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Disclaimer:
This outline is meant to be an overview or introduction and is not intended to be thoroughly
descriptive or instructional with regard to an particular separation process contained herein.
Adsorption is a process that occurs when a gas or liquid solute accumulates on the
surface of a solid or a liquid (adsorbent), forming a molecular or atomic film (the adsorbate). It is
different from absorption, in which a substance diffuses into a liquid or solid to form a solution. The
term sorption encompasses both processes, while desorption is the reverse process.
Activated carbon adsorption explained:
The best example of activated carbon adsorption explained as a process is in a wastewater
treatment plant where millions of gallons of polluted, unfiltered water enters into a system and must
be treated before being transported to nearby water systems. Activated carbon has an affinity for
various organics and is use for organic contaminant removal from water supplies. There are several
forms of activated carbon: powdered activated carbon (PAC) is ground loose carbon. Adsorption
isotherm tests are run to determine the effectiveness on the substance and its concentration in the
surrounding solution at equilibrium. Granular Activated Carbon (GAC) is typically used in drinking
water systems where liquid flows through stationary beds through the adsorpbent. In generic terms,
the carbon is heavier in weight than water and traps contaminants into its material.
Adsorption is indicative in most natural physical, biological, and chemical systems, and is widely
used in industrial applications such as activated charcoal, synthetic resins and water purification.
Adsorption, ion exchange and chromatography are sorption processes in which certain adsorptives
are selectively transferred from the fluid phase to the surface of insoluble, rigid particles suspended
in a vessel or packed in a column.
Centrifugation is a process that involves the use of the centripetal force for the
separation of mixtures, used in industry and in laboratory settings. More-dense components of the
mixture migrate away from the axis of the centrifuge, while less-dense components of the mixture
migrate towards the axis. In chemistry and biology, increasing the effective gravitational force on a
test tube so as to more rapidly and completely cause the precipitate ("pellet") to gather on the
bottom of the tube. The remaining solution is properly called the "supernate" or "supernatant liquid".
Since "supernatant" is an adjective, its usage alone is technically incorrect, although many
examples can be found in scientific literature. The supernatant liquid is then either quickly decanted
from the tube without disturbing the precipitate, or withdrawn with a Pasteur pipette.
Common Centrifugation Vocabulary and Formulas:
S-value: the sedimentation coefficient is a number that gives information about the molecular
weight and shape of the particle. S-value is expressed in Svedberg units. The larger the Svalue, the faster the particle separates.
Pelleting time: time taken to pellet a given particle. T = K/S where T= pellet time in hours. K
= K-factor of the rotor, and S = sedimentation coefficient.
Chromatography terms
Plotted on the x-axis is the retention time and plotted on the y-axis a signal (for example
obtained by a spectrophotometer, mass spectrometer or a variety of other detectors)
corresponding to the response created by the analytes exiting the system. In the case of an
optimal system the signal is proportional to the concentration of the specific analyte
separated.
Types of Chromatography
Planar Chromatography
Original location
spot of the mixture.
Column chromatography
As the mobile phase (solventlike liquid in this case) passes
downward through the
column, the different parts of
the mixture travel through at
different rates. The different
rates of travel result in a
separation of the mixture.
Column chromatography is a separation technique in which the stationary bed is within a tube. The
particles of the solid stationary phase or the support coated with a liquid stationary phase may fill
the whole inside volume of the tube (packed column) or be concentrated on or along the inside tube
wall leaving an open, unrestricted path for the mobile phase in the middle part of the tube (open
tubular column). Differences in rates of movement through the medium are calculated to different
retention times of the sample.
In 1978, W. C. Still introduced a modified version of column chromatography called flash column
chromatography (flash). The technique is very similar to the traditional column chromatography,
except for that the solvent is driven through the column by applying positive pressure. This allowed
most separations to be performed in less than 20 minutes, with improved separations compared to
the old method. Modern flash chromatography systems are sold as pre-packed plastic cartridges,
and the solvent is pumped through the cartridge. Systems may also be linked with detectors and
fraction collectors providing automation. The introduction of gradient pumps resulted in quicker
separations and less solvent usage.
Gas chromatography
Gas chromatography (GC), also sometimes known as Gas-Liquid chromatography, (GLC), is a
separation technique in which the mobile phase is a gas. Gas chromatography is always carried out
in a column, which is typically "packed" or "capillary" .
Gas chromatography (GC) is based on a partition equilibrium of analyte between a solid stationary
phase (often a liquid silicone-based material) and a mobile gas (most often Helium). The stationary
phase is adhered to the inside of a small-diameter glass tube (a capillary column) or a solid matrix
inside a larger metal tube (a packed column). It is widely used in analytical chemistry; though the
high temperatures used in GC make it unsuitable for high molecular weight biopolymers or proteins
(heat will denature them), frequently encountered in biochemistry, it is well suited for use in the
petrochemical, environmental monitoring, and industrial chemical fields. It is also used extensively
in chemistry research.
Liquid chromatography
Liquid chromatography (LC) is a separation technique in which the mobile phase is a liquid. Liquid
chromatography can be carried out either in a column or a plane. Present day liquid
chromatography that generally utilizes very small packing particles and a relatively high pressure is
referred to as high performance liquid chromatography (HPLC).
In the HPLC technique, the sample is forced through a column that is packed with irregularly or
spherically shaped particles or a porous monolithic layer (stationary phase) by a liquid (mobile
phase) at high pressure. HPLC is historically divided into two different sub-classes based on the
polarity of the mobile and stationary phases. Technique in which the stationary phase is more polar
than the mobile phase (e.g. toluene as the mobile phase, silica as the stationary phase) is called
normal phase liquid chromatography (NPLC) and the opposite (e.g. water-methanol mixture as the
mobile phase and C18 = octadecylsilyl as the stationary phase) is called reversed phase liquid
chromatography (RPLC). Ironically the "normal phase" has fewer applications and RPLC is
therefore used considerably more.
Affinity chromatography
Affinity chromatography is based on selective non-covalent interaction between an analyte and
specific molecules. It is very specific, but not very robust. It is often used in biochemistry in the
purification of proteins bound to tags. These fusion proteins are labelled with compounds such as
His-tags, biotin or antigens, which bind to the stationary phase specifically. After purification, some
of these tags are usually removed and the pure protein is obtained.
Reversed-phase chromatography
Reversed-phase chromatography is an elution procedure used in liquid chromatography in which
the mobile phase is significantly more polar than the stationary phase.
Two-dimensional chromatography
In some cases, the chemistry within a given column can be insufficient to separate some analytes.
It is possible to direct a series of unresolved peaks onto a second column with different physicochemical (Chemical classification) properties. Since the mechanism of retention on this new solid
support is different from the first dimensional separation, it can be possible to separate compounds
that are indistinguishable by one-dimensional chromatography.
Countercurrent chromatography
Countercurrent chromatography (CCC) is a type of liquid-liquid chromatography, where both the
stationary and mobile phases are liquids. It involves mixing a solution of liquids, allowing them to
settle into layers and then separating the layers.
Chiral chromatography
Chiral Chromatography involves the separation of stereoisomers. In the case of enantiomers, these
have no chemical or physical differences apart from being three dimensional mirror images.
Conventional chromatography or other separation processes are incapable of separating them. To
enable chiral separations to take place, either the mobile phase or the stationary phase must
themselves be made chiral, giving differing affinities between the analytes. Chiral chromatography
HPLC columns (with a chiral stationary phase) in both normal and reversed phase are commercially
available.
The crystal growth is the subsequent growth of the nuclei that succeed in achieving the critical
cluster size. Nucleation and growth continue to occur simultaneously while the supersaturation
exists. Supersaturation is the driving force of the crystallization, hence the rate of nucleation and
growth is driven by the existing supersaturation in the solution. Depending upon the conditions,
either nucleation or growth may be predominant over the other, and as a result, crystals with
different sizes and shapes are obtained (control of crystal size and shape constitutes one of the
main challenges in industrial manufacturing, such as for pharmaceuticals). Once the
supersaturation is exhausted, the solid-liquid system reaches equilibrium and the crystallization is
complete, unless the operating conditions are modified from equilibrium so as to supersaturate the
solution again.
Drying is a mass transfer process resulting in the removal of water moisture or moisture
from another solvent, by evaporation from a solid, semi-solid or liquid (hereafter product) to end in a
solid state. To achieve this, there must be a source of heat, and a sink of the vapor thus produced.
In the most common case, a gas stream, e.g., air, applies the heat by convection and carries away
the vapor as humidity. Other possibilities are vacuum drying, where heat is supplied by contact
conduction or radiation (or microwaves) while the produced vapor is removed by the vacuum
system. Another indirect technique is drum drying, where a heated surface is used to provide the
energy and aspirators draw the vapor outside the room.
Freeze drying or lyophilization is a drying method where the solvent is frozen prior to drying and is
then sublimed, i.e., passed to the gas phase directly from the solid phase, below the melting point
of the solvent. Freeze drying is often carried out under high vacuum to allow drying to proceed at a
reasonable rate. This process avoids collapse of the solid structure, leading to a low density, highly
porous product, able to regain the solvent quickly. In biological materials or foods, freeze drying is
regarded as one of the best if not the best method to retain the initial properties. It was first used
industrially to produce dehydrated vaccines, and to bring dehydrated blood to assist war casualties.
Now freeze drying is increasingly used to preserve some foods, especially for backpackers going to
remote areas. The method may keep protein quality intact, the same as the activity of vitamins and
bioactive compounds.
In turn, the mechanical extraction of the solvent, e.g., water, by centrifugation, is not considered
"drying". The ubiquitous term dehydration may mean drying of water-containing products as foods,
but its meaning is more vague, as it is also applied for water removal by osmotic drive from a salt or
sugar solution. In medicine, dehydration is the situation by which a person loses water by
respiration, sweating and evaporation and does not incorporate, for whatever reason, the "makeup" water required to keep the normal physiological behavior of the body.
Drying may be either a natural or an intentional process. The process of extreme drying is called
desiccation.
Laboratory distillation set-up: 1: Heat source 2: Still pot 3: Still head 4: Thermometer/Boiling point
temperature 5: Condenser 6: Cooling water in 7: Cooling water out 8: Distillate/receiving flask 9:
Vacuum/gas inlet 10: Still receiver 11: Heat control 12: Stirrer speed control 13: Stirrer/heat plate
14: Heating (Oil/sand) bath 15: Stirrer bar/anti-bumping granules 16: Cooling bath.
Evaporation this separation technique removes a volatile liquid solvent from its
dissolved or suspended components.
Extraction Processes
Leaching is the process of extracting a substance from a solid by dissolving it in a liquid.
Brewing of tea and coffee can similarly be considered as leaching process.
Flocculation is a process where a solute comes out of solution in the form of floc or
"flakes." The term is also used in colloid chemistry to refer to the process by which fine particulates
are caused to clump together into floc. The floc may then float to the top of the liquid, settle to the
bottom of the liquid, or can be readily filtered from the liquid. Flocculation and sedimentation are
widely employed in the purification of drinking water as well as sewage treatment, stormwater
treatment and treatment of other industrial wastewater streams.
Filtration
Mesh, bag and paper filters are used to remove large particulates suspended in fluids, eg. fly
ash,
membrane processes including microfiltration, ultrafiltration, nanofiltration, reverse osmosis,
dialysis (biochemistry) utilising synthetic membranes can separate micrometre-sized or
smaller species.
Oil-water separation - gravimetric separator used to remove suspended oil droplets from
wastewaters in oil refineries, petrochemical and chemical plants, natural gas processing
plants and similar industries.