ELECTRIPHOREISIS AND ELECTROCHEMISTRY Flashcards by Unknown Unknown

→ A molecule that contains both acidic and basic groups

Ampholyte

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The rate of migration of a charged solute in an electric
field, expressed per unit field strength.

● Electrophoretic Mobility

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Preferential movement of water in one direction through
electrophoresis medium due to selective binding of one
type of charge on the surface of the medium.

Endosmosis

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→ The migration of charged macromolecules.

Electrophoretogram

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The migration of small charge ions.

Iontophoresis

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The migration of charged macromolecules.

Zone Electrophoresis

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is acetylated to form cellulose acetate by treating it
with acetic anhydride

Cellulose acetate

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Different fractions of lipoproteins
based on their density

High-density lipoproteins (HDL)
▪ Low-density lipoproteins (LDL)
▪ Intermediate-density lipoproteins
(IDL)
▪ Very low-density lipoproteins (VLDL)

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negatively charged

Cathode

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positively charged

Anode

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Versatile and powerful analytical technique capable of
separating and analyzing a diverse range of ionized analytes.

ELECTROPHORESIS

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Macromolecules of interest:

Proteins in serum
→ Urine
→ Cerebrospinal fluid (CSF)
→ Erythrocytes and tissue and,
→ Other biologic body fluids

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Separates by electrical charge
● A highly purified uncharged polysaccharide derived from agar
● Neutral; therefore, doesn’t produce electroendosmosis

Agarose gel

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CONVENTIONAL ELECTROPHORESIS (4)

→ Separation
→ Staining
→ Detection
→ Quantification

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Referred to as PAGE
Involves separation of protein on the basis of charge and
molecular size
● Layers of gel with different pore sizes are used

Polyacrylamide gel

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Used to separate macromolecules on the basis of both
surface charge and molecular charge

Starch

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Power supplies operating at either constant current or
constant voltage are available commercially.

DRIVING FORCE (ELECTRICAL POWER)

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is a molecule whose net charge can be either
positive or negative

AMPHOLYTE

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Serves as a multifunctional component in the electrophoretic
process

BUFFER

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 Serum protein separation,
 Poor resolution, weak buffer

Barbitone buffer –
(around 8.0 pH)

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Enzyme separation,
Low buffering capacity, high
conductivity

Phosphate buffer
– (around 7.0 pH)

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Nucleic acid separation
 Good resolution, high buffering
capacity, low conductivity

Tris – borate –
EDTA buffer
(TBE) – (pH
around 8.0)

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Nucleic acid separation
 High resolution, high buffering
capacity, low conductivity

Tris – acetate –
EDTA buffer
(TAE) – (pH
around 8.0)

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Protein separation
 High buffering capacity, low
conductivity

Tris – glycine
buffer – (pH more
than 8.0)

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Based on refractivity (the ability of the substance to bend light)

REFRACTOMETRY

Involves the measurement of electrical signals associated with chemical system that are incorporated into an electrochemical cell

ELECTROCHEMISTRY

Converts chemical energy into electrical energy ● The redox reaction is spontaneous and is responsible for the production of electrical energy

GALVANIC CELL

Also known as a salt bridge are required to complete the circuit between the reference and without contaminating anything

Liquid junction

Measurement of a cell potential (voltage) under equilibrium conditions

A. POTENTIOMETRY

give me the two types of electrodes

Reference Electrode Indicator Electrode

→ Produces constant potential

Reference Electrode

→ Responds to changes in the activity of solution → Measuring electrode

Indicator Electrode

→ Mercury/mercurous chloride → Contains a known concentration of potassium chloride → Mercury ions react with pure sample components than do silver ions

a. Calomel electrode

→ Overall better and faster → Consists of a silver electrode immersed in a solution of potassium chloride that has been saturated with silver chloride → Can be used in temperatures greater than 60°C whereas calomel electrodes cannot

b. Silver/silver chloride

c. Normal hydrogen electrode

2 Types of ISE

1. Direct ISE 2. Indirect ISE

Glass aluminum silicate

Sodium

Valinomycin gel

Potassium

Organic liquid membrane ion exchanges

Calcium and Lithium

Gas and enzyme electrodes

Selective for the detection of hydrogen ions

pH electrode

pH electrode contained within a plastic jacket

pCO2 electrode

Is an electrochemical titration in which the titrant is electrochemically generated and the endpoint is detected by amperometry

COULOMETRY

The measurement of the electrical current at a single applied potential ● The measurement of the current flow produced by an oxidation-reduction reaction

AMPEROMETRY

→ Used to determine the partial pressure of oxygen in blood incorporated in blood gas analyzers uses an amperometric or current-sensing electrolytic cell as the indicator electrode

pO2 gas-sensing electrode

→ Employed in glucometer that determines a random determination of glucose by glucose oxidase principle

Glucose electrode

Based on the current-potential relationship in an electrochemical cell when the potential is applied

VOLTAMMETRY

for Lead and Iron Testing

Anodic stripping voltammetry

Voltammetry Three Electrodes:

a. Working Electrode b. Reference Electrode c. Auxiliary Electrode

→ Makes contact with the analyte → Facilitate the transfer of charge to and from the analyte

Working Electrode

→ A half cell with a known reduction potential

b. Reference Electrode

→ To sustain electrolysis → Process by which electric current is passed through a substance to effect a chemical change (substance loses or gains an electron)

c. Auxiliary Electrode

Measurement of the osmolality of an aqueous solution such as serum, plasma and urine; measurement of the concentration of dissolve solute particles in a solution

OSMOMETRY

Are those properties of a solution that are only a function of the concentration (molality) of the particles in solution. Ideally, the size of the particles, the mass of the particles, and the type of particles do not affect the colligative properties.

COLLIGATIVE PROPERTIES

● Minimum pressure which needs to be applied to a solution to prevent the inward flow of its pure solvent across a semipermeable membrane

Osmotic Pressure

→ The tendency of a fluid, usually water, to pass through a semipermeable membrane into a solution where the solvent concentration is higher, thus, equalizing the concentration of the materials on the other side of the membrane

Osmosis

Measure of the tendency of a material to change into the gaseous or vapor state ● Increases with temperature

Vapor Pressure

Measurement is based on the change in electrical resistance across an aperture when a particle in conductive liquid passes through this aperture.

ELECTRICAL IMPEDANCE

All of the following increases as the osmolality increases, EXCEPT: a. Osmotic pressure b. Vapor pressure c. Boiling point

2. What is the test used for coulometry? a. Sodium Test b. Potassium Test c. Chloride Test

3. Which of the following describes an agarose gel? a. Used for DNA or protein separations b. Polysaccharide extracted from sea weed c. Staining can be done after pouring the gel

b. Polysaccharide extracted from sea weed

Buffer used for separating proteins and has high buffering capacity, low conductivity: a. Tris – glycine buffer b. Tris – acetate – EDTA buffer (TAE) c. Tris – borate – EDTA buffer (TBE)

. Tris – glycine buffer

The migration of charged macromolecules: a. Zone Electrophoresis b. Iontophoresis c. Electrophoretogram

First statement: Cathode is negatively-charged Second statement: Anode is postiviely-charged a. Only the 1st statement is correct. b. Only the 2nd statement is correct. c. Both statements are correct. d. Both statements are incorrect.

β-Globulin/s: a. Immunoglobulin b. Transferrin c. Both a and b

b. Transferrin

All of the following affect the charge of ampholytes, EXCEPT: a. pH b. Ionic strength c. Molecular mass

c. Molecular mass

Internal reference electrode for pH electrode: a. Silver b. Silver chloride c. Silver nitrate d. Two of the choices are correct. e. None of the choices are correct

Ethidium Bromide and and Sybr Gold are dyes used to visualize what analyte? a. Protein b. DNA c. Hemoglobin