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
e.g. Separatory Funnel Hot Water Extraction

Solvent Extraction
Liquid-Liquid Extraction

Phase 2 (organic solvent) Immiscible = dont mix much! Phase I (water) S = solute

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

Solvent Extraction
Partition Coefficient K = AS2 AS1 K =
m = moles of S q = fraction of S in Phase 1 1-q = fraction of S in Phase 2

Solvent Extraction
Partition Coefficient K = AS2 AS1 Fraction in Phase 1 = (after one extraction) Fraction in Phase 1 (after n extractions) = q

[S]2 [S]1

[S]2 [S]1 = V1 V1 + KV2

n

(1-q)m/V2 qm/V1

qn

V1 V1 + KV2

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

Solid-Phase Extraction
Solvent 2

Phase 1

(in Solvent 1)

K
Phase 2

Distribution = Coefficient (D)

Total Concentration in Phase 2 Total Concentration in Phase 1 D = = = [B]2 [B]1 + [BH+]1 K Ka Ka + [H+] K B

KK 21 K K21 S

Example: Extraction of base (B)

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

S S S S S

KK 21 K2 K1 S

Solid-Phase Microextraction (SPME)

Solvent (Eluant)

Chromatography
= Stationary Phase

Z
Mixture (with analyte)

Mobile Phase (solvent, gas)

Column Packing (sorbent)

Eluate

Chromatography
Packed Open Tubular

Elution

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

Chromatography
Column Chromatography

Chromatography
High-Performance Liquid Chromatography (HPLC)

Chromatography

Chromatography

Chromatography
Flow Rate
Length Radius (r)

1 cm V = "r2 (1 cm) e.g. r = 0.46 cm V = "(0.046 cm)2(1 cm) = 0.665 mL*

*1 cm3 = 1 mL

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

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

The Chromatogram
a graph showing the detector response as a function of elution time Vr = tr uv

tr = Retention Time

uv = Volume Flow Rate

Detector Response

tm

tr = Adjusted Retention Time = tr - tm

Unretained Mobile Phase (solvent or gas)

Time

The Chromatogram
Relative Retention! = tr2/tr1 > 1 tr2
Detector Response

The Chromatogram
Capacity Factor:! k = tr tm = tr tm tm

tr = tr - tm Detector Response

tr1

tm

Time

Time

Retention Time and Partition Coefficient

k = = time in the stationary phase time in the mobile phase moles of solute in stationary phase moles of solute in mobile phase k =
Cs/Cm = partition coefficient

Efficiency of Separation
Resolution = tr = Vr = 0.589tr wav wav w1/2av
#h

w1/2 =2.35 w = 4

CsVs CmVm Vs K Vm = = tr tm k2 k1 = K2 K1

Detector Response

k =

tr2 tr1

Relative retention

Time

Peak Width

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

Flux (J) = -D dc (mol/m2 s) dx D = diffusion coefficient

(see Table 23-1)

x c

x + dx d - dc

J (Flux)

dx

Efficiency of Separation

c=

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

2

c = concentration (mol/m3) t = time x = distance from center of band x=0

2Dt

2 = 2Dt

Efficiency of Separation
Plate Height x = ux t t = x/ux 2 = 2Dt 2 = 2D x ux 2 = 2D x ux 2 = H x H = plate height

Efficiency of Separation
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) = tB

= separation factor = uA uB Resolution N L

tA

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