Theoretical Plate

Theoretical plates represent a hypothetical division of

chromatographic columns, and each plate represents an
equilibrated partitioning of the solute between the stationary
and mobile phases.

From:
Encyclopedia of Spectroscopy and Spectrometry (Third Edition),
2017

Related terms:

Facilitated Diffusion, High-Performance Liquid Chromatography,

Capillary Electrophoresis, Electrophoresis, Elution, Liquid Chromatography,

Mobile Phase Composition, Ultra High Performance Liquid Chromatography,

Solution and Solubility, Diffusion Coefficient

Gas Chromatography and Gas

Chromatography—Mass Spectrometry
Eric Stauffer, ... Reta Newman, in Fire Debris Analysis, 2008

8.2.4 Efficiency
The efficiency of a column is measured by the number of theoretical
plates (N), or more often, the height equivalent to a theoretical plate
(HETP or H). The theoretical plate number (N) usually is expressed by one
of two equations [9]:

2
𝑁 = 16 ⋅ (𝑤𝑡𝑅𝑏 ) (Equation 8-2)

or

𝑡 2 (Equation 8-3)
𝑁 = 5.545 ⋅ (𝑤𝑅ℎ )

where tR refers to the retention time of the peak and Wb refers to the
peak width at baseline in Equation 8-2 and Wh its width at half-height in
Equation 8-3. Figure 8-3 illustrates the values used with these equations.
The higher the plate number N, the greater the efficiency of the column.
One can understand quickly that the narrower the peak (low W), the
higher N, and thus the efficiency. Efficiency is thus a measure of peak
broadening; an efficient system will result in narrow peaks, whereas a
less efficient column will contribute to band broadening.

FIGURE 8-3. The different values used to calculate the plate number (N).
The height equivalent to a theoretical plate (HETP) can be calculated
when both N and the column length (L) are known [10, 11]:

𝐻𝐸𝑇𝑃 = 𝐿 (Equation 8-4)

𝑁

As a result, the lower the HETP, the better the resolution and the more
efficient the separation. Efficiency is optimized when N is maximized and
HETP is minimized. The van Deemter equation is another way of
expressing efficiency; it takes into account three factors that, along with
the mobile phase linear velocity, will affect efficiency and, therefore, the
HETP value [13]. These factors are represented by the variables A, B, and C
in the van Deemter equation [10]:

𝐻𝐸𝑇𝑃 = A + 𝜇𝐵 + C ⋅ 𝜇 (Equation 8-5)

Theoretical Plate Concept

Historically, the concept of the number of plates as a measure of

efficiency is based upon separation by distillation. The ability to
separate by distillation was reflected in the number of plates, within
each of which distinct equilibria occurred. The greater the number of
plates, the better the potential for separation. Imagine a distillation
column with two plates, then one with 100 plates (refer to Figure 7-4,
which shows a simplified schematic of a distillation tower with only
five plates). Clearly, more plates allow for improved separation as a
more refined gradient of temperatures can be used. This concept was
adapted to chromatography by Martin and Synge in 1941 [12]:

The behaviour of a column consisting of a number of “theoretical plates”,

within each of which perfect equilibrium between the two phases occurs,
can be described with great simplicity. Peters [1922] showed that the
continuous or packed type of distillation column (in which equilibrium is
not established at any point) could be divided up into a number of layers
each of which was equivalent to one theoretical plate, and the height of
such a layer was called the H.E.T.P. or “height equivalent to one theoretical
plate”. For the present purpose the H.E.T.P. is defined as the thickness of the
layer such that the solution issuing from it is in equilibrium with the mean
concentration of solute in the non-mobile phase throughout the layer. It
can be shown from diffusion arguments that the H.E.T.P. is a constant
through a given column except when the ratio of the concentrations of the
solution entering and leaving the plate differs greatly from unity [cf.
Sherwood, 1937]. It may be taken as constant for the chromatogram
without serious error.

The theoretical plate concept is a good way to express relative

efficiency, although it is important to understand that this model has
limitations. When using the term “theoretical plates” one must
remember that there are not actual distinct equilibria being achieved.
Another concern with the theoretical plate model is that it does not
adequately account for band broadening. However, the number of
theoretical plates or the HETP is still used today as a means for
expressing the efficiency of a chromatographic system.

in which A represents the Eddy diffusion phenomenon2 in the column, B

represents molecular (or axial) diffusion, C relates to the resistance to
mass transfer, and μ is the linear velocity of the mobile phase through the
chromatographic system [10]. The linear velocity is calculated based on
the following equation [15]:

𝜇= 𝐿 (Equation 8-6)
t𝑀

where L is the length of the column [cm] and tM is the retention time[s] of
an unretained analyte (dead time). The mobile phase linear velocity, μ,
therefore is expressed in cm/s.

Using the van Deemter equation, one can plot the efficiency of a carrier
gas in terms of HETP as the dependent variable (y-axis) against the linear
velocity as the independent variable (x-axis). This plot is referred to as a
van Deemter plot, and can be used to compare the efficiency of one
carrier gas to another (see Figure 8-4).
FIGURE 8-4. A van Deemter plot showing the effect of each variable from
the corresponding equation. Note that the Eddy diffusion is independent
of the linear velocity, thus it is constant. Because the resistance to mass
transfer (C) is multiplied by the linear velocity, it increases with μ. The
invert occurs with the molecular diffusion (B).
All three parameters (A, B, and C) lead to band broadening: the spreading
apart of a group of identical analytes throughout the system. By
minimizing band broadening, identical analytes stick together and a
better separation is obtained because peaks are narrow rather than broad.
With Figure 8-4, one can quickly appreciate the influence of the mobile
phase flow rate on each parameter. The A term in the van Deemter
equation is relevant only to packed column systems and thus is constant
for a given column. Nevertheless, it is independent of the linear velocity μ
as indicated by Equation 8-5 and Figure 8-4. The B term represents the
natural tendency of the analytes to redistribute themselves from a region
of high concentration to a region of lower concentration in the mobile
phase [2]. Finally, the C term, resistance to mass transfer, represents the
fact that the transfer of an analyte from the mobile to the stationary
phase and vice versa is not instantaneous and has a certain inertia. As a
result, the analyte concentration profile of the stationary phase is slightly
behind the equilibrium position and the analyte concentration profile of
the mobile phase is slightly ahead of the equilibrium position [2]. As
expected, the C term increases with the flow rate as shown in Figure 8-4.

Since the Eddy diffusion does not apply to capillary columns, it may be
useful to become familiar with the Golay equation, in which the A term
has been removed and the C term has been expended [8]:

𝐻𝐸𝑇𝑃 = 𝜇𝐵 + (𝐶𝑠 + 𝐶𝑚 ) ⋅ 𝜇 (Equation 8-7)

where Cs is the mass transfer from the stationary phase to the mobile one
and Cm is the mass transfer from the mobile phase to the stationary one.

Another important chromatographic concept is that of resolution. The

resolution, R, is a measure of the true separation of two consecutive peaks
[10]. One can calculate the resolution of a chromatographic system from
two consecutive peaks using the following equation:
𝑅 = 𝑊12+⋅ 𝑑𝑊2 (Equation 8-8)

where d is the separation between two peaks (measured from top to top)
and W1 and W2 are the widths at baseline of the two peaks. This is
illustrated in Figure 8-5.

FIGURE 8-5. The measures used to calculate the resolution (R) of a

chromatographic system.

The goal in chromatography is to avoid coelution. This is an issue because

two coeluting peaks will mix together in the detector. For example, each
compound's contribution to the overall structural information provided
by the mass spectrometer will not be distinguishable, thus their
identification will be hampered. This is the reason why it is important to
achieve baseline resolution (R = 1.5) as the ultimate goal for all peaks. In
fire debris analysis, this goal can never be achieved in practice, however it
is important to adjust the chromatographic parameters to approximate it
as closely as possible. A resolution below 1.5 will not provide complete
separation and a resolution above 1.5 does not offer any extra advantages
to the separation. Figure 8-6 shows the peak configurations for three
different values of R.

FIGURE 8-6. The effect of resolution (R) on the separation of two

consecutive peaks.

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Bioseparation Engineering
Oliver Kaltenbrunner, ... Shuichi Yamamoto, in
Progress in Biotechnology, 2000

4.2 Selection of test velocity

Comparison of the effects of test velocity on HETP should be independent
of particle size. Using Equation 3 The dimensionless flow velocity can be
expressed as

𝑑𝑝 ⋅ 𝜀 ⋅ 𝑢 𝑣 𝑢 0 ⋅ 𝑑𝑝 Equation 4
Re ⋅ Sc = 𝑣 ⋅ 𝐷𝑚 = 𝐷𝑚

and HETP data can be made dimensionless by eqn. 4. The optimal flow
velocity for the test is when the ratio between the a term and the total h is
a maximum. This ensures the greatest contribution is from the packing
effects and minimizes the contribution from diffusion and mass transfer
limitations. Figure 4 shows a plot of the ratio a/h from Equation 3 versus
the flow velocity ReSc. Testing a column at a linear velocity of ReSc < 5
cannot give reliable information on the quality of the column packing.
HETP values in this range primarily reflect diffusion of tracer ions and is
strongly influenced by temperature and flow rate variations Conversely,
the higher the linear flow velocity, the greater is the contribution from
mass transfer limitations (c-term) and the higher the back pressure.
Hence, the best operating range is at linear velocities of 5 < ReSc < 15 which
ensures a high contribution of the a term.

Figure 4. Contribution of the a term of Equation 3 to the total broadening

h.
Figure 3. Column integrity test results for different particle size resins as a
function of flow velocity. The data are fitted using Equation 3.
For the proposed test method, with DNaCl, water = 1.5 10- 5 cm2s- 1, the
superficial velocity (u0) can be estimated as

3000 < 𝑢0 < 8000 Equation 5

𝑑𝑝 𝑑 𝑝

where u0 is in cm/h when dp is in μm.

For small particles, the optimum test region is restricted by back pressure
constraints. Applying the Kozeny Carman equation (12)

3 𝑑𝑝
2
(6)
𝑢0 = 𝜀
⋅ Δp
270 ⋅ 1 − 𝜀 𝜇 𝐿
2

allows for an estimate of the maximum column length (Lmax) for a given
dp and a back pressure (Δpmax) limitation. When equations 6 and 4 are
combined, Lmax to allow ReSc = 5 at Δpmax is

𝛥𝑝max 𝑑𝑝 𝜀
3
3
(7)
𝐿max = 𝜇 ⋅ 1350 ⋅ 𝐷𝑚 ⋅ 1 − 𝜀2

For the test method with injections of 1 M NaCl in a 0.1 to 0.2 M NaCl
equilibration buffer at 20 °C (μ0.2M Nacl, 20°C = 1.024 cP) and assuming ε = 0.4
and Dm = 1.5 10- 5 cm2s- 1, the equation simplifies to

𝛥𝑝max ⋅ 𝑑𝑝
3
(8)
𝐿max = 6
1.25 ⋅ 10

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Unit Operations
Pauline M. Doran, in Bioprocess Engineering Principles, 1995

10.7.3 Theoretical Plates in Chromatography

The concept of theoretical plates is often used to analyse zone broadening
in chromatography. The idea is essentially the same as that described in
Section 10.4 for an ideal equilibrium stage. The chromatography column is
considered to be made up of a number of segments or plates of height H;
the magnitude of His of the same order as the diameter of the resin
particles. Within each segment equilibrium is supposed to exist.
As in adsorption operations, equilibrium is not often achieved in
chromatography so that the theoretical-plate concept does not accurately
reflect conditions in the column. Nevertheless the idea of theoretical
plates is applied extensively, mainly because it provides a parameter, the
plate height H, which can be used to characterise zone spreading. Use of
the plate height, which is also known as the height equivalent to a
theoretical plate (HETP), is acceptable practice in chromatography design
even though it is based on a poor model of column operation. HETP is a
measure of zone broadening; in general, the lower the HETP value the
narrower is the solute peak.

HETP depends on various processes which occur during elution of a

chromatography sample. A popular and simple expression for HETP takes
the form:

𝐻 = 𝐴𝑢 + 𝐵𝑢 + 𝐶 (10.44)

where H is plate height, u is linear liquid velocity, and A, B and C are

experimentally-determined kinetic constants. A, B and C include the
effects of liquid–solid mass transfer, forward and backward axial
dispersion, and non-ideal distribution of liquid around the packing. As
outlined in Section 10.6.4, overall rates of solute adsorption and
desorption in chromatography depend mainly on mass-transfer steps.
Values of A, B and C are reduced by improving mass transfer between
liquid and solid phases, resulting in a decrease in HETP and better column
performance. Eq. (10.44) and other HETP models are discussed further in
other references [16, 17].

HETP for a particular component is related to the elution volume and

width of the solute peak as it appears on the chromatogram. If, as shown
in Figure 10.18(a), the pulse has the standard symmetrical form of a
normal distribution around a mean value¯𝑥, the number of theoretical plates
can be calculated as follows:
Figure 10.18. Parameters for calculation of: (a) number of theoretical
plates, and (b) resolution.

𝑁 = 16(𝑉𝑤e )
2 (10.45)

where N is number of theoretical plates, Ve is the distance on the

chromatogram corresponding to the elution volume of the solute, and w
is the base line width of the peak between lines drawn tangent to the
inflection points of the curve. Eq. (10.45) applies if the sample is
introduced into the column as a narrow pulse. Number of theoretical
plates is related to HETP as follows:

𝑁 = 𝐻L (10.46)

where L is the length of the column. For a given column, the greater the
number of theoretical plates the greater is the number of ideal
equilibrium stages in the system and the more efficient is the separation.
Values of H and N vary for a particular column depending on the
component being separated.

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CHROMATOGRAPHY | Principles
V.R. Meyer, in Encyclopedia of Analytical Science (Second Edition),
2005

Theoretical Plate Number and Plate Height

A visual idea of a theoretical plate is presented in Figure 1. The
mathematical expression of the number of theoretical plates N in a
chromatographic system is obtained from the width of a peak in relation
to its retention:

𝑡 2 𝑡 2 ℎ 𝑡 2 [8]
𝑁 = 16 𝑤R = 5.54𝑤1R/ 2 = 2𝜋 𝐴P PR

where w1/2 is the peak width at half height of the peak; hP is the peak
height; and AP is the peak area.

The greater the plate number of a chromatographic system, the more

difficult the separation problems that can be solved. In principle the
number of plates can be increased by using a longer column. Practical
problems, mainly the pressure drop across the column and difficulties in
manufacturing long packings or capillaries, set a limit at ∼105 plates in all
chromatographic techniques.

To describe the quality of a chromatographic system the height equivalent

to a theoretical plate, HETP or H, is more useful than N:

𝐿
𝐻=𝑁 [9]

where L is the length of the chromatographic bed.

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DISTILLATION | Packed Columns: Design

and Performance
L. Klemas, J.A. Bonilla, in Encyclopedia of Separation Science, 2000

System properties
Numerous investigations have tried to correlate experimental HETP data
with the distillation system fluid physical properties. The best and most
consistent correlations tend to confirm that the HETP is proportional to
reference HETPo and a factor proportional to the system physical
properties:

𝑛 𝑛 [15]
HETP = HETPo (𝜇𝛼 / 𝑆g 𝛿) / (𝜇𝛼 / 𝑆g 𝛿)
0
when the n exponent best fit is between 0.15 and 0.21. Replacing HETPo
from eqn [14] into eqn [15], and using the reference system we obtain:

f
HETP=(2.0𝐾p / 𝐹p )(𝜇𝛼 / 𝑆g 𝛿)
0.2 [16]

In addition, theoretical considerations suggest that the HETP is related to

λ=m/(L/V), the ratio of the slopes of the equilibrium line and operating
line, by the correlation:

HETP=𝜆ln(𝜆) / (𝜆 − 1)HTU

where HTU is the height of a transfer unit. Then:

f
HETP=(2.0𝐾p / 𝐹p )(𝜇𝛼 / 𝑆g 𝛿)
0.2
𝑓(𝜆) [17]

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Capillary Zone Electrophoresis

Robert Weinberger, in
Practical Capillary Electrophoresis (Second Edition), 2000

2.9 OPTIMIZING THE CAPILLARY LENGTH

The efficiency of the separation (theoretical plates) is directly
proportional to the capillary length (38), provided the field strength is
kept constant. The limitation here is available voltage. Most instruments
produce a maximum of 30 kV. Once the capillary length reaches a certain
point, the field strength must be reduced and no further gains in
efficiency are realized (6). Based on Eq. (2.16), the electrophoretic
resolution depends on the square root of the number of theoretical plates
and, thus, on the square root of the capillary length (38). Increasing the
capillary length beyond the limits imposed by the voltage maximum
lengthens the separation time without any substantial benefits. These
effects are illustrated in Figure 2.22, where the number of theoretical
plates is linear with the capillary length until 30 kV is reached. Note that
the resolution increase is proportional to the square root of the number of
theoretical plates.
FIGURE 2.22. Impact of capillary length on the number of theoretical
plates and resolution.
Most chemists overly rely on the length of the capillary to perform their
separations. This results in lengthy separations. Since diffusion is time
related, the sensitivity of the method declines as well. If more time is
spent optimizing the separation chemistry, shorter capillaries can be
employed, with obvious benefits.

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Methods
A. Shallan, ... M. Breadmore, in
Encyclopedia of Forensic Sciences (Second Edition), 2013

Background Electrolytes
The composition of the electrolytes inside the capillary is typically what
defines the mode of separation that occurs and the order in which peaks
move past the detector. The solution chemistry requirements for the
mode of separation are discussed in more detail below, but within each
separation mode, variation of the composition will change the selectivity.
Taking zone electrophoresis, for example, the exact composition of the
electrolyte will influence the sensitivity, resolution, and separation time,
as depicted in Figure 4. For example, changing the pH of the electrolyte
will change the net charge of weak acids and bases which will impact
upon the μep leading to a change in resolution and separation time. pH
may also change the surface charge and zeta potential of the capillary
thus changing the EOF, which will also impact upon the same criteria.
Changing the pH of the electrolyte will also change the ionic strength,
which will impact on both μep and EOF, thus also influencing the
separation. Similarly, adding an organic solvent such as methanol will
change the solvation of the ions and the viscosity of the electrolyte, which
will again influence mobility and EOF.

Figure 4. Factors affecting separation in CE.

The complexity of electrophoresis can often be daunting to the
electrophoresis newcomer, but there are a number of general principles
and conditions that can be used to rapidly identify the feasibility of a
specific separation, which can then be further refined as necessary.
Phosphate buffers at pH 2 and 7, and borate at pH 9, at a concentration of
10–50 mM are suitable electrolytes to start with as they provide a
reasonable ionic strength and have excellent transmission properties for
UV–vis detection. For MS detection, 1 M formic acid and acetic acid
provide low pH options, while at high pH, 10–100 mM ammonium
acetate/formate is the most popular option, while conductivity detection
typically uses 20–50 mM HIS/MES. It can also be simpler when starting to
perform separations in a co-EOF manner, or in a suppressed EOF
environment. Unmodified fused silica capillaries are good for cations and
for anions at low pH where the EOF is suppressed. If a high pH is required
for anion separations, then reversing the EOF is easily done by
incorporating a cationic surfactant (such as CTAB) into the electrolyte or
by coating the capillary as discussed above. If the analytes are neutral,
then a suitable additive such as SDS must be added to perform an
electrokinetic chromatography (EKC) separation.

Once conditions for a basic separation have been established, there are
then a myriad of electrolyte additives that can be used to enhance the
separation in one way or another. This may be simply changing the type
of salts used in the electrolyte (through for example, differences in ion-
association interaction), changing the solvent (organic solvents and mixed
solvents can provide uniquely different selectivity), and the addition of
additives to alter charge or size through secondary equilibria, and various
combinations of these. It is this ability to vary the position of a peak
within a separation and to adjust the conditions to manipulate the system
to achieve the desired outcome that is one of the reasons that
electrophoresis is such a powerful separation technique. The other is the
high efficiency.

Maximizing Efficiency
It is misleading to discuss theoretical plates in electrophoresis;
nevertheless, it is a convenient concept to describe analyte peak shape
and for comparison with other separation techniques. Efficiency is
described by the number of theoretical plates (N) and is related to the
height equivalent to the theoretical plates (Hth) by

𝐿
𝑁 = 𝐻d [7]
th

where Ld is the effective length of the capillary. The theoretical plate

number can be determined directly from an electropherogram by

2
𝑁 = 5.54𝑤1𝑡/ 2 = µapp × 2𝐷𝑉m [8]

where t is the migration time, w½ is temporal peak width at half-height,

and Dm is the diffusion coefficient of the analyte.

CE has much higher efficiencies than can be achieved by HPLC, with

typical plate numbers from 100 000 to 500 000 plates/m. This is primarily
because CE does not have many of the sources of inefficiency that HPLC
does and because of the flat plug-like profile of the EOF. Ideally in CE the
zone dispersion is only due to longitudinal diffusion. In reality, inherent in
all electrophoretic separations is electromigration dispersion. This arises
when there is a difference in electrophoretic mobility of the analyte and
the electrolyte co-ion (Figure 5). If the analyte ion has a higher μep than
that of the electrolyte co-ion, then the peak will be fronting, if μep are
equal then the peak will be symmetrical, and if the analyte has a lower
μep then the peak will be tailed. Electromigration dispersion can be
decreased by matching the mobilities of the buffer constituent to the
sample mobility or by maintaining a running buffer concentration
approximately two orders of magnitude higher than that of the sample.
Figure 5. Peak distortions due to differences in mobilities between sample
and BGE.
During any electrophoretic separation heat will be generated. The
temperature increase depends on the power and is determined by the
capillary dimensions, conductivity of the buffer, and the applied voltage.
If the heat is not dissipated efficiently, temperature gradients can develop
across the capillary with ions in the center of the capillary having a higher
μep than those at the capillary wall and band broadening will be observed.
This can be a significant issue when performing separations in plastic
microchips that have a lower thermal conductivity than glass and cannot
dissipate the heat as effectively. There are a number of methods that
indicate excessive heat generation and possible temperature gradients.
These phenomena may be indicated if efficiency is reduced as the voltage
is increased. And, a disproportionate increase in current with voltage,
Ohm's law, indicates temperature gradients. Measures to control Joule
heating may include reducing the capillary inner diameter, active
temperature control, or the use of low-mobility buffers which contain
large, minimally charged ions, such as Tris, borate, and histidine.

One other major source of band broadening in CE is that of wall

interaction. Depending on the extent of interaction, peak tailing and even
total adsorption of the solute can occur. The primary causes of adsorption
to the fused silica walls are electrostatic interactions between charged
analytes and the charged wall, and hydrophobic interactions. Significant
adsorptive effects have been observed especially for large peptides and
proteins, and it is imperative to minimize these to obtained highly
efficient separations. The use of zwitterionic buffer systems and pH and
ionic strength extremes can be useful for overcoming these issues, but the
most prominent approach is to modify the capillary wall to limit solute
adsorption.

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CHROMATOGRAPHY | Convective
Transport in Chromatographic Media
A.E. Rodrigues, in Encyclopedia of Separation Science, 2000

Conventional Packings
The column performance can be assessed in terms of HETP as a function
of bed superficial velocity following the classic van Deemter analysis. The
van Deemter equation for linearly retained species in conventional
packings of sphere geometry is:

𝜀p (1 − 𝜀b )𝑏
2
[2]
HETP = 𝐴 + 𝑢𝐵0 + 15
2
2 𝜏d 𝑢 0
[𝜀b + 𝜀p (1 − 𝜀b )𝑏]

where εp is the intraparticle porosity, εb is the bed porosity and b=1+

{(1−εp)/εp}K is the adsorption equilibrium parameter for a linear isotherm
with slope K. In a condensed form the van Deemter equation is:

HETP = 𝐴 + 𝑢𝐵0 + 𝐶𝑢0

with:

2
𝜀p (1 − 𝜀b )𝑏
𝐶 = 15
2
2 𝜏d
[𝜀b + 𝜀p (1 − 𝜀b )𝑏]

For protein separation the B term is negligible and the plot HETP versus u0
in HPLC is a straight line in most of the domain when conventional
supports are used. Moreover, the HETP increases with the square of the
particle size.

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Bioseparation Engineering
E.K. Lee, S.J. Ahn, in Progress in Biotechnology, 2000

5.1 Column Packing

For ion exchange column, HETP (height equivalent to a theoretical plate)
is measured usually by conductivity spiking method and its value is
compared against the mean bead diameter [5]. If it is lower than
approximately twice the mean diameter, a column is considered well
packed. HETP needs to be measured after each packing. It is essential to
use the same HETP measurement method each time and to trace its value
after each cycle. It should be noted that HETP can only indicate how well
packed the bed is and not the condition of the resin [7].

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GAS CHROMATOGRAPHY | High-Speed

Techniques
P.J. Marriott, R.A. Shellie, in
Encyclopedia of Analytical Science (Second Edition), 2005

Some Fundamental GC Theory and Routes

toward High-Speed Analysis
In practical GC, it is important that the number of theoretical plates
required (Nreq) in order to provide a desired resolution (Rs) for a given pair
of components (defined by α and k) can be calculated. This can be
achieved by using the Purnell equation (eqn [1]):

2
𝑁req = 16𝑅s 𝛼𝛼− 1
2 𝑘 + 12 [1]
𝑘

′ ′
where the separation factor 𝛼 = 𝑡R2 / 𝑡R1 , and retention factor k refers to
the last eluting peak in the pair. Rs is measured experimentally as

𝑅s =
Δ𝑡R [2]
4𝜎

Equation [1] can be applied to assist in the selection of the appropriate

chromatographic conditions to provide the minimum resolution required
for the analysis task. Baseline resolution is achieved at Rs>1.5, and will be
sufficient to provide accurate quantitative data in most applications. In
many cases solutes tend to be over-separated (Rs ≫2), and excess
resolution equates to excessive analysis time.

By extending this theory it is possible to determine the minimum time in

which sufficient resolution of a critical pair of peaks can be achieved (eqn
[3]):

𝑡R = 16𝑅
2 𝐻 1 + 𝑘3 𝛼 2 [3]
𝑢¯ 𝑘 𝛼−1
2

Equation [3] reveals that the column temperature (because this influences
k), carrier gas linear velocity (which will change the plate duration (the
Purnell criterion) 𝐻 / 𝑢¯ ), and column selectivity (affects k; α/α − 1) are
important considerations for reducing analysis time.

Since it is likely that the critical pair of components will not be the last
eluting peaks, they do not provide the best indication of the total analysis
time; however, this can be given by eqn [4]:

𝑡R = (𝐿 / 𝑢¯ )(𝑘𝑛 + 1) [4]
where L is the column length, 𝑢¯ the average linear velocity of the carrier
gas, and kn the retention factor of the last eluting compound in the
sample.
Equation [4] is very useful for deducing the general operating
requirements of high-speed analysis. The use of a short column, a higher
than usual carrier gas velocity, and relatively small retention factors
(which can be easily achieved by using high temperature and/or thin film
columns), can reduce analysis times significantly. There will, however, be
a penalty associated with reducing the analysis time by using these
straightforward steps, namely a reduction in the resolving power due to
reduced peak capacity and increased band broadening. Equation [4] does
not implicitly allow the effective resolution of the system to be evaluated.

Column selection and stationary phase selectivity can play a major role in
reducing the requirement for very high plate numbers. For example, a
relatively new PLOT capillary column, which uses a polar adsorbent as the
stationary phase, has unique selectivity toward oxygenated compounds.
The capability of such a stationary phase column to provide excellent
separation of oxygenates in the presence of hydrocarbons is illustrated in
Figure 1. Stationary phase selectivity can be manipulated to achieve better
separation of target compounds in a shorter time than would normally be
required using a nonselective column, but stationary phase optimization
will be limited to the analysis of low-complexity samples, i.e., those
containing fewer than say 50 components, or to mixtures containing a
small number of target analytes. This is supported by statistical overlap
theory, in which the extent of peak overlap in gas chromatograms of
complex mixtures can be predicted.

Figure 1. The use of a selective stationary phase column for the high-
speed analysis of ethanol in unleaded petrol.

Other specialized high-speed GC alternatives include the use of

multicapillary columns, packed columns, and flash-GC. Multicapillary
columns are made by combining some 900 capillaries, each with a dc of
40 μm into a bundle of about 1 m length. These multicapillary columns
allow the use of high flow rates and are characterized by a high sample
capacity. The 1980s saw some promising work in the area of fast-GC using
micropacked columns; for example, the separation of a mixture of
alkanes was performed using a short 320 μm i.d. column, packed with
10 μm particles. Flash-GC devices are based upon resistively heated metal
capillary columns, allowing extremely high heating rates and very fast
cooling times. These latter approaches cannot be considered mainstream
technologies, and no further comment shall be made in the present
article.

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URL: https://www.sciencedirect.com/science/article/pii/B0123693977007287

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