Analytical Chemistry

Lecture # 20 Analytical Separations

Chapter 23

Steps in Chemical Analysis

1. Formulating the Question
2. Selecting the Analytical Procedure
3. Sampling
4. Sample Preparation
5. Analysis
6. Interpretation and Reporting
7. Drawing Conclusions

Solvent Extraction

Solvent Extraction
Liquid-Liquid Extraction
e.g. Separatory
Funnel
Phase 2
(organic solvent)

Hot Water
Extraction

Immiscible
= dont mix
much!

S = solute

Phase I
(water)

S
S S
S S
S S
S
S
S
S
S SS
S S
S S S

Solvent Extraction

Solvent Extraction

Partition Coefficient

Partition Coefficient

AS2

AS1
K

[S]2

[S]1
(1-q)m/V2
qm/V1

AS2

AS1

[S]2
[S]1

Fraction in Phase 1 =
(after one extraction)

V1
V1 + KV2

Fraction in Phase 1
(after n extractions)

qn

V1
V1 + KV2

m = moles of S
q = fraction of S in Phase 1
1-q = fraction of S in Phase 2

Solvent Extraction

Solid-Phase Extraction
Solvent 2

pH and Solvent Extraction

a neutral species is more soluble in an organic solvent and a
charged species is more soluble in aqueous solution

Distribution
=
Coefficient (D)

Total Concentration in Phase 2

Total Concentration in Phase 1

K = [B]2/[B]1
Ka = ([H+][B]1)/[BH+]1

[B]2
[B]1 + [BH+]1

K Ka
Ka + [H+]

B = Ka/(Ka + [H+])

K
Phase 2

KK
21

Example: Extraction of base (B)

D

Phase 1

(in Solvent 1)

KK2
1 S
S

KK
21
K2

K1

S
S
S S

K B

Solid-Phase Microextraction (SPME)

Chromatography
= Stationary Phase

Solvent (Eluant)

Elution

Mobile Phase (solvent, gas)

Mixture
(with
analyte)

Column Packing
(sorbent)

Eluate

Chromatography
Packed

Open Tubular

Chromatography
Types of Chromatography (see Chapter 26)
1.
2.
3.
4.
5.

Adsorption Chromatography
Partition Chromatography
Ion-Exchange Chromatography
Molecular Exclusion Chromatography
Affinity Chromatography

Chromatography
Adsorption Chromatography
Mobile Phase (solvent or gas)
Stationary Phase
e.g. SiO2

Adsorbed

Chromatography
Partition Chromatography
Mobile Phase (gas)

Liquid Stationary
Phase

Solid
e.g. SiO2

Chromatography
Ion-Exchange Chromatography
+

Mobile Phase (solvent/water)

Stationary Phase
(= Resin with
cations or anions)

+
++

+ +

+
+
+
+

+ + + +

Chromatography
Molecular Exclusion
= Gel Filtration or Gel Permeation

Chromatography

Chromatography
Column Chromatography

Affinity Chromatography
Mobile Phase (solvent/water)
Stationary Phase
with Covalent
Ligand
(e.g. Antibody)

Chromatography

Chromatography

High-Performance Liquid Chromatography (HPLC)

Chromatography

Chromatography
Flow Rate
Length
Radius (r)

1 cm
V = r2 (1 cm)
e.g. r = 0.46 cm

If 20% of
column volume
is solvent, then
0.133 mL/cm

V = (0.046 cm)2(1 cm)

= 0.665 mL*
*1 cm3 = 1 mL

The Chromatogram

Chromatography

a graph showing the detector response as a

function of elution time
Vr = tr uv

Flow Rate
tr = Retention Time

e.g. 1 mL/min
Linear Flow Rate (ux): how many centimeters are traveled in 1
minute by the solvent
e.g. 1 mL/min / (0.133 mL/cm) = 7.52 cm/min

Detector Response

Volume Flow Rate (uv): how many milliliters of solvent per minute

tm

uv = Volume Flow Rate

tr = Adjusted
Retention Time
= tr - tm

Unretained Mobile Phase

(solvent or gas)

Time

The Chromatogram

The Chromatogram

Relative Retention
= tr2/tr1 > 1

Capacity Factor:

tr1

tm

Time

Time

Retention Time and Partition Coefficient

k =

time in the stationary phase

time in the mobile phase

Resolution = tr = Vr = 0.589tr
wav wav
w1/2av

moles of solute in stationary phase

moles of solute in mobile phase

Cs/Cm = partition coefficient

Efficiency of Separation

Relative retention

k =

CsVs
CmVm

k =

Vs K
Vm

tr2
tr1

= tr
tm
k2
k1

K2
K1

w1/2
=2.35
w = 4

Detector Response

tr tm = tr
tm
tm

tr = tr - tm
Detector Response

Detector Response

tr2

k =

Peak Width

Time

Efficiency of Separation
3
Resolution = 0.75

6
Resolution = 1.5

Efficiency of Separation

a band of solute broadens as it moves through

a chromatography column

x
c

Flux (J) = -D dc
(mol/m2 s)
dx
D = diffusion coefficient
(see Table 23-1)

x + dx
d - dc

J (Flux)

dx

Efficiency of Separation

c=

m e-x /(4Dt)
4Dt
2

c = concentration (mol/m3)
t = time
x = distance from center of band

x=0

2Dt

2 = 2Dt

Efficiency of Separation

Efficiency of Separation

Plate Height
x = ux t

2 = 2Dt

t = x/ux

2 = 2D x
ux
2 = 2D x
ux
2 = H x
H = plate height

Plate Height
2 = H x
H = 2/x
Number of Plates (N) = Length of Column (L)
H
= w/4
2
N= L=
Lx = L = 16L2
2
2
2
H

w
N = 16 tr2 = tr2
w2
2

N = 5.55 tr2
w1/22

Efficiency of Separation
Plate Height and Resolution

Resolution =

N
4

( 1)

= separation factor = uA
uB
Resolution

tA

= tB

e.g. Doubling L increases resolution by

Assigned Problems in Chapter 23:

Problems: 23-1, 23-8, 23-11, 23-19, 23-21, 23-27