexcited, ground state.

A

Atomic Absorption Spectrometry (AAS)

153
Q

Atomic Absoprtion Spectrophotometry (AAS)

Light Source

A

Hollow Cathode Tube

154
Q

Atomic Absoprtion Spectrophotometry (AAS)

Interferences

A

Chemical, matrix, and ionization

155
Q

____ is used for measurement of unexcited trace metals such as Ca and Mg.

A

Atomic Absorption Spectrophotometry (AAS)

156
Q

An ____ is used to convert ions to atom.

A

Atomizer

157
Q

In AAS, a ____ is used to modulate the light source.

A

chopper

158
Q

In AAS, ____ is added to samples to form stable complexes with phosphate.

A

Lanthanum or Strontium chloride

159
Q

Volumetric Principle

A

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

160
Q

Principle: The unkown sample is made to react with a known solution in the presence of an indicator.

A

Volumetric

161
Q

Examples of Volumetric Method

A
  • Schales and Schales Method
  • EDTA Titration Method
162
Q

____ is for measuring abundant large particles and bacterial suspension.

A

Turbidimetry

163
Q

Principle of Turbidimetry

A

Determines the amount of light blocked by a particulate matter

164
Q

Principle: Determines the amount of light blocked by a particulate matter.

A

Turbidimetry

165
Q

Turbidimetry depends on ____.

A

specimen concentration and particle size

166
Q

In turbidimetry, the solutions are measured using ____.

A

visible photometers or spectrophotometers

167
Q

____ is used for protein measurements, bacterial growth, antibiotic sensitivity, and clot formation.

A

Turbidimetry

168
Q

____ is more sensitive for protein measurement than turbidimetry.

A

Nephelometry

169
Q

____ is for measuring the amount of antigen-antibody complexes.

A

Nephelometry

170
Q

Nephelometry Principle

A

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

171
Q

Principle: Determines the amount of scattered light by a particulate matter suspended in a turbid solution.

A

Nephelometry

172
Q

Light scattering depends on ____.

A

wavelength and particle size

173
Q

Light scatered is measured at an angle typically, ___.

A

15 to 90 degrees

174
Q

Components of Nephelometry

A
  • Light source
  • Collimator
  • Monochromator
  • Sample cuvette
  • Stray light trap
  • Photodetector
175
Q

The ____ is proportional to concentration.

A

detector (PM tube) output

176
Q

____ is the migration of charged particles in an electric field.

A

Electrophoresis

177
Q

____ separates proteins on the basis of their electric charge densities.

A

Electophoresis

178
Q

During electrophoresis, proteins are ___.

A

negatively charged

179
Q

____ is the migration of small charged ions.

A

Iontophoresis

180
Q

____ is the migration of charged molecules.

A

Zone electrophoresis

181
Q

____ means that the net charge can either be positive or negative.

A

Amphoteric

182
Q

____ is the movement of buffer ions and solvent that is relative to a fixed support.

A

Electroendosmosis or Endosmosis

183
Q

Components of Electrophoresis

A
  • Electric power
  • Support medium
  • Buffer
  • Sample
  • Detecting system
184
Q

Supporting media for Electrophoresis

A
  • Paper electrophoresis
  • Starch Gel
  • Cellulose Acetate
  • Agarose Gel
  • Polyacrylamide Gel
185
Q

____ employs a filter paper strips soaked in a buffer solution, usually diethyl barbituric acid.

A

Paper electrophoresis

186
Q

What is the usual buffer solution in paper electrophoresis?

A

Diethyl barbituric acid

187
Q

____ separates by surface charge and molecular size.

A

Starch gel

188
Q

____ separates by molecular size.

A

Cellulose acetate

189
Q

____ is neutral; separates by electrical charge; does not bind to protein.

A

Agarose gel

190
Q

____ neutral; separates on the basis of charge and molecular size.

A

Polyacrylamide gel

191
Q

____ is a detector of electrophoretic machine.

A

Densitometer

192
Q

____ measures the absorbance of the stain.

A

Densitometer

193
Q

____ scan and quantify electrophoretic pattern.

A

Densitometer

194
Q

____ reads gel and cellulose acetate membrane.

A

Densitometer

195
Q

____ is directly proportional to net charge, and inversely proportional to molecular size and viscosity.

A

Electrophoretic mobility

196
Q

Electrophoretic mobility is ____ to net charge, and ____ to molecular size and viscosity.

A

directly proportional; inversely proportional

197
Q

At pH ____, the gamma globulins move toward the cathode, despite the fact that they are negatively charged.

A

8.6

198
Q

After electrophoresis, the gel is treated with a mild fixative, such as ____, that precipitates the proteins at the positions to which they have migrated.

A

acetic acid

199
Q

____ precipitates the proteins at the positions to which they have migrated.

A

Mild fixative (acetic acid)

200
Q

Frequently, gel shows ____ in which samples at the center of the gel migrate further than those at the edges.

A

smile artifact

201
Q

In ____, separating molecules migrate through pH gradient.

A

Isoelectric focusing

202
Q

____ uses a constant gradient.

A

Isoelectric focusing

203
Q

How is the pH gradient created in isoelectric focusing?

A

By adding acid to the anodic area of the electrolyte cell, and then adding base to the cathode area.

204
Q

____ is ideal for separating proteins of identical sizes but with different net charges.

A

Isoelectric focusing

205
Q

Supporting media for Isoelectric Focusing

A
  • Agarose gel
  • Polyacrylamide gel
  • Cellulose acetate
206
Q

Advantages:
* Ability to resolve mixtures of proteins
* Detect isoenzyme
* Identification of genetic variants
* Detect CSF oligoclonal band

A

Isoelectric focusing

207
Q

In ____, samples molecules are separated by electro-osmotic flow.

A

Capillary electrophoresis

208
Q

(+) charged ions move ____.

A

faster

209
Q

(-) charged ions move ____.

A

slower

210
Q

Uses:
* Separation, quantitation, and determination of MW of proteins
* Analysis of PCR
* Analysis of organic and inorganic substances and drugs

A

Capillary Electrophoresis

Clinical Chemistry 1 (5 decks)

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