Centrifugation

How is relative centrifugal force (RCF) calculated

1.12 x R x (RPM/1000)^2

What is column chromatography

Column packed with appropriate stationary phase and equilibrated with mobile phase, sample loaded on top and flushed through mobile phase, if substances have different rates of migration they are separated within the column and eluted at different times

What is high performance liquid chromatography

Liquid sample injected into tube packed with particles (stationary phase), individual components of sample are moved down the packed column with a liquid (mobile phase) forced through column by high pressure delivered by pump

What is gas chromatography

Mobile phase is gas, requires sample to be volatile or converted to volatile derivative, injected through septum into heated port and rapidly evaporated and pulled through column with carrier gas

What is thin layer chromatography

Mobile phase is solvent and stationary phase is glass plate with silica layer, less sensitive

What g-force do large particles require

Smaller g-force

What are the essential components of chromatographic separation

Stationary phase, chromatographic bed, mobile phase, delivery system and detection system

What is the criteria for successful isopycnic centrifugation

Density of sample particle must fall within the limits of gradient densities, any gradient length is acceptable, run time must be sufficient for the particles to band, excessive run times have no adverse effects

What is the criteria for successful rate-zonal centrifugation

Density of the sample must be less than the lowest density portion of the gradient and vice versa, pathlength of the gradient must be sufficient for the separation to occur

What are the two types of centrifugation

Differential centrifugation and density gradient centrifugation

What are the steps of centrifugation

Homogenisation through sound, pressure, sheering or a mild detergent and placing in a centrifuge

What is the order of organelles in diameter

Nuclei, mitochondria, lysosomes and ribosomes

What is isopycnic separation

Particles separate based on their density, particle size only effects the rate at which they move to their matching density

What are the four major separation modes that are used to separate compounds in HPLC

Reversed phase chromatography, normal phase and adsorption chromatography, ion exchange chromatography and size exclusion chromatography

What is the order of organelles in density

Ribosomes, nuclei, mitochondria and lysosomes

What is the sedimentation coefficient

S = vt/a

What is chromatographic separation

Separation of individual components of a sample on the basis of difference in their physical characteristics

What are the three main types of chromatographic separation

Thin layer chromatography, column chromatography and gas chromatography

What is centrifugation

Use of centripedal force for the separation of mixtures, allowing separation of particles based on their size, density and viscosity of the medium

What is rate zonal centrifugation

particles move at different rates depending on their size and mass