Its very effective presentation of HPLC technique with VWD detectors.we also demonstrate the Force degradation Study,Stability indicating Methods, in Different Environment Conditions.

1. PROJECT ON
ESTMATION AND FORCED DEGRADATION STUY OF
THIAZOLE DERIVATIVE BY HPLC TECHNIQUE
Presented By:- Mr. Dinkar Kamkhede
MSc.II.CHE.IV,Sem.2014
Vidyabharati Mahavidyalaya,
Amravati.

2. ESTMATION AND FORCED
DEGRADATION STUY OF THIAZOLE
DERIVATIVE BY HPLC TECHNIQUE
High Performance Liquid Chromatography
HPLC is characterized by the use of high pressure to push a
mobile phase solution through a column of stationary phase
allowing separation of complex mixtures with high resolution.

3. HPLC
Types of Analysis Qualitative&Quantitatve
Stationary Phase 3-dimentional column
Instrumention Much! With many adjestable
parameters
Sample Apllication Injection
Rheodyne Injector
Mobile Phase Movement High pressure solvent delivery
Viusalization Result ‘online’ detection
Variable (UV/Vis)
Form of Result Peaks,Rt. retention time

4. HPLC
Chromatograms
Rt = 3.0 min.
faster moving
less retained
Rt = 5.2 min.
slower moving
more retained
0 1 2 3 4 5 6 7
Time (minutes)
Absorbance
Approximation
of peak area by
triangulation
Area =
base x height
2
base
height
Peak A Peak B

5. Chromatography Stationary Phases
relatively polar surface
O O O
| | |
OSiOSiOSiOH
| | |
O O O
| | |
OSiOSiOSiOH
| | |
O O O
bulk (SiO2)x surface
relatively nonpolar surface
Silica Gel
O O O
| | |
OSiOSiOSiOR
| | |
O O O
| | |
OSiOSiOSiOR
| | |
O O O
bulk (SiO2)x surface
Derivatized Silica Gel
Where R = C18H37
hydrocarbon chain
(octadecylsilyl deriv.
silica or “C18”)
“normal phase” “reversed phase”

6. Normal vs. Reversed Phase Chromatography
1.In this column type, the retention is governed by the
interaction of the polar parts of the stationary phase
and solute. For retention to occur in normal phase, the
packing must be more polar than the mobile phase
with respect to the sample
2.In this column the packing material is relatively nonpolar and
the solvent is polar with respect to the sample. Retention is the
result of the interaction of the nonpolar components of the
solutes and the nonpolar stationary phase. Typical stationary
phases are nonpolar hydrocarbons, waxy liquids, or bonded
hydrocarbons (such as C18, C8, etc.) and the solvents are polar
aqueous-organic mixtures such as methanol-water or
acetonitrile-water.

7. What is a stability indicating
method?
What is a stability indicating method?
A stability indicating method is a quantitative test method that can detect
possible degradants and impurities of drug substance (API) and drug products,
normally using High Performance Liquid Chromatography (HPLC). Stability
information is needed for regulatory submissions such as IND (Investigational
New Drug Application) and NDA (New Drug Applications) and to set expiration
dates for the API or drug product.
Forced Degradation
The forced degradation studies are another very important part of the
validation of the stability indicating method. In forced degradation
studies, samples are stored under extreme conditions (acid, base,
peroxide, heat, light, humidity etc) in order to rapidly screen drug
product stabilities.
Stability-indicating methods are traditionally performed using gradient
elution, in order to ensure that degradants of various chemical
compositions are all detected.

8. Summary of the work
Forced degradation studies are indispensable in the development of stability-
indicating and degradant monitoring methods as part of a validation protocol.
Forced degradation studies also provide invaluable insight in investigating
degradation products and pathways of drug substances and products.
it is strongly recommended these studies be started as early as possible to be
able to provide valuable information that can be used to assess the inherent
stability of a drug, and to improve formulations and the manufacturing process.
Given that no specific set of conditions will be applicable to all drug
substances and products.

9. CONCLUSION
The method development and validation are continuous and interrelated
processes that are conducted throughout the drug development process. The
analytical validation verifies that a given method measures a parameter as
intended and establishes the performance limits of the measurement.
Reproducible quality HPLC results can only be obtained if proper attention
has been paid to the method development, validation and system’s suitability
to carry out the analysis. The validated methods produce results within known
uncertainties that are helpful to continuing drug development and provide
emerging knowledge supporting the product. The time and effort that is
devoted into developing scientifically sound and robust analytic methods
should be aligned with the drug development stage.

10. REFERENCES
 Shewiy, D. H, Kaale, E,Risha,P. Dejaegher, G. B. Verbeke, J. S. Heyden, Y. V. Pharmaceut.
J. Biomed. Anal 2012, 66, 11–23.
 Rockville, M. D. General Tests, Chapter 621 – Chromatography System Suitability, United
States Pharmacopeial Convention (USP), USP 31 (2009)
 parate, detect, and quantify the various drug-related degradants that can form on storage or
manufacturing, Julia T, Mena AJ, Aucoin MG, Kamen AA. Development and validation of
a HPLC method for the quantification of baculovirus particles. J Chromatogr B 2011; 879:
61–68.quantify any drug-related impurities that may be introduced during synthesis. Forced
degradation studies (chemical and physical stress
 Khan MC, Reddy NK, Ravindra G, Reddy KVSRK, Dubey PK.Development and validation
of a stability indicating HPLC method for simultaneous determination of four novel
fluoroquinolone dimers as potential antibacterial agents. J Pharmaceut Biomed Anal 2012;
59:162–16.,

11. Thank
You....