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CLINICAL CHEMISTRY 1 > PRELIM LEC: ANALYTICAL TECHNIQUES AND INSTRUMENTATION (TRANSES) > Flashcards

PRELIM LEC: ANALYTICAL TECHNIQUES AND INSTRUMENTATION (TRANSES) Flashcards

TRANSES AND NOTES

1
Q

-are described as photons of
energy traveling in waves

A

Electromagnetic Radiation

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2
Q

is the highest point while trough is the lowest point

A

Crest or peak

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3
Q

is the distance of two peaks/crest or troughs
when light travels in a wavelike manner

A

Wavelength

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4
Q

Units:

A

Angstrom (A)Millimicrons (mu) Nanometer (nm)

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5
Q

Conversion factors:

A

1 nm = 10 A; 1 nm = 1 mu; 1 A = 10 mu

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6
Q

is the number of waves that pass an observation point in a unit of time

A

Frequency

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7
Q

is the distance between two adjacent peak and
trough

A

Amplitude

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8
Q

is inversely proportional to the frequency of the light wave

A

Wavelength

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9
Q

is inversely proportional to the wavelength of light

A

Energy

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10
Q

 The shorter the wavelength,

A

the larger number of photons will
be contained in a given distance

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11
Q

 More photons represents more energy, hence,

A

shorter wavelengths represent higher energy

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12
Q

 Wavelength or frequency of the electromagnetic waves are
perceived as

A

color or hue

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13
Q

shorter wavelength = high frequency color

A

bluish colors

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14
Q

long wavelength = low frequency colors

A

reddish color

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15
Q

 Height or amplitude of the electromagnetic waves are
perceived as

A

intensity or brightness

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16
Q

great amplitude=

A

bright colors

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17
Q

small amplitude =

A

dull colors

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18
Q

two kinds of wavelengths:

A

Visible Spectra
Invisible Spectra

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19
Q

can be observed at 340-700nm

A

Visible Spectra

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20
Q

wavelength of less
than 400nm

Invisible Spectra

A

ultraviolet

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21
Q

wavelength of greater than
700 nm

Invisible Spectra

A

Infrared

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22
Q

Other types of electromagnetic energy includes:

A

Cosmic Rays, Gamma Rays, X-rays and Appliances (Radio, TV, Microwave, Etc.)

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23
Q

Red (650-700)

Complementary Color

A

Green

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24
Q

Orange (600-650)

Complementary Color

A

Blue

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25
Q

Yellow (550-600)

Complementary Color

A

Indigo

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26
Q

Green (500-550)

Complementary Color

A

Red

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27
Q

Blue (450-500)

Complementary Color

A

Orange

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28
Q

Indigo (400-450)

Complementary Color

A

Yellow

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29
Q

Violet (350-400)

Complementary Color

A

Yellow

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30
Q

 Principle: Measures the amount of light transmitted to determine the concentration of the light-absorbing substance in the solution; the measurement of the light transmitted by a solution to determine the concentration of the light-absorbing substance in the solution

A

Spectrophotometry

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31
Q

states that the concentration of substance is directly proportional to the amount of light absorbed but inversely proportional to the logarithm of transmitted light

A

Beer-Lamberts Law

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32
Q

ratio of the radiant energy transmitted, divided by the radiant energy incident on the sample

A

%T

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33
Q

amount of light
absorbed

A

Absorbance (OD-optical density)

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34
Q

provides electromagnetic radiation as visible, infrared, or UV light

A

Light source

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35
Q

TYPE OF LIGHT SOURCE:

A

1.Tungsten/Tungsten-iodide lamp
2.Mercury Vapor Lamp
3.Deuterium Discharge Lamp
4.Infrared Energy Source
5.Quart Halide Lamp
6.Hollow Cathode Lamp

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36
Q

ideal for emission of light within the visible region (iodide prolongs stability of Tungsten); produces energy wavelength from 340-700nm (visible region); used for moderately diluted solution

A

Tungsten/Tungsten-iodide lamp

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37
Q
  • ]can emit UV ligh
  • exists narrow bands of energy at well defined places in the spectrum UV and visible light
A

Mercury Vapor Lamp

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38
Q

energy wavelength UV
range (down to 165nm)

A

Deuterium Discharge Lamp

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39
Q

above 800 nm

A

Infrared Energy Source

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40
Q

contains small amt of halogen such as iodine to prevent the decomposition of vaporized tungsten

A

Quart Halide Lamp

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41
Q

consists of a gas-tight chamber containing anode, a cylindrical cathode and insert gas such as helium

A

Hollow Cathode Lamp

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42
Q

reduces stray light and prevents scattered
light from entering the monochromator

A

Entrance Slit

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43
Q

solates the specific wavelength of choice

A

Monochromator

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44
Q

TYPES OF MONOCHROMATOR:

A
  1. Prism
  2. Gratings
  3. Colored Filters
  4. Interference Filters
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45
Q

wedge-shaped pieces of glass, quartz, or sodium chloride that allows transmission of light wherein each side of the prism has different thickness allowing selection of wavelength of light

A

Prism

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46
Q

disperses white light into a continuous spectrum of colors based on variation of refractive index for different wavelength

A

Prism

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47
Q

has small grooves cut at such an angle that each grooves behave like a very small prism and the wavelengths are bent as they pass a sharp corner

A

Gratings

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48
Q

separate white light into various color comp

A

Gratings

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49
Q

made of glass that absorbs some portion of the electromagentic spectrum and transmit others wherein light energy is absorbed by dye components on the class and is dissipated as heat

A

Colored Filters

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50
Q

band pass is 35-50nm or more

A

Colored Filters

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51
Q

enhances desired wavelength by constructive
interference and eliminates others by destructive interferences

A

Interference Filters

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52
Q

utlizes the wave cx of light to enhance the intensity of the desired wavelength by constructive interference and eliminates others by destructive interference and reflections

A

Interference Filters

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53
Q

band pass is 10-20nm

A

Interference Filters

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54
Q

where the light from the monochromator will pass through

A

exit slit

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55
Q

used to hold the solution in the instrument
whose concentration is to be measured

A

Analytical Cell

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56
Q

TYPES OF ANALYTICAL CELLS:

A
  1. Borosilicate Glass
  2. Quartz or Plastic
  3. Aluminum Silica Glass
  4. Soft glass
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57
Q

alkaline solution that do not etch
glass

A

Borosilicate Glass

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58
Q

best for wavelength below 320nm
PLASTIC CELLS - BOTH VISIBLE AND UV RANGE

A

Quartz or Plastic

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59
Q

best for visible light

A

Aluminum Silica Glass

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60
Q

best for acidic solution

A

Soft glass

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61
Q
  • converts transmitted light energy into an equivalent amount of electrical energ
A

Detector

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62
Q

TYPES OF DETECTOR:

A
  1. Barrier layer cells
  2. Photoemission tube
  3. Photomultiplier tube
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63
Q

composed of film of light sensitive
material; no power source needed

A

Barrier layer cells

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64
Q

has photosensitive material that gives off electron when light energy strikes it; requires
an outside voltage for operation

A

Photoemission tube

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65
Q

used a series of electrodes to internally amplify the photosignal before leaving the tube

A

Photomultiplier tube

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66
Q

simplest method of displaying output of the detection system

A

Meter (read out device)

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67
Q

Principle: measures the light emitted when electrons in an atom become excited by heat energy produced by the flame

A

Flame Emission Spectrophotometry

68
Q

Measures electrolytes with a 1+ charge:

69
Q

Excited atoms return to ground state by emitting light energy that is characteristic of that atom

A

Flame Emission Spectrophotometry

70
Q

Sodium filter- transmits only

A

yellow light (589 nm)

71
Q

Potassium filter- transmits only

A

violet light (367 nm)

74
Q

Lithium filter- transmits only

A

red light (767nm)

75
Q

-breaks the chemical bonds to produce atoms
-source of energy that will be absorbed by the atoms to enter the excitation state

76
Q

using a mixture of hydrogen and oxygen gas (acetylene, propane or natural gas)

77
Q

breaks up the solution into finer droplets so that the atom will absorb heat energy from the flame and get excited

A

Atomizer or Burner

78
Q

aspirate sample directly into
flame

A

Total Consumption Burner

80
Q

involves the gravitational feeding of solution

A

Premix Burner

81
Q

involves the gravitational feeding of solution

FLAME EMISSION SPECTOPHOTOMETRY

A

Interference Filter

82
Q

serves as photodetector

FLAME EMISSION SPECTOPHOTOMETRY

83
Q

is the preferred internal standard which also acts as a radiation buffer

84
Q

Criteria for Internal Standard

A

a. Concentration should be precisely the same in all samples and standards
b. Energy must be close to the required amount to excite the measured element
c. Normally found in solution being analyzed

85
Q

Principle: measures concentration of the element by detecting absorption of electromagnetic radiation by atoms

A

Atomic Absorption Spectrophotometry

86
Q

The elements are not excited but are dissociated from their chemical bonds and placed in the unionized, unexcited ground state

A

Atomic Absorption Spectrophotometry

87
Q

Measures electrolytes with a 2+ charge:

Atomic Absorption Spectrophotometry

A

Ca2+, Mg2+

88
Q

hollow cathode lamp, which produces a
wavelength of light that is specific for the kind of metal in the cathode

Components of AAS

A

Light Source

89
Q

modulates light beam
coming from the light source

Components of AAS

A

Mechanical Rotating Chopper

90
Q
  • uses flame to dissociate the chemical bonds and
    form free unexcited atoms

Components of AAS

91
Q

ADV: flame is more concentrated and hotter
DISADV: produces large droplets in the flame; noisier

Components of AAS

A

Total Consumption Burner

92
Q

gases are mixed; sample is atomized before entering the flame

Components of AAS

A

Premix Burner

93
Q

ADV: greater absorption and sensitivity; less noisy; large droplets go to the waste
DISADV: flame is less ho

Components of AAS

A

Premix Burner

94
Q

selects the desired wavelength from a spectrum of wavelength

Components of AAS

A

Monochromator

95
Q

Interferences:
situation at which the flame could not
dissociate the sample into neutral atoms

Components of AAS

96
Q

Interferences:
situation at which atoms in the flame
become excited and emits energy

Components of AAS

A

Ionization

97
Q

Principle: Unknown samples are made to react with a known solution in the presence of an indicator

A

Volumetry/Titrimetry

98
Q

Volumetry/Titrimetry Sample tests:

A

Schales & Schales method;EDTA Titration

99
Q

Principle: it is the isolation of the pure form of the sample and its derivatives and the determination of its dry weight

A

Gravimetry

100
Q

Gravimetry Sample tests:

A

Lipid determination

101
Q

Principle: Measures the amount of light blocked by a suspension of particular matter as light passes through the cuvette

A

Turbidimetry

102
Q

Turbidimetry Factors affecting measurement:

A

o Size and number of particles
o Depth of the tube
o Cross-sectional area of each particle

103
Q

Principle: Measures the amount of light scattered by small particles at an angle to the beam incident on the cuvette

A

Nephelometry

104
Q

Factors affecting measurement same as turbidimetry

A

Nephelometry

105
Q

Principle: measures multiple properties of cells suspended in a moving fluid medium

A

Flow Cytometry

106
Q

Flow Cytometry Process:

A

a. All cells pass a single-file through a sensing point, where they are intercepted by a laser beam
b. Cell suspensions are introduced into the flow chamber.
c. As the cells will pass through the flow chamber, they are surrounded by low-pressure sheath that creates laminar flow forcing the specimen into the center.
d. Each of the cells is intersected by light
e. The laser light excites the dye which emits a color of light that is detected by the photomultiplier tube, or light detector.
f. The cells are then sorted based from their electrical charge
g. As the drop forms, an electrical charge is applied to the stream to form a charge
h. This charged drop is then deflected left or right by charged electrodes and into waiting sample tubes.
i. Drops that contain no cells are sent into the waste tube. The end result is three tubes with pure subpopulations of cells.
j. The number of cells is each tube is known and the level of fluorescence is also recorded for each cell.

107
Q

INTERPRETATION:
cell size

Flow Cytometry

A

Forward light scatter

108
Q

INTERPRETATION:
cell granularity and nuclear irregularity

Flow Cytometry

A

90° angle scatter

109
Q

Principle: involves the separation of a mixture on the basis of specific differences of the physical and chemical characteristics of the different components on a supporting
medium

A

Chromatography

110
Q

the constituents of the mixture are separated by a continuous redistribution between two phases:

Chromatography

A

mobile phase
stationary phase

111
Q

mobile phcarries the complex mixture

Chromatography

A

mobile phase

112
Q

where the mobile phase flows

Chromatography

A

stationary phase

113
Q

Types of Chromatorgraphy:

A
  1. Paper Chromatography
  2. Thin Layer Chromatography
  3. Liquid-Liquid Chromatography
  4. Ion Exchange Chromatography
  5. Column Chromatography
  6. Gel Chromatography
  7. Gas Chromatography
114
Q

Principle: A spot of the substance fractioned is placed on the paper just above the solvent level

A

Paper Chromatography

115
Q

Basis of Separation:

Paper Chromatography

A

o Rate of diffusion
o Solubility of solute
o Nature of the solvent

116
Q

Same principle as paper chromatography but differs in
the sorbent used

A

Thin Layer Chromatography

117
Q

thin plastic plates impregnated to a layer of
silica gel, alumina, polyacrylamide gel or starch gel

Thin Layer Chromatography

118
Q

Principle: separation of substances according to their solubility in an organic/non-polar solvent and in an aqueous/polar solvent

A

Liquid-Liquid Chromatography

119
Q

“Like Dissolves Like”

Liquid-Liquid Chromatography

A

o highly polar substance = more soluble in a
highly polar solvent (water)
o lesser polar substance = more soluble in a
less polar solvent (organic substance)

120
Q

Clinical use:

Liquid-Liquid Chromatography

A

fractionation of barbituates and lipids

121
Q

Principle: the use of a resin (the stationary solid phase) is used to covalently attach anions or cations onto it

A

Ion Exchange Chromatography

122
Q

Principle: adsorption of the solutes of a solution through a stationary phase and separates the mixture into individual components

A

Column Chromatography

123
Q

Basis of Separation:

Column Chromatography

A

o difference in pH
o polarity of solven

124
Q

Clinical use:

Column Chromatography

A

Fractionation of sugars

125
Q

Principle: the use of a resin (the stationary solid phase) is used to covalently attach anions or cations onto it

A

Gel Chromatography

126
Q

Basis of Separation:

Gel Chromatography

A

o Molecular weight & size
o Charge of ions
o Hydrophobicity of the molecules

127
Q

Hydrophilic gels

Gel Chromatography

A

soluble in aqueous medium
ex. dextran, agarose, polyacrylamide

128
Q

Hydrophobic gels

Gel Chromatography

A

soluble in organic solvents
ex. methylatede sephadex, polystyrene beads

129
Q

Clinical use:

Gel Chromatography

A

fraction of polysaccharides, NA, proteins, enzymes, isoenzymes

130
Q

Principle: separating and measuring nanograms and pictogram amounts of volatile substance

A

Gas Chromatography

131
Q

Kinds of GC:
sorbent is solid
w/ a large surface

Gas Chromatography

A

Gas-Solid Chromatography

132
Q

Kinds of GC:
sorbent is a
non-volatile liquid

Gas Chromatography

A

Gas-Liquid Chromatography

133
Q

Basis of Separation:

Gas Chromatography

A

o sample volatility
o rate of diffusion into liquid layer of the column packing
o solubility of sample in the liquid layer

134
Q

Clinical use:

Gas Chromatography

A

drug screening and drug analysis
fractionation of steroids, lipids, barbituates, blood alcohol and other toxicologic substances

135
Q

Principle: measures the difference in voltage at a constant
current

A

Potentiometry

136
Q

relationship between the measured voltage
and the unknown concentration

Potentiometry

A

Nerst Equation

137
Q

Principle: measurement of difference in current at a
constant voltage

A

Polarography

138
Q

relationship between the difference in
current and voltage

Polarography

A

Ilkovic Equation

139
Q

Principle: measures the current flow between two nonpolarizable electrodes between a known electrical potential is
established

A

Conductometry

140
Q

Principle: measures the amount of current that flows when constant voltage is applied the measuring electrode

A

Amperometry

141
Q

Principle: measures the amount of electricity (coulombs) at a fixed potential

A

Coulometry

142
Q

1 coloumb =

Coulometry

A

1 amper per second

143
Q

number of coulombs consumed can be
directly related to the concentration of the unknown

Coulometry

A

Faraday’s Law

144
Q

Principle: Measures the fluorescence or the energy emission that occurs when a certain compound absorbs electromagnetic radiation, become excited and then return to an energy state that is usually higher than their original level

A

Fluorimetry

145
Q

Emitted light has longer wavelength than the incident/excited light due to the loss of energy during collision

A

Fluorimetry

146
Q

energy is equal to or lower than the absorbed energy (rarely happens)

Fluorimetry

A

Phosphorescence

146
Q

Main problem w/ fluorescence:

Fluorimetry

147
Q

Principle:migration or movement of charged particles in an
electric field

A

Electrophoresis

148
Q

Factors affecting rate of migration:

Electrophoresis

A
  1. Net electric charge
  2. Size & Shape of Molecule
  3. Electric Fluid Strength
149
Q

higher electric charge = faster migration

Electrophoresis

A

Net electric charge

150
Q

bigger molecules = slower migration

Electrophoresis

A

Size & Shape of Molecule

151
Q

higher ionic strength = slower movement

Electrophoresis

A

Electric Fluid Strength

152
Q

more voltage = more movement

Electrophoresis

A

Temperature

152
Q

*Problem with increased temp:

Electrophoresis

A

oDenaturation of Proteins
oEvaporation of solvent increases ionic strength

153
Q

earliest support media
Disadv: paper is fragile and easily damaged staining of protein

Electrophoresis

A

Paper Electrophoresis

153
Q

Nature of Supporting Media:

Electrophoresis

A
  1. Paper Electrophoresis
  2. Starch Gel Electrophoresis-
  3. Cellulose Acetate Electrophoresis
  4. Agarose Electrophoresis
  5. Polyacrylamide Gel Electrophoresis
154
Q

good for large samp
Disadv: fragile and unable to store results permanently

Electrophoresis

A

Starch Gel Electrophoresis

155
Q

strip with a clear plastic backing w/ a coating of cellulose acetate particles attached to it
Disadv: becomes brittle when dried

Electrophoresis

A

Cellulose Acetate Electrophoresis

156
Q

Disadv: electric neutrality; separation is strictly on the basis of electric charge and uniformity of material size

Electrophoresis

A

Agarose Electrophoresis

156
Q

uses protein size as the major factor in the separation process and the net charge of proteins

Electrophoresis

A

Polyacrylamide Gel Electrophoresis

157
Q

Clinical Use:

Electrophoresis

A

analysis of proteins (serum) that can provide
quick and useful information regarding the presence or absence of disease entities

158
Q

Specimens:

Electrophoresis

A

Serum, Urine, Cerebrospinal Fluid

159
Q

1.

Advantages:
o rapid results
o increases number of tests performed
o saves time and effort
o eliminates the need for more staff
o economical
o reduces errors in calculation and transcription
o better precision and accuracy

A

Automation

160
Q

Basic Approaches in Automation:

Automation

A
  1. CONTINUOUS FLOW ANALYZER
  2. DISCRETE ANALYZER
  3. CENTRIFUGAL ANALYZER
161
Q

-Sequential analysis
-Uniformity in test performance

Automation

A

CONTINUOUS FLOW ANALYZER

162
Q

-Separate analysis
-Most popular and versatile analyzer

Automation

A

DISCRETE ANALYZER

163
Q

-Batch analysis
-Centrifugal force moves the reagents and sample to a mixing chamber, into a cuvette, passing a light beam and measuring the absorbance

Automation

A

CENTRIFUGAL ANALYZER

CLINICAL CHEMISTRY 1 (12 decks)

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