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Clinical Chemistry > Analytical Method > Flashcards

Analytical Method Flashcards

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1
Q

transmitted by electro waves that are characterized by their frequency and wavelength

A

energy

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2
Q

distance between 2 successive peaks and it is expressed in nanometer (nm)

A

wavelength

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3
Q

number of vibration of wave motion per second

A

frequency

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4
Q
  • technique used to determine the concentration of colored compounds in solutions
  • principle that the concentration of a substance is proportional to the intensity
    of the color of the solution
A

colorimetry

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5
Q

primary analytical utility of spectrophotometry or filter photometry is the isolation of discreet portions of the spectrum for purposes of measurement

A

photoelectric colorimetry

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6
Q

measurement of light intensity in a narrower wavelength.

A

spectrophotometric measurement

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7
Q

measurement of light intensity

A

Photometric measurement

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8
Q

involves the measurement of the light transmitted by a solution to determine the
concentration of the light-absorbing in the solution

A

Spectrophotometry

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9
Q
  • simplest type of absorption spectrometer
  • designed to make one measurement at a time at one specified wavelength
  • absorption maximum of the analyte must be known in advance when a single instrument is used
A

Single beam spectrophotometer

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10
Q
  • an instrument is this the monochromatic light in the two components - one through the sample, one through a reference solution or blank
  • additional beam corrects for variation in light source intensity
  • The absorbance of the sample can be recorded via electrical
    the output of the sample beam
A

Double beam spectrophotometer

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11
Q
  • an analytical instrument used to quantify the analytes in a given sample using a light beam
  • absorption of light by the sample
A

spectrophotometer

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12
Q

2 photodetectors, for the
sample beam and reference
beam

A

double beam in space

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13
Q

one photodetector and alternately passes the monochromator through the sample cuvet and the reference cuvette using a
chopped rotating sector mirror

A

double beam in time

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14
Q

provides the initial light that will pass through the sample

A

Radiant Energy/Light source

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15
Q

emits radiation that changes in intensity; widely used in the laboratory

A

Continuum source

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16
Q

most commonly used light source in the lab

A

Tungsten

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17
Q

used to provide UV radiation in analytic specimens

A

Deuterium

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18
Q

discharge lamp, produces a continuous source of radiation, which covers
both the invisible and visible range

A

Xenon lamps

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19
Q

emits limited radiation and wavelength, line source emits discrete lines widely used
in atomic absorption and fluorescent spectroscopy

A

Line source

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20
Q
  • minimizes unwanted or stays light and prevents the entrance of scattered light into the monochromator system
A

Entrance slit

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21
Q

most common cause of loss of linearity at high analyte concentration

A

ERROR

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22
Q

it isolates specific or individual wavelengths of light from a broad spectrum produced by the light source

A

Monochromator

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23
Q
  • wedge-shaped pieces of glass, quartz or sodium chloride
  • can be rotated, allowing only the desired wavelength to pass through an exit slit.
  • A narrow light focused on a prism is refracted as it enters the more dense glass
A

prisms

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24
Q
  • most commonly used; better resolution than prism
  • made by cutting grooves (parallel grooves) or slits into an aluminized surface of a flat piece of crown glass
  • wavelengths are bent as they pass a sharp corner
A

Diffraction gratings

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25
Q
  • simple, least expensive, not precise but useful
  • made by placing semi-transparent silver films on both sides of a dielectric such as magnesium fluoride
  • usually pass a wide band of radiant energy and have a low transmittance of the selected
    wavelength
A

filters

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26
Q

optical components used in spectrometers and other optical devices to disperse light into
its component wavelengths

A

Holographic gratings

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27
Q
  • controls the width ofthe light beam (bandpass)
  • narrow fraction of the spectrum to reach the sample cuvette
  • Spectral purity of the spectrophotometer is reflected by the bandpass, that is, the narrower the bandpass, the greater the resolution.
A

exit slits

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28
Q
  • also called absorption/analytical/sample cell
  • holds the solution whose concentration is to be measured
A

cuvettes

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29
Q

kinds of cuvettes: most commonly used (can be used in 350-2000nm)

A

Alumina silica glass

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30
Q

used for measurement of solution requiring visible and ultraviolet spectra

A

Quartz/plastic

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31
Q

detects and converts transmitted light into photoelectric energy. It detects the amount of
light that passes through the sample in the cuvet

A

Photodetector

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32
Q

kind of detector:
- simplest detector; least expensive; temperature-sensitive
- used in filter photometers with a wide bandpass.

A

barrier layer cell

33
Q

kind of detector:
- contains cathode and anode enclosed in a glass case
- photosensitive material that gives off electron when light energy strikes it
- requires an external voltage for operation

A

Phototube

34
Q

kind of detector:
- most commonly used detector - measures visible and UV regions
- excellent sensitivity and has a rapid response - detects very low levels of light

A

Photomultiplier tube (PMT)

35
Q

kind of detector:
- not as sensitive as PMT but with excellent linearity.
- measures light at a multitude of wavelengths - detects less amount of light.
- lower dynamic range and higher noise compared to PMT
- most useful as a simultaneous multichannel detector.

A

Photodiode

36
Q

kind of detector:
- displays output of the detection system

A

Meter or read-out device

37
Q
  • states that the concentration of the unknown substance is directly proportional to the absorbed
    light and inversely proportional to the amount of transmitted light
  • mathematically establishes the relationship between concentration and absorbance.
A
  • Beer’s Law
38
Q
  • amount of light absorbed
  • proportional to the inverse log of
    transmittance
A

absorbance

39
Q

ratio of the radiant energy transmitted (T) divided by the radiant energy incident on the
sample

A

Percent Transmittance

40
Q
  • blank contains serum but without the reagent to complete the assay
  • blank corrects absorbance caused by the color of the reagents
  • measures absorbance of the sample and reagent in the absence of the end of product, and corrects the measurement for optical interference (like hemoglobin) absorbing the wavelength of measurement
A

Blanking Technique

40
Q

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

A

Flame Emission Photometry (FEP)

41
Q

measures the light absorbed by atoms dissociated by heat.

A

Atomic Absorption Spectrophotometry (AAS)

41
Q

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

A

volumetric

42
Q
  • measuring abundant large particles (proteins) and bacterial suspensions.
  • depends on specimen concentration and particle size. The measurement of reduction of light is due to particle formation
A

TURBIDIMETRY

43
Q
  • measuring the amount of antigen-antibody complexes (proteins)
  • It determines the amount of scattered light by a particulate matter suspended in a
    turbid solution
A

nephelometry

44
Q
  • migration of charged particles in an electric field.
  • separates proteins on the basis of their electric charge densities.
A

electrophoresis

45
Q

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

A

Amphoteric

46
Q

movement of buffer ions and solvent relative to the fixed
support

A

Electroendosmosis/Endomosis

47
Q

migration of small charged ions

A

lontophoresis

48
Q

migration of charged macromolecules

A

Zone electrophoresis

49
Q
  • separates molecules by migration through a pH gradient
  • ideal for separating proteins of identical sizes but with different net charges.
A

Isoelectric Focusing

50
Q
  • sample molecules are separated by electro-osmotic flow (EOF)
  • utilizes nanoliter quantities of specimens.
A

capillary electrophoresis

51
Q

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

A

CHROMATOGRAPHY

52
Q
  • used for fractionation of sugar and amino acid.
  • Sorbent (stationary phase) - Whatman paper
A

paper chromatography

53
Q
  • a semiquantitative drug screening test
  • Sample components are identified by comparison with standards on the same plate
A

Thin layer chromatographty

54
Q
  • used for separation of steroids, barbiturates, blood, alcohol and lipids.
  • useful for compounds that are naturally volatile or can be easily converted into a volatile
    form
A

gas chromatography

55
Q
  • based on the fragmentation and ionization of molecules using a suitable source of energy
  • can also detect structural information and determination of in molecular weight.
A

Mass Spectroscopy (MS)

56
Q
  • gold standard for drug testing
  • also used for xenobiotics, anabolic steroids, and pesticides.
  • uses an electron beam means to split the drug emerging from the column into its component ions -
    drugs are detected fragments neans of the presence of decomposition fragmerts which arise after degradation of the analytes.
A

Gas Chromatography-Mass Spectroscopy (GC-MS)

57
Q
  • based on the distribution of solutes between a liquid mobile phase and a stationary phase
  • HPLC is the most widely used liquid chromatography
A

Liquid Chromatography

58
Q
  • uses pressure for fast separations, controlled temperature, in-line detectors and gradient elution technique
A

High Performance Liquid Chromatography (HPLC)

59
Q
  • detecting nonvolatile substances in body fluids.
  • utilized to confirm positive results from screening of illicit drug
  • a complementary method to GC-MS
A

Liquid Chromatography-Mass Spectroscopy (LC-MS)

60
Q

separates molecules based on differences in their size and shape

A

Gel/Gel Permeation/Gel Filtration/Size Exclusion /Molecular Sieve Chromatography

61
Q

for separation of enzymes, antibodies and proteins - example: dextran and agarose

A

Hydrophilic gel (Gel Filtration)

62
Q

separation of triglyceride and fatty acid - example: sephadex

A

Hydrophobic gel (Gel Permeation)

63
Q

exchange of sample ions and mobile phase ions with the charge groups of the stationary phase.
- Used to separate amino acids,
proteins, nucleic acids.

A

Ion exchange chromatography

64
Q
  • separation of compounds are based on their partition between the liquid mobile phase in the
    liquid stationary phase coated in solid support
  • used to separate therapeutic drugs and their metabolites.
A

Partition chromatography

65
Q
  • uses immobilized biochemical ligands as the stationary phase.
  • separation uses the lock-key binding which is present widely in biologic systems.
  • used to separate lipoproteins, carbohydrates and glycated hemoglobins; antibodies
A

Affinity chromatography

66
Q
  • separation is based on the differences between the adsorption and the desorption of solutes at the surface of a solid particle
  • compounds are adsorbed to a solid support such a silica or alumina.
A

Adsorption chromatography

67
Q
  • measures the amount of light intensity present over a zero background
  • determines the amount of light emitted by a molecule after excitation by electromagnetic radiation
A

FLUOROMETRY/MOLECULAR
LUMINESCENCE

68
Q
  • refers to the process by which the intensity of fluorescence is reduced due to various interactions between the fluorescent molecules and other substances or environmental
    factors
  • can affect the accuracy and reliability of fluorescence-based assays, which are commonly used to measure the concentration of analytes in biological sample
A

Quenching

69
Q
  • a surface phenomenon in which molecules, ions, or particles from a fluid (gas or liquid) adhere to the surface of a solid or a liquid
  • process differs from absorption, where the substance permeates or is uniformly distributed throughout the bulk of the solid or liquid
A

Adsorption

70
Q
  • 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
  • The chemical reaction yields an electronically excited compound that emits light as it returns to its ground state, or that transfers its energy to another compound, which then produces emission
A

CHEMILUMINESCENCE

71
Q
  • the measurement of the osmolality of an aqueous solution such as serum, plasma, or
    urine.
  • based on measuring changes in the colligative properties of solutions that occur owing to variations in particle concentration
A
  • OSMOMETRY
72
Q
  • measurement of electrical potential due to the activity of free ions - change in voltage
    indicates activity of each analyte
  • measurement of differences in voltage (potential) at a constant current
A

Potentiometry

73
Q
  • measurement of the amount of electricity (in coulombs) at a fixed potential
  • electrochemical titration in which the titrant is electrochemically generated and the
    endpoint is detected by amperometry
A

Coulometry

74
Q
  • measurement of the current flow produced by oxidation-reaction.
A

Amperometry

75
Q
  • measurement of differences in current at a constant voltage
A

Polarography

76
Q
  • measurement of current after which a potential is applied to an electrochemical cell
  • allows sample to be preconcentrated, thus utilizing minimal analyte
A

Voltammetry

Clinical Chemistry (4 decks)

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