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There

are two theories to explain chromatography

Plate theory - older;


developed by Martin & Synge in 1941
Rate theory - currently in use
Proposed by van Deemter in 1956
Accounts for the dynamics of the separation

View

column as divided into a number (N)


of adjacent imaginary segments called
theoretical plates
Within each theoretical plate analyte(s)
completely equilibrate between stationary phase
and mobile phase
Column

Theoretical plate

Chromatographic principle
Sample
mixture

Mobile phase
Equilibrium
establishes at each
point (ideally)
Stationary phase

The molecules of the


mixture interact with the
molecules of the Mobile
and Stationary Phase
Each molecule
interacts differently
with MP and SP

Retardation of rate
of movement of
molecules
Different distribution
coefficients and different
net rates of migration

Greater

separation occurs with:


greater number of theoretical plates (N)
as plate height (H or HETP) becomes
smaller
L = N H or H = L / N
where L is length of column, N is number
of plates, and H is height of plates or
height equivalent to theoretical plate
(HETP)

The

number of theoretical plates that


a real column possesses can be
found by examining a
chromatographic peak after elution
by various methods like
- half-height method
- USP method

N is a ratio of tR and of Wb which is 4


N = 5.55 tR2/ w1/22 = 16 tR2/ w2
where:
tR is retention time
w1/2 is width at h0.5
w is width measured at
baseline

Where:
N = Number of theoretical plates
Ve = elution volume or retention time (mL, sec, or
cm)
h = peak height
w1/2 = width of the peak at half peak height (mL,
sec, or cm)

Nmax = 0.4 * L/dp


where:
Nmax - maximum column efficiency
L - column length
dp - particle size
So, the smaller the particle size the
higher the
efficiency!

Band spreading - the width of bands


increases as their retention time
(tR) or retention volume (VR)
increases

A band exhibiting a width of 4 mL and


a
retention volume of 49 mL, is eluted
from a column. What width is expected
for a band with a retention volume of
127 mL eluting from the same analyte
mixture on the same column?
ANS: 10.4 mL

The smaller HETP, the narrower the


eluted peak

It is not unusual for a chromatography


column
to have millions of theoretical plates
Columns often behave as if they have
different numbers of plates for different
solutes present in same mixture

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