01 ANALYTICAL TECHNIQUES AND INSTRUMENTATION Flashcards
Spectrophotometry (give the absorbed color and observed color):
400-435
Violet
Green yellow
Spectrophotometry (give the absorbed color and observed color):
435-480
Blue
Yellow
Spectrophotometry (give the absorbed color and observed color):
480-490
Green blue
Orange
Spectrophotometry (give the absorbed color and observed color):
490-500
Blue green
Red
Spectrophotometry (give the absorbed color and observed color):
500-560
Green
Purple (magenta)
Spectrophotometry (give the absorbed color and observed color):
560-580
Yellow green
Violet
Spectrophotometry (give the absorbed color and observed color):
580-595
Yellow
Blue
Spectrophotometry (give the absorbed color and observed color):
595-605
Orange
Green blue
Spectrophotometry (give the absorbed color and observed color):
605-650
Red orange
Blue green
Spectrophotometry (give the absorbed color and observed color):
650-750
Red
Blue
Most common light source for spectrophotometry
Tungsten
This part of the spectrophotometry minimizes unwanted stray of light and prevents entrance of scattered light on the monochromator system
Entrance slit
The most common cause of lost linearity at high analyte concentration
Stray light
This isolates specific or individual wavelength of light
Monochromator
This controls the width of the light beam
Exit slit
Can be described as the total range of wavelengths transmitted
Exit slit
Holds the solution whose concentration is to be measures
Cuvette
This detects and converts transmitted light into photoelectric energy
Photodetector
This measures light emitted by one atom burned in a flame
Flame emission photometry
Color of flame emitted:
Sodium
Yellow
Color of flame emitted:
Potassium
Purple
Color of flame emitted:
Calcium
Orange
Color of flame emitted:
Magnesium
White
Color of flame emitted:
Copper I
Blue
Color of flame emitted:
Copper II
Green
Color of flame emitted:
Lithium
Red
Color of flame emitted:
Cesium
Violet
Color of flame emitted:
Zinc
Yellow
Color of flame emitted:
Strontium
Red
Measures the light absorbed by one atom burned in a dissociated by heat
Atomic absorption spectrophotometry
This determines the amount of scattered light by a particulate matter suspended in a turbid solution
Nephelometry
The light scattering during nephelometry depends on which factor/s?
Particle size
Wavelength
This measures the amount of antigen-antibody complexes
Nephelometry
Nephelometry measures at what angle?
15-90
Nephelometry uses which principle?
Mie Scatter
This measures reduction in light transmission by one particle formation
Turbidimetry
Turbidimetry uses which principle?
Raleigh scatter
This is the process of separating this charged constituents of a sample by means of an electrical current
Electrophoresis
Migration of charged macromolecules in the presence of an electrical power through porous supports
Zone electrophoresis
Porous supports for zone electrophoresis
Paper
Cellulose acetate
Agarose gel
Has a net charge that can either be positive or negative depending on pH conditions
Amphoteric
Movement of buffer ions and solvent relative to the fixed support
Electroendosmosis/endosmosis
Migration of small charged ions
Iontophoresis
This support media is specific for serum proteins
Cellulose acetate
This support media is used for DNA separation with 10-15 bands
Agarose gel
This support media is used for isoenzymes
Polyacrylamide gel
Planar form chromatography:
fractionation of sugar and amino acids through whattman paper sorbent
Paper chromatography
Planar form chromatography:
Used for TDM to separate drug molecules through plastic plates sorbent
Thin layer chromatography
Column form chromatography:
elution of volatile compounds based on boiling point, used to separate steroids, lipids, alcohols
Gas chromatography
Column form chromatography:
Differences in absorption of gases at solid phase surfaces
Gas solid chromatography
Column form chromatography:
Differences in solute partitioning between gaseous mobile phase vs liquid stationary phase
Gas liquid chromatography
Column form chromatography:
distribution of solute between liquid mobile phase vs liquid stationary phase
Liquid chromatography
Column form chromatography:
Most widely used LC, uses pressure for fast separation, fractionation of drugs, hormones, and Hgb variants
HPLC
Column form chromatography:
Separate non-volatile substances in human body fluids, complementary method to GC-MS as it is used to confirm positive results from screening illicit drugs
LC-MS
Instrumentation technique:
Fragmentation and ionization of molecules using suitable energy
Mass spectroscopy
Gold standard for forensic science and drug screening
GC-MS
Gold standard for protein identification
MS/MS
Instrumentation technique:
Determine the structure of organic compound
Nuclear magnetic resonance spectroscopy
Separation technique:
Migration is based on electrical charge
electrophoresis
Separation technique:
Migration is based on physical/chemical properties
chromatography
Separation technique:
Migration is through a pH gradient
isoelectric focusing
Separation technique:
Migration is through an electro-osmosis flow
capillary electrophoresis
Instrumentation technique:
amount of light emitted by a molecule after excitation by electromagnetic radiation
Fluorometry
Instrumentation technique:
Measures amount of light intensity present over a dark background
Fluorometry
Light sources for fluorometry
Mercury, xenon, uv lights
The reduction or limitation of a particle in an excited state
quenching
Factors affecting quenching
pH, temp, uv, chemical changes
Instrumentation technique:
measurement of luminescence produced by chemical reaction producing light emission
Chemiluminescence
Instrumentation technique:
Measures amount of light emission based on chemical or electrochemical reaction
Chemiluminescence
Instrumentation technique:
No excitation radiation is required, and no monochromators are needed because the chemiluminescene arises from one species
Bishop
The instrument that measures chemiluminescence
Luminometer
Instrumentation technique:
A simplified spectrophotometer that does not require a light source or monochromator since light is produced in the chemical reaction
Wendy
In Wendy, the ___, ___, and ___ are the main components of a luminometer
sample cuvette
photodetector
readout device
Instrumentation technique:
changes in colligative property of solutions that occur due to variations in particle concentrations
Osmometry
Example of osmotic particles
Glucose, BUN, sodium
What are the four colligative properties?
Boiling point, freezing point, osmotic pressure, vapor pressure
Electrochemistry technique:
Measurement of electric potential due to the activity of free ions
Potentiometry
Reference electrodes for potentiometry
Calomel, silver-silver chloride, H+ electrode
Parameters measured by potentiometry
pH and pCO2
Potentiometry uses ___ equation
Nernst
Electrochemistry technique:
Electrochemical transducer capable of responding to one given ion
Ion selective electrode
Electrochemistry technique:
Measurement of the amount of electricity fixed at potential in coulombs
Coulometry
In coulometry, the end point is detected by ___
amperometry
Electrochemistry technique:
Electrochemical titration in which the titrant is electrochemically generated
coulometry
Parameters measured by coulometry
Chloride tests - CSF, Serum, Sweat
Coulometry uses ___ law
Faraday’s
Electrochemistry technique:
Current flow produced by oxidation reaction
Amperometry
Electrochemistry technique:
Measurement of differences in current at a constant voltage
Polarography
Electrochemistry technique:
Measurement of current after a potential is applied to an electrochemical cell
Voltametry
Parameters measured by amperometry
pO2, glucose, chloride, peroxidases
Parameters measured by Anodic stripping voltametry
lead and iron studies
Polarography uses ___ equation
Ilkovic