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Tailing Fronting
a b
When smaller quantities are moving faster than the larger quantities
Fronting peak
When smaller quantities of the solute are retained strongly than the
larger quantities Tailing peak
• Please note that the theory and definitions are based on
the fact that we assume all chromatographic peaks to
have a Gaussian distribution, i.e. “tailing” and “fronting”
are assumed to be minimal
Quantitative description of column efficiency
• Quantitative measures of column efficiency:
– Plate height (H) or height equivalent to a theoretical
plate (HETP)
– Plate count or number of theoretical plates, N
• The two are related by the equation:
𝐿
𝑁=
𝐻
where L is the length of the column
N is directly proportional to L
• Longer column provides higher N (efficiencies)
N is inversely proportional to H
• Smaller H provides higher N
Theoretical plates:
• Martin and Synge defined these terms,
by looking at the processes in a
chromatographic column as if it were
similar to a distillation column which
consists of numerous discrete, but
adjacent narrow layers called
theoretical plates.
• At each plate, equilibration of the
solute between mobile and stationary
phase take place
• Movement of the solute was then
treated as a stepwise transfer of
equilibrated mobile phase from one
plate to the next
N and H
• The plate count (N) and the plate height (H) are
widely used in literature and by experts in the field
of chromatography
• They are very useful quantities for comparing
separation power of different separations and
efficiencies among columns
• The plate theory successfully accounts for the
Gaussian shape of chromatographic peaks and
their rate of movement down a column
Definition of plate height, H
• If we assume a Gaussian peak,
then the extent of band
broadening is given by its
standard deviation, s, or its
variance, s2
• The efficiency of a column can
be defined as the variance per
unit length of column:
𝜎2
𝐻=
𝐿
• The plate height can be
thought of as the length of
𝐿
column that contains a fraction 𝐼𝑓 𝑤𝑒 𝑠𝑢𝑏𝑠𝑡𝑖𝑡𝑢𝑡𝑒 𝐻 𝑖𝑛𝑡𝑜 𝑡ℎ𝑒 𝑁 =
𝐻
of analyte that lies between L 𝐿 2
and L - s 𝑡ℎ𝑒𝑛 𝑁 = 2
𝜎
See Skoog et al, Appendix 1 for more information on standard deviation etc.
Calculating the number of theoretical plates
Column Chromatogram
(cm/s)
Multiple flow paths or eddy dispersion term (A)
A-term
𝐻=𝐴 + 𝐵𝑢 + 𝐶𝑢
Plate height
Broadening due to multiple flow paths
Independent of the flow rate
Theoretically: A = 2dp ; = packing Mobile phase velocity
Plate height
Broadening due to diffusion of analyte
in the mobile phase
Diffusion: movement of molecules
Mobile phase velocity
from the region of high concentration
to a region of low concentration
Theoretically: B = 2DM ; DM = diffusion
coefficient of analyte in the mobile
phase
Inversely proportional to the flow rate
Faster mobile phase velocity (u) leads
to smaller B/u less time available for
diffusion to take place
Smaller DM leads to smaller B
Resistance to mass transfer term (Cu)
𝐵
𝐻 =𝐴+ 𝑢
+ 𝐶𝑢
Slow
Stat. phase equilibration Stat. phase
Mobile phase Mobile phase
Resistance to mass-transfer term (Cu)
• The mass transfer term, Cu, is
actually the sum of two terms: Cu = CSu + Cmu
– the stationary phase mass-
transfer term, CSu, which is
linear, is
𝐶𝑆 𝑢 ∝ 𝑑𝑓2 𝐷𝑆
– and the mobile phase mass-
transfer term, Cmu, which bears
a complex dependency on
mobile phase velocity, is
– For packed columns:
𝐶𝑀 𝑢 ∝ 𝑑𝑝2 𝐷𝑀
– For open tubular columns:
𝐶𝑀 𝑢 ∝ 𝑑𝑐2 𝐷𝑀
where dc is the column diameter
Interpretation of a van Deemter curve
uopt : optimal mobile phase
velocity
Hmin 𝐿
Recall: 𝑁 =
𝐻
(cm/s)
uopt
Ideally, one should work at Uopt i.e. the velocity which
delivers the minimum plate height (Hmin) the
contribution of each term is minimal, i.e. higher N
(efficient separation)
Example: the effect of particle size dp on H
References
• D. A. Skoog, F. J. Holler and S. R. Crouch, Principles of
Instrumental Analysis, Chapter 26
• Analytical Chemistry 2.0 by David Harvey (electronic revision
of Modern Analytical Chemistry by David Harvey)
Further reading:
• D. C. Harris, Quantitative Chemical Analysis
• H. H. Willard, L. L. Merritt, Jr., J. A. Dean, F. A. Settle, Jr.,
Instrumental Methods of Analysis