( chromatography)

(mobile phase)
(stationary phase)

(
)

()
(
)

20Tswett

HPLC


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t : Retention time

(GC)

(GSC)
(GLC)
(LC)

(LSC)
(LLC)
(SFC)

(Column Chromatography)

(Planar Chromatography)

(Paper Chromatography)

(Thin Layer Chromatography)

Affinity

Wh/2
W

Wh/2
W

( dead time, tM )

( retention time, tR )

(tR )

Wh/2
W

(VM)

(relative retention value)

(selectivity factor)

(partition coefficient)
(capacity factor)
(separation factor)

(resolution)

(partition coefficient)
(capacity factor)

(
)
K = CL/CM

k = CL VL/CM VM = KVL/VM = (tR- tM)/tM = tR/tM

KY kY ' (t R )Y tM

K X k X ' (t R ) X tM

N: number of theoretical plates

N=16(tR/W)2
H: height equivalent of a theoretical plates
()
H=LW2/16tR2
L: column length

L=NH N=L/H


Resolution
Rs=Z/(WA/2+WB/2)
RS

2 t R B t R A
WA WB

N 1 kB '

4 1 kB '


(N)

RS

()

1.0 1.001 1.01 1.1

2 t R B t R A
WA WB

N 1 kB '

4 1 kB '

1.5

2.0

(-1)/ 0 0.001 0.01 0.091 0.38 0.50

(GC)(HPLC)

k
0.5 1.0

3.0 5.0 8.0 10.0 30 50

k/(1+ k) 0.33 0.50 0.75 0.83 0.89 0.91 0.97 0.98


(eddy diffusion)
(longitudinal diffusion)
(nonequilibrium mass
transfer)

van Deemter equation

H = A + B/u + Cu
A()
A=2dp
B()
C

van Deemter
equation
Partition Packing
Packed Columns

Carrier Gas

Liquid Phase
Solid Support
Very porous with very high surface area

Capillary Columns

Wall Coated Open Tubular (WCOT)

Liquid Phase

van Deemter equation

0.004

(M)

0.003
0.002
0.001
0
0

0.01

0.02

0.03

0.04

0.05

(M/sec)

0.06

0.07

0.08

GC-MS, FT-IR

a. b. c.

fi = mi/Ai mi = fi x Ai
fi = mi/Ai, fs = ms/As
fi = (mi/Ai/) (ms/As) = miAs/msAi

(normalization method)

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()

Helium


Carrier and Detector Support Gases

Chosen with the consideration of the type of

detector used
Inert
Dry
Pure
Using Compressed Gas
Safely
Obtain safety information from your
company's safety department or
from your local gas supplier.

/ Split/Splitless Inlet

Split Mode for

Pulsed Split

Splitless Mode for

Pulsed Splitless

Major Component Analyses

Allows for larger injection volume

Trace Component Analyses

Allows for larger injection volume

Split Flow Diagram: PreInjection

SEPTUM

(SEPTUM)
PURGE VENT

2 mL/min
TOTAL FLOW

50 mL/min

SPLIT VENT
48 mL/min

(INLET)
PURGE VALVE

47.4 mL/min

= CARRIER GAS

LINER

0.6 mL/min

SEAL
COLUMN

Column Types

Open (Capillary)

Packed

Wall Coated
Open Tube

LENGTH (meters)
I.D. (mm)

PACKED

SERIES 530

NARROW BORE

.5-10

5-100

5-100

2-4

.530

.1-.25

1.5 3 m
2 4 mm

( 60100)

15 100 m
0.2 0.75 mm

Gas Solid Chromatography (GSC)

10% of Applications
Adsorbent
Carrier Gas
Packed Columns

Adsorbent Packing (GSC)

Porous with large surface area

Capillary - Porous Layer Open Tubular (PLOT)

Carrier Gas

Gas Liquid Chromatography

(GLC) Partition Packing
Packed Columns

Carrier Gas

Liquid Phase
Solid Support
Very porous with very high surface area

Capillary Columns

Wall Coated Open Tubular (WCOT)

Liquid Phase

Gas Liquid Chromatography

(GLC)
(90% Of Applications)
Separation by partitioning or differential solubility in
stationary phase
Components are separated based on differences in polarity
(interaction of dipole forces)
Generally
"Like dissolves like"
or
Polar interacts with polar

Example:

Alcohols are polar

Carbowax is polar liquid phase

Summary of column
parameters on Resolution
Parameter

effect on R

down sides

Column length the longer the better

longer elution time

Film thickness

smaller sample loads

the thinner the better

poor retention for

extremely volatile
compounds
Inner diameter the smaller the better smaller sample loads


Comparison of Column Types
HP Column Evaluation Sample
Packed Column Analysis:

1
2
4
5

5% OV101
on 80/100 Chromosorb

Capillary
30m X 0.53mm X .88m

Conventional
30m X 0.32mm X .25m

7
9
1
1
&
1
2
1
0

1
2

7
&
8
5
46

1
2
1
0
1
1

(Sensitivity)
(Stability)(Reproducibility)
(Linear dynamic range)

(Response time)

Thermal Conductivity Detector (TCD)

Flame Ionization Detector (FID)

Electron Capture Detector (ECD)

Nitrogen Phosphorus Detector (NPD)

Flame Photometric Detector (FPD)



Electrolytic Conductivity Detector (ELCD, HALL)

Photoionization Detector (PID)

Mass Selective Detector (MSD)

Infrared Detector (IRD)

Atomic Emission Detector (AED)

HPLC

HPLC Sample Injector