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analytical techniques Flashcards

1
Q

is polarized and coherent and has narrow spectral width and small cross section area with low divergence.

A

-Laser light

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2
Q

Single beam spectrophotometer

A

Light Source> Entrance Slit> Monochromator>Exit Slit>Sample Cuvet>PM tube>A/D > Display

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3
Q

ELECTROPHORESIS

A
  • Migration of charged solutes/particles in an electrical field.
  • Five components: Driving force, support medium, buffer, sample, detecting system
  • Procedure:
  • support materials:cellulose acetate, agarose gel, polyacrylamide gel, starch gel
  • treatment and application of sample
  • detection and quantitation
  • electroendosmosis
  • isoelectric focusing
  • capillary electrophoresis
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4
Q

*Group of techniques used to separate complex mixtures on basis of different physical interactions

A

CHROMATOGRAPHY

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5
Q

measures light transmitted by a solution to determine concentration of light-absorbing substance in a solution

A

spectrophotometric instruments

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6
Q

*Migration of charged solutes/particles in an electrical field.

A

ELECTROPHORESIS

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7
Q

-Applications of Mass Spectrophotometry in clinical lab

A

Structural information and molecular weight determination

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8
Q

Components of spectrophotometer

A

Light source
Monochromators
Sample cell
photodetectors

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9
Q
  • chloride ISEs have largely replaced cuilometric titrations

- anodic stripping voltametry was widely used for analysis of lead.

A

•Cuolometric chloridometers and anodic stripping voltametry

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10
Q

Spectrophotometer quality assurance

A

Wavelength accuracy
stray light
linearity

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11
Q

detector of mass spectrophotometry

A

Use of an electron multiplier to detect ions

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12
Q

measure light intensity without consideration of wavelength

A

photometric instruments

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13
Q

Concentration of a substance is directly proportional to right amount of light absorbed or inversely proportional to logarithm of transmitted light

A

beer’s law

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14
Q

most common mass analyzer used today

A

Quadrupole:

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15
Q

routinely used to measure concentration of trace metals that are not easily excited

A

Atomic Absorption Spectrophotometer

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16
Q

*Basic Components of Chromatography:

A

Mobile phase
Stationary phase
Column holding stationary phase
Separated components

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17
Q
  • atmospheric pressure ionization
A

> electrospray ionization

>Atmospheric pressure chemical ionization

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18
Q

-sample introduction and ionization

MASS SPECTROPHOTOMETRY

A
  • Electron ionization

* atmospheric pressure ionization

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19
Q

LASER stands for?

A

-Light Amplification by Stimulated Emission of Radiation

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20
Q

*Procedure of the electrophoresis:

A
  1. Sample is soaked in hydrated support for 5 minutes.
  2. Support is put into electrophoresis chamber filled with buffer.
  3. Constant voltage or current is applied for a specific time.
  4. Support is removed and placed in fixative or rapidly dried.
  5. Zones are stained with appropriate dye.
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21
Q

measures light emitted by excited atoms

A

flame photometry

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22
Q

*modes of separation of chromatography

A

Adsorption
Partition
Steric exclusion
Ion-exchange chromatography

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23
Q

no longer routinely used, replaced by ion selective electrodes

A

flame photometry

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24
Q

*Chromatographic Procedures

A
  • Thin layer chromatography (TLC)
  • HPLC high Performance Liquid Chromatography
  • gas chromatography
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25
basic instrumentation of fluorometry
Filter fluorometers measure concentrations of solution that contain fluorescing molecules source emits short-wavelength high-energy excitation light mechanical attenuator controls light intensity
26
* A variant of column chromatography * thin layer of sorbent is uniformly coated on glass/plastic plate * samples are applied as spots near edge of plate * one edge of plate is placed in solvent
-Thin layer chromatography (TLC)
27
used to determine concentration of Na, Li, or K
Flame photometry
28
-when coupled with other techniques, has powerful analytical capabilities with widespread clinical applications
MASS SPECTROPHOTOMETRY | ns
29
measures concentration by detecting absorption of electromagnetic radiation by atoms rather than by molecules sensitive and precise
Atomic Absorption Spectrophotometer
30
-used as a detector to identify samples eluting from gas chromatographic or HPLC colum
MASS SPECTROPHOTOMETRY
31
-based on interaction of radiant energy and suitably excited atoms or molecules
Laser application
32
ELECTROCHEMISTRY
* Galvanic and Electrolytic Cells * Half-cells. * Ion-Selective Electrodes. * pH electrodes * Gas-sensing electrodes * enzymes electrodes * Cuolometric chloridometers and anodic stripping voltametry
33
•components: of hplc
pumps, columns, sample injectors, detectors, recorders
34
-Part of chemical energy generated produces excited intermediates that decay to a ground state with emission of photons.
Chemiluminescence
35
* solvent migrates up thin layer by capillary action | * After solvent reaches predetermined height, plate is removed and dried.
-Thin layer chromatography (TLC)
36
similar to turbidity except different angle of measurement.
*nephelometry-
37
- chloride ISEs have largely replaced cuilometric titrations | - anodic stripping voltametry was widely used for analysis of lead.
•Cuolometric chloridometers and anodic stripping voltametry
38
•uses pressure for fast separations, controlled temperature, inline detectors, and gradient elution techniques.
-HPLC high Performance Liquid Chromatography
39
Can serve as source of incident energy in spectrophotometer or nephelometer
laser application
40
INSTRUMENTATION FOR PROTEOMICS
* Proteomics - Two-Dimensional electrophoresis - MALDI-TOF and SELDI-TOF Mass spectrophotometry.
41
advantage and disadvantage of fluorometry
advantage: greater specificity and sensitivity disadvantage : very sensitive to environmental changes
42
-Two-Dimensional electrophoresis
1. First dimension: Proteins are resolved according to their isoelectric points using immobilized pH gradients 2. second Dimension: proteins are separated according to size.
43
-unlike fluorescence, no excitation radiation or monochromators are required.
Chemiluminescence
44
can be used for: determination of structure, identification of samples, and diagnosis.
-laser spectrophotometry
45
-Oxidation reactions of luminol, arcidinium esters & dioxetanes
Chemiluminescence
46
-An ISE covered by immobilized enzymes that can catalyze a specific chemical reaction
•enzymes electrodes
47
basic filter fluorometer
source> attenuator > primary filter> sample holder> secondary filter> detector(photo multiplier) > readout
48
* Proteomics - investigation of protein products encoded by genes - important for disease detection
*Proteomics
49
*Five components of electrophoresis:
Driving force, support medium, buffer, sample, detecting system
50
-Shotgun approach to discovery of new biochemical markers
*Proteomics
51
measures concentration of particulate matter in sample.
*turbidity-
52
•used to separate mixtures of compounds that are volatile or can be made volatile
-gas chromatography
53
Laser application
-Light Amplification by Stimulated Emission of Radiation -based on interaction of radiant energy and suitably excited atoms or molecules -Laser light is polarized and coherent and has narrow spectral width and small cross section areawith low divergence. Can serve as source of incident energy in spectrophotometer or nephelometer -laser spectrophotometry can be used for: determination of structure, identification of samples, and diagnosis.
54
OSMOMETRY AND ANALYTICAL TECHNIQUES FOR POCT
* Osmometry | * Analytic Techniques for Point-of- Care Testing POCT
55
*support materials of electrophoresis:
cellulose acetate, agarose gel, polyacrylamide gel, starch gel
56
• components of gas chromatography
: columns, detectors
57
Chemiluminescence
- Part of chemical energy generated produces excited intermediates that decay to a ground state with emission of photons. - unlike fluorescence, no excitation radiation or monochromators are required. - Oxidation reactions of luminol, arcidinium esters & dioxetanes
58
>sample in a small tube is lowered into a chamber with cold refrigerant circulating from cooling unit > Thermistor is immersed in sample. > wire is used to stir sample until it is cooled to several degrees below its freezing point.
*Freezing point osmometer
59
*Measures concentration of solute particles in a solution
•Osmometry
60
>sample in a small tube is lowered into a chamber with cold refrigerant circulating from cooling unit > Thermistor is immersed in sample. > wire is used to stir sample until it is cooled to several degrees below its freezing point.
*Freezing point osmometer
61
>POCT devices are widely used for clinical application
• Analytic Techniques for Point-of- Care Testing POCT

Analytical Chemistry (2 decks)

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