SUPER CRITICAL FLUID CHROMATOGRAPHY

SUPERCRITICAL FLUID
CHROMATOGRAPHY
Seminar Presentation
PRESENTED BY:
DARSHAN N U
M. PHARM 2nd SEMESTER
DEPARTMENT OF PHARMACEUTICAL CHEMISTRY
PRESENTED TO:
Dr. T. YUNUS PASHA.
HEAD OF THE DEPARTMENT
DEPARTMENT OF PHARMACEUTICAL CHEMISTRY
1
SUPERCRITICAL FLUID CHROMATOGRAPHY
(SFC)

CONTENT
1. Introduction
2. Principle
3. Instrumentation
4. Advantages
5. Applications
6. Previously asked Examination Questions
7. References
2
(SFC)

HISTORY
1) Use of Super Critical Fluid Chromatography (SFC) Was First
Proposed In 1958 By J. Lovelock.
2) First Reported Use Was In 1962 by klesper et.al., Who Separated
Thermally Liable Porphyrins
(SFC) 3

INTRODUCTION
1. Supercritical fluid chromatography (SFC) is a column chromatographic
technique in which a supercritical fluid is used as a mobile phase.
2. A supercritical fluid is a gas or liquid brought to a temperature and a
pressure above its critical point.
3. In SFC, the mobile phase is delivered by a high-pressure pump. The
sample is usually injected as a solution by means of a high-pressure
injection valve.
4. The column may either be a packed column, comparable to a high-
performance liquid chromatographic (HPLC) column, or an open
capillary column, comparable to a capillary gas chromatographic (GC)
column, but with somewhat smaller internal diameters (50-100μm).
4
(SFC)

CONTINUED…
5. Detection is performed either on-line, (i.e., UV-VIS) or after the
expansion of the fluid [i.e., flame ionization detection (FID) or mass
spectrometry (MS)].
6. The mobile phase is kept as a supercritical fluid by means of a
restrictor until either on-line detection has been performed or just
before the expansion into a gas phase detector.
5
(SFC)

SUPERCRITICAL FLUID
(SFC) 6

SUPERCRITICAL FLUID
7
(SFC)

Phase diagram for Carbon dioxide
(SFC) 8

PRINCIPLE
It is combination of GC and HPLC, the Principle is based on Adsorption
and partition Chromatography.
The principle is based on Supercritical Fluid.
(SFC) 9

INSTRUMENTATION
1. Mobile phase
2. Stationary Phase
3. Pump
4. Injection System
5. Oven
6. Column
7. Detector
10
(SFC)

WATERS METHOD STATION II.
(SFC) 11

SOFTWARE:THAR SFC SUPERCHROM
(SFC) 12

INSTRUMENTATION
13
(SFC)

Block diagram of an instrument for SFC
(SFC) 14

MOBILE PHASE
There are a number of possible fluids which may be used in SFC as the
mobile phase. Mostly CO2 is used. The main disadvantage of carbon
dioxide is its inability to elute very polar compounds. This can be
overcome by adding a small portion of a second fluid called a Modifier
fluid. (Alcohols, cyclic ethers, acetonitrile and chloroform) The addition
of the modifier fluid improves the solvating ability of the supercritical
fluid and sometimes enhances selectivity of the separation.
15
(SFC)

STATIONARY PHASE
Silica/Alumina
1. Useful for non-polar compounds.
2. Lead to irreversible adsorption of some polar solute.
16
(SFC)

PUMPS
1. Flow controlling is the vital function of pumping systems.
2. In contrast to HPLC pumping systems pressure rather than flow control is
necessary and pulseless operation is more important.
3. In general type of high pressure pump used in super critical fluid is
determined by column type.
4. For packed columns for easier blending of mobile phase or introduction
of modifier fluids reciprocating pumps are generally used.
5. For capillary super critical fluid syringe pumps are most commonly
employed.
6. Reciprocating pumps allow easier mixing of mobile phase or introduction
of modifier fluids.
7. Syringe pumps provide consistent pressure for neat mobile phase.
17
(SFC)

OVEN
A thermostatic column oven is required for precise
temperature control of the mobile phase. Conventional
GC or LC ovens are generally used
18
(SFC)

BACK PRESSURE REGULATOR (BPR)
This is a device which is used to maintain desired pressure in column by
pressure adjustable diaphragm or controlled nozzle, so that, same
column outlet pressure is maintained irrespective of mobile phase pump
flow rate.
It keeps mobile phase super critical throughout separation.
19
(SFC)

INJECTORS
• Injection in SFC is usually achieved by switching of the content of a
sample loop into the carrier fluid at the column entrance by means of a
suitable valve.
• For packed column SFC, a conventional HPLC injection system is
adequate, but for the capillary column SFC, the sample volume
depends on column diameters and small sample volumes must be
quickly injected into the column, therefore pneumatically driven
valves are used.
20
(SFC)

COLUMN
• Once the sample is injected into the supercritical stream it is carried
into the analytical column. The column contains a highly viscous
liquid (called a stationary phase) into which the analytes can be
temporarily adsorbed and then released based on their chemical
nature.
• This temporary retention causes some analytes to remain longer in the
column and it allows the separation of the mixture.
21
(SFC)

COLUMNS
• There are two types of analytical columns used in SFC,
A). Packed columns contain small deactivated particles to which the
stationary phases adheres.
The columns are made of stainless steel.
B). Capillary columns are open tubular columns of narrow internal
diameter made of fused silica, with the stationary phase bonded to the
wall of the column.
22
(SFC)

DETECTOR
Different types of detectors are used in conjunction with SFC. Some of
the commonly used detectors are as follows:
1. Flame ionization
2. Ultraviolet
3. Fluorescence
4. Mass spectrometer
23
(SFC)

ADVANTAGES
1. Supercritical fluid chromatography has several main advantages
over other conventional chromatographic techniques(GC and
HPLC).
2. Compared with HPLC, SFC provides rapid separations without the
use of organic solvents.
3. With the desire for environmentally conscious technology, the use of
organic chemicals as used in HPLC could be reduced with the use of
SFC.
4. Because SFC generally uses carbon dioxide collected as a by-
product of other chemical reactions or is collected directly from the
atmosphere, it contributes no new chemicals to the environment.
24
(SFC)

ADVANTAGES
5. SFC separations can be done faster than HPLC separations. because
the diffusion of solutes in supercritical fluids is about ten times
greater than that in liquids (and about three times less than in gases).
This results in a decrease in resistance to mass transfer in the column
and allows for fast high resolution separations.
6. Compared with GC, capillary SFC can provide high resolution
chromatography at much lower temperatures. This allows fast
analysis of thermolabile compounds.
25
(SFC)

APPLICATIONS
• SFC finds use in industry primarily for separation of chiral molecules,
and uses the same columns as standard HPLC systems. SFC is now
commonly used for a chiral separations and purifications in the
pharmaceutical industry.
• Fossil Fuels and Hydrocarbons
• Agrichemicals
• Drugs and Pharmaceuticals
• Polymers
• Explosives and Propellants
• Separations of Enantiomers
26
(SFC)

APPLICATIONS
27
1. Separation of enantiomers
Separation of metoprolol by HPLC (a) and SFC (b) on a chiralcel OD
stationary phase.
In (a) the mobile phase was 20% 2 – propanol in Hexane with 1%
diethyl amine; selectivity α = 2.67 and resolution Rs = 4.8
in (b), CO2 was used containing 20% methanol with 0.5% isopropyl
amine; α = 2.77 and Rs = 12.7
(SFC)

28
(SFC)

PREVIOUSLYASKED EXAMINATION QUESTIONS.
1. Write a note on supercritical fluid chromatography. RGUHS-2018
2. Discuss the Phase diagram of Carbon dioxide.
29
(SFC)

REFERENCES
1. Chromatographic Analysis of Pharmaceuticals; edited by John A.
Adomovics Page no: 239 to 269.
2. Super critical fluid chromatography “SKOOG” instrumental analysis
Page no: 935 to 940.
30
(SFC)

31
(SFC)
THANK YOU

SUPER CRITICAL FLUID CHROMATOGRAPHY

Recommended

Recommended

More Related Content

What's hot

What's hot (20)

Similar to SUPER CRITICAL FLUID CHROMATOGRAPHY

Similar to SUPER CRITICAL FLUID CHROMATOGRAPHY (20)

More from Sri Adichunchanagiri College of Pharmacy

More from Sri Adichunchanagiri College of Pharmacy (16)

Recently uploaded

Recently uploaded (20)

SUPER CRITICAL FLUID CHROMATOGRAPHY