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Clinical Chemistry (CC 1) > (P) Lec 5: Analytical Techniques in CC (P2) > Flashcards

(P) Lec 5: Analytical Techniques in CC (P2) Flashcards

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1
Q

Refers to migration of charged particles in an electric field

A

Electrophoresis

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2
Q

Separates proteins based on their electric charge densities

A

Electrophoresis

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3
Q

Charge of proteins

A

Negative

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4
Q

Buffer used in electrophoresis and its pH

A

Barbital/Veronal (pH = 8.6)

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5
Q

Terms

Has a net charge of that can either be positive or negative depending on the pH condition

A

Amphoteric

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6
Q

Terms

Movement of buffer ions and solvent relative to the fixed support (gel)

A

Electroendosmosis/Endosmosis

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7
Q

Terms

Migration of small charged ions

A

Iontophoresis

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8
Q

Terms

Migration of charged macromolecules

A

Zone Electrophoresis

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9
Q

Familiarize the components of electrophoresis

A
  • Electrical power
  • Support medium
  • Buffer
  • Sample
  • Detecting system
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10
Q

What’s the detecting system of electrophoresis in MolBIo

A

UV light or radiophotography

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11
Q

What are the factors affecting the rate of Migration?

A

Net electric charge of the molecule
Size and shape of the molecule
Electric field strength
Nature of the supporting medium
Temperature of the operation

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12
Q

What are the supporting media in electrophoresis?

A
  1. Cellulose acetate
  2. Agarose gel
  3. Polyacrylamide gel
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13
Q

Familiarize the stains for electrophoresis bands

A

Amido Black
Ponceau S
Oil Red O
Sudan Black
Fat Red 7B
Coomassie Blue
Gold/Silver Stain

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14
Q

Visualization for separating lipoproteins with fats (lipids + proteins)

A

Oil Red O
Sudan Black
Fat Red 7B

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15
Q

Staining protein found in CSF for staining during electrophoresis

A

Coomassie Blue

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16
Q

Staining for smaller ions in electrophoresis

A

Gold/silver stain

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17
Q

Measures electrophoretic bands
Measures the absorbance of the stain
Scans and quantitates electrophoretic pattern

A

Densitometry

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18
Q

Types of electrophoresis

A

Isoelectric focusing & Capillary Electrophoresis

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19
Q

Isoelectric electrophoresis separate by migration through what?

A

pH gradient

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20
Q

Ideal for separating proteins of identical sizes but different net charges
Proteins move in electric field until they reach a pH equal to their isoelectric point

A

Isoelectric focusing

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21
Q

Advantages of isoelectric electrophoresis

The ability to resolve mixture of _____ (identical in sizes)
Detects _______
identify genetic variants of ______
Detects CSF _______

A

protein
isoenzyme
genetic variants of proteins
oligoclonal banding

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22
Q

In this type of electrophoresis, sample molecules are separated by Electro-Osmotic Flow

A

Capillary electrophoresis

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23
Q

Capillary electrophoresis uses which unit to express quantity of specimen

A

Nanoliter

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24
Q

Capillary Electro

Which charged ions in the specimen emerge early at the capillary outlet because of EOF and the ion movements are in the same directions?

A

Positively charged

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25
Q

Capillary Electro

Which charged ions in the specimen towards the capillary outlet but at a slower rate?

A

Negatively charged

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26
Q

Refers to separation of soluble components in a solution by specific differences in the physical and chemical characteristics of the different constituents

A

Chromatography

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27
Q

Familiarize basis of separation of chromatography

A

Rate of diffusion
Solubility of the solute
Nature of the solvent
Sample volatility/solubility
Distribution between 2 liquids
Molecular size
Hydrophobicity of molecule
Ionic attraction
Differential distribution between 2 immiscible liquids
Selective separation of substances
Differences in the absorption and desorption of solutes

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28
Q

Two forms of chromatography

A

Planar and column

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29
Q

Paper chromatography is used for the fractionation of which two components?

A

Sugar and amino acid

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30
Q

Refers to
Semi-quantitative drug screening test
Sample components are identified by comparison with standards on the same plate

A

Thin layer chromatography (TLC)

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31
Q

T or F: TLC is quantitative drug screening test

A

False (semi)

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32
Q

T or F: Extraction of drug in TLC is pH independent

A

False (pH dependent)

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33
Q

In TLC, this is defined as thin plastic plates impregnated with a layer of silica or alumina

A

Sorbent

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34
Q

Type of chromatography that uses a tube

A

Column

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35
Q

Used for the separation of steroids, barbiturates, blood, alcohol, and lipids

A

Gas chromatography

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36
Q

Useful for compounds that are naturally volatile or can be easily converted into a volatile form

A

Gas chromatography

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37
Q

Samples are introduced into the gas chromatography column using a _________________

A

hypodermic syringe or an automated sample

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38
Q

Elution order of volatiles in gas chromatography are based on what?

A

Boiling point

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39
Q

Mobile phase of samples in gas chromatography

A

Nitrogen
Helium
Hydrogen
Argon (Inert gasses)

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40
Q

In mass spectroscopy, the separation is based on what?

A

Fragmentation and ionization

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41
Q

MS can also detect _______ and determination of ______________

A

Structural information, molecular weight

42
Q

T or F: Compound cannot be detected by MS if it isn’t separated first by GC

A

True

43
Q

Refers to double spectroscopy

A

Tandem Mass Spectroscopy

44
Q

The tandem of GC and MS is called what?

A

Gas-Chromatography-Mass Spectroscopy

45
Q

This can detect 20 inborn errors of metabolism from a single blood spot

A

Tandem Mass Spectroscopy

46
Q

Gold standard for drug testing

A

GC-MS

47
Q

Identify which is false about GC -MS

a.) Gold standard for drug testing
b.) Also used for xenobiotics, anabolic steroids, and pesticides
c.) Qualitative measurement of drug can be performed by selective ion monitoring
d.) Uses an electrode beam to split the drug emerging from the column into its component ions

A

C (guess why nalang)

48
Q

GC-MS

The position of the parent-molecule ion and degradation products give rise to _____________ that which will provide the final identity of the drug of interest

A

Fingerprint patterns

49
Q

Chromatography based on the distribution of solutes between a liquid phase and a stationary phase

A

Liquid chromatography

50
Q

What is the fixed component and moving component in Liquid Chromatography?

A

Stationary phase: fixed component
Liquid phase: Moving component

51
Q

This uses high pressure for fast separations, controlled temperature, in line detectors and gradient elution technique

A

High Performance Liquid Chromatography (HPLC)

52
Q

Used in separation and quantification of various hemoglobin associated with specific diseases

A

HPLC

53
Q

E2 eong c0nfiMatOrY miTod nG gc-ms, an0 i2????

A

koriqUe! liqUiD chRoma - Mas sPectRo

54
Q

Also used in therapeutic drug monitoring, toxicology, and studies of drug metabolites
Requires interface methods to convert non-volatile to volatile compounds

A

LC-MS

55
Q

What are the interface methods used in LC-MS?

A

Electrospray (ES)
Atmospheric Pressure Chemical Ionization (ARCI)

56
Q

Familiar the separation mechanisms in Liquid Chroma

A

Gel/Gel Permeation/Gel Filtration/Size Exclusion/Molecular Sieve Chromatography
Ion exchange chromatography
Partition Chromatography
Affinity Chromatography
Adsorption Chromatography

57
Q

determines the light emitted by a molecule after excitation by electromagnetic radiation

This is the principle of which mechanism/process?

A

FLUORIMETRY/MOLECULAR LUMINESCENCE

58
Q

Light source of Fluorimetry/Molecular Luminescence

A

Mercury or Xenon Lamp

59
Q

Light detector of Fluorimetry

A

Photomultiplier Tube or Phototube

60
Q

This is used to measure porphyrins, magnesium, calcium, and catecholamines

A

Flourinetry

61
Q

How many monochromators are used in fluorimetry?

A

Two

62
Q

Assoc type, which is true about Fluorimetry

a. The incident light is prevented from striking the photodetector by the primary filter
b. The wavelength that is best absorbed by the solution to be measured is selected by the second filter

A

D (u rationalize it for more challenge, amen)

63
Q

T or F: Fluorimetry is 100x more sensitive than spectrophotometer

A

False (1000x)

64
Q

This is affected by quenching

A

Fluorimetry

65
Q

Refers to the loss of fluorescence and radiation

A

Quenching

66
Q

Familiarize the parameters affected by the quenching effect of fluorimetry

A

pH
Temperature changes
Chemical contaminants
UV light changes

67
Q

Emission of light created by your sample after a chemical ог electrochemical reaction and not from absorption of electromagnetic energy

A

Chemiluminescence

68
Q

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

This is the principle of which mechanism/analytical technique?

A

Chemiluminescence

69
Q

Use of chemiluminescence

A

Immunoassays (serology)

70
Q

Photodetector of chemiluminescence

A

Photomultiplier tube (Luminometer)

71
Q

Which is more sensitive between fluorescence and chemiluminescence?

A

Chemiluminescence

72
Q

T or F:

In chemiluminescence, no excitation radiation is required and no monochromator are needed because the chemiluminescence arises from one species

A

True

73
Q

Measurement of OSMOLALITY of an aqueous solution such as serum, plasma or urine

A

Osmometry

74
Q

Based on measuring changes in the colligative properties of solutions that occur owing to variations in particle concentrations

This is the principle of which mechanism/analytical technique?

A

Osmometry

75
Q

What are the four colligative properties involved in osmometry?

A

Osmotic pressure
Boiling point
Freezing point
Vapor pressure

76
Q

These are the solutions measured in osmometry?

A

serum, plasma or urine

77
Q

These are used on osmotic particles that when added, will increase the osmolality reading on the osmometer

A

Glucose
Urea
Sodium

78
Q

Which of the colligative properties increases when osmolality is increased?

A

Osmotic pressure
Boiling point

79
Q

Which of the colligative properties decreases when osmolality is increased?

A

Freezing point
Vapor pressure

80
Q

Which is true about Freezing Point Depression Osmometry:

a.) Most commonly used method for measuring the changes in the colligative properties of a solution
b.) Based on the principle that addition of solute molecules lowers the temperature at which a solution freezes.

A

C

81
Q

A 1.0 mOsm/kg solution has a freezing point depression at __________when compared with a pure solvent (usually water)

A

0.00186°C

82
Q

Freezing point of abd osmolality of plasma?

A

Osmolality: 285 mOsm/kg
Freezing point: -53 ° C

83
Q

What are measured in electrochemistry techniques?

A

Current and voltage

84
Q

Electrochemistry

Measurement of electrical potential due to the activity of free ions
Follows the NERNST EQUATION
A change in the voltage indicates activity of each analyte

A

Potentiometry

85
Q

Reference electrodes of potentiometry

A

Calomel and Silver chloride

86
Q

Potentiometry uses which tests

A

pH and pCO2 tests

87
Q

Electrochemical transducer capable of responding to one given ion
VERY SENSITIVE and SELECTIVE for the ion it measures.

A

Ion selective electrode

88
Q

Ionic selectivity in Ion selective electrode depends on the ___________ used

A

membrane/barrier composition

89
Q

T or F: Ion selective electron is selective, sensitive, and specific

A

False (not specific)

90
Q

In Ion selective electrode, this measures the electrolytes dissolved in the fluid phase of the sample

A

ISE analyzers

91
Q

ISE membrane

Glass membrane electrode measures what analyte?

A

Sodium

92
Q

ISE membrane

Liquid membrane + valinomycin measures what analyte?

A

Potassium

93
Q

ISE membrane

Silver membrane electrodes measures what analyte?

A

Chloride

94
Q

T or F: Ion selective electrode does not discriminate between ions causing voltage differences between the measuring electrode and the standard electrode

A

Truew

95
Q

Electrochemistry

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 at amperometry
Follows FARADAY’s LAW

A

Coulometry

96
Q

Test used in coulometry

A

Chloride test

97
Q

Electrochemistry

Measurement of the current flow produced after by an oxidation reaction

A

Amperometry

98
Q

Interferences in coulometry

A

Bromide
Cyanide`
Cysteine

99
Q

Uses of amperometry

A

pO2
glucose
chloride
peroxidase determinations

100
Q

Measurement of differences in current at a constant voltage
Follows the ILKOVIC EQUATION

A

Polarography

101
Q

Electrochemistry

Measurement of current after which a potential is applied to an electrochemical cell
Allows the sample to be pre-concentrated, thus utilizing minimal analytes

A

Voltammetry

102
Q
A

Clinical Chemistry (CC 1) (7 decks)

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