dinkar kamkhede hplc presentation 2014-2015

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.

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.

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

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)

Ab

sorb

ance

Approximation

of peak area by

triangulation

Area = base x height

2

base

height

Peak A Peak B

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.

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.

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.

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.

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.,

Thank

You....

dinkar kamkhede hplc presentation 2014-2015

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