Analytical Methods and Instrumentation Flashcards
(210 cards)
1
Q
____ provide the foundation for all measurements made in a modern clinical chemistry laboratory.
A
Analytic Techniques and Instrumentation
2
Q
Four Basic Disciplines
A
- Spectrometry
- Luminescence
- Electroanalytic Methods
- Chromatography
3
Q
Methods under Spectrometry
A
- Spectrophotometry
- Atomic Absoption Spectrometry (AAS)
- Mass Spectrometry (MS)
4
Q
Methods under Luminescence
A
- Fluorescence
- Chemiluminescence
5
Q
Methods under Electroanalytic Methods
A
- Electrophoresis
- Potentiometry/Amperometry
6
Q
Methods under Chromatography
A
- Gas
- Liquid
- Thin-layer
7
Q
Other Analytical Methods
A
- Colorimetry
- Volumetric (Titrimetric)
- Turbidimetry
- Nephelometry
- Fluorometry
- Osmometry
- Electrochemistry Techniques
8
Q
The primary goal of ____ is the isolation of discreet portions of the sprectrum.
A
Photocolorimetry
9
Q
Photocolorimetry is used for measuring ____ in the laboratory.
A
analytes
10
Q
Measurements under Photocolorimetry
A
- Spectrophotometric measurement
- Photometric measurement
11
Q
____ is the measurement of light intensity in a narrower wavelength.
A
Spectrophotometric measurement
12
Q
____ is the measurement of light intensity without consideration of the wavelength.
A
Photometric measurement
13
Q
____ involves the measurement of the light transmitted by a solution to determine the concentration of the light-absorbing substance.
A
Spectrophotometry
14
Q
Two Types of Spectrophotometer
A
- Single Beam
- Double Beam
15
Q
____ is the simplest type of spectrophotometer.
A
Single Beam Spectrophotometer
16
Q
____ is designed to make one measurement at a time at one specified wavelength.
A
Single Beam Spectrophotometer
17
Q
In ____, the absorpition maximum of the analyte must be known in advanced.
A
Single Beam Photometer
18
Q
In single beam spectrophotometer, the ____ must be known in advanced.
A
maximum absorption
19
Q
____ splits the monochromatic light into two components.
A
Double Beam Spectrophotometer
20
Q
In ____, one beam passes throughout the sample and the other beam passes through reference solution or reagent blank.
A
Double Beam Spectrophotomer
21
Q
What is the purpose of the additional beam in Double Beam Spectrophotometer?
A
To correct for variation of light source intensity
22
Q
In double beam spectrophotometry, absorbance of the sample can be recorded directly as the ____ of the sample beam.
A
electrical output
23
Q
2 Types of Double Beam Spectrophotometer
A
- Double-beam in space
- Double-beam in time
24
Q
____ uses two photodetectors for the sample and reference beam.
A
Double-beam in space spectrophotometer
25
Using ____, the beam is split into two paths and measured by matched detectors.
double-beam in space spectrophotometer
26
____ uses one photodetector, and alternately passes the monochromatic light using a chopper or rotating sector minor.
Double-beam in time spectrophotometer
27
Double-beam in time spectrophotometer uses a ____ to alternately pass the monochromatic light through the sample cuvette and reference cuvette.
chopper or rotating sector minor
28
Using ____, the beam is split into two but is measured by the same detector.
double-beam in time spectrophotometer
29
6 Basic Components of Single or Double-Beam Configuration Spectrophotometer
1. Stable source of radiant energy
2. Filter
3. Sample holder
4. Radiation detector
5. Signal processor
6. Readout device
30
____ isolates the specific region of the EFS.
Filter
31
____ is also called ***"photomultiplier".***
Radiation detector
32
Visible light travels at a speed of ____.
300,000 km/s
33
Visible light can be broken down into ____ colors.
7
34
Colors of the visible spectrum
* Red
* Orange
* Yellow
* Green
* Blue
* Indigo
* Violet
35
Visible spectrum range
350 nm - 700 nm
36
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 350-430
**Color Absorbed:**
**Complementary Color:** Yellow - Blue
Violet
37
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 350-430
**Color Absorbed:** Violet
**Complementary Color:**
Yellow - Blue
38
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:**
**Color Absorbed:** Violet
**Complementary Color:** Yellow - Blue
350 - 430
39
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 431-475
**Color Absorbed:**
**Complementary Color:** Yellow
Blue
40
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 431-475
**Color Absorbed:** Blue
**Complementary Color:**
Yellow
41
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:**
**Color Absorbed:** Blue
**Complementary Color:** Yellow
431 - 475
42
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 476-495
**Color Absorbed:**
**Complementary Color:** Orange
Green - blue
43
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 476-495
**Color Absorbed:** Green-blue
**Complementary Color:**
Orange
44
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:**
**Color Absorbed:** Green-blue
**Complementary Color:** Orange
476 - 495
45
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 496-505
**Color Absorbed:**
**Complementary Color:** Red
Blue - green
46
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 496-505
**Color Absorbed:** Blue-green
**Complementary Color:**
Red
47
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:**
**Color Absorbed:** Blue-green
**Complementary Color:** Red
496 - 505
48
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 506-555
**Color Absorbed:**
**Complementary Color:** Purple
Green
49
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 506-555
**Color Absorbed:** Green
**Complementary Color:**
Purple
50
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:**
**Color Absorbed:** Green
**Complementary Color:** Purple
506 - 555
51
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 556-575
**Color Absorbed:**
**Complementary Color:** Violet
Yellow - green
52
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 556-575
**Color Absorbed:** Yellow-green
**Complementary Color:**
Violet
53
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:**
**Color Absorbed:** Yellow-green
**Complementary Color:** Violet
556 - 575
54
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 576-600
**Color Absorbed:**
**Complementary Color:** Blue
Yellow
55
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 576-600
**Color Absorbed:** Yellow
**Complementary Color:**
Blue
56
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:**
**Color Absorbed:** Yellow
**Complementary Color:** Blue
576 - 600
57
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 601-650
**Color Absorbed:**
**Complementary Color:** Green-blue
Orange
58
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 601-650
**Color Absorbed:** Orange
**Complementary Color:**
Green - blue
59
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:**
**Color Absorbed:** Orange
**Complementary Color:** Green-blue
601 - 650
60
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 651-700
**Color Absorbed:**
**Complementary Color:** Blue-green
Red
61
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:** 651-700
**Color Absorbed:** Red
**Complementary Color:**
Blue - green
62
# **Colors and the Complementary Colors of the Visible Spectrum**
**Wavelength:**
**Color Absorbed:** Red
**Complementary Color:** Blue-green
651 - 700
63
____ provides polychromatic light and must generate sufficient energy or power to measure the analyte of interest.
Light source or Radiant source
64
To give accurate absorbance measurements throughout its absorbance range, the response to change of light source in light intensity must be ____.
linear
65
Types of Light or Radiant Source
* Continuum
* Line
66
____ emits radiation that changes in intensity.
Continuum source
67
Which type of light or radiant source is widely used in the laboratory?
Continuum source
68
Examples of Continuum Source
* Tungsten
* Deuterium
* Xenon
69
____ emits limited radiation and wavelength.
Line Source
70
Examples of Line Source
* Mercury and Sodium vapor lamps
* Hallow cathode tube
* Laser
71
____ minimizes unwated or stray light and prevents the entrance of scattered light into the monochromator system.
Entrance slit
72
Entrance slit minimizes ____ and prevents the entrance of ____.
unwatanted/stray light; scattered light
73
____ is any wavelength outside the band transmitted by the monochromator.
Stray light
74
____ limits the maximum absorbance that a spectrophotometer can achieve.
Stray light
75
____ is the most common cause of linearity at high analyte concentration.
Stray light
76
Stray light is the most common cause of ____ at high analyte concentration.
linearity
77
____ isolates specific or individual wavelength of light.
Monochromator
78
Kinds of Monochromator
* Prisms
* Diffraction gratings
* Filters
* Holographic gratings
79
____ are wedge-shaped pieces of glass, quartz, or sodium chloride.
Prisms
80
____ can be rotated, allowing only the desired wavelength to pass through an exit slit.
Prisms
81
A narrow light focused on a prism is ____ as it enters a more dense glass.
refracted
82
____ are most commonly used for it has a better resolution than prisms.
Diffraction gratings
83
____ are made by cutting grooves or slits into an illuminated surface of a flat piece of crown glass-wavelengths.
Diffraction gratings
84
Grooves of diffraction gratings
15,000 to 30,000 per inch
85
____ is simple and less expensive, but is not precise.
Filter
86
____ are made by placing semi-transparent silver films on both sides.
Filters
87
Filters are made by placing ____ on both sides.
semi-transparent silver films
88
____ produces monochromatic light based on the principle of constructive interference of waves.
Filter
89
Filters produce ____ based on the principle of constructive interference of waves.
monochromatic light
90
Filters produce monochromatic light based on the ____.
principle of constructive interference of waves
91
____ controls the width of the light beam.
Exit slit
92
Width is also called as ____.
bandpass
93
____ is the total range of wavelength transmitted.
Bandpass
94
____ allows only a narrow fraction of the spectrum to reach the cuvette.
Exit slit
95
The narrower the bandpass, the ____ the resolution.
greater
96
The ____ the bandpass, the greater the resolution.
narrower
97
The ____ holds the solution whose concentration is to be measured.
cuvette
98
Other names for cuvette
* Absorption cell
* Analytical cell
* Sample cell
99
Kinds of Cuvette
* Alumina silica glass
* Quartz/Plastic
* Borosilicate glass
* Soft glass
100
____ is the most commonly used cuvette.
Alumina silica glass
101
____ is used for the measurement of solution requiring visible and ultraviolet spectra.
Quart/Plastic Cuvette
102
____ transmit light effectively at a wavelength greater or equal to 220.
Silica Cuvette
103
The path length of a cuvette is ____.
1 cm
104
To increase sensitivity, some cuvettes are designed to have a path length of ____.
10 cm
105
____ detects and converts transmitted light into photoelectric energy.
Photodetector
106
Photodetector detects and converts transmitted light into ____.
photoelectric energy
107
Kinds of Photodetector
* Barrier Layer Cells/Photocell/Photovultaic Cell
* Phototube
* Photomultiplier tube
* Photodiode
108
____ is used in filter photometers with a wide bandpass.
Barrier Layer Cells/Photocell/Photovultaic Cell
109
____ is a basic photo transducer that is used for detecting and measuring radiation in the visible region.
Barrier Layer Cells/Photocell/Photovultaic Cell
110
____ is the simplest detector, least expensive, temperature sensitive, and non-linear.
Barrier Layer Cells/Photocell/Photovultaic Cell
111
____ has a photosensitive material that gives off electron when light energy strikes it.
Phototube
112
____ contain cathodes and anodes enclosed in a glass case.
Phototube
113
____ requires external voltage.
Phototube
114
Phototube requires ____.
external voltage
115
____ has an excellent sensitivity and has a rapid response.
Photomultiplier Tube
116
____ detects very low level of light.
Photomultiplier Tube
117
____ is the most commonly used detector for visible and UV region.
Photomultiplier Tube
118
____ can detect and amplify radiant energies.
Photomultiplier Tube
119
____ contains dynodes.
Photomultiplier Tube
120
____ are a series of anodes with high positive voltage.
Dynodes
121
____ is 200x more sensitive than phototube.
Photomultipler Tube
122
____ is not as sensitive as PMT but with excellent linearity.
Photodiode
123
____ measures light at a multitude of wavelength.
Photodiode
124
____ detects less amount of light.
Photodiode
125
____ is most useful as simultaneous multichannel detector.
Photodiode
126
____ displays output of the detection system.
Meter or read-out device
127
____ states that the concentration of the uknown substance is directly proportional to the absorbed light and inveresely related to the transmitted light.
Beer's Law
128
According to Beer's Law, the concentration of the unknown substance is ____ to the absorbed light.
directly proportional
129
According to Beer's Law, the concentration of the unknown substance is ____ to the transmitted light.
inversely proportional
130
____ is the amount of light absorbed.
Absorbance
131
Absorbance cannot be measured directly by ____.
Spectrophotometer
132
____ states how much of the light the sample absorb.
Absorbance
133
Absorbance is also known as ____.
optical density
134
____ is the fraction of the incident light which is transmitted.
Transmittance
135
____ is the amount of light that successfully passes through the substance and comes out the other side.
Transmittance
136
____ contains serum but without the reagent to complete the assay.
Blank
137
____ corrects absorbance caused by the color of the reagent.
Reagent blank
138
Measures not effective in some cases of turbidity, ____ may be necessary.
ultracentrifugation
139
To correct for artefactual absorbance reading, ____ may be used.
blanking procedures or dual wavelength method
140
____ means that the wavelength indicated on the control dial is the actual wavelength of light passed by the monochromator.
Wavelength accuracy
141
____ is demonstrated when a change in concentration results in a straight line calibration curve.
Linearity
142
____ measures the light transmitted by a single atom burned in a flame.
Flame Emission Photometry (FEP)
143
Principle of Flame Emission Photometry (FEP)
Excitation of electrons from lower to higher state of energy
144
**Principle:** Excitation of electrons from lower to higher state of energy.
Flame Emission Photometry (FEP)
145
# **Flame Emission Photometry (FEP)**
Light source
Flame
146
# **Flame Emission Photometry (FEP)**
Method
Indirect Internal Standard Method
147
# **Flame Emission Photometry (FEP)**
Internal standard
Lithium or Cesium
148
____ is used for the measurement of excited atoms such as Na and K.
Flame Emission Photometry (FEP)
149
# **Flame Emission Photometry (FEP)**
____ indicates changes in the fuel reading of the instrument.
Flickering of light
150
____ measures the light absorbed by atoms dissociated by heat.
Atomic Absorption Spectrometry (AAS)
151
Principle of Atomic Absorption Spectrophotometry (AAS)
Element is not excited but merely dissociated from its chemical bond and placed in an unionized, unexcited, ground state.
152
**Principle:** Element is not excited but merely dissociated from its chemical bond and placed in an unionized, unexcited, ground state.
Atomic Absorption Spectrometry (AAS)
153
# **Atomic Absoprtion Spectrophotometry (AAS)**
Light Source
Hollow Cathode Tube
154
# **Atomic Absoprtion Spectrophotometry (AAS)**
Interferences
Chemical, matrix, and ionization
155
____ is used for measurement of unexcited trace metals such as Ca and Mg.
Atomic Absorption Spectrophotometry (AAS)
156
An ____ is used to convert ions to atom.
Atomizer
157
In AAS, a ____ is used to modulate the light source.
chopper
158
In AAS, ____ is added to samples to form stable complexes with phosphate.
Lanthanum or Strontium chloride
159
Volumetric Principle
The unknown sample is made to react with a known solution in the presence of an indicator.
160
**Principle:** The unkown sample is made to react with a known solution in the presence of an indicator.
Volumetric
161
Examples of Volumetric Method
* Schales and Schales Method
* EDTA Titration Method
162
____ is for measuring abundant large particles and bacterial suspension.
Turbidimetry
163
Principle of Turbidimetry
Determines the amount of light blocked by a particulate matter
164
**Principle:** Determines the amount of light blocked by a particulate matter.
Turbidimetry
165
Turbidimetry depends on ____.
specimen concentration and particle size
166
In turbidimetry, the solutions are measured using ____.
visible photometers or spectrophotometers
167
____ is used for protein measurements, bacterial growth, antibiotic sensitivity, and clot formation.
Turbidimetry
168
____ is more sensitive for protein measurement than turbidimetry.
Nephelometry
169
____ is for measuring the amount of antigen-antibody complexes.
Nephelometry
170
Nephelometry Principle
Determines the amount of scattered light by a particulate matter suspended in a turbid solution
171
**Principle:** Determines the amount of scattered light by a particulate matter suspended in a turbid solution.
Nephelometry
172
Light scattering depends on ____.
wavelength and particle size
173
Light scatered is measured at an angle typically, ___.
15 to 90 degrees
174
Components of Nephelometry
* Light source
* Collimator
* Monochromator
* Sample cuvette
* Stray light trap
* Photodetector
175
The ____ is proportional to concentration.
detector (PM tube) output
176
____ is the migration of charged particles in an electric field.
Electrophoresis
177
____ separates proteins on the basis of their electric charge densities.
Electophoresis
178
During electrophoresis, proteins are ___.
negatively charged
179
____ is the migration of small charged ions.
Iontophoresis
180
____ is the migration of charged molecules.
Zone electrophoresis
181
____ means that the net charge can either be positive or negative.
Amphoteric
182
____ is the movement of buffer ions and solvent that is relative to a fixed support.
Electroendosmosis or Endosmosis
183
Components of Electrophoresis
* Electric power
* Support medium
* Buffer
* Sample
* Detecting system
184
Supporting media for Electrophoresis
* Paper electrophoresis
* Starch Gel
* Cellulose Acetate
* Agarose Gel
* Polyacrylamide Gel
185
____ employs a filter paper strips soaked in a buffer solution, usually diethyl barbituric acid.
Paper electrophoresis
186
What is the usual buffer solution in paper electrophoresis?
Diethyl barbituric acid
187
____ separates by surface charge and molecular size.
Starch gel
188
____ separates by molecular size.
Cellulose acetate
189
____ is neutral; separates by electrical charge; does not bind to protein.
Agarose gel
190
____ neutral; separates on the basis of charge and molecular size.
Polyacrylamide gel
191
____ is a detector of electrophoretic machine.
Densitometer
192
____ measures the absorbance of the stain.
Densitometer
193
____ scan and quantify electrophoretic pattern.
Densitometer
194
____ reads gel and cellulose acetate membrane.
Densitometer
195
____ is directly proportional to net charge, and inversely proportional to molecular size and viscosity.
Electrophoretic mobility
196
Electrophoretic mobility is ____ to net charge, and ____ to molecular size and viscosity.
directly proportional; inversely proportional
197
At pH ____, the gamma globulins move toward the cathode, despite the fact that they are negatively charged.
8.6
198
After electrophoresis, the gel is treated with a mild fixative, such as ____, that precipitates the proteins at the positions to which they have migrated.
acetic acid
199
____ precipitates the proteins at the positions to which they have migrated.
Mild fixative (acetic acid)
200
Frequently, gel shows ____ in which samples at the center of the gel migrate further than those at the edges.
smile artifact
201
In ____, separating molecules migrate through pH gradient.
Isoelectric focusing
202
____ uses a constant gradient.
Isoelectric focusing
203
How is the pH gradient created in isoelectric focusing?
By adding acid to the anodic area of the electrolyte cell, and then adding base to the cathode area.
204
____ is ideal for separating proteins of identical sizes but with different net charges.
Isoelectric focusing
205
Supporting media for Isoelectric Focusing
* Agarose gel
* Polyacrylamide gel
* Cellulose acetate
206
**Advantages:**
* Ability to resolve mixtures of proteins
* Detect isoenzyme
* Identification of genetic variants
* Detect CSF oligoclonal band
Isoelectric focusing
207
In ____, samples molecules are separated by electro-osmotic flow.
Capillary electrophoresis
208
(+) charged ions move ____.
faster
209
(-) charged ions move ____.
slower
210
**Uses:**
* Separation, quantitation, and determination of MW of proteins
* Analysis of PCR
* Analysis of organic and inorganic substances and drugs
Capillary Electrophoresis
