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CLINICAL CHEMISTRY > ANALYTICAL METHODS AND INSTRMENTATION - PART 2 > Flashcards

ANALYTICAL METHODS AND INSTRMENTATION - PART 2 Flashcards

1
Q

It measures the light emitted by a single atom burned in a flame

A

Flame Emission Photometry (FEP)

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2
Q

what is the principle behind the flame emission photometry

A

Excitation of electrons from lower to higher energy

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3
Q

what is the light source for the flame emission photometry

A

Flame (also serves as the cuvette)

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4
Q

what is the method used in flame emission photometry

A

Indirect internal Standard Method

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5
Q

internal standard used for flame emission photometry

A

Lithium/Cesium

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6
Q

what is the purpose of Lithium/Cesium in flame emission photometry

A

corrects variations in flame and
atomizer characteristics

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7
Q

It is used for the measurement of excited ions (sodium and
potassium)

A

Flame Emission Photometry (FEP)

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8
Q

in FEP, “ Flickering light indicates __.

A

changes in the fuel reading of the
instrument

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9
Q

Purpose of Flame in FES

A

Breaks the chemical bond to produce atoms

Source of energy absorbed by the atoms to enter an
excited state

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10
Q

in FEP, it’s used to 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

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11
Q

interference filters as monochromator used in FLAME EMISSION PHOTOMETRY

A

NA, K , LITHIUM

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12
Q

what color do NA produced as an interference filter in FEP

A

transmit yellow light (589 nm)

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13
Q

what color do K produced as an interference filter in FEP

A

transmit violet light (767 nm)

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14
Q

what color do LITHIUM produced as an interference filter in FEP

A

transmit red light (761nm)

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15
Q

QUALITY CONTROL IN FES

referred internal standard; also acts as a radiation buffer

A

lithium

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16
Q

Reasons why lithium is preferred:

A

→ Its emission characteristics are similar to those of Na+ and K+
+ → Normally present as a trace element in human tissues and does
not present interferences in the determination

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17
Q

purpose of quality control in FEp

A

to achieve stability where there is fluctuations caused by
changes in fuel of air pressure which affects flame temperature and
rate of sample aspiration

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18
Q

It measures the light absorbed by atoms dissociated by heat.

A

Atomic Absorption Spectrophotometry
(AAS)

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19
Q

WHAT IS THE PRINCIPLE OF AAS

A

Element is NOT EXCITED by merely dissociated from its chemical bond and
place in an unionized, unexcited, ground state.

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20
Q

what is the light source of atomic absorption spectophotometry

A

Hollow-cathode lamp

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21
Q

what are the interferences for atomic absorption spectophotometry

A

chemical, matrix (differences in viscosity) and ionization

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22
Q

It is used for measurement of unexcited trace metals (calcium and magnesium)

A

atomic absorption spectophotometry

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23
Q

It is more sensitive than FEP; it is accurate, precise and very specific

A

Atomic Absorption Spectrophotometry
(AAS)

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24
Q

what is the internal standard used for atomic absorption spectophotometry

A

Internal standard is not needed - changes in aspiration have little effect on the
number of ground state atom

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25
Q

what is the atomizer used for the atomic absorption spectophotometry

A

nebulizer/graphite furnace is used to convert ions to atoms

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26
Q

what is used in atomic absorption spectrophotometry to modulate the light source

A

chopper

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27
Q

in AAS;

is added to samples to form stable complexes with phosphate

A

lanthanum

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28
Q

what is the principle behind the volumetric or titrimetric

A

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

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29
Q

examples of volumetric or titimetric

A

❑Schales and Schales method: chloride test
❑EDTA Titration Method: Calcium Test

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30
Q

For measuring abundant large particles (proteins) and bacterial
suspension

A

turbidimetry

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31
Q

what is the principle behind the turbidimetry

A

It determines the amount of LIGHT BLOCKED (reduction of
light) by a particulate matter in a TURBID SOLUTION

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32
Q

2 things that can affect the turbidimetry

A

specimen concentration
particle size

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33
Q

The measurement of reduction of light is due to particle formation.

A

TURBIDIMETRY

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34
Q

Solutions requiring quantitation by turbidimetry are measured
using____

A

visible photometers or visible spectrophotometers.

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35
Q

what are the uses of turbidimetry

A
  • protein measurements (CSF and urine)
  • to detect bacterial growth in broth cultures
  • antimicrobial test (broth method)
  • to detect clot formation
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36
Q

For measuring the amount of antigen-antibody complexes (proteins).

A

NEPHELOMETRY

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37
Q

what is the principle for NEPHELOMETRY

A

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

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38
Q

2 things that can affect nephelometry

A

Light scattering depends on WAVELENGTH and PARTICLE SIZE

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39
Q

IN NPHELOMETRY,

Light scattered by particles is measured at an angle, typically ___
degrees to the beam incident on the cuvet.

A

15-90

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40
Q

Is the migration of charged particles in an electric field. it
separates proteins on the basis of their electric charge and
densities.

A

ELECTROPHORESIS

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41
Q

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

A

Amphoteric

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42
Q

is the movement of
buffer ions and solvent relative to the fixed support

A

Electroendosmosis/Endosmosis

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43
Q

is the migration of small charged

A

iontophoresis

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44
Q

is the migration of charged
macromolecules

A

Zone electrophoresis

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45
Q

Factors Affecting Rate of Migration:

A
  1. Net electric charge of the molecule
  2. Size and charge of the molecules
  3. Electric field strength
  4. Nature of the supporting medium
  5. Temperature of operation
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46
Q

what are the supportng media

A

Cellulose acetate
Agarose gel
Polyacrylamide Gel

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47
Q

a supporting media that separates by molecular size

A

cellulose acetate

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48
Q

a supporting media that separates by electrical charge; it does
not bind protein

A

Agarose gel

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49
Q

a supporting media that separates on the basis of
charge and molecular size; separates proteins into 20
fractions;

A

polyacrylamide gel

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50
Q

supporting media that is used to study isoenzymes

A

Polyacrylamide Gel

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51
Q

Stains for Visualization of Fractions
(Bands):

fats fats fats
O S F

A

OIL RED O
SUDAN BLACK
FAT RED 7B

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52
Q

Stains for Visualization of Fractions
(Bands):

protein
protein

AB
PS

A

Amido black
Ponceau s

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53
Q

Stains for Visualization of Fractions
(Bands):

CSF PROTEIN

A

Coomassie Blue

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54
Q

Stains for Visualization of Fractions
(Bands):

very sensitive even to nanogram
quantities of proteins

A

Gold/Silver stain

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55
Q

It measures the absorbance of stain - concentration of
the dye and protein fraction.

A

densitometry

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56
Q

It scans and quantitates electrophoretic pattern

A

Densitometry

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57
Q

It separates molecules by migration through a pH gradient.

A

Isoelectric Focusing

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58
Q

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

A

Isoelectric Focusing

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59
Q

In isoelectric focusing,

pH gradient is created by adding acid to the ___ of
the electrolyte cell and adding base to the ___

A

anodic area; cathode area

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60
Q

in isoelectric focusing, Proteins move in the electric field until they reach a pH equal to their ___.

A

isoelectric point

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61
Q

what are thr supporting media for isoelectric focusing

A

agarose gel, polyacrylamide gel and
cellulose acetate

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62
Q

what are the advantages of isoelectric focusing

A

+ 1. The ability to resolve mixture of proteins.
+ 2. To detect isoenzymes of ACP, CK and ALP in serum.
+ 3. To identify genetic variants of proteins such as alpha-1 antityrpsin.
+ 4. To detect CSF oligoclonal bandin

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63
Q

In this method, sample molecules are separated by electro-osmotic
flow (EOF)

A

Capillary Electrophoresis

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64
Q

It utilizes nanoliter quantities of specimens.

A

Capillary Electrophoresis

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65
Q

In capillary electrophoresis,

__ ions in the specimen emerge early at the capillary
outlet because the EOF and the ion movement are in the same
direction.

___ ions in the specimen move towards the capillary
outlet but at a slower rat

A

Positively charged; Negatively charged

66
Q

what are the uses of the capillary electrophoresis

A

separation, quantitation and determination of molecular
weights of proteins and peptides; analysis of Protein products;
analysis of organic and inorganic substances and drugs

67
Q

It involves separation of soluble components in a
solution by specific differences in physical-chemical
characteristics of the different constituents

A

CHROMATOGRAPHY

68
Q

what are the bases of separation

A

+ 1. Rate of Diffusion
+ 2. Solubility of the solute
+ 3. Nature of the solvent
+ 4. Sample volatility/solubility
+ 5. Distribution between 2 liquid phases
+ 6. Molecular Size (molecular sieving)
+ 7. Hydrophobicity of the molecule
+ 8. Ionic attraction
+ 9. Differential distribution between two immiscible liquids
+ 10. Selective separation of substances
+ 11. Differences in adsorption and desorption of solutes

69
Q

2 Forms of Chromatography

A

+A. Planar
+B. Column

70
Q

It is used for fractionation of sugar and amino acid

A

Paper chromatography

71
Q

paper used for paper chromatography

A

Sorbent (stationary phase) - Whatman paper

72
Q

It is a semiquantitative drug screening test

A

Thin Layer Chromatography (TLC)

73
Q

Sample components are identified by comparison with
standards on the same plate

A

Thin Layer Chromatography (TLC)

74
Q

Extraction of the drug is ____ - which means the ph must be
adjusted to reduce the solubility of the drug in the aqueous
phase.

(dependent, independent)

A

pH dependent

75
Q

in the thin layer chromatography,

When all drug spots including the standards have migrated with
the solvent front, it is cause by ___

A

incorrect aqueous to nonaqueous
solvent mixture.

76
Q

what are the samples we can use for thin layer chromatography

A

Biological samples such as blood, urine and gastric fluid can be used for the test

77
Q

what is the sorbent used for thin layer chromatography

A

thin plastic plates impregnated with a layer of silica gel
or alumina.

78
Q

It is used for separation of steroids, barbiturates, blood, alcohol and
lipids.

A

Gas Chromatography (GC)

79
Q

It is useful for compounds that are naturally volatile or can be easily
converted into a volatile form.

A

Gas Chromatography (GC)

80
Q

in gas chromatography,

If the molecule of interest is not volatile enough for direct injection, it is
necessary to derivatize it into a more volatile form.

true or false

A

true

81
Q

biological samples used in gas chromatography

A

urine and blood

82
Q

urine and blood are introduced in the GC column using

A

hypodermic syringe or an automated sampler

83
Q

In GC column, The specimens are _ and _onto the column.

A

vaporized; swept

84
Q

what is the detector used in gas liquid chromatography GLC

A

Flame ionization

85
Q

Separation occurs based on differences in absorption at
the solid phase surfaces

A

Gas Solid Chromatography (GSC)

86
Q

Separation occurs by differences in solute partitioning
between the gaseous mobile phase and the liquid stationary
phase.

A

Gas Liquid Chromatography (GLC)

87
Q

it is based on the fragmentation and ionization of
molecules using a suitable source of energy

A

Mass Spectroscopy (MS)

88
Q

it can also detect structural information and
determination of molecular weight.

A

Mass Spectroscopy (MS)

89
Q

Before a compound can be detected and quantified by
mass spectroscopy, it must be separated by ___

A

Gas chromatography

90
Q

It is the gold standard for drug testing

A

Gas Chromatography-Mass
Spectroscopy (GC-MS

91
Q

It is also used for xenobiotics, anabolic steroids and pesticides.

A

Gas Chromatography-Mass
Spectroscopy (GC-MS)

92
Q

in this method, quantitative measurement of drug can be performed
by selective ion monitoring

A

Gas Chromatography-Mass
Spectroscopy (GC-MS)

93
Q

It uses an electron beam to split the drug emerging from the column into
its component ions - drugs are detected by means of the presence of
decomposition fragments which arise after degradation of the analytes.

A

Gas Chromatography-Mass
Spectroscopy (GC-MS)

94
Q

In GC - MS,

The position of the parent molecule-lon and degradation products give
rise to fingerprint patterns which will provide the final identity of the drug
of interest

TRUE OR FALSE

A

TRUE

95
Q

Every drug has its own fingerprint pattern which is compared to a
computer library of known fragmentation patters

true or false

A

true

96
Q
  • can detect 20 inborn errors of metabolism from a
    single blood spot
A

Tandem mass spectroscopy (MS/MS)

97
Q

It is based on the distribution of solutes between a
liquid mobile phase and a stationary phase.

A

Liquid Chromatography

98
Q

_____is the most widely used liquid chromatography.

A

HPLC (High Performance Liquid
Chromatography)

99
Q

It uses pressure for fast separations, controlled temperature, in
line detectors and gradient elution technique.

A

High Performance Liquid
Chromatography (HPLC)

100
Q

what are the uses of High Performance Liquid
Chromatography (HPLC)

A

fractionation of drugs, hormones, lipids, carbohydrates
and proteins; separation and quantitation of various
hemoglobins associated with specific diseases (e.g.,
thalassemia); rapid HbA1c test (within 5 minutes)

101
Q

In reverse phase HPLC, the mobile phase is ____ than
stationary phase.

A

more polar

102
Q

It is for detecting nonvolatile substances in body fluids

A

Liquid Chromatography-Mass
Spectroscopy (LC-MS)

103
Q

It is utilized to confirm positive results from screening of elicited
drugs - it is a complementary method to GC-MS

A

Liquid Chromatography-Mass
Spectroscopy (LC-MS)

104
Q

It is also used in therapeutic drug monitoring, toxicology and
studies of drug metabolites

A

Liquid Chromatography-Mass
Spectroscopy (LC-MS

105
Q

It requires interface methods to convert nonvolatile to volatile
compounds.

A

Liquid Chromatography-Mass
Spectroscopy (LC-MS)

106
Q

whta are the interface methods in Liquid Chromatography-Mass
Spectroscopy (LC-MS)

A

Electrospray (ES) and Atmospheric Pressure
Chemical Ionization (APC

107
Q

An ____ is used in HPLC and GC methods to
compensate for variation in extraction

A

internal standard

108
Q

The mechanism in this type of chromatography is the
exchange of sample ions and mobile-phase ions with the
charged group of the stationary phase

A

Ion Exchange Chromatography

109
Q

Is for separation of amino acids, proteins and nucleic
acids.

A

Ion Exchange Chromatography

110
Q

Separation of nucleic acids and proteins depends primarily
on the charge and ionic charge density

A

Ion Exchange Chromatography

111
Q

Separation of compounds is based on their partition
between a liquid mobile phase and a liquid stationary
phase coated on a solid support.

A

Partition Chromatography (Liquid-Liquid
Chromatography)

112
Q

Is for separation of therapeutic drugs and their
metabolites.

A

Partition Chromatography (Liquid-Liquid
Chromatography)

113
Q

It uses immobilized biochemical ligands as the
stationary phase to separate a few solutes from other
unretained solutes.

A

Affinity Chromatography

114
Q

This type of separation uses the so-called lock-and-key
binding that is widely present in biologic systems.

A

Affinity Chromatography

115
Q

affinity chromatogaphy ❑Is for separation of___

A

lipoproteins, carbohydrates and
glycated hemoglobins; antibodies.

116
Q

Separation is based on the differences (competition)
between the adsorption and desorption of solutes at the
surface of a solid particle

A

Adsorption Chromatography (Liquid
Solid Chromatography)

117
Q

in adsorption chromatography, LIQUID - SOLID

The compounds are adsorbed to a solid support such ___

A

as
silica or alumina

118
Q

It measures the amount of light intensity present over a zero
background.

A

FLUOROMETRY /MOLECULAR LUMINESCENCE

119
Q

what is the principle behind FLUOROMETRY /MOLECULAR LUMINESCENCE

A

It determines the amount of light emitted by a molecule
after excitation by electromagnetic radiation.

120
Q

whta is the light detector used in FLUOROMETRY /MOLECULAR LUMINESCENC

A

Photomultiplier tube or phototube

121
Q

minerals or elements for which the FLUOROMETRY /MOLECULAR LUMINESCENCE is used

A

: porphyrins, magnesium, calcium and catecholamines

122
Q

fluorometry or molecular luminescence,

It uses 2 monochromators____

A

(either filters, prisms or gratings) -

123
Q

Is about 1000x more sensitive than spectrophotometer - emitted
radiation is measured directly

A

FLUOROMETRY /MOLECULAR LUMINESCENCE

124
Q

fluorometry/molecular luminescence

It is affected by ___- pH and temperature changes, chemical
contaminants, UV light change

A

quenching

125
Q

light source for fluorometry or molecular luminescence

A

Mercury arc lamp, Xenon lamp

126
Q

It differs from fluorescence and phosphorescence in that
the emission of light is created from a chemical or
electrochemical reaction, and not from absorption of
electromagnetic energy

A

CHEMILUMINESCENCE

127
Q

what is the principle behind the CHEMILUMINESCENCE

A

The chemical reaction yields an electronically
excited compound that emits light as it is ground state,
or that transfers its energy to another compound, which
then produces emission

128
Q

what is the use of chemiluminecence

A

Immunoassays

129
Q

photodetector used for CHEMILUMINESCENCE

A

Photomultiplier tube (luminometer

130
Q

It Is more sensitive than fluorescence

A

CHEMILUMINESCENCE

131
Q

chemiluminescence

In this method, no excitation radiation is required and no monochromators
are needed because the chemiluminescence arises from one species.

true or false

A

true

132
Q

The excited products formed in the oxidation reaction produce
___ on return to the single state

A

chemiluminescence

133
Q

It is the measurement of the osmolality of an aqueous solution such as serum,
plasma, or urine.

A

OSMOMETRY

134
Q

what is the principle behind the osmometry

A

It is based on measuring changes in the colligative properties of solutions
that occur owing to variations in particle concentration

135
Q

osmotic particles in osmometry

A

glucose, urea nitrogen and sodium

136
Q

Colligative properties of the solution

A

osmotic pressure, boiling point, freezing point,
and vapor pressure

137
Q

As the osmolality of a solution increases the following reactions occur:

what will happen to the osmotic pressure, boiling point, freezing point, and the vapor pressure

A

osmotic pressure increases
boiling point is elevated

freezing point is depressed; and the
vapor pressure is also depressed

138
Q

Is the most commonly used method for measuring the
changes in colligative properties of a solution

A

Freezing-point depression osmometry

139
Q

It is based on the principle that addition of solute molecules
lowers the temperature at which a solution freezes.

A

Freezing-point depression osmometry

140
Q

what is the freezing point of a 1.0 mOsm/kg solution

A

0.00186° C

141
Q

what is the freezing point of a blood plasma that has an osmolality of 285 mOsm/k

A

-0.53° C.

142
Q

The measurement of current or voltage
generated by the activity of a specific ion.

A

ELECTROCHEMISTRY TECHNIQUES

143
Q

It is the measurement of electrical potential due to the activity of
free ions - change in voltage indicates activity of each analyte.

A

POTENTIOMETRY

144
Q

It is also the measurement of differences in voltage (potential) at
a constant current.

A

POTENTIOMETRY

145
Q

potentiometry It follows the ___ or which law

A

Nernst equation.

146
Q

Concentration of ions in a solution can be calculated from the
measured potential difference between the two electrodes.

A

POTENTIOMETRY

147
Q

what are the reference electrodes of potentiometry

A

calomel and silver-silver chloride

148
Q

whata re the uses of potentiometry

A

pH and pCO2, tests

149
Q

It is an electrochemical transducer capable of responding to one given
ion

A

Ion Selective Electrode (ISE)

150
Q

It is very sensitive and selective for the ion it measures - it measures
the activity of one ion much more than other ions present in the sample

A

Ion Selective Electrode (ISE)

151
Q

It is the measurement of the amount of electricity (in
coulombs) at a fixed potential.

A

Coulometry

152
Q

It is an electrochemical titration in which the titrant is
electrochemically generated and the end point is detected by
amperometry

A

Coulometry

153
Q

Coulometry follows which law

A

Faraday’s law

154
Q

uses of coulometry

A

Chloride test in
csf, serum and sweat

155
Q

interference in coulometry

A

bromide, cyanide and cysteine

156
Q

It is the measurement of the current flow produced by an
oxidation-reaction

A

Amperometry

157
Q

uses of Amperometry

A

pO2, glucose, chloride and peroxidase determination

158
Q

→ It is the measurement of differences in current at a constant voltage

A

Polarographyt

159
Q

it follows the ilkovic equation

A

Amperometry

160
Q

The measurement of current after which a potential is
applied to an electrochemical cell

A

Voltammetry

161
Q

It allows sample to be preconcentrated, thus utilizing
minimal analyte.

A

Voltammetry

162
Q

___ voltametry - for lead and iron.

A

Anodic stripping

CLINICAL CHEMISTRY (14 decks)

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