Outline

Separation Techniques Phase Separations

p
What are Phase Separations? Partition Equilibrium Theory Competing Equilibria and the Distribution Ratio Extraction Efficiency Separation Efficiency Solid Phase Extraction
www.chem.unsw.edu.au/UGNotes/Guilhaus/
CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1 1

Vaq

KD

M. Guilhaus UNSW 1999 - All rights reserved.

Vorg

D
q

What are Phase Separations?

Analytes often present in low concentration in a complex sample matrix . Phase separations are used to extract the analyte preferentially from a complex or interfering matrix into a cleaner solvent matrix.
M. Guilhaus UNSW 1999 - All rights reserved.

They rely on the analyte being more soluble in an immiscible or immobilised phase. Liquid-liquid and liquid-solid phase extractions are important in: Sample clean-up Sample preconcentration

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

Partition Equilibrium Theory

When a solute is soluble in two immiscible phases that are in contact, it partitions according to the Distribution Coefficient Kd Non-ionic compounds dissolve readily in non-polar organic solvents such as: - Chloroform - Hexane - Carbon disulphide - Diethylether etc.

M. Guilhaus UNSW 1999 - All rights reserved.

C6H5COOH(aq)
example

C6H5COOH(org)

Kd =

[C 6 H 5COOH]org [C 6 H 5COOH]aq

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

Competing Equilibria
Solutes often participate in other equilibria in both phases. Consider benzoic acid (C 6H5COOH = HA)

HA(org) + HA(org) Kd HA(aq) + H2O

Kdim

M. Guilhaus UNSW 1999 - All rights reserved.

HA2(org)

Ka

A-(aq) + H3O+

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

Distribution Ratio
The distribution ratio (D) measures the relative molar concentration of analyte (in all its forms) in the two phases
Consider the previous example of benzoic acid (C6H5COOH = HA)

D=

Corg Caq [HA]org + 2[(HA )2 ]org + [A ]org [HA]aq + 2[( HA )2 ]aq + [ A ]aq
M. Guilhaus UNSW 1999 - All rights reserved.

D=

HA(org) + HA(org) Kd HA(aq) + H2O

Kdim

(HA)2(org)

Ka

A-(aq) + H3O+

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

Distribution Ratio
Which concentration terms in the D equation will be negligible? Under what pH conditions will the value of D be highest in this example?

D=

Corg Caq [HA]org + 2[(HA )2 ]org + [A ]org [HA]aq + 2[( HA )2 ]aq + [ A ]aq
M. Guilhaus UNSW 1999 - All rights reserved.

D=

HA(org) + HA(org) Kd HA(aq) + H2O

Kdim

(HA)2(org)

Ka

A-(aq) + H3O+

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

Distribution Ratio

The distribution ratio is a quantitative measure of how a solute partitions between two immiscible solvents.
M. Guilhaus UNSW 1999 - All rights reserved.

Its value depends on the solution conditions (e.g., pH).

D is not an equilibrium constant.

D must be determined for each set of solvent pair and experimental conditions.

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

Efficiency of Extraction
It is important to know and control how much analyte will be recovered in an extraction. Normally we aim at 99.9% or more for quantitative analysis For a single extraction it is easy to predict efficiency as a function of D and the volumes of the organic and aqueous phases.
M. Guilhaus UNSW 1999 - All rights reserved.

CorgVorg
EQUILIBRIUM

CinVaq

CaqVaq

p q
8

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

Efficiency of Extraction

p q

p is the fraction of analyte (any form) transferred to the organic phase after equilibration q is the fraction of analyte (any form) left in the aqueous phase after equilibration

p=

D D + (Vaq / Vorg ) 1 1 + D(Vorg / Vaq )

M. Guilhaus UNSW 1999 - All rights reserved.

q=

CorgVorg
EQUILIBRIUM

CinVaq

CaqVaq

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

Efficiency of Extraction - Example

Cl2 has D=10 between water and CCl4. Calculate the %extracted from 100 mL of aqueous Cl 2 into : (a) 50mL of CCl4

100 mL 1 0 50 mL M. Guilhaus UNSW 1999 - All rights reserved. q = 0.17 p = 0.83

p=0.83 (83% Cl 2 transferred to organic phase) q=0.17 (17% Cl 2 remains in aqueous phase)
(b) 500mL of CCl4

100 mL 1 q = 0.02

p=0.98 (98% Cl 2 transferred to organic phase) q=0.02 (2% Cl 2 remains in aqueous phase)
500 mL

p = 0.98 0

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

10

Efficiency of Extraction - Example

Increasing the volume of organic phase has certainly increased the recovery of the analyte Cl2, but .. Has the cost changed? Is there an environmental cost? Is there an impact on the occupational health and safety of the lab? What about the cost of the equipment? Compare the concentration of Cl 2 in the CCl4 in both cases. Which case is better in this regard?
500 mL 100 mL 1 q = 0.02

100 mL 1 0 50 mL M. Guilhaus UNSW 1999 - All rights reserved. q = 0.17 p = 0.83

p = 0.98 0

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

11

Efficiency - Large Vorg

Using large amounts of solvent is: Expen$ive Bad for the environment
M. Guilhaus UNSW 1999 - All rights reserved.

Bad for health and safety of lab staff Usually a fire hazard Usually impractical because or sze and weight of apparatus.

What is a better way to improve efficiency without using large amounts of solvent?
CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1 12

Efficiency - Multiple Extractions

Consider using several equal smaller amounts of solvent, using each to reextract the aqueous phase, collecting the combined organic phases. 0 1 0

1 2 3

q pq q2

M. Guilhaus UNSW 1999 - All rights reserved.

p + pq

Successive extractions improve recovery efficiency with smaller total amount of solvent

pq2 p+pq+pq2

q2
0

pqn-1

qn-1

qn

p+pq+pq2++pqn-1
13

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

Efficiency - Multiple Extractions Example

Cl2 has D=10 between water and CCl4. Calculate the %extracted from 100 mL of aqueous Cl2 with two separate extractions each with 25 mL of CCl4

Calculate p and q p = 0.0.714 q = 1-0.714 = 0.286 First Extraction

p= p=

D D + (Vaq / Vorg ) 10 10 + (100 / 25)

M. Guilhaus UNSW 1999 - All rights reserved.

71.4% (100p) Cl2 removed to organic phase 28.6% (100q) remains in aqueous

Second Extraction
71.4% of 28.6% = 20.4% of Cl2 extracted into organic phase Total extracted = 91.8% (71.4% + 20.4%) c.f. 83% for single 50 mL step
CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1 14

Achieving a Specified Efficiency

A Typical Question (in the real world and in the exam!) Knowing D, the volume ratio gives p and q. If we need to achieve,say, 99% recovery, how many extractions are required? Example

D = 3.5, Vaq=10, Vorg=10

99% or more must be recovered Let n=number of extractions Calculation of p gives value of 0.78 so q=0.22 Fraction remaining in aqueous is qn Fraction in collected organic phases is 1-qn 0.99 = 1-qn 0.22n = 0.01
M. Guilhaus UNSW 1999 - All rights reserved.

nlog(0.22) = log(0.01) n = 3.04

How many extractions are required?

15

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

Achieving a Specified Efficiency

In the previous example n = 3.06 so 4 extractions were needed. How could you adjust the method to achieve 99% recovery in exactly 3 extractions?

M. Guilhaus UNSW 1999 - All rights reserved.

Clue: change the volume ratio Clue: Change the value of D by adjusting the pH

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

16

Separating Solutes
We have seen how successive extractions improve recovery efficiency. What if we have two solutes that extract so that one has a much higher D than the other. Will successive extractions help us to remove the more extractable solute from the less extractable one?

Example:

M. Guilhaus UNSW 1999 - All rights reserved.

Solution has equal concentrations of A and B

DA=100 DB=0.1 Vaq=Vorg

Lets see what happens in 2 successive extractions

17

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

Separating Solutes
Example (contd):
Extraction 1: 99% A extracted; 9.1% B extracted A is 100[B]/[A] = 9.2% contaminated with B Extraction 2: 99.99% of A extracted; 17% B extracted
M. Guilhaus UNSW 1999 - All rights reserved.

A is 100[B]/[A] = 17% contaminated with B

Successive extractions improve recovery but reduce selectivity

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

18

Using Countercurrent Fractionation

The organic extract is back-washed with fresh aqueous phase. From previous example 1st Extraction removes 99% A and 9.1% B Now shake extract with fresh Vaq
M. Guilhaus UNSW 1999 - All rights reserved.

98% (99% of 99%) of A remains in organic phase 0.83% (9.1% of 9.1%) of B remains in organic phase Extract of A is now (100[B]/[A] = 100(0.83)/(98) = 0.85% contaminated with B.

Countercurrent fractionation improves separation (selectivity) but some recovery efficiency is lost
CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1 19

Solid Phase Extraction

The organic phase is bound (immobilised) on a porous solid support. For example: a C-18 hydrocarbon bonded to a support material. Aqueous solution is passed through the solid phase and analytes dissolve in bonded phase. Analytes released by passing an organic solvent or adjusting the pH of a solvent to ionise the extracted compound and elute them from the cartridge. System used in automated analyses.

M. Guilhaus UNSW 1999 - All rights reserved.

CHEM2801 Analytical & Physical Chemistry for Food Science - Separations -1

20