DEVELOPMENT AND VALIDATION OF RP HPLC METHOD FOR THE DETERMINATION OF ANTIHYPERTENSIVE DRUGS

DISSERTATION PROTOCOL

SUBMITTED TO THE

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

BANGALORE, KARNATAKA.

BY

FULETRA JIGNESHKUMAR DINESHBHAI

M.PHARM PART-I

DEPARTMENT OF QUALITY ASSURANCE

M.M.U. COLLEGE OF PHARMACY

RAMANAGARAM-562159

KARNATAKA

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA, BANGALORE

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / NAME OF THE CANDIDATE
AND ADDRESS (IN BLOCK LETTERS) / FULETRA JIGNESHKUMAR DINESHBHAI
S/O Mr.DINESH BHAI
31,VRUNDAVAN DOUPLEX,
NR SATYAMEV HOSPITAL,CHANDKHEDA-382424
AHMEDABAD,GUJARAT.
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 PHARMCY IN
QUALITY ASSURANCE
4. / DATE OF ADMISSION OF COURSE / 01-10-2010
5. /

TITLE OF TOPIC

/ DEVELOPMENT AND VALIDATION
OF RP HPLC METHOD FOR THE DETERMINATION OF ANTIHYPERTENSIVE DRUGS
6. / BRIEF RESUME OF THE
INTENDED 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 or animal? 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 / (FULETRA JIGNESHKUMAR DINESHBHAI)
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. MOHAMED KHALEEL
PRINCIPAL
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 ROADRAMANAGARAM-571511
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
Hypertension is a cardiovascular disorder. It is most common disease so that development andprocess validation of HPLC method for antihypertensive drugs hold importance. As there are lot of side effects associated with several dosage forms,it is necessary to examine the amount of therapeutically active component in a particular dosage form.These antihypertensive drugs are used to lower the blood pressure in hypertension.
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 number of separation methods were reported.HPLC technique is a simple, rapid, and precise method for the analysis of antihypertensive 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. Oza.CK, A et al. had developed rp-hplc method for the determination of losartan potassium and perindopril erbumine in combined tablet dosage form. The method involved by using HiQSil-C-18W ODS, 5 /m column and mobile phase was ACN: water in proportion of 50:50 v/v, pH adjusted to 3.2 ± 0.1 with 1 % o-phosphoric acid. The flow rate was 1.0 mLmin-1 and effluent was monitored at 210 nm. The retention time of losartan potassium and perindopril erbumine were eluted at 6.7 min and 4.5 min respectively.
  1. Kumar.GVS A et al. worked ondevelopment and validation of reversedphase hplc method for simultaneous estimation of telmisartan and amlodipine in tablet dosage form.The chromatographic separation was achieved on analytical column with mobile phase consisting mixture of Potassium dihydrogen phosphate (0.02M, pH 3.0 adjusted with ortho‐phosphoric acid) and acetonitrile in ratio (60:40 v/v) at flow rate of 1.5ml/min and detector wavelength 237 nm.The retention time of Amlodipine and telmisartan was found to be 3.5 and 8.1 min respectively.
  1. Sankar.GD Aet al. performed development and validation of a rapid RP-HPLC method for the determination of Venlafaxine Hydrochloride in pharmaceutical dosageformsusingexperimentaldesign.Themethodutilizesa5μmVarian® Microsorb-MV 100 C18 column (250 mm x 4.6 mm) at ambient temperature. A 23 factorial design consisting of 3 factors at 2 levels was set up to standardize the chromatographic conditions. The optimum mobile phase consisted of acetonitrile, 0.04 M potassium dihydrogenphosphate buffer and methanol (45:25:30, v/v), with pH adjusted to 5.5 using 10% phosphoric acid solution. The mobile phase was delivered isocratically at a flow rate of 1 mL/min with UV detection at 224 nm.Venlafaxine hydrochloride was eluted at 3.43 min.
  1. Kumar.NS A et al. worked on analytical method development and validation of amlodipine and hydrochlorothiazide in combined dosage form by rp-hplc. The method involved C18 column (Phenomenex C18, 5μ, 250mm x 4.6mm). The sample was analysed using Triethylamine: Acetonitrile: Methanol in the ratio of 50:25:25(pH adjusted to 3.0 with Orthrophosphric acid) as a mobile phase at a flow rate of 2.0ml/min and detection at 235nm. The retention time for Amlodipine and Hydrochlorothiazide was found to be 6.631 and 2.183 min respectively, and recoveries from combined dosage form were between 98 and 102%.
  1. Chouhan.K A et al. had developed validated rp-hplc method for simultaneous determination ofatorvastatin and ramipril and its application in drug formulation.Chromatographywas carried out on a Phenomenex – Luna, C18 (250 x 4.6 mm i.d.,5μ) column. using filtered and degassed mixture of acetonitrile ,water and methanol (55:40:5) as mobile phase at a flow rate of
1.0ml/min and effluent was monitored at 237nm.
  1. Pachauri.S A et al. performed development & validation of hplc method for analysis of someantihypertensive agents in their pharmaceutical dosage forms.The quantitative determination of analyte(s) was performed on PUROSPHERE STAR RP 18e analytical column(250×4.6 mm) with 0.2 %v/v TEA buffer (pH: 3.0): ACN as mobile phase, at a flow rate of 1.0 mL min-1. Detectionwas made by extracting PDA spectra at 215 nm respectively.
  1. Boopathy.D A et al. worked on analytical method development and validation oflosartan potassium and atenolol in combineddosage form by rp-hplc. The method utilisesC18 column (Phenomenex C18, 5μ, 250mm x 4.6mm). The sample was analysed using Triethylamine: Acetonitrile: Methanol in the ratio of 50:30:20(pH adjusted to 4.0 with phosphric acid) as a mobile phase at a flow rate of 1.2ml/min and detection at 235nm. The retention time for Losartan Potassium and Atenolol was found to be 3.767 min and 2.210 min respectively, and recoveries from combined dosage form were between 98 and 102%.
ENCLOSURE-III
6.3 Objectives 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 antihypertensive drug in tablet dosage forms as well assubsequent validation of developed method according to ICH guidelines.
7. MATERIALS AND METHODS:
Materials:
Antihypertensive drug,distilled water,methanol,ethanol,propanol etc.
Methods:
  • All experiments will be carried out in the Department of Quality Assurance, M.M.U College of Pharmacy, Ramanagaram.
  • Pure samples of antihypertensive drugs shall be procured from industries involved in manufacture of these drugs.
  • Sample (tablets) shall be procured from market.
  • New HPLC (Model- SPD-M20A 230V, made in Japan) method shall be developed and validated for the estimation of antihypertensive drugs.
The developed stability indicating HPLC method shall be applied for the determination of antihypertensive 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 antihypertensive 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 antihypertensive 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 antihypertensive 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
LIST OF REFERENCES
  1. Oza.CK, Prajapati.JP, Vyas.AJ, and Mehta.P. RP-HPLC method for the determination of losartan potassiumand perindopril erbumine in combined tablet dosage form. Int. J. Pharm. and Bio. Sci. 2011;2(01):709-715.
  1. Kumar .GVS, Prasad.R. Development and validation of reversedphaseHPLC method forsimultaneous estimation of telmisartan and amlodipine in tablet dosageform. Int. J. Pharm. and Ph’cal. Sci. 2010;2(03):128-131.
  1. Sankar.GD, Somasekhar.V, Kumar Shiva.HN. Development and validation of a rapidRP-HPLC method for the determination ofvenlafaxine hydrochloride in pharmaceuticaldosage forms using experimental design. E-J. Chem. 2009;6(04):1091-1102.
  1. Kumar.NS, Safeer.K, Anbarasi.B. Analytical method development and validation of Amlodipine and Hydrochlorothiazide in combined dosage form by RP-HPLC. Int. J. Chem.Tech. Res. 2010;2(01):21-25.
  1. Chouhan.K, Agrawal.S, raj.C, Jain.S, Subramaniam.BA and Raj.NR. Validated RP-HPLC method for simultaneous determination of Atorvastatin and Ramipril and its application in drug formulation. Schr. Res. Lbry. Der Pharmacia Lettre. 2011; 3(02): 194-202.
  1. Pachauri.S, Paliwal.S, Srinivas.KS, Singh.Y, Jain.V. Development & validation of HPLC method for analysis of some Antihypertensive agents in their pharmaceutical dosage forms. J. ph’cal. sc. and res. 2010;2(08):459-464.
  1. Boopathy.D, Abdus.SK, Perumal.P. Analytical method development and validation of Losartan Potassium and Atenolol in combined dosage form by RP-HPLC. Int. J. PharmTech Res. 2010;2(01):471-474.
  1. Singh.B, Patel.DK, Ghosh.SK. Development of Reverse-Phase HPLC method for simultaneous analysis of Metoprolol Succinate and Hydrochlorothiazide in a tablet formulation. Trop.J. ph’cal res. 2009;8(06):539-543.
  1. Chabukswar.RA, Kuchekara.SB, Jagdalea.CS, Mehetrea.MD, Morea.SAand Lokhandeb.DP. Development and validation of a RP-HPLC method for simultaneous estimationof Olmesartan Medoxomil and Amlodipine Besylate in tablet dosage form. Schr. res. lbry. 2010;2(04):307-312.
  1. Valarmathy.J, Samueljoshua.L, Rathinavel.G, Thanuja.SCand Sivakumar.T. RP-HPLC method development and validation for assay of Levetiracetam in tablet dosage form. Res. J. pharm. and tech. 2008;1(03):394-397.