Separations

Extractions and
Chromatography
 Separations
Take a mixture of chemical species and
separate it into individual chemical
compounds (physical & chemical
properties come into play).

Analyze the separated components (off-line

or on-line).
Extractions, precipitations, chromatography,
electrophoresis, etc.
Extractions
Typically, from water into an organic solvent, or vice versa. Extraction
efficiency depends on the difference in intermolecular forces in solvent solute
interactions.

[A]org

[A]aq
 Extraction Equation Derivation

What if you do it twice?

Or three times?
 Extractions (Partition)
A solute partitions between
two immiscible liquids
(2 phases)

m = moles S
Non-covalent forces involved?
q = fraction of S in phase 1
[S]2
V1 = volume of phase 1
V2 = volume of phase 2
[S]1
K=
S 2 (1 − q )m / V2
=
S 1
Van der waals forces, H-bonding
Etc. ( “like dissolves like” ) qm / V1
 Extractions (Partition)
K =
S 2
=
(1 − q )m / V2
S 1 qm / V1

Rearrange to obtain….
V1
q=
V1 + KV2
[S]2 Our goal is to make q small
(does the equation make sense?)

•n extraction stepsn
 V1 
[S]1 q n
=  
 V1 + KV2 
Large K, big V2 and multiple extractions help
(3 extractions with 10 mL better than one with 30 mL)
 Example 24.1
The distribution constant for iodine between an organic solvent and
H2O is 85. Find the concentration of I2 remaining in the aqueous layer
after extraction of 50.0 mL of 1.00 x10–3 M I2 with the following
quantities of the organic solvent:
(a) 50.0 mL; (b) two 25.0-mL portions; (c) five 10.0-mL portions.
Extractions Meet Acid-Base Chemistry
1. Neutral compounds
primarily dissolve in the
organic phase.
2. Charged compounds
Organic
only dissolve in the
aqueous phase.
[S]2 B 3. Like dissolves like

Aqueous Consider:
[S]1 BH+
Some B B + H2O  BH+ + OH-
Extractions Meet Acid-Base Chemistry
B + H2O  BH+ + OH-

D=
B2
B1 + BH + 1
Remember, from CH 10:

Organic

K=
B 2
Ka =
H  B 1
+

[S]2 B B 1 and
BH 
+
1

Aqueous
[S]1 BH+ K  Ka
D=
Ka + H
Effect of pH ??
+
 
Transfer ions into the organic phase with complexing agents:

D depends on pH…
 Chromatography Basics
Definition – Separation technique based on the differential distribution of
solutes between a moving mobile phase and a stationary phase.
 Chromatography Basics
Definition – Separation technique based on the differential distribution of
solutes between a moving mobile phase and a stationary phase.

Important elements of phase equilibria, mass transport, and kinetics

are involved
Forms→

Adsorption Partition (or absorption)

Size Exclusion Chromatography
There are a lot of other forms:

Exchange cations for H +

 Distribution Coefficients
Ca2+(aq) + 2H+(res) Ca2+ (res) + 2H+(aq)

Typically, H+ dominates
in both media, and
changes minimally.

Monovalent cations: Ag> Cs> Rb> K> NH4> Na> H> Li

Divalent cations: Ba> Pb> Sr> Ca> Ni> Cd> Cu> Co> Zn> Mg> UO2
 The General Chromatographic Process

Plug injection

Phase dis-
tribution, &
separation

Elution &
detection
 The Chromatogram

HPLC Plot of the response of a

10 pesticides Detector at end of column
UV absorbance
Quantitate by peak area

(220 nm) Versus

The length of time it takes
to elute from column

Notice Gaussian profile

(many random processes
shape the band)

Qualitative info in retention time

 Adjusted Retention Time
& Selectivity

1 2 3 4 5 6  suppose V (mL)

t r 2
t r = t r − t m =
α or ɣ are changed by t r1  = 1.0 → no separation.
How much separation is
changing the phases needed depends on how
or conditions. sharp the peaks are.
Retention Factor
(also called capacity factor)

K is an equilibrium
k’ is found from
constant (i.e., a
the chromatogram
tr − tm thermodynamic
k =
for each peak
parameter) → so is k’
tm
K = [ ]stationary / [ ]mobile & k’ = amtstat / amtmob
Vmob is easy to find k’ = K Vstat / Vmob
Vstat is not Capacity terms
Theoretical Plates
Terms borrowed from
fractional distillation.

Industrial petroleum
distillation columns have
plates and extraction points
that remove components
based on their boiling point.

Early theory treated a

chromatography column as a
series of (maybe microscopic)
plates. More plates ~ better
separation.
 Separation Efficiency
w1/2 w

2 2
16t 5.55t H =
L
N= = 2 r r

w 2
w1 / 2 N

N = number of “theoretical plates”. H is the height of a “theoretical plate”.

Big N and small H means efficient separation (sharp peaks).

A longer column will increase N
Resolution (the main objective)
= peak separation/width

R = Δtr/wav = (√N/4)(γ-1)

Ɣ = t2/t1 N = tr2/σ2
 Resolution (the main objective)
Mistaken as one peak

Almost good enough

Note: chromatography
Done for analysis & to isolate band width at baseline = 4 
 Resolution (the main objective)

Equation 24-20 in the textbook.

R = 1.0 good
R = 1.5 excellent
R > 2.0 only good efficiency selectivity capacity
 Resolution vs Time
The goal is high resolution, in the shortest
amount of time. These two things can be
incompatible.

Equation 24-22 in the textbook.

 Example 24.2
Substances A and B have retention times of 16.40 and 17.63 min,
respectively, on a 30.0-cm column. An unretained species passes
through the column in 1.30 min. The peak widths (at base) for A and B
are 1.11 and 1.21 min, respectively.
Calculate (a) the column resolution, (b) the average number of plates in
the column, (c) the plate height, (d) the length of column required to
achieve a resolution of 1.5, and (e) the time required to elute substance
B on the column that gives an R value of 1.5.

a) R = Δtr/wav d)
16t r2 5.55t r2
. =
b) N 2
= 2
w w1 / 2
L
c) H = e)
N
 ??Questions??
tr − tm

What is fraction not extracted

tm

What are Van der Waals forces

k’ = amtstat / amtmob
=

Definition of the technique of chromatography

(GLC, GSC, HPLC, TLC, etc.)
t r 2
Define (via eq & from a chromatogram) =
selectivity factor t r1
retention factor (about capacities)
16t r2 L
plate height & number N = H =
w2 N
resolution (Rs)
How can you improve resolution? tN
R= =r
wav 4
 How to Resolve B and C
Increase peak separation through better selectivity;
α (or γ) a matter of thermodynamics

This means changing the stationary and/or

mobile phase. It is better not to have to do
that every time.
 How to Resolve B and C
Increase peak sharpness through efficiency;
A matter of kinetics (mass transport)

This approach is more

generally useful

This usually means using a longer column, narrower

column, which improves resolution for every analysis.
Temperature programming or solvent programming can
be easily adjusted each time, as needed.
 What Causes Peak Broadening?
B
H = A + + Cu
Make
H u is flow rate

u
Small

A is the Eddy C is the Mass Transfer

Diffusion (Multiple B is the Longitudinal Term (actually has two
Path) Term Diffusion Term components)

Packed (particles) A concentrated band of If the analytes move down

columns result in analytes diffuses as it the column and
broadening since traverses the column. equilibrium is not
different analyte The slower the established, peaks
molecules can take band moves the more it broaden. The faster the
different paths down diffuses (inverse u analytes move the less
the column dependence) time they have to
equilibrate (u dependence)
 What Causes Peak Broadening?

A
Term -
eddy
diffusion
- With smaller particles, more decisions → averages out
- Open tubular columns do not have this problem

4

B
Term -
axial
diffusion
104 – 105
- Standard deviation of Gaussian band  = (Dm t)1/2 less in liquids
- Use high flow rates than in gases.
See Table 22-1
 What Causes Peak Broadening?

C
Terms -
Cm & Cs;
Mass transfer
 What Causes Peak Broadening?

Van Deemter Plot

C
Terms -
Cm & Cs

Use slow
Flow rate
An Expanded van Deemter Equation:

Particle size effect

A + B + Cs + Cm

Another version:

A
Carrier gas effect