Development and Validation of Rp- Hplc Method Forthe Estimation of Oral Hypoglycemicdrug

DEVELOPMENT AND VALIDATION OF RP- HPLC METHOD FORTHE ESTIMATION OF ORAL HYPOGLYCEMICDRUG

DISSERTATION PROTOCOL

SUBMITTED TO

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

BANGALORE, KARNATAKA.

BY

ASHWANI

M.PHARM, PART-1

DEPARTMENT OFQUALITY ASSURANCE

M.M.U. COLLEGE OF PHARMACY

RAMANAGARAM-562159

KARNATAKA

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE-560041 KARNATAKA,

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION

1. / NAME OF THE CANDIDATE AND ADDRESS (IN BLOCK LETTERS) / ASHWANI
S/O Mr. RABINDRA PRASHAD SINGH
Q.NO:- 3067, SECTOR- 11C, BOKARO STEEL CITY
JHARKHAND
2. / NAME OF THE INSTITUTION / M.M.U COLLEGE OF PHARMACY
K.K DODDI, RAMADEVERA BETTA ROADRAMANAGARAM-562159
KARNATAKA
3. /

COURSE OF STUDY AND SUBJECT

/ MASTER OF PHARMACY IN
QUALITY ASSURANCE
4. / DATE OF ADMISSION OF COURSE / 01-10-2010
5. /

TITLE OF TOPIC

/ DEVELOPMENT AND VALIDATION OF RP- HPLC METHOD FORTHE ESTIMATION OF ORAL HYPOGLYCEMIC DRUG
6. / BRIEF RESUME OF THEINTENDED WORK
6.1 Need for the study
6.2 Review of the literature
6.3 Objectives of the study / ENCLOSURE-I
ENCLOSURE-II
ENCLOSURE-III
7. /

MATERIALS AND METHODS

7.1 Source of data
7.2 Method of collection of data
7.3 Does study require any Investigations or interventions to be conducted on patients or Other human oranimal? If so, Please describe briefly.
7.4 Has ethical clearance been obtained from your institution in case of 7.3 / ENCLOSURE-IV
ENCLOSURE-V
ENCLOSURE-VI
ENCLOSURE-VI
8. / LIST OF REFERENCES / ENCLOSURE-VII
9. / SIGNATURE OF CANDIDATE
10. / REMARKS OF GUIDE / RECOMMENDED FOR THE DISSERTATION WORK
11 / NAME AND DESIGNATION OF
11.1 Guide
11.2 Signature of guide
11.3 Co guide (if any)
11.4 Signature of co guide
11.5 Head of department
11.6 Signature of head of department / PROF. NIRMAL.T. HAVANNAVAR HEAD OF THE DEPARTMENT (QUALITY ASSURANCE)
M.M.U COLLEGE OF PHARMACY
K.K DODDI, RAMADEVERA BETTA ROADRAMANAGARAM-562159
KARNATAKA
Not applicable
Not applicable
PROF. NIRMAL.T. HAVANNAVAR HEAD OF THE DEPARTMENT (QUALITY ASSURANCE)
M.M.U COLLEGE OF PHARMACY
K.K DODDI, RAMADEVERA BETTA ROAD RAMANAGARAM-562159
KARNATAKA
12. / 12.1 Remarks of the
Chairman and principal
12.2 Signature / FORWARDED AND RECOMMENDED FOR FAVOURABLE CONSIDERATION
(PROF. MOHAMED KHALEEL)
6 / BRIEF RESUME OF THE INTENDED WORK
ENCLOSURE-I
6.1 Need for the study
Diabetes is one of the most common disease so that development andprocess validation of HPLC method for antidiabetic drugs hold importance, as there is lot of side effects associated with several dosage formit is necessary to examine the amount of therapeutically active component in a particular dosage form.
Although there is spectacular advancement in the instrumental methods of analysis, the success or failure of such method largely depends upon how pure is the sample for such analysis. This is because a light impurity in analyte will lead to erroneous results. So large no of separation methods were reported.HPLC technique is a simple, rapid, and precise method for simultaneous analysis of drugs and it is the most widely used analytical technique for the quantitative analysis of pharmaceuticals, biomolecules, polymers, of organic compounds etc.
The high recovery and low co-efficients of variation confirm the suitability of this method for analysis of the drugs. By statistical analysis it will prove that this method is specific, accurate, reliable and reproducible for the analysis of bulk drug and in pharmaceutical formulation without any interference from the excipients.There are certain drugs for which analytical method except HPLC are reported. Hence, there is need to develop simple, accurate, precise, sensitive analytical method for those drugs.
ENCLOSURE II
6.2Review of literature
1.Venkata R A et al.developed and validate LC method for the estimation of Glipizide in pharmaceutical dosage form and serum. Optimization of mobile phase was performed based on asymmetric factor and peak area obtained.
2.Sheikh R A et al. worked on RP-HPLC method for the analysis of Glipizide in pharmaceutical dosage forms. RP-C18 column with UV detectors. With HPLC assay method the peak area of standard solution and sample solution was recorded and analyte concentration of the drugs was quantified using the regression time obtained for the pure sample.
3.RathinavelG. A et al. simultaneously estimated Rosiglitazone and Gliclazide in tablet dosage form by RP-HPLC method. Chromatography was carried out on Phenomenix Gemini C18 column using the mixture of ammonium phosphate buffer, Acetonitrile and methanol in the ratio 50:35:15 v/v with flow rate of mobile phase 1mL min-1 and effluent was monitored at 254 nm.
4.Havaldar FH.A et al. had done simultaneous estimation of Glimepiride, Rosiglitazone and Pioglitazone Hydrochloride in pharmaceutical dosage form.Separation was achieved with a nucleodur C–18 column having 250x4.6 mm i.d. with 5 μm particle size and water HPLC grade adjusted to pH 3.0 using diluted orthophosphoric acid and acetonitrile (80:20 v⁄v) with gradient program as eluent at a constant flow rate of 0.8 ml per min. UV detection was performed at 215 nm. The retention time of glimipiride, rosiglitazone and pioglitazone hydrochloride were about 17.9 min, 6.31 min and 8.24 min.
5.Bhamare PC.A et al. Performed ondevelopment and validation of a precise singlestability indicating HPLC method for determinations of Metformin hydrochloride andFenofibrate, in Pure form and in Pharmaceutical tablets.The HPLC method was carried out on Inertsil octadecylsilane C18 (250 mm x 4.6 mm i.d., 5 μm particle size) column. A mobile phase composed of acetonitrile - water (adjusted to pH 3 using orthophosphoric acid) in proportion of 70:30 v/v, at flow rate of 1 ml/min was used for the separation. Detectionwas carried out at 250nm.
6.Rao NM.A et al. haddevelopment and validation for estimation of Voglibose in bulk and tablet dosage forms by RP-HPLC method. The HPLC separation was carried out by reverse phase chromatography on RP-18e, Hibar RT column (250×4.6mm) with a mobile phase composed of 0.025M potassium dihydrogen phosphate pH 2.5 : acetonitrile: methanol (40:55:5) in isocratic mode at a flow rate of 1ml/min. The detection was monitored at 282nm.
7.Reddy BP. Aet al. haddeveloped and validate of simultaneous determination of Pioglitazone and Glimepiride in pharmaceutical dosage form by RP-HPLC. A phenomenex Luna C18 column (4.6X150mm) was used for saperation.The mobile phase was acetonitirle: KH2PO4 buffer (60:40%v/v) (Ph6) at the flow rate of 1.5 ml/min with detection at 230 nm. Several mobile phase composition were tried to resolve the peak of Pioglitazone and Glimepiride.
8.KarthikA. A et al.worked onSimultaneous Determination of Pioglitazone and Glimepiride in bulk drug and pharmaceutical dosage form by RP-HPLC method. The quantification was carried out using Inertsil ODS (250 x 4.6 mm) column and mobile phase comprised of acetonitrile and ammonium acetate (pH 4.5; 20mM) in proportion of 60:40 (v/v). the flow rate was 1.0 ml/min and effluent was monitored at 230 nm.
ENCLOSURE-III
6.3Objectives of the study

• The main objective of the present investigation involves development of RP-HPLC method using simple mobile phase which is sensitive and rapid for quantification of oral hypoglycemic drug in tablet dosage forms as well as subsequent validation of developed method according to ICH guide lines.
• To apply the stability indicating HPLC method for theestimation of Oral hypoglycemic drugs in pharmaceutical formulations.

7.MATERIALS AND METHODS:
Materials:
Oral hypoglycemic drug, HPLC grade distilled water, solvent like ethanol, methanol etc.
Methods:

• All experiments will be carried out in the Department of Quality Assurance, M.M.U College of Pharmacy, Ramanagaram.
• Pure samples of oral hypoglycemic drug shall be procured from industries involved in manufacture of these drugs.
• Sample (tablets) shall be procured from market.
• New HPLC (Model–SPD-M20A, made in Japan) method shall be developed and validated for simultaneous estimation of oral hypoglycemic drugs.

The developed stability indicating HPLC method shall be applied for simultaneous Determination of oral hypoglycemic drugs, degraded products in pharmaceutical formulation.
ENCLOSURE-IV
7.1. Source of Data
1) Review of literature from :
a. Journals : such as
- Indian journal of pharmaceutical sciences.
- International journal of comprehensive pharmacy.
- International journal of pharmaceutical research.
- European journal of pharmaceutical sciences.
- Indian journal of chemistry and technology.
- Asian journal of chemistry.
b. Internet browsing.
2) Library: M.M.U College of pharmacy.
3) Laboratory based studies.
ENCLOSURE-V
7.2. Method of Collection of Data

1. Procurement of the drug sample and chemicals.
2. Determine the solubility of oral hypoglycemic drugs in various solvents and buffers.
3. Scanning the solution in UV-Visible region and selecting the solvents for various analytical studies.
4. Preparation of standard calibration curve
5. Develop and validate analytical method for oral hypoglycemic drugs by UV-Visible spectrophotometer using,

a)Standard absorptivity value.
b)Calibration graph.
c)Single point and double point standardization.

1. Develop HPLC method to estimate oral hypoglycemic drugs in bulk and pharmaceutical formulation.
2. Validation of developed analytical method as per ICH guidelines.

ENCLOSURE-VI
7.3. Does the study require any investigation or intervention to be conducted on patients or other humans or animals? If so, please mention briefly.
-NO-
7.4. Has ethical clearance been obtained from your institution in case of 7.3?
-NOT APPLICABLE-
ENCLOSURE-VII
8. LIST OF REFERENCES
1.RayanamV,Rao AL.and RamanaMV.Development and validation of LC method for the estimation of Glipizide in pharmaceutical dosage form and serum.Int. J. Res. Pharm. & chem. 2011; 1(01): 50-53.
2.SheikhRandAsifK.Reverse phase high performance liquid chromatographic method for the analysis of Glipizide in pharmaceutical dosage form. Inter. J. Res. Pharm. Chem. 2010; 1(02): 455-458.
3.RathinavelG,UmaUN.ValarmathyJ,SamueljoshuaL,Thanuja CS,GaneshM. A et a., RP-HPLC Method for the simultaneous estimation of Rosiglitazone and Gliclazide in Tablets E-J. Chem. 2009;6(4): 1188-1192.
4.Freddy,HavaldarH,andVairalDL.Simultaneous Estimation ofGlimepiride,Rosiglitazone and Pioglitazone Hydrochloride in the pharmaceutical dosage formin the Pharmaceutical Dosage Form.E-J. Chem. 2010;7(4):1326-1333.
5.BhamarePC,BariSB,NatarajanS,PatilAA,PatilSHandShirodePT.Development and Validation of a precise singlestability indicating HPLC method fordeterminations of Metformin hydrochloride andFenofibrate, in Pure form and in PharmaceuticalTablets. Inte. J. PharmTech Res. 2011; 3(01): 505-515.
6.Raw MN, Konda RK, Bagyalakshmi, Ravi TK, andMogili R. RP-HPLC method development and validation for estimation of Voglibose in bulk and tablet dosage forms Int. J. Res. Pharm. Sci. 2010; 1(2): 190-194.
7.ReddyBP,BoopathyD,MathewB,PrakashM,andPerumalP.Methoddevelopment and validation of simultaneous determination of Pioglitazone and Glimepiride in pharmaceutical dosage form by RP-HPLC.Inte.J.ChemTech Res. 2010; 2(1): 50-53.
8.RayanamV, RaoAL,andRamanaMV. Development and Validation of LC method for the estimation of Rosiglitazone in pharmaceutical dosage form and Serum.Inte. J. Adv. Ph’cl.sci. 2011; 2(1):46-49.
9.AKarthik,RaoMC,KrishnamurthyB,RanjithkumaA,MusmadiP.Aetal.Simultaneous Determination of Pioglitazone and Glimepiride in bulk drug and pharmaceutical dosage form by RP-HPLC method.Pak.J.Pharm.Sci. 2008;21(4):421-425.