Void Volume

Definition - What does Void Volume mean?

Void volume is the volume of the pores or space between particles in:

Ion exchanger

Filter media

Other granular material

This is often expressed as a percentage of the total volume occupied by the material.
Void volume refers specifically to the volume of liquid phase contained inside a column. The same
term is sometimes also used informally to refer to the volume of a cavity in the column/tubing or
fittings.
Void volume is also known as dead volume.
Corrosionpedia explains Void Volume
Void volume is the volume of mobile phase (Vm or V0) in a column. In an ideal case, it is equal to the
mobile phase hold-up volume. For example, if the stationary phase occupies 40% of the total column
volume, the void volume would be 60% of the total column volume. Consider a column that is 25 cm
long with an inner diameter of 1 cm. The total column volume is 19.6 mL. If the mobile phase
occupies 60% of the column volume, the void volume is 11.8 mL.
In column chromatography, the volume of the mobile phase is the total bed volume of the column
minus the volume occupied by the support particles.
In gel chromatography or exclusion volume, it is the volume of the mobile phase passing through the
gel required to elute a molecule that never entered the stationary phase.
Various methods to estimate the total void volume include:

Pycnometry

Minor disturbance method

Tracer pulse method

Hold-up volume based on unretained compounds

Accessible volume based on Martin's rule and its descendants

In post-tensioned ducts, where voids are discovered or suspected, a simple pressure test using
compressed air can be used to determine the approximate void volume. It is the presence of
corrosion, which may occur in the void, which is a cause for concern.
Void volume is an important carbon black structure property. A profile of void volume as a function of
applied pressure provides a means to assess carbon black structure at varying levels of density and
aggregate reduction.

Void Volume
The void volume is the volume of mobile phase (V m or V0) in a column. For example,
if the stationary phase occupies 40% of the total column volume, the void volume
would be 60% of the total column volume.

Consider a column that is 25 cm long with an inner diameter of 1 cm. The total
column volume is 19.6 mL (V = r2L = 3.14 * 0.52 * 25 cm). If the mobile phase
occupies 60% of the column volume, the void volume is 11.8 mL.

PUMP SYSTEM
In the last module you appreciated the crucial role played by a detector in the HPLC system. In this
module you will be introduced to the pump which provides continuous and consistent flow of mobile
phase through the HPLC system. A pump can be compared to the human heart which continuously
pumps blood throughout the body but though the human heart can withstand changes in blood
pressure within specified limit due to stress and strain the HPLC pump is required to deliver flow of
mobile phase at constant pressure and flow rate. Changes in both these parameters can lead to
errors in the results. In simple language the HPLC pump has to have ruggedness and at the same
time should be able to provide reproducible flow characteristics run after run. The operational
pressure limits have a vast range depending upon analysis requirements. In normal analytical
operation the pressure can vary between 2000 5000 psi but in applications covered under UHPLC
mode operating pressure can be as high as 15000 18000 psi.
An ideal pump should have the following desirable characteristics

Solvent compatibility and resistance to corrosion

Constant flow delivery independent of back pressure

Convenience of replacement of worn out parts

Low dead volume for minimum problems on solvent changeover

Three commonly used pump types are Syringe type pumps, Constant pressure pumps and
Reciprocating piston pumps.
Constant pressure pumps provide consistent continuous flow rate through the column with the use
of pressure from a gas cylinder. Valving arrangement allows rapid refill of solvent chamber. A low
pressure gas source is needed to generate high liquid pressures.
Syringe Type Pumps are suitable for small bore columns. Constant flow rate is delivered to column
by a motorized screw arrangement. Solvent delivery rate is set by changing voltage on the motor.
These pumps deliver pulseless flow independent of column backpressure and changes in viscosity
but major disadvantages are limited solvent capacity and limitation on gradient operation
Reciprocating Piston pumps deliver solvent(s) through reciprocating motion of a piston in a
hydraulic chamber. On the back stroke the solvent is sucked in and gets delivered to the column in
the forward stroke. Flow rates can be set by adjusting piston displacement in each stroke. Dual and
triple head pistons consist of identical piston chamber units which operate at 180 0 or 1200 phase
difference. The solvent delivery of reciprocating pump systems is smooth because while one pump is
in filling cycle the other is in the delivery cycle. High pressure output is possible at constant flow rate
and gradient operation is possible. However, pulse dampening is required for further elimination of
pressure pulses.
We now have a fair understanding of different pumps and their operation principles and our next post
will deal with injectors for injection of samples into the flowing mobile phase stream.

High Performance Liquid Chromatography Methods,

Benefits and Applications
By G.P. Thomas

Topics Covered
What is HPLC?
How Does HPLC Work?
Manufacturers of HPLC Systems
Benefits of HPLC
Applications of HPLC
References

What is HPLC?
High-performance liquid chromatography or high-pressure liquid chromatography (HPLC) is
a chromatographic method that is used to separate a mixture of compounds in analytical
chemistry and biochemistry so as to identify, quantify or purify the individual components of
the mixture.

Reversed phase HPLC/UHPLC chromatography is a commonly used separation mode. It

provides dynamic retention of compounds possessing hydrophobic and organic functionality.
A combination of hydrophobic and van der Waals type interactions between all the target
compound and both the stationary and mobile phases enables the retention of these
compounds by reversed phase.

How Does HPLC Work?

In very small amounts, the sample mixture to be separated and tested is sent into a stream
of mobile phase percolating via the column. There are different types of columns available
with sorbents of varying particle sizes and surfaces.
The mixture moves through the column at varying velocities and interact with the sorbent,
also known as the stationary phase. The velocity of each component in the mixture depends
on its chemical nature, the nature of the column and the composition of the mobile phase.
The time at which a specific analyte emerges from the column is termed as its retention
time. The retention time is measured under specific conditions and considered as the
identifying characteristic of a given analyte.
Sorbent particles might be hydrophobic or polar in nature. The commonly used mobile
phases include any miscible combination of water and organic solvents such as acetonitrile
and methanol. Water-free mobile phases can also be used.
The aqueous component of the mobile phase might contain acids like formic, phosphoric or
trifluoroacetic acid or salts to enable the separation of the sample components. The
composition of the mobile phase is either maintained as a constant or as varied during the
chromatographic analysis. The constant approach is effective for the separation of the
sample components that are not very dissimilar in their affinity for the stationary phase. In
the varied approach, the composition of the mobile phase differs from low to high eluting
strength. The eluting strength of the mobile phase is reflected by analyte retention times
where high eluting strength produces fast elution.
The composition of the mobile phase is chosen based on the intensity of interactions
between several sample components and the stationary phase.
The HPLC partitioning process is quite similar to the liquid-liquid extraction process except
that the former is a continuous process unlike the latter which is a step-wise process. It is
recommended that trial partitioning processes be performed to determine the exact HPLC
method that would provide adequate separation.

Manufacturers of HPLC Systems

There are a wide range of HPLC options in the market today. The following is a list of various
HPLC system producers with a brief introduction of their products:

Dionex, a company based on the science of chromatography, manufactures the

UltiMate3000 Rapid Separation LC system, the UltiMate 3000 RSLCnano system, and
the Coronafamily of universal charged aerosol detectors.

Jasco Analytical Instruments provide a range of HPLC systems. The LC-2000 Plus
Series HPLC Systems is uniquely configurable. With high performance and low cost,
these HPLC systems can be adapted to virtually any requirement from simple isocratic
QA to advanced multi-solvent/multi-column method development. The Isocratic LC2000plus-Iso is provided with a pump, degasser, autosampler, and UV/Vis detector. The
pump is designed with flow rates from 1L to 10mL/min at pressures up to 500 bar for
use with 2, 3, 4, 6 and up to 10mm ID columns. The Binary Gradient LC-2000plus-HPG
is similar to the isocratic system but is provided with a two-solvent gradient pump
configuration. The pumps flow rates are compatible with 1mm columns. The
Quaternary Gradient LC-2000plus-LPG is also similar to the isocratic system; however,
this model is provided with a quaternary gradient pump configuration for maximum
solvent flexibility.

The Preparative HPLC System from Gilson has a wide flow range that can be adapted
for both semi-preparative and preparative separations. Its high pumping power allows a
wide range of preparative column sizes. The system is designed with a new rinse
station system comprising a flowing jet wash, and allows the use of up to two different
rinse solvents.

Sharp HPLC Systems from AAPPTec are provided with pumps that are
microprocessor controlled, thus enabling highly precise and highly accurate flow rates,
which can help obtain excellent analytical reproducibility and highest measurement
accuracy.

Benefits of HPLC
The key benefits of HPLC systems are as follows:

Controls and automates chromatography instrumentation

Provides data management, security features, and reporting and instrument
validation.
Powerful and adaptable
Increases productivity by managing all the areas of analysis - from sample to
instrument, and from separation to reporting results.
Affordable

Applications of HPLC
HPLCs can be used in the following applications:

Water purification

Preconcentration of trace components

Ligand-exchange chromatography

Ion-exchange chromatography of proteins

High-pH anion-exchange chromatography of carbohydrates and oligosaccharides