HPLC Method Development & Validation for Dissolution Method of Drug X ( Hypolipidaemic) in Tablet Dosage form

Supervisor
Dr. Asif Husain
Snr. Assistant Professor Deptt.of Pharm.Chemistry Jamia Hamdard

Co-supervisor
Dr. M.Mumtaz Alam Assistant Professor Dept. of Pharm.Chemistry Jamia Hamdard Mr. Moloy Mitra Associate Director, AR, R&D-I Ranbaxy Research Lab. Gurgaon

Presented by:
MD. SABIR AZIM

3rd Semester M.Pharm (Pharm. Analysis) Jamia Hamdard

High-performance liquid chromatography (HPLC) is a chromatographic technique that can separate a mixture of compounds and is used in biochemistry and analytical chemistry to identify, quantify and purify the individual components of the mixture. HPLC typically utilizes different types of stationary phases, a pump that moves the mobile phase and analyte through the column, and a detector that provides a characteristic retention time for the analyte.

HPLC

Instrumentation
A) B) C) High pressure pump Column / injector system Mobile phase

D)

Detector

Drug dissolution testing is an analytical technique used to assess release profiles of drugs from pharmaceutical products, generally solid oral products. Drug dissolution testing plays an important role as a routine quality control test, for characterizing the quality of the Product. Dissolution from the dosage form involves mainly two steps: liberation of the drug from the formulation matrix (disintegration), dissolution of the drug (solubilization of the drug particles) in the liquid medium. In case of low solubility & high permeability drugs, drug dissolution may be rate limiting step for drug absorption. A dissolution profile is recommended for drug products in this category.

Why is HPLC important?

Very Sensitive. Better & faster separation technique. Greater reproducibility. Continuous & quantitative detection. Higher accuracy & precision. A variety of stationary phases are readily available. This technique is free from the problems of sample volatility & thermal instability. Ideally suited for separation of macromolecules & ionic species of biomedical interest.

 Drug X is a very effective hypolipidaemic drug. It is a newer drug of this category. It has fewer side effect in comparison to other drug of this class. Its small dose(10 mg) is enough for treating Hypercholesterolemia. Marketed demand is very high.

OBJECT /AIM OF STUDY

To develop and validate a highly specific, reliable and cost effective HPLC method for Dissolution method of drug X in Tablet dosage form. Optimization of chromatographic conditions. Validation of developed method.

RATIONALE OF STUDY

There is no effective HPLC method available for dissolution of the Drug X. Although only few methods have been reported in literature, but all of them use large amount of reagents and consume lot of time, our aim is to shorten the run time and make it cost effective. Drug X is a very effective hypolipidaemic drug. it has relatively fewer side effects. It is a newer drug of this class. The market is rising for this drug and it has wide scope in both in developed and developing countries.

DRUG PROFILE (Drug X)

Category Structure : Hypolipidaemic Drug. :

Description Solubility

: An off White to white powder. : Freely soluble in methanol, ethanol and Acetone. Practically insoluble in water. Melting Point : 164166 C (327331 F) BCS Class : Class II ( low solubility, high permeability) Protein Binding : 90% Molecular mass : 409.4 g mol-1 Mechanism of Action : Drug X localizes at the brush border of the small intestine , where it inhibits the absorption of cholesterol from the intestine

PLAN OF WORK
Literature Review : Collection of all pertinent literature regarding Drug Profile (Drug X) Analytical Methodology: Identification of the drug:1) DSC Analysis 2) IR Analysis 3) UV Analysis

Analytical Method Development for Dissolution method of Drug X in tablet dosage form and Optimization of method development parameters. Validation of the developed method.

DSC Analysis

IR Spectra

Melting point of the drug was found to be 165C Conclusion On the basis of above performed analysis it was found that the drug sample was pure and authentic

UV Analysis of Drug X

STANDARD in Methanol ( max = 232 nm)

SAMPLE in Methanol ( max = 232 nm)

Standard Curve in Methanol

0.9 0.8 0.7 0.6 Abssorbance 0.5 0.4 0.3 0.2 0.1 0 0 5 10 15 conc. (g/ml) 20 25 30

y = 0.032x R = 0.997

max 238nm

HPLC Method development and validation for dissolution method of Drug X in tablet dosage form.

As per FDA guideline: Sod. Acetate Buffer (pH 7.0) + 0.5 % w/v SLS Preparation of Dissolution Media ( As per USP): Dissolve about 13.8 gm of monobasic sodium phosphate and 50 gm SLS in 10 Liter of distilled water. Mix vigorously till the SLS is dissolved. Adjust pH 7.0 + 0.05 with 1N NaOH Solution. Preparation of 1 N NaOH Solution : Weigh accurately about 4 gm NaOH and dissolve in 100 ml of distilled water.

Method Development

Method development usually requires selecting the method requirements and deciding on what type of instrumentation to utilize and why.

It involves selection of: Best buffer & its pH Best mobile phase Best diluents Best column (S. phase) Best column temperature Best detector

Method Validation

Validation is the process of determining the suitability of a given methodology for providing useful analytical data. Validation is required for any new method to ensure that it is capable of giving reproducible and reliable results, when used by different operators employing the same equipment in the same or different laboratories. Parameters (as defined by USP) for Dissolution Method Specificity Linearity System Precision Method Precision Accuracy Stability in analytical solution.

REFERENCES

L.R Snyder,J.J Kirkland; Introduction to Modern Liquid Chromatography,2nd edition, Wiley Interscience Publication,1979 ICH, Q2 (R1) Validation of analytical procedures. International Conference on Harmonization: June. 1994 The Merck index. An encyclopedia of chemical drugs and biological. 13th ed. Merck Research laboratories division of merck & co., INC. 2001; P.958, 1163, 1567. Willard HH, Dean AJ. Instrumental methods of analysis. 7th ed. CBS Publishers and distributors; 1986. P. 513-515, 580-604. Sethi PD. High Performance Liquid Chromatography quantitative analysis of pharmaceutical formulations. 1st ed. CBS Publishers & Distributors; 2001. P. 71-94. Tripathi KD. Essential of pharmacology. 6th ed. Jaypee brothers medical publisher Ltd. 2008.P.614-618.

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sabirazim@gmail.com