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CLINICAL CHEMISTRY > Analytical Techniques and Automation > Flashcards

Analytical Techniques and Automation Flashcards

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1
Q

Describes the measurement principles used in the clinical chemistry laboratory

A

Analytic Techniques

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2
Q

Examples of analytic techniques

A

Spectrophotometry
Electrochemistry
Electrophoresis
Chromatography

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3
Q

Analytic techniques often used to determine concentrations of analytes in the CC lab

A

Spectrophotometry
Electrochemistry

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4
Q

Described as photons of energy traveling in waves

A

Electromagnetic radiation

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5
Q

Most recognizable forms of electromagnetic radiation

A

Light and radiant energy

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6
Q

Other types of electromagnetic radiation

A

Gamma rays
X-rays
Microwaves
Ultraviolet radiation
Radiofrequency
Radiation

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7
Q

The linear distance between any two equivalent points on a successive wave

A

Wavelength

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8
Q

Unit used in the visible spectrum

A

Nanometer nm

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9
Q

The relationship between wavelength and energy (E) is described by

A

Planck’s formula

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10
Q

Planck’s formula

A

𝐸 = h𝑣

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11
Q

Planck’s constant value

A

6.62 x10^-27 erg/sec

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12
Q

Unit of energy in the centimeter gram second system unit

A

erg

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13
Q

Is erg an SI unit?

A

No

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14
Q

h in the Planck’s formula

A

Planck’s constant

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15
Q

v in Planck’s in formula

A

Frequency

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16
Q

Number of oscillations of the waveform in a second

A

Frequency

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17
Q

Changes that occur in a period of time. Movement of waveform

A

Oscillation

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18
Q

Relationship between wavelength and frequency

A

Inversely proportional

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19
Q

Relationship between energy of electromagnetic radiation and wavelength

A

Inversely proportional

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20
Q

Wavelength of visible region

A

400-700nm

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21
Q

Wavelength of ultraviolet region

A

<400nm

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22
Q

Wavelength of infrared region

A

> 700nm

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23
Q

Visible region falls in between

A

Color violet (set at 400nm) and red (at 700nm)

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24
Q

States that the concentration of a substance is directly proportional to the amount of light absorbed or inversely proportional to the algorithm of the transmitted light

A

Beer’s Law

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25
Q

Converts the radiant energy into equivalent electrical energy

A

Photodetector

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26
Q

Function of sample cuvette

A

Contain the sample

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27
Q

Function of light source

A

Strikes the sample in the cuvette

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28
Q

Formula for % transmittance

A

% transmittance = T/I x 100

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29
Q

T in % transmittance formula

A

Radiant energy transmitted/ transmitted light

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30
Q

I in % transmittance formula

A

Radiant energy incident of the sample/ incident light

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31
Q

Amount of energy absorbed by the sample

A

Incident light (I)

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32
Q

Radiant energy that strikes photodetector

A

Transmitted light (T)

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33
Q

It refers to the amount of light absorbed

A

Absorbance

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34
Q

Formula for absorbance

A

𝐴 = 𝜀 ×𝑏 × 𝑐

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35
Q

𝜀 in absorbance formula

A

Molar absorptivity

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36
Q

b in absorbance formula

A

Length of the light path through the solution

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37
Q

c in absorbance formula

A

Concentration of absorbing molecules

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38
Q

Relationship between absorbance and concentration of the absorbing molecules or analyte of interest

A

Directly proportional

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39
Q

Equipment used to measure the light transmitted by a solution

A

Spectrophotometer

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40
Q

Function of spectrophotometer

A

Determine the concentration of the light absorbing substance in the solution

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41
Q

Components pf spectrophotometer

A

light source
Monochromator
Sample cell or cuvet
Photodetector
Meter or read-out device

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42
Q

Function of light source

A

Provides polychromatic light

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43
Q

What is polychromatic light

A

Light of several wavelengths. Variation in the color formation

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44
Q

Light source for visible and near-infrared region use

A

Incandescent tungsten or tungsten-iodide lamp

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45
Q

Light source for UV region use

A

Dueterium lamp and memory arc lamp

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46
Q

Light source will depend on the wavelength. True or False?

A

True

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47
Q

2 types of light source

A

Continuum
Line

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48
Q

Type of light source that emits radiation that changes in intensity

A

Continuum

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49
Q

Examples of continuum light source

A

Tungsten (visible region)
Dueterium (UV region)
Xenon (visible and UV region)

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50
Q

Type of light source that emits a few discrete lines or bands of radiation

A

Line

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51
Q

Examples of line light source

A

Mercury and sodium vapor
lamps (UV and visible)
Hollow cathode lamp (atomic absorption spectroscopy/ spectrophotmetry)

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52
Q

Function of monochromator

A

Isolates individual wavelengths of light

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53
Q

When light source strikes the monochromator, light source will produce ______

A

Polychromatic lights

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54
Q

Wavelength in nanometers (nm) at peak transmittance

A

Nominal wavelength

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55
Q

Range of wavelength about one half peak transmittance

A

Spectral bandwidth (FWHM or full width at half peak maximum)

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56
Q

Area from one point of a wave to another

A

Bandpass

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57
Q

Total length of wavelength

A

Bandpass

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58
Q

Shape and material for sample cuvette

A

Round or square; made of material that is transparent to radiation

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59
Q

Path length of sample cuvette

A

1 cm

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60
Q

Types of materials used for cuvette and their respective region of use

A

Fused silica or quarts: UV region
Alumina-silicate glass: 350-2000 nm
wavelength
Plastic cuvette: visible region

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61
Q

Double-beam spectrophotometers has how many cuvette?

A

Two cuvets (one for the sample and one for the solvent)

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62
Q

Function of photodetector

A

Converts the transmitted radiant energy into an equivalent amount of electrical energy

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63
Q

Types of photodetector

A

Barrier-layer cell or photocell (selenide cell)
Phototube
Photomultiplier tube (PMT)
Phototransistors and Photodiode

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64
Q

The least expensive; temperature sensitive photodetector

A

Barrier-layer cell or photocell

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65
Q

Composition of Barrier-layer cell or photocell

A

Selenium on a plate of iron

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66
Q

Barrier-layer cell or photocell is mainly used in _____

A

Filter photometers

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67
Q

Photodetector that contains cathode and anode enclosed in a glass tube

A

Phototube

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68
Q

Photodetector that has photosensitive material that gives off electrons when light energy strikes it

A

Phototube

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69
Q

Most common type of photodetector

A

Photomultiplier tube (PMT)

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70
Q

Characteristics of Photomultiplier tube (PMT)

A

200 times more sensitive than the phototube
Highly sensitive to UV and visible radiation

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71
Q

Characteristics of Photodiode

A

Not as sensitive as PM tube but has excellent linearity and speed

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72
Q

Beam of light that strikes the photodetector reflect the amount of analyte present in the sample

A

Linearity

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73
Q

Concentration of analyte can immediately be displayed in the digital meter because it can immediately convert amount of radiant energy into an equivalent energy

A

Speed

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74
Q

Function of meter or read-out device

A

Displays output of the detection system

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75
Q

Examples of meter or read-out device

A

Digital meters
d’Arsonval meters
Recorders
Light-emitting diodes (LEDs)
Cathode-ray tubes (CRTs)
Liquid crystal displays (LCDs)

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76
Q

Simplest type of absorption spectrometers

A

Single-beam spectrophotometer

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77
Q

Function of single-beam spectrophotometer

A

Measure one measurement at a time at one specified wavelength

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78
Q

Components of single-beam spectrophotometer

A

Light source
Monochromator
Sample cuvette
Photodetector (PM tube)
Read out device or meter

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79
Q

Components of double-beam spectrophotometer

A

Light source
Monochromator
Sample cuvette
Reference Cuvette
Photodetector (PM tube)
Read out device or meter

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80
Q

Spectrophotometer design that uses 2 photodetectors, 2 sample cuvettes

A

Double beam in space

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81
Q

Spectrophotometer design that uses 1 photodetector; chopper is used to pass the monochromatic radiation through the sample cuvette and then to the reference cuvette

A

Double-beam in time

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82
Q

A device that rotates or breaks up radiation beams so the beam of light can pass through the photodetector

A

Chopper

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83
Q

This implies that a photometer is measuring at the wavelength that it is set to

A

Wavelength or photometric accuracy

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84
Q

Material used to measure wavelength or photometric accuracy

A

Special glass-type optical filters:
Didymium glass (600 nm)
Holmium oxide (360 nm)

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85
Q

A test using glass filters or solutions that have known absorbance values for a specific wavelength

A

Absorbance check

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86
Q

The ability of a photometric system to yield a linear relationship between the radiant power incident upon its detector and the concentration

A

Linearity

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87
Q

Linearity is monitored using _____

A

Optical filters or solutions

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88
Q

Any light that impinges upon the detector that does not originate from a polychromatic light source (an interference)

A

Stray light

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89
Q

The presence of stray light is checked using _____

A

Special cut-off filters

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90
Q

Measures concentration by detecting the absorption of electromagnetic radiation by atoms rather than by molecule

A

Atomic Absorption Spectrophotometer

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91
Q

When using AAS, sample should be ___

A

Atomized

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92
Q

Light source for AAS

A

Hollow-cathode lamp
Electrodeless discharge lamp (new edition)

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93
Q

Function of chopper in AAS

A

Modulate the light beam that would strike the sample cell that comes in flame

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94
Q

Photodetector for AAS

A

Photomultiplier tube

95
Q

Application of AAS

A

To measure concentration of trace metals ( lead, mercury, cadmium)

96
Q

Components of single-beam AAS

A

Light Source
Chopper
Sample
Monochromator
PM tube (Photomultiplier tube)
Readout/ Meter

97
Q

AAS that does not require a burner to produce a flame but uses electric current

A

Flameless AAS

98
Q

Method used to measure light emitted by excited atoms

A

Ion Selective Electrode

99
Q

Application of ion selective electrode

A

To measure concentrations of sodium, potassium, and lithium

100
Q

Measures the concentrations of solutions that contains fluorescing molecules

A

Fluorometry

101
Q

Light source for fluorometry

A

Mercury (for filter fluorometers)
Xenon arc (for spectrofluorometers)

102
Q

Fluorometer that use filter as monochromator (primary and secondary filter)

A

Filter fluorometers

103
Q

Fluorometer that use prisms or gratings

A

Spectrofluorometers

104
Q

Photodetector for fluorometry

A

Photomultiplier tube

105
Q

Basic component of Filter fluorometer

A

Source
Attenuator
Primary filter
Sample holder
Secondary filter
Detector (PM)
Readout

106
Q

Filters used in filter fluorometer and their function

A

Primary filter: selects wavelength of light that is best absorbed by sample
Secondary filter: passes the longer wavelength of light to photodetector

107
Q

Advantages of fluorometry

A

Specific and sensitive

108
Q

Disadvantage of fluorometry

A

Sensitive to environmental changes resulting to Quenching (decrease in fluorescence due to these changes)

109
Q

Environmental changes causing quenching

A

Use of contaminated chemicals
Change in solvents
Use of UV light which causes photochemical changes
Change in temperature

110
Q

The emission of light as a result of chemical reaction

A

Chemiluminescence

111
Q

How does chemiluminescence differ from fluorescence?

A

No excitation radiation is required and no monochromators are needed

112
Q

It is the production of electromagnetic radiation when a chemical reaction yields an excited product

A

Chemiluminescence

113
Q

Advantages of chemiluminescence

A

Subpicomolar detection limits, speed, ease of use, and simple instrumentation

114
Q

Disadvantage of chemiluminescence

A

Impurities can degrade sensitivity and specificity

115
Q

Principle of turbidimetry and nephelometry

A

Based on the scattering of radiation by particles in suspension

116
Q

Applications of turbidimetry and nephelometry

A

Measurement of antigen-antibody reactions, prealbumin, and other serum proteins

117
Q

It is the measurement of the light scattered by a particulate solution

A

Nephelometry

118
Q

3 types of light scatters

A

Rayleigh theory
Mie theory
Rayleigh-Debye theory

119
Q

This theory states that when the wavelength of light is greater than the diameter of the particle, then there is symmetrical distribution of light scattering

A

Rayleigh theory

120
Q

According to rayleigh theory, the minimum light scatter occurs at how many degrees to the incident light?

A

90 degrees

121
Q

This theory states that if the wavelength of light < the particle diameter (d > 0.1 λ), then the light scatters forward

A

Mie theory

122
Q

This theory states that if the wavelength of light is approximately the same as the particle size, more light scatters in the forward direction than in other direction

A

Rayleigh-Debye theory

123
Q

Device used to measure a concentration of a solution using the analytical technique nephelometry

A

Nephelometer

124
Q

Components of nephelometer

A

Light source
Collimator
Monochromator
Sample cuvette
Photodetector

125
Q

Light source for nephelometer

A

Mercury-arc lamp
Tungsten- filament lamp
Light-emitting diode
Laser

126
Q

Function of collimator

A

Narrows or control a beam of light

127
Q

Determines the amount of light blocked by a suspension of particles

A

Turbidimetry

128
Q

Applications of turbidimetry

A

It is used in microbiology analyzers, coagulation analyzers, and is used to quantify protein concentration in biologic fluids such as urine and CSF

129
Q

What is the difference between nephelometry and turbidimetry?

A

Nephelometry detects (right-angle or forward) scattered light, and turbidimetry measures a reduction of light transmitted in the forward direction

130
Q

Electrochemistry techniques

A

Potentiometry
Coulometry
Amperometry
Voltammetry

131
Q

Involves the measurement of the current or voltage generated by the activity of specific ions

A

Electrochemistry

132
Q

Most widely used technique in clinical measurements

A

Potentiometry

133
Q

Measurement of potential (voltage) between two electrodes in a solution

A

Potentiometry

134
Q

The two electrodes in potentiometry

A

Reference Electrode
Indicator Electrode

135
Q

Electrode with a constant voltage

A

Reference Electrode

136
Q

Most used reference electrode in the laboratory

A

Calomel and Silver/Silver chloride

137
Q

Measuring electrode

A

Indicator Electrode/Analytical Electrode

138
Q

Can be calculated from the measured potential difference between 2 electrodes

A

Concentration of Ions

139
Q

Replaced the Flame Photometry

A

Ion-Selective Electrode

140
Q

Membrane used to measure sodium

A

Glass aluminum silicate

141
Q

Membrane used to measure potassium

A

Valinomycin Gel

142
Q

Membrane used to measure calcium and lithium

A

Organic Liquid Ion Exchangers

143
Q

Membrane used to measure carbon dioxide and ammonia

A

Gas Electrodes

144
Q

Membrane used to measure urease and glucose oxidase

A

Enzyme Electrodes

145
Q

Two type of ISE

A

Direct ISE
Indirect ISE

146
Q

ISE that does not require sample dilution

A

Direct ISE

147
Q

ISE that requires sample dilution before the analysis phase

A

Indirect ISE

148
Q

Used to measure hydrogen ion (concentration) activity

A

pH electrode

149
Q

Internal Reference Electrode

A

Silver/silver chloride

150
Q

External Reference Electrode

A

Calomel electrode

151
Q

A pH electrode within a plastic jacket (has sodium bicarbonate buffer and gas-permeable membrane)

A

pCO2

152
Q

Measures the quantity of electricity (in coulombs) needed to convert an analyte to a different oxidation state

A

Coulometry

153
Q

Applications of coulometry

A

To measure chloride ion in serum, plasma, CSF, and sweat samples

154
Q

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

A

Amperometry

155
Q

Applications of amperometry

A

To measure chloride ion in serum, plasma, CSF, and sweat samples; pO2 electrode blood gas analyzers

156
Q

Gas sensing electrode

A

pO2 Electrode

157
Q

Application of pO2 electrode

A

To measure the partial pressure of oxygen in the blood

158
Q

it is a method in which a potential (voltage) is applied to an electrochemical cell and the resulting current is measured

A

Voltammetry

159
Q

Used to measure heavy metals such as lead

A

Anodic Stripping Voltammetry

160
Q

Measurement of the number of dissolved particles in a fluid

A

Osmometry

161
Q

Osmotically active particles

A

Glucose
Urea Nitrogen/ Blood Urea Nitrogen (BUN)
Sodium

162
Q

Effects of increased osmolality

A

Osmotic pressure increases
Boiling point increases
Freezing point decreases
Vapor pressure decreases

163
Q

Is used to measure the concentration of solute particles in a solution

A

Osmometer

164
Q

The process of separating the charged constituents of a sample by means of an electric current

A

Electrophoresis

165
Q

Electrophoresis is the separation of charged compounds based on their

A

Electrical charge

166
Q

A substance that can either have a negative, zero or positive charge depending on the conditions

A

Amphotheric

167
Q

Compounds that when dissolved in water can act either as acid or as a base

A

Ampholytes

168
Q

Negatively charged ions

A

Anion

169
Q

Positively charged ions

A

Cation

170
Q

Ions that are neutral and have both positive and negative charges at different locations throughout the molecule

A

Zwitterions

171
Q

Negatively charged electrode

A

Cathode

172
Q

Positively charged electrode

A

Anode

173
Q

Factors affecting the mobility of particles

A

Net charge of the particle
Size and shape of the particle
Strength of the electric field
Chemical and physical properties of the medium
Electrophoretic temperature

174
Q

Migration of small ions

A

Iontophoresis

175
Q

Migration of charged macromolecules in a porous support medium

A

Zone Electrophoresis

176
Q

Components of electrophoresis

A

Power supply
Buffer
Support Medium
Sample
Detecting System

177
Q

Function of power supply

A

Supplies constant current or voltage in the system

178
Q

Known as the driving force in electrophoresis

A

Power supply

179
Q

Why does the power supply named as the driving force in electrophoresis?

A

Because this drives the molecules through the support medium

180
Q

Function of buffer

A

Provide ions that will enable the movement of current and migration of particles
Maintain the pH at a relatively constant value

181
Q

A mixture of proton-donating and proton-accepting substances that functions to maintain the pH at a constant value

A

Buffer

182
Q

Barbital (veronal) pH

A

8.6

183
Q

Tris-boric EDTA pH

A

8.7

184
Q

Cation migrates to the

A

Cathode

185
Q

Anion migrates to the

A

Anode

186
Q

pH will influence the charge of the analyte. True or False?

A

True

187
Q

Relationship between ionic strength and mobility

A

LOW I.S = more charge will be carried = faster mobility
HIGH I.S = less charge will be carried = slower mobility

188
Q

A network of interacting fibres or a polymer that is solid but traps large amount of solvent in its pores or channels inside

A

Support media

189
Q

The support media must not interact with the analyte, it is just supposed to support the analyte to pass through it. True or False?

A

True

190
Q

Examples of support media

A

Cellulose Acetate
Agarose Gel
Polyacrilamide Gel

191
Q

Cellulose that is acetylated from cellulose acetate by treating it with acetic anhydride

A

Cellulose acetate

192
Q

Cellulose acetate separates serum proteins into how many bands

A

5 bands

193
Q

5 bands produced when using cellulose acetate

A

Albumin
Alpha-1
Alpha-2
Beta
Gamma

194
Q

The support media of choice

A

Cellulose acetate

195
Q

Useful in doing electrophoresis with proteins that can be separated with 5 bands

A

Isoelectric Focusing

196
Q

Used as a purified fraction of agar that comes from red algae

A

Agarose gel

197
Q

Unique features of agarose gel

A

It is neutral and does not produce electroendosmosis

198
Q

Agarose gel separates proteins into how many bands?

A

10-15 bands

199
Q

Support medium used to separate protein based on charge and molecular size

A

Polyacrilamide gel

200
Q

Polyacrilamide gel separates serum proteins into how many bands?

A

20 or more bands

201
Q

Result of electrophoresis consisting of separated strands of a macromolecule

A

Electrophoretogram

202
Q

Detecting system that can already be conducted if the samples are already dyed or stained

A

Direct observation

203
Q

The simplest way of detection in electrophoretic system

A

UV visualization

204
Q

A device that measures the degree of darkness of a photographic or semitransparent material or of a reflecting surface

A

Densitometer

205
Q

Examples of electrophoretic detecting system

A

Electrophoretogram
Direct observation
Staining
Radioactive dye
UV visualization
Densitometer

206
Q

Applications of electrophoresis

A

DNA Fractionation
Isoenzyme Determination
Protein Fractionation

207
Q

Process of forming electric cloud that prevents analytes or particles from migrating into the support medium

A

Electroendosmosis

208
Q

Separation is determined through the speed of the analyte’s migration

A

Isoenzyme determination

209
Q

Separate complex mixture on basis of different physical interactions between individual compound and stationary phase of the system

A

Chromatography

210
Q

Basic components of chromatography and their functions

A

Mobile Phase: carries complex mixture
Stationary Phase: through which mobile phase flows
Column: holds the stationary phase
Eluate: separated component

211
Q

Modes of separation in chromatography

A

Adsorption
Partition
Steric Exclusion
Ion exchange

212
Q

2 classifications of chromatography based on stationary phase

A

Planar chromatography
Column chromatography

213
Q

Classification of chromatography where the stationary phase is coated with a sheet of paper or bound to glass or plastic plate

A

Planar chromatography

214
Q

Classification of chromatography where the stationary phase is packed into tube or coated onto the inner surface of the tube/column

A

Column chromatography

215
Q

Examples of Planar chromatography

A

Paper chromatography

216
Q

Examples of Column chromatography

A

Thin layer chromatography
Gas chromatography
Liquid chromatography

217
Q

Application of Paper chromatography

A

Fractionation of sugar and amino acid

218
Q

Application of Thin-Layer chromatography

A

Drug screening

219
Q

Application of Gas chromatography

A

Separate mixture of compounds that are volatile made or can be made volatile

220
Q

Application of Liquid chromatography

A

Uses pressure for fast separation of thermolabile substance

221
Q

Why do we need to separate thermolabile substances immediately?

A

Thermolabile substances must be separated immediately, because when it is exposed to high temp. they will be destroyed and goes unstable that can cause us not to recover anything

222
Q

How can we differentiate a gas chromatography to a liquid chromatography?

A

We can differentiate them based on their mobile phase

223
Q

How can we differentiate planar and column chromatography?

A

We can differentiate them based on their stationary phase

224
Q

Forces the mobile phase through the column

A

Pumps

225
Q

Holds the stationary phase

A

Columns

226
Q

Introduce the sample into the mobile phase

A

Sample injectors

227
Q

Produce an electronic signal proportional to the concentration of separated component

A

Detectors

228
Q

Most common photodetector used in spectrophotometer

A

Photomultiplier tube

229
Q

Save the measurement of elutions

A

Recorders

230
Q

Elution strength of the mobile phase is constant

A

Isocratic elution

231
Q

2 distinct portion of mass spectrophotometer

A

Fragmentation
Ionization

232
Q

Separate the components of a mixture

A

Fragmentation

233
Q

Methods use to ionize samples

A

Electron Spray Ionization (ESI)
Matrix Assisted Laser Desorption Ionization (MALDI)

234
Q

Analyzers used to measure mass-to-charge ratio

A

Quadrupole mass analyzers
Iron trap analyzers
Time of flight analyzer

CLINICAL CHEMISTRY (10 decks)

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