Supercritical fluid chromatography is a technique that uses supercritical fluids like carbon dioxide as the mobile phase. Supercritical fluids exist above the critical temperature and pressure of a substance and have properties between gases and liquids, making them able to dissolve non-volatile compounds. SFC offers advantages over HPLC like faster analysis times and advantages over GC like the ability to analyze non-volatile and thermally-labile compounds. SFC has been applied to analyze compounds in various areas like pharmaceuticals, natural products, foods, polymers and more.
HPLC[ HIGH PERPROMANCE LIQUID CHROMATOGRAPHY OR HIGH PRESSURE LIQUID CHROMAT...Dr. Ravi Sankar
GASSCHROMATOGRAPHY[GC], ADVANCED STUDY OF THE FOLLOWING AND THEIR APPLICATIONS, INTRODUCTION, THEORY, COLUMN OPERATION,INSTRUMENTATION AND DETECTION,APPLICATIONS AND ADVANTAGES OF GC,PRINCIPLE OF SEPARATION IN GC, HOW GC MECHINE WORKS? COLUMN, DETECTORS.
BY P.RAVISANKAR, VIGNAN PHARMACY COLLEGE, VADLAMUDI, GUNTUR, A.P, INDIA.
Reviews the SFC technique along with applications and case studies.
Use of animations disturbs the view. For full ppt, drop me a mail at varadbende96@gmail.com
This Powerpoint presentation helps us to know the basic working principles, instrumentation an advantage of super critical fluid chromatography.
Contact Details:
Anbu Dinesh Jayakumar
M.Pharmacy ( Pharmaceutical Chemistry)
Sri Ramakrishna Institute of Paramedical Sciences, Coimbatore
Mobile : 8838404664 / 8608890121( Whatsapp)
Email: anbudinesh007@gmail.com
Gas chromatography head points:
Invention of Chromatography
original chromatography Experiment
Common types of chromatography
Paper and Thin layer chromatography
How does chromatography work?
Theoretical Plate
gas chromatography
schematic of GC
carrier gas-supply
Injection port
sample Injection system
split/spitless Injection
sample valves
GC columns
open tubular columns
Temperature Control
Solid Support Materials
Particle size of Supports
The stationary Phase
Detection systems
Characteristics of the Ideal Detector
Flame Ionization Detectors
Thermal Conductivity Detector
Electron-capture Detectors
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Chromatography is an analytical method in which compounds are physically separated and measured.
The main purpose of chromatography is to separate and quantify the target sample.
The Chromatography technique used to separate a mixture of compounds in pharmaceutical sciences , analytical analytical Chemistry with the purpose of identifying, quantifying and purifying the individual components of the mixture.
Supercritical fluid extraction and Supercritical fluid chromatography are techniques which use supercritical fluids as solvent for both extraction and separation respectively.
The properties such as density, viscosity and diffusion constant of the supercritical fluids are intermediate between those of a substance in gaseous and liquid state.
This helps in efficient extraction and chromatographic separation compared to other techniques.
HPLC[ HIGH PERPROMANCE LIQUID CHROMATOGRAPHY OR HIGH PRESSURE LIQUID CHROMAT...Dr. Ravi Sankar
GASSCHROMATOGRAPHY[GC], ADVANCED STUDY OF THE FOLLOWING AND THEIR APPLICATIONS, INTRODUCTION, THEORY, COLUMN OPERATION,INSTRUMENTATION AND DETECTION,APPLICATIONS AND ADVANTAGES OF GC,PRINCIPLE OF SEPARATION IN GC, HOW GC MECHINE WORKS? COLUMN, DETECTORS.
BY P.RAVISANKAR, VIGNAN PHARMACY COLLEGE, VADLAMUDI, GUNTUR, A.P, INDIA.
Reviews the SFC technique along with applications and case studies.
Use of animations disturbs the view. For full ppt, drop me a mail at varadbende96@gmail.com
This Powerpoint presentation helps us to know the basic working principles, instrumentation an advantage of super critical fluid chromatography.
Contact Details:
Anbu Dinesh Jayakumar
M.Pharmacy ( Pharmaceutical Chemistry)
Sri Ramakrishna Institute of Paramedical Sciences, Coimbatore
Mobile : 8838404664 / 8608890121( Whatsapp)
Email: anbudinesh007@gmail.com
Gas chromatography head points:
Invention of Chromatography
original chromatography Experiment
Common types of chromatography
Paper and Thin layer chromatography
How does chromatography work?
Theoretical Plate
gas chromatography
schematic of GC
carrier gas-supply
Injection port
sample Injection system
split/spitless Injection
sample valves
GC columns
open tubular columns
Temperature Control
Solid Support Materials
Particle size of Supports
The stationary Phase
Detection systems
Characteristics of the Ideal Detector
Flame Ionization Detectors
Thermal Conductivity Detector
Electron-capture Detectors
https://www.linkedin.com/in/preeti-choudhary-266414182/
https://www.instagram.com/chaudharypreeti1997/
https://www.facebook.com/profile.php?id=100013419194533
https://twitter.com/preetic27018281
Please like, share, comment and follow.
stay connected
If any query then contact:
chaudharypreeti1997@gmail.com
Thanking-You
Preeti Choudhary
Chromatography is an analytical method in which compounds are physically separated and measured.
The main purpose of chromatography is to separate and quantify the target sample.
The Chromatography technique used to separate a mixture of compounds in pharmaceutical sciences , analytical analytical Chemistry with the purpose of identifying, quantifying and purifying the individual components of the mixture.
Supercritical fluid extraction and Supercritical fluid chromatography are techniques which use supercritical fluids as solvent for both extraction and separation respectively.
The properties such as density, viscosity and diffusion constant of the supercritical fluids are intermediate between those of a substance in gaseous and liquid state.
This helps in efficient extraction and chromatographic separation compared to other techniques.
fluid chromatography (SFC) can be used on an analytical
scale.
It is a combination of High performance liquid chromatography (HPLC)
and Gas chromatography (GC).
It can be used with non-volatile and thermally labile analytes.
It can be used with the universal flame ionization detector.
It is important to producing narrower peaks due to rapid diffusion.
It is important for the chiral separations and analysis of high-molecularweight
hydrocarbons.
Supercritical fluids are suitable as a substitute for organic solvents in a
range of industrial and laboratory processes.
Instrumentation of HPLC, principle by kk sahuKAUSHAL SAHU
INTRODUCTION
Instrumentation of HPLC
TYPES OF HPLC
PARAMETERS
APPLICATION
CONCLUSION
REFERENCE
High-performance liquid chromatography ( HPLC) is a specific form of column chromatography generally used in biochemistry and analysis to separate, identify, and quantify the active compounds.
HPLC mainly utilizes a column that holds packing material (stationary phase), a pump that moves the mobile phase(s) through the column, and a detector that shows the retention times of the molecules.
All you need to know about the HPLC Chromatography!
HPLC (High-performance liquid chromatography) also referred as high-pressure liquid chromatography, is a technique used in analytical labs to separate, identify, and quantify each component in a mixture. HPLC separates and purifies compounds according to their polarity. I have tried to simplify the whole process of HPLC Chromatography and explain in simple terms. Let’s look at the main components involved in HPLC in this video.
I discussed about Normal Phase Chromatography, Reverse Phase Chromatography and their comparison and differences. Watch this video for more details.
If you have any doubts: Please leave a comment in the comments section below.
High- performance Liquid Chromatography”/
(High- pressure Liquid Chromatography) is a powerful tool in analysis, it yields High Performance and high speed compared to traditional columns chromatography
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4. History
The use of a supercritical fluid
mobile phase in chromatography
was first proposed in 1958 by J.
Lovelock.
The first actual report use of this in
a chromatographic system was in
1962 by Klesper et al, who used it
to separate thermally-labile
porphyrins.
5. S.F.C
•
S.F.C is a column chromatographic technique in which
supercritical fluid is used as a mobile phase.
•
Cost efficient
•
User friendly
•
Better resolution
•
Faster analysis
6. What is a supercritical fluid?
Supercritical Fluid exists at
temperatures and pressures
above its critical temperature
and pressure and have
densities, viscosities and
other properties that are
intermediates between those
of the substance in its
gaseous and liquid state.
7. Properties of Supercritical Fluid
•
High densities so they have a remarkable ability to dissolve
large, non-volatile molecules .
• Lower viscosities relative to liquid solvents.
• Inexpensive
• Ecofriendly
• Non-toxic.
• Higher diffusion coefficients and lower viscosities
relative to liquids.
8.
SFC Advantages vs HPLC
Supercritical fluids have low viscosities
- faster analysis (5 to 10 X faster).
- the use of open tubular columns is feasible.
Column lengths from 10 to 20 m are used.
Resolving power is ~5X that of HPLC.
SFC Advantages vs GC
Can analyze non-volatile, polar, or adsorptive solutes without
derivatization.
Can analyze thermally labile compounds.
Can analyze solutes of much higher molecular weight.
9. Theory :
based on the density of the supercritical fluid which
corresponds to solvating power.
As the pressure in the system is increased, the
supercritical fluid density increases and correspondingly
its solvating power increases.
Therefore, as the density of the supercritical fluid
mobile phase is increased, components retained in the
column can be made to elute.
This is similar to temperature programming in GC or
using a solvent gradient in HPLC.
12. Gas supply or Mobile Phase:
Liquid CO2 Pump
Cost
interference with chromatographic
detectors
physical properties like
nontoxic nature
nonflammable
are considered while selecting a
mobile phase.
14. Carbon dioxide is the ideal to satisfy all the above properties.
Safe to use
nontoxic
nonflammable
noncorrosive
inert.
Detector compatible
• The main disadvantage of it is very polar or ionic compounds
are not able to be eluted.
• This can be overcome by adding a small portion of a second
fluid called a Modifier fluid. (alcohols, cyclic ethers, acetonitrile
and chloroform).
15. Modifiers
CO 2 is not a very good solvent for high molecular
weight, ionic and polar analytes
This can be overcome by adding a small portion of a second
fluid called modifier fluid
This is generally an organic solvent, which is completely
miscible with carbon dioxide
methanol, acetonitrile, ethanol and 1-propanol.
16. Stationary Phase
Same as those for GC and LC, with some modification.
• Silica/Alumina
Useful for non-polar compounds
Lead to irreversible adsorption of some polar solutes
• Widely used polar Stationary Phase are
Polysiloxanes:- stable, flexible Si--O bond lead to
good diffusion.
Substituted with chemical groups for selective
interaction with analyte
Polymethylsiloxanes:- increase efficiency in
separating closely related polar analytes
Cyanopropyl polysiloxanes:-useful for compounds
with -COOH
17. Pumps:
Here mainly flow control is necessary.
So, syringe pumps are used for capillary
SFC for consistent pressure.
For Packed columns for easier blending
of the mobile phase or introduction of
modifier fluids reciprocating pumps are
used.
Syringe Pump
18. Injector
•
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.
Injector Volumes
Open tubular columns
◦ Injection volumes >96nL
◦ Greater volume affects resolution
Packed columns
◦ Injection volumes >1uL
20. SFC Columns
•
Open tubular (derived from GC)
-Large theoretical plates ~X500
-Easier to control pressure (low P drop)
•
Packed (derived from HPLC)
- Faster analysis
- Higher flow rates
- Higher sample capacity
21. Detectors
The choice of detectors will
depend upon
•
•
•
•
mobile phase composition
column type
flow rate
ability to withstand the high
pressures of SFC.
22. Contd..
•
•
•
It is compatible with both HPLC and GC detectors.
Flame photometric detectors
Flame ionization detectors
•
liquid-phase detectors like RID, ultraviolet-visible
spectrophotometric detectors and
•
light scattering detectors have been employed for SFC.
23. Back-Pressure Device or Restrictor
•
This is a device, which is used to maintain desired pressure in
the column by
- a pressure-adjustable diaphragm or
- controlled nozzle
so that the same column-outlet pressure is maintained
irrespective of the mobile phase pump flow rate.
• It keeps the mobile phase supercritical throughout the
separation and often must be heated to prevent clogging.
• The pressure restrictor is placed either after the detector or at
the end of the column.
24. Applications of SFC
By now SFC has been applied to wide variety of materials.
natural products
drugs
foods
pesticides
herbicides
surfactants
polymers and polymer additives
chiral compound
25. Natural products
•
separation of underivatized triterpene acids
estimation of caffeine from tea and conjugated bile acids
analysis of panaxadiol / panaxatriol in ginseng .
Pesticides
•
analysis of pesticide residues in canned foods, fruits and
vegetables wherein pyrethroids, herbicides, fungicides and
carbamates have been tested .
26.
27. Surfactants
Separation of the oligomers in a sample of the nonionic
surfactant Triton X100 .
Lipids
for the analysis of high molecular weight lipids like
triacylglycerols.
Separation of fatty acid methyl esters, biosynthetic
polyunsaturated fatty acids (PUFA) 37 , nonsaponifiable
lipids , cholesterol and its esters in human serum and food
samples
28. Drugs
Separation of drugs like
phenothiazine antipscychotics,
beta blockers
felodipine
clevidipine
methylated betacyclodextrins
vasodialators like isosorbide mononitrate, isosorbide
dinitrate, cyclandelate, nimodipine, amlodipine , oestrogens
combinations of various nonsteroidal antiinflammatory
drugs like flufenamic acid, mefenamic
acid, fenbufen, indomethacin mixtures, acetyl salicylic
acid, ketoprofen and fenbufen
29. Chiral compounds
SFC has been applied to separation of a large number of
enantiomers, diasterioisomers and geometrical isomers like
achiral and chiral analysis of camazepam and its
metabolites,
chiral separation of 1,3 dioxolane derivatives
Organometallics
Separation of metal chelates and organometals of thermally
labile category, chelates of transition metals, heavy
metals, organometallic compounds of lead, mercury and tin
has been carried out by SFC.
30. Conclusion
In the overall ranking of chromatographic techniques, it has
been judged that SFC falls somewhere between HPLC and
GC as the chromatographic method of choice. Lately SFC has
found a niche in the field of pharmaceutical chemistry and has
gained much support in the field of bioanalytical applications