HPLC Method Development & HPLC Method Development & Validation for Dissolution Method Validation for Dissolution Method of Drug X of Drug X ( Hypolipidaemic) in ( Hypolipidaemic) in Tablet Dosage form 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 3 rd Semester M.Pharm (Pharm. Analysis) Jamia Hamdard
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HPLC Method Development & Validation for HPLC Method Development & Validation for Dissolution Method of Drug X Dissolution Method of Drug X
( Hypolipidaemic) in Tablet Dosage form ( Hypolipidaemic) in Tablet Dosage form
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.
InstrumentationA) High pressure pump
B) Column / injector system
C) Mobile phase
D) Detector
HPLCHPLC
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?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 OBJECT /AIM OF STUDYSTUDY
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 STUDYRATIONALE 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.
Category : Hypolipidaemic Drug. Structure :
Description : An off White to white powder. Solubility : Freely soluble in methanol, ethanol and Acetone.
Practically insoluble in water. Melting Point : 164–166 °C (327–331 °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
DRUG PROFILE DRUG PROFILE (Drug X)(Drug X)
PLAN OF WORKPLAN 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.
IR Spectra
ConclusionOn the basis of above performed analysis it was found that the drug sample was pure and authentic
Melting point of the drug was found to be 165̊<C
DSC Analysis
UV Analysis of Drug X
STANDARD in Methanol (λ max = 232 nm) SAMPLE in Methanol (λ max = 232 nm)
λmax 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 DevelopmentMethod 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 ValidationMethod 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.
REFERENCESREFERENCES 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.
Introduction To Analytical Method Development For Pharmaceutical Formulations. [Internet]. html [Accessed 22 July, 2008]. Available from: http:// www.pharmainfo.net/ reviews/ introduction-analytical-method-development-pharmaceutical-formulations
Furman WB, Layloff TP, Tetzlaff RF. Validation of computerized liquid chromatographic systems. J. AOAC Int. 1994;77(5):1314-1318.
www.fda.gov.in United State Pharmacopoeia 2011, vol-2, 455, 511& 711-714 Scott PWR. Liquid chromatography for the analyst. Marcel
Dekker Inc. New York USA; 1994. P. 1-10. (vol.67). Validated Analytical Methods for Determination of Active
Ingredients from Bulk Drugs and Pharmaceutical Dosage Forms. Rev 2008; 6(1). html [Accessed 20th Dec. 2009]. Available from: http://www.pharmainfo.net/reviews/validated-analytical-methods-determination-active-ingredients-bulk-drugs-and pharmaceutical.
