that my own creation on gpc

1. GEL PERMEATION CHROMATOGRAPHY
(GPC)
Presented By
Md. Amzad Hossain
M.Sc. Student
ID: 0417032702
Department of Chemistry
Bangladesh University of
Engineering & Technology
Course Coordinator
Dr. Chanchal Kumar Roy
Assistant Professor
Department of Chemistry
Bangladesh University of
Engineering & Technology
1

2. Contents
• What is gel permission chromatography (GPC)?
• Why GPC is important ?
• Basic difference with other techniques ?
• Basic theory of GPC
• Working Principle of GPC
• Instrumentation
• Calibration
• Advantage of GPC
• Disadvantage of GPC
• Application
• Conclusion
• References

3. What is Gel Permeation Chromatography
It is a subset of a very common
technique called size exclusion
chromatography (SEC), which involves
the separation of molecules by their
hydrodynamic volume in solution,
commonly referred to as the molecular
size

4. Why is GPC important?
GPC can determine several important parameters.
# Number average molecular weight,
# Weight average molecular weight,
# Z weight average molecular weight, and
# Molecular weight distribution.

5. Differences with other techniques
• When an aqueous solution is used to transport the sample through
the column the technique is known as gel filtration
chromatography.
• The name gel permeation chromatography is used when an organic
solvent is used as a mobile phase. GPC must operate free of
interactions to assure separation by size only. Only the entropy
effects should influence the separation.
• HPLC methods rely on interactions between sample and stationary
phase;
those interactions result from ion exchange, bio-affinity, or
chirality.

6. Basic theory of GPC

7. FIGURE : Schematic representation of a gel permeation chromatograph.

8. Instrumentation

9. FIGURE : Typical gel permeation chromatogram. Dotted lines represent volume “counts.”
Elution volume (Vr) (counts)
BaselineDetector
response

10. FIGURE . Universal calibration for gel permeation chromatography. THF, tetrahydrofuran.
Log([η]M)
109
108
107
106
105
18 20 22 24 26 28 30




Polystyrene (linear)
Polystyrene (comb)
Polystyrene (star)
Heterograft copolyner
Poly (methyl methacrylate)
Poly (vinyl chloride)
Styrene-methyl methacrylate graft copolymer
Poly (phenyl siloxane) (ladder)
Polybutadiene

















Elution volume ()5 ml counts, THF solvent)

11. FIGURE . Typical semilogarithmic calibration plot of molecular weight versus retention volume.
Retention volume (Vr) (counts)
106
105
104
103
Molecularweight(M)

12. Universal calibration method
[η]1M1 = [η]2M2
logM2 = (
1 + a2
1
)log(
K2
K1
) + ( 1 + a2
1 + a1
)logM1
Fromm the Mark-Houwink- sakurada Relationship
η1= K1M1ª
η2= K2M2 ª
Combining these equation

13. Advantages
• Has well defined separation
time
• Can provide narrow bands
• Low chance for analyte loss
• Determination of MW of
polymers
• Less time of analysis
• Non interactive mode of
seperation

14. Disadvantages
@ Limited number of peaks that can be resolved
@ Peak detection is difficult.
@Requires at least 10% difference in MW for reasonable
resolution of peaks
@Pre-filtration of sample

15. Applications
• Proteins fractionation
• Purification
• Molecular weight determination.
• Separation of sugar, proteins, peptides, rubbers
and others on the basis of their size.
• This technique can be use to determine the
quaternary structure of purified proteins

16. Conclusion
• It is non reactive techniques
• Mobile phase is polymer
• Rapid technique
• Most convenient for molecular weight
distribution

17. reference
• Book reference
I. Willard H H, Merritt l l, Dean J A, Settle F A.
Instrumental Methods of Analysis.2012:7:644-648
II. Sharma B K. Instrumental Methods of Chemical
Analysis.2004:pg161-170
• Web references
I. http://chemcatalog.waters.com/publication/frame.
php?i=142735&p=292&pn=&ver=flex
II. http://www.waters.com/waters/en_US/GPC---Gel-
Permeation-
Chromatography/nav.htm?cid=10167568&locale=en
_US