STABILITY INDICATING HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF ATORVASTATIN CALCIUM AND PIOGLITAZONE IN PHARMACEUTICAL FORMULATION

DISSERTATION PROTOCOL

SUBMITTED TO

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES

BANGALORE, KARNATAKA.

BY

CHARAN RAJUM

M.PHARM, PART-I

DEPARTMENT OF QUALITY ASSURANCE

NARGUNDCOLLEGE OF PHARMACY

BANGALORE-85

(2010-2012)

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES,

BANGALORE,KARNATAKA.

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / NAME OF THE CANDIDATE
AND ADDRESS (IN BLOCK LETTERS) / CHARAN RAJU M
NO. 8/326, I FLOOR, 5THCROSS, OPP.
Dr. NARASHIMAMURTHY HOUSE, RAMAKRISHNA ROAD, HOSKOTE,BANGALORE-562114,
KARNATAKA.
2. /

NAME OF THE INSTITUTION

/ NARGUNDCOLLEGE OF PHARMACY,
DATTATREYANAGAR, II MAIN,
100 FEET RING ROAD,
BSK III STAGE,
BANGLORE-85,
KARNATAKA.
3. /

COURSE OF STUDY AND SUBJECT

/ MASTER OF PHARMACY IN
QUALITY ASSURANCE
4. / DATE OF ADMISSION OF COURSE / 25stNOV2010
5. /

TITLE OF TOPIC

/ “STABILITY INDICATING HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF ATORVASTATIN CALCIUM AND PIOGLITAZONE IN PHARMACEUTICAL FORMULATION”
6.
6.1
6.2
6.3
7.
7.1
7.2
7.3
7.4
8. / BRIEF RESUME OF INTENDED WORK:
NEED FOR THE STUDY:
The dyslipidemia associated with insulin resistance and type 2 diabetes is characterized by elevated triglycerides and decreased HDL cholesterol. Although LDL cholesterol may not be elevated in type 2 diabetes, an increase in the proportion of small, dense, and potentially more atherogenic LDL cholesterol particles is observed. In addition to LDL cholesterol, elevated triglyceride levels and reduced HDL cholesterol levels are both risk factors for coronary heart disease (CHD). Compared with nondiabetic individuals, patients with type 2 diabetes have a two- to fourfoldhigher risk of CVD, and dyslipidemia is an important contributor to the increased risk in this population [1].
Many medications used to treat type 2 diabetes or hyperlipidemia can be administered once per day. In this situation, adding a second drug that is also taken once per day may lead to reduced adherence.In order to increase patient compliance of type 2 diabetes and hyperlipidemia, fixed-dose combination products (FDCP) of Pioglitazone and Atorvastatin calcium are available.
Atorvastatin Calcium is HMG-CoA reductase inhibitor. It competitively inhibits conversion of 3-Hydroxy-3-methyl glutaryl Coenzyme A (HMG-CoA) to mevalonate by the enzyme HMG-CoA reductase.Therapeutic doses reduce CH synthesis[2]
Pioglitazone is hypoglycemic agent which is related to thiazolidinediones, it is selective agonist for the nuclear peroxisome proliferators–activator, which enhances the transcription of several insulin responsive genes.Hepatic gluconeogenesiss is also suppressed [3].
Analysis of FDC during process and in final product is quite exigent job for pharmaceutical manufacturers in adherence with strict rules and regulations about the safety and quality of manufactured products in the pharmaceutical industry. These FDC products can present daunting challenges to the analytical chemist responsible for the development and validation of analytical methods. Chromatographic techniques are most accepted analytical tool while the individual components in FDC can be simultaneously measured without the need for separation and extraction in presence of their degraded products.
Literature survey reveals that there is no stability indicating RPHPLC method reported for simultaneous estimation of Atorvastatin Calcium and Pioglitazone in pharmaceutical formulation, hence stability indicating RPHPLC method development of Atorvastatin calcium and Pioglitazone in combination has been found interested and selected for the study.
REVIEW OF LITERATURE :
Pioglitazone is chemically [(±)-5-[[4-[2-[5-ethyl -2- pyridinyl) ethoxy] phenyl]-Methyl]-2, 4-] thiazolidinedionemonohydrochloride. Molecular formula is C19H20N2O3S•HCl andmolecular weight is 392.90. It is sparingly soluble in water, freely soluble in methanol and chloroform. Chemical structure of Pioglitazoneisas below. [4]

Atorvastatin ischemically[R-(R*, R*)]-7-[2-(4-Fluorophenyl)-5-(1-methylethyl)-3-phenyl-4-(phenylaminocarbonyl)-1H-pyrrol-1-yl]-3,5-dihydroxy-heptanoic acid calcium salt.Molecular formula is(C33H34FN2O5).Caand molecular weight is 1155.36.It isvery soluble in water, soluble in alcohol, practically insoluble in methylene chloride. Chemical structure of Atorvastatin Calciumas shown below. [5]

  • R. VIJAYAMIRTHARAJ,et al (2010) developed and validatedRPHPLC method for simultaneous estimation of Telmisartan and Atorvastatin Calcium in tablet dosage formsusing UV detector. Selected mobile phase was a combination of Acetonitrile: Buffer (0.01M Potassium dihydrogen phosphate) 65:35(v/v), pH 4.00 (adjusted with Orthophosphoric acid) and the wavelength selected was 250nm. The flow rate was kept at 2.0 ml/min, and the injection volume was 10μl. The separation was performed on Phenomenex C18 column (250x4.6mm i.d, 5μm) at ambient temperature.[6]
  • SASMITA KU. ACHARJYA, et al (2010) developed and validated RPHPLC method for the quantitation of Atorvastatin Calcium and Amlodipine Besylate in combined tablet dosage forms. Quantitation was achieved using a reversed-phase Hypersil silica BDS (250x4.6mm with 5 μmparticle size) column at ambient temperature with mobile phase consisting of 0.05M ammonium acetate buffer (pH-4) and acetonitrile in the ratio (40: 60, v/v). The flow rate was maintained at 1.0 mL/min and measurements were made at a wavelength 240.0nm. [7]
  • SATHEESH K. SHETTY,et al (2010)reported a sensitive and rapid stability-indicating HPLC method for the simultaneous quantitative determination of Aspirin (ASP), Atorvastatin (ATV), Atenolol (ATL) and Losartan potassium (LST) in a polypill form in the presence of degradation products. Efficient chromatographic separation was achieved on a C18 stationary phase with mobile phase combination of buffer (0.1% Orthophosphoric acid, pH 2.9) and Acetonitrile,delivered in a gradient mode at flow rate of 1.0 mL/min. The quantitation was carried out using ultraviolet detection at 230 nm. The combination of drugs are exposed to thermal, acid/base hydrolytic, humidity and oxidative stress conditions, and the stressed samples were analyzed by proposed method. The method was validated with respect to ICH guidelines.[8]
  • KRISHNA R.GUPTA,et al (2009) developed and subsequently validated a stability indicating reverse-phase HPLC method for simultaneous estimation of Atorvastatin (ATR) and Nicotinic acid (NTA) from their combined dosage form. The proposed RP-HPLC method utilizes a Phenomenex C18, 5 μm, 250 mm X 4.6 mm i.d. column, at ambient temperature andthe mobile phase consisted of acetonitrile and 50mm potassium dihydrogen phosphate buffer (68:32, v/v), apparent pH adjusted to 4.5±0.1 with phosphoric acid solution.The effluent flow rate was monitored at 0.8 mL/ min, and UV detection at 247 nm. The drug products are exposed to thermal, acid/base hydrolytic, humidity and oxidative stress conditions, and the stressed samples were analyzed by proposed method.[9]
  • MEETA A JILADIA, et al (2009)proposed simple and sensitive HPTLC method for quantitative estimation of Pioglitazone in bulk and tablet dosage forms. Pioglitazone was chromatographed on silica Gel 60 F254 TLC plate using Toluene: methanol: ammonia (7:3:0.1 v/v) as mobile phase. Pioglitazone showed Rf value 0.50±0.03 and scanned at 268 nm using Camag TLC Scanner 3. The method was validated in terms of linearity (200–1200 µg/spot). [10]
  • THAMAKE SL,et al (2009)worked on analytical method development and validation of simultaneous estimation of Atorvastatin Calcium and Ramipril from capsule dosage form by first order derivative spectroscopy.For this method linearity observed in 5-40 μg/ml for ATR and 2-20 μg/ml for RAM.[11]
  • KARTHIKA, et al (2008) quantitatively determined Pioglitazone and Glimpiride simultaneously in bulk drug and pharmaceutical dosage from by RP- HPLC method. The quantification was carried out using Inertsil ODS (250 × 4.6 mm, 5μ) 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 the effluent was monitored at 230 nm. [12]
  • LINCY JOSEPHet al (2008)developed simultaneous estimation of Atorvastatin and Ramipril by RP-HPLC and Spectroscopy method. Individual λ-max for Atorvastatin was observed as 247 nm and that of Ramipril was 208 nm . simultaneous estimation of these drugs were carried out using Vierodt’s equation . While in HPLC method, mobile phase used was the mixture of 50% acetonitrile and 50% buffer (1 gm sodium perchlorate in 500 ml milli-Q-water, adjusted the pH to 2.5 with dilute phosphoric acid) and Column used was Intersil ODS (250 x 4.6 mm, 5μm.) Flow rate was maintained at 1.2 ml/minute and detection was done at 215 nm. Column temperature was kept at 40ºC.[13]
OBJECTIVES OF THE STUDY:
Review of literature revealed that there are few analytical methods for estimation of drug or in the combination with other drugs and there is only one liquid chromatographic method reported for the simultaneous estimation of AtorvastatinCalcium andPioglitazone by RPHPLCin capsule dosage form. So the objective of the present research work are:
To obtain the stress degraded products of Atorvastatin Calcium andPioglitazone by exposing a formulation which is under study for different stress conditions like acid, base, oxidative, reductive and neutral media.
To develop a HPLC method for simultaneous estimation of AtorvastatinCalcium andPioglitazone in presence of separate degraded products.
To validate the method using parameters like accuracy, precision, linearity and range for the estimation of these drugs in pharmaceutical dosage form.
To study the stress degradation behavior ofAtorvastatinCalcium andPioglitazone by analyzing the different products obtained after degradation using HPLC method.
To apply the stability indicating HPLC method for the simultaneous estimation of Atorvastatin Calcium andPioglitazoneand degraded product in a pharmaceutical formulation.
MATERIALS AND METHODS:
Materials
Atorvastatin Calcium,Pioglitazone, HPLC grade solvents and chemicals like Methanol, Acetonitrile, Water, Orthophosphoric acid, Triethylamine, Potassium dihydrogen phosphate, Dipotassium hydrogen phosphate and other analytical regents.
SOURCE OF DATA:
Data will be obtained from Science Direct, and other internet facilities, literature search and related articles from library of Nargund College of Pharmacy, Digital Library of RGUHS, Bangalore, etc.
Journals
Indian Journal of Pharmaceutical Sciences
Journal of Pharmaceutical and Biomedical Science
International Journal of Pharmacy and Pharmaceutical Science
Asian Journal of Pharmaceutics
Journal of Chromatography B
Asian Journal of Research Chemistry
Journal of Wisconsin Medical
Pakistan Journal of Pharmaceutical Science
Internet Browsing
  • india.com
MethodofCollectionofData (IncludingSampling Procedures, IfAny)
Procurement of drug samples and marketed formulations.
Development of HPLC method for estimation of Atorvastatin Calcium and
Pioglitazone.
Validation of all developed analytical methods as per ICH guidelines.
Does the study require any investigation or intervention to be conducted on patients or other humans or animals? If so, please mention briefly.
- NOT APPLICABLE -
Has ethical clearance been obtained from your institution in case of 7.3?
- NOT APPLICABLE-
LIST OF REFERENCES:
  1. Ronald B, Goldberg MD. Diabetes Care.Diabetes Care 2005, 28(7),1547–54.
  2. McDonagh M, Peterson K, Thakurta SG, Dana, T. Drug class review on fixed
dose combination drug products for the treatment of type 2 diabetes and hyperlipidemia, 2007 ( ( access on 11-06-2011).
  1. on 11-06-2011).
  2. (access on 11-06-2011).
  3. (access date 13-04-2011).
  4. Vijayamirtharaj R, RameshJ, Jayalakshmi B,Hashim HB. Development and validation of RP-HPLC method for simultaneous estimation of telmisartan and atorvastatin calcium in tablet dosage forms. Pharmacie Globale (IJCP). 2010;01, 4 (03): 1-4 (Available online at
  5. Sasmita A, Annapurna M, Aailaja K.Liquid chromatographic method for simultaneous estimation of atorvastatin calcium and amlodipine besylate inpharmaceutical dosage forms. Int J Pharma Bio Sci.2010;1(4): 161-70. (Available online at
  6. Satheesh KS, Koduru VS, Radhakrishnanand P, Borkar RM, Devrukhakar PS, Jogul J, Tripathi UM. Quantitative application to apolypill by the development of stability indicating LCmethod for the simultaneous estimation of aspirin, atorvastatin, atenolol and losartan potassium.American J Anal Chem.2010; 2: 59-69.
  7. Krishna R, Gupta, Askarkar SS, Wadodkar SG.Stability indicating RP-HPLC method for simultaneous determination of atorvastatin and nicotinic acid from their combined dosage form. Eurasian J Anal Chem. 2009;4(3): 294-303.
  8. Meeta A Jiladia, SS Pandya, and Viidyasagar G. A simple and sensitive HPTLC method for estimation of pioglitazone in bulk and tablet dosage forms. Asian J Res Chem. 2009;2(2): 207-9. (Available from
  9. Thamake SL, Jadhav SD, Pishawikar SA.Development and validation of method for simultaneous estimation of atorvastatin calcium and ramipril from capsule dosage form by first order derivative spectroscopy.Asian J Res Chem. 2009; 2(1): 52-3.
  10. Karthik A, Subramanian G, Rao MC, BhatK, Ranjithkumar A, Musmade P, Surulivelarjan M, Karthikeyan K and Udupa N. Simultaneous determination of pioglitazone and glimpiride in bulk drug and pharmaceutical dosage from by RP- HPLC method. Pak J Pharm Sci. 2008; 21(4):421-5.
  11. Joseph L, GeorgeM and Rao VRB. Simultaneous estimation of atorvastatin and ramipril by RP-HPLC and spectroscopy. Pak J Pharm Sci. 2008; 21(3): 282-4.

9. / Signature of the candidate / (CHARAN RAJU M)
10. / Remarks of the Guide / RECOMMENDED FOR THE DISSERTATION WORK.
11. / Name & Designation of
(in block letters)
11.1Guide
11.2 Signature / DR. J. N. NARENDRA SARATH CHANDRA
PROFESSOR, HEAD OF THE DEPARTMENT
DEPARTMENT OF QUALITY ASSURANCE
NARGUNDCOLLEGE OF PHARMACY
Dr. J. N. Narendra Sarath chandra
11.3 Head of the department
11.4 Signature / DR. J. N. NARENDRA SARATH CHANDRA
PROFESSOR, HEAD OF THE DEPARTMENT
DEPARTMENT OF QUALITY ASSURANCE
NARGUNDCOLLEGE OF PHARMACY
Dr. J. N. Narendra Sarath chandra
12. / 12.1 Remarks of Principal
12.2 Signature / FORWARDED AND RECOMMENDED FOR FAVOURABLE CONSIDERATION.