“DEVELOPMENT AND VALIDATION OF QUANTITATIVE ANALYTICAL METHOD FOR AMILORIDE AND HYDROCHLORTHAIZIDE IN COMBINATION IN PHARMACEUTICAL DOSAGE FORMS”
DISSERTATION PROTOCOL
SUBMITTED TO
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BANGALORE, KARNATAKA.
BY
PATEL RONAK VASANTBHAI
M.PHARM, PART-I
DEPARTMENT OF QUALITY ASSURANCE
NARGUND COLLEGE OF PHARMACY
BANGALORE-85
(2010-2012)
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA.
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / NAME OF THE CANDIDATEAND ADDRESS (IN BLOCK LETTERS) / PATEL RONAK VASANTBHAI
NARGUND COLLEGE OF PHARMACY,
DATTATREYA NAGAR, II MAIN,
100 FEET RING ROAD,
BSK III STAGE,
BANGALORE-85,
KARNATAKA.
2. /
NAME OF THE INSTITUTION
/ NARGUND COLLEGE OF PHARMACY,DATTATREYA NAGAR, II MAIN,
100 FEET RING ROAD,
BSK III STAGE,
BANGLORE-85,
KARNATAKA.
3. /
COURSE OF STUDY AND SUBJECT
/ MASTER OF PHARMACY INQUALITY ASSURANCE
4. / DATE OF ADMISSION OF COURSE / 1st JULY 2010
5. /
TITLE OF TOPIC
/ “ DEVELOPMENT AND VALIDATION OF QUANTITATIVE ANALYTICAL METHOD FOR AMILORIDE AND HYDROCHLORTHAIZIDE IN COMBINATION IN PHARMACEUTICAL DOSAGE FORMS ”6.
6.1
6.2
6.3
7.
7.1
7.2
7.3
7.4
8. / BRIEF RESUME OF THE INTENDED WORK:
NEED FOR THE STUDY:
Hypertension is the most common cardiovascular disease where elevated arterial pressure causes pathological changes in the vasculature and hypertrophy of the left ventricle. As consequence, hypertension is the principal cause of stroke, leads to disease of coronary arteries with myocardial infarction and sudden cardiac death, and is a major contributor to cardiac failure, renal insufficiency, and dissecting aneurysm of the aorta [1].
Amiloride is potassium sparing diuretic acts by affecting Na+ reabsorption in the cortical collecting duct. It specifically blocks the apical membrane epithelial Na+ channel (ENaC). The reduced rate of Na+ reabsorption diminishes the gradient that facilitates the K+ secretion. In addition to their effect on distal Na+ and K+ transport, it inhibits H+ secretion by the late distal tubule and late collecting duct [2].
Hydrochlorothiazide is thaizide diuretic consisting benzothaidiazine ring acts in the distal convoluted tubule where they block Na+ - Cl- cotransport. Na+ - Cl- cotransport occurs in luminal surface of the distal convoluted tubules. Thus, to exert its diuretic action hydrochlorothiazide must reach the luminal surface of the distal convoluted tubules. At higher dose sometimes it exerts carbonic anhydrase inhibition [2].
High blood pressure and water retention are the causes of the congestive heart failure. Hence the goal of medical treatment should be to decrease blood volume and fluid retention which can lower the blood pressure. Therapeutic options available are diuretics, rennin inhibitors, ganglion blockers, vasodilators. Diuretics are the drugs which act by changing the electrolyte level in blood. The major side effects of diuretics are imbalance in electrolyte level in blood like hypokalemia. Combination of Hydrochlorothiazide and Amiloride is the diuretic therapy that work together to keep electrolyte level in balance. Hydrochlorothiazide, while effective, often causes low potassium level in blood. On the other hand Amiloride is “potassium sparing diuretic” which can increase potassium level in the blood. Thus this combination levels the electrolyte balance [3].
There are few analytical methods reported for simultaneous estimation of Amiloride and Hydrochlorothiazide. Investigation of simple analytical methods is in need for the quantitative estimation of these drugs in combination in pharmaceutical dosage form.
REVIEW OF LITERATURE :
Ø Chemically Amiloride is 3,5-diamino-6-chloro-N-(diaminomethylene)pyrazine-2-carboxamide. Molecular formula is C6H8ClN7O and molecular weight is 229.627 g/mol. It is freely soluble in hot water, DMSO [4,5].
Ø Chemical structure of Amiloride [4].
Amiloride
( Ι )
Ø Chemically Hydrochlorothiazide is 6-chloro-1,1-dioxo-3,4-dihydro-2H-1,2,4-benzothiadiazine-7-sulfonamide. Molecular formula is C7H8ClN3O4S, molecular weight is 297.74 g/mol. It is slightly soluble in water, freely soluble in alcohol [6,7].
Ø Chemical structure of Hydrochlorothiazide [6].
Hydrochlorothiazide
( ΙΙ )
Ø Kakde RB, Kotak VH, Barsagade AG, Chaudhary NK, Kale DL developed spectrophotometric method for estimation of Amlodipin besylate and Bisoprolol fumarate in pharmaceutical preparation using 10 % methanol as a solvent at two wavelengths 222 nm and 365 nm by simultaneous equation method [8].
Ø Maitreyi Z, Amit K proposed validated RP-HPLC method for simultaneous estimation of Atenolol and Hydrochlorothiazide in pharmaceutical dosage forms by using Zorbax SB-CN (250 x 4.6 mm), 5μm column in isocratic mode with mobile phase, water: buffer: methanol (50:35:15). The flow rate was 1.2 ml/min. The peak purity of Atenolol and Hydrochlorothiazide were 0.999 and 1.000 respectively. The percentage relative standard deviation was below 2.0 % [9].
Ø Pratik D, Das SN, Mahapatra SK, Annapurana MM developed UV spectrophotometric method for the simultaneous estimation of Hydrochlorothiazide and Ramipril in bulk and its pharmaceutical dosage forms. UV spectra were measured at 212 and 226 nm for Hydrochlorothiazide and Ramipril. That obeyed linearity at range of 1.25-40 mg/ml and 0.5-16 mg/ml [10]
Ø Nagavalli D, Vaidhyalingum V, Shantha A, Shankar AS, Divya O proposed simultaneous spectrophotometric determination of Losartan potassium, Amlodipin besylate and Hydrochlorothiazide in pharmaceuticals by chemometric method and calibration were constructed using the absorption data matrix to the concentration data matrix. The range of measurement was 230.5-350.4 nm. The linearity range was found to be 8-40, 1-5 and 3-15mg ml-1 for Losartan potassium, Amlodipin besylate and Hydrochlorothiazide respectively [11].
Ø Ankit BC, Rakesh KP, Sunita AC, Krupa VG developed estimation of Valsartan and Hydrochlorothiazide in pharmaceutical dosage forms by absorption ratio method and linearity was found to be in the range of 2-20 μg/ml. The molar absorptivity for both the drugs were calculated at two wavelengths 270.5 nm and 231.5 nm respectively [12].
Ø Sridharan D, Thenmozhi A, Rajamanikam V, Sundarnandavalli S, Planikumar B developed simultaneous estimation of Irbesartan and Hydrochlorothiazide by UV spectroscopy using multicomponant mode of analysis by concept of standard addition. The linearity of Irbesartan and Hydrochlorothiazide was found in the range of 10-26 and 5-13µg/ml at wavelength 234 and 272 nm respectively [13].
Ø Vrushali T, Vijya V, Ujjwala K, Sashikant D developed novel UV spectrophotometric method for estimation for Ramipril and Hydrochlorothiazide by simultaneous equation and area under curve method. The first method include the measurement of absorbance at 210 and 270 nm for Ramipril and Hydrochlorothiazide and other method of AUC includes measurement of AUC under wavelength range 202-237.5 and 247-289 nm for ramipril and hydrochlorothiazide [14].
Ø Toral M, Stefnaia P, Silvia Q, Pablo R developed simultaneous determination of Amiloride and Furosamide by first digital derivative spectroscopy method. The determination was carried out at 241.4 and 343.6 nm for furosamide and amiloride respectively. The determination range was found to be of 6.9 × 10-8 to 16× 10-5 and 6.8 × 10-8 to 8× 10-5 respectively for furosamide and amiloride [15].
Ø Sane RT, Bale VR, Nayak VG and Ladge KD worked on the development of RP-HPLC method for separation of Hydrochlorothiazide, Amiloride hydrochloride and Atenolol using C-18(10 µm , 300×4.6 mm) column and methanol, water, glacial acetic acid, triethylamine (22+78+01+0.1 v/v) as a mobile phase. The flow rate was maintained 1 ml/min and detection was carried out at 273 nm[16].
Ø Jain R amd Jain CL developed RP-HPLC method for quantitative analysis of Hydrochlorothiazide and Enalapril maleate in pharmaceutical dosage form. Composition of mobile phase used was water, methanol, phosphoric acid (90-10-0.1 v/v) and C-18 (10 µm , 300×3.9 mm) column was used. Detection was done at 215 nm initially and 272 nm after 5 minutes or 205 nm throughout the analysis[16].
OBJECTIVES OF THE STUDY:
Review of literature revealed that there is few analytical method for the simultaneous estimation of Amiloride and Hydrochlorothiazide in pharmaceutical dosage form.
Hence the objectives of the present work are:
Ø To develop different analytical methods to estimate Amiloride and Hydrochlorothiazide in combined form by UV-Visible spectrophotometer.
Ø To develop and optimize HPLC method to estimate Amiloride and Hydrochlorothiazide in combined dosage form.
Ø To validate the developed methods by ICH guidelines.
MATERIALS AND METHODS:
Materials
Amiloride, Hydrochlorothiazide, Amiloride and Hydrochlorothiazide Formulation, Distilled Water, Methanol, Ethanol, Isopropanol, DMSO, Sodium hydroxide.
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.
Ø Journals
Indian Journal of Pharmaceutical Sciences
International Journal of Pharmaceutical Research
International Journal of Chem. Tech Research
E- Journal of Chemistry
International Journal of Chemistry Research
International Journal of Pharmacy and Pharmaceutical Sciences
International Journal of Applied Biology and Pharmaceutical Technology
J Pharm Pharmaceut Sci
ACTA Chromatographica
Journal of Pharmacy Research
American journal of health-system pharmacy.
Open Medicine.
Ø Text Books and Pharmacopoeia
· Beckett AH, Stenlake JB. Practical Pharmaceutical Chemistry. 4th ed.
Delhi: CBS Publisher and Distributors; 1997.
· Sethi PD. Quantitative Analysis of Drugs in Pharmaceutical Formulation.
3rd ed. Delhi: CBS Publisher and Distributors.
· Higuchi T and Brochman E, Hanseen H. Pharmaceutical Analysis. Delhi: CBS Publisher and Distributors; 2005.
· Mendham J, Denney RC, Barnes JD, Kthomas MJ. Vogel’s text Book of Quantitative Chemical Analysis. 6th ed. Pearson education pvt Ltd; 2002.
· Indian Pharmacopoeia, Government of India, Ministry of Health and Family Welfare, Published by The Indian Pharmacopoeia commission Gaziabad, 2; 2007.
Ø Internet Browsing
· www.sciencedirect.com
· www.google.com
· www.rxlist.com
· www.medlineindia.com
· www.google scholar.com
· www.emedtv.com
Method of Collection of Data (Including Sampling Procedures, If Any)
Ø Procurement of drugs sample and marketed formulations.
Ø Solubility determination of Amiloride and Hydrochlorothiazide in various solvents and buffers.
Ø Studying the spectra of both the drugs in UV–Visible region in different solvents/buffers and selecting the solvents for various analytical studies.
Ø Development of analytical methods by simultaneous equation, absorption ratio, derivative spectroscopy, etc for the estimation of Amiloride and Hydrochlorothiazide in combined form by UV-visible spectrophotometer.
Ø Development of HPLC method, optimization of the method for estimation of Amiloride and Hydrochlorothiazide.
Ø 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) Goodman and Gilman’s. The pharmacological basis of therapeutics. 10th ed. United Stats: McGraw Hill companies; 2001. p. 871-33.
2) Charles RC, Robert ES. Modern pharmacology with clinical application. 6th ed. p. 245-8.
3) http://hypertension.emedtv.com/moduretic/moduretic.html.
4) http://en.wikipedia.org/wiki/Amiloride/ (access date sep 15,2010)
5) http://sigma aldrich.com/productinformation/Amiloride
6) http://en.wikipedia.org/wiki/Hydrochlorothiazide/ (access date Sep 15, 2010).
7) http:// formulation%20emixt%20updates.htm
8) Kakde RB, Kotak VH, Barsagade AG, Chaudhary NK, Kale DL. Spectrophotometric method for estimation of Amlodipin besylate and Bisoprolol fumarate in pharmaceutical preparations. J Pharm Tech Res. 2008; 1(4): 513-5.
[Available from http://rjptonline.org/RJPT_%20Vol1(4)/48.pdf ]
9) Maitreyi Z., Amit K. Development and validation of RP-HPLC for the simultaneous estimation of Atenolol and Hydrochlorothiazide in pharmaceutical dosage forms. J Pharm Res H Care. 2(3): 248-52.
[Available from http://www.arjournals.org/ijaps1/ijaps.2010.0976.1055.01020.pdf]
10) Pratik D, Das SN, Mahapatra SK, Kar SK, Annapurana MM. UV spectrophotometric method for the simultaneous estimation of hydrochlorothiazide and ramipril in bulk and its pharmaceutical dosage forms. Int J Pharm tech. 2009; 2(3): 25-29.
[Available from http://ijpst.com/files/IJPST,%20Vol-3,%20Art-5,09.pdf]
11) Nagavalli D, Vaidhyalingum V, Shantha A, Shankar AS, Divya O. Simultaneous spectrophotometric determination of Losartan potassium, Amlodipin besylate and hydrochlorothiazide in pharmaceuticals by chemometric methods. Acta Pharm Res. 2010: 141-52.
12) Ankit BC, Rakesh KP, Sunita Ac, Krupa VG. Estimation of valsartan and hydrochlorothiazide in pharmaceutical dosage forms. Int J applied bio. 2010; 1(2): 455-64.
[Available from http://ijabpt.com/pdf/1233-VAL%20HCTZ%20paper.pdf]
13) Sridharan D, Thenmozhi A, Rajamanikam V, Sundarnandavalli S, Planikumar B. Simultaneous estimation of irbesartan and hydrochlorothiazide by UV spectroscopy using multicomponant mode of analysis. Int J Chem Tech Res. 2010; 2(2): 876-9.
[Available from http://sphinxsai.com/s_v2_n2/CT_V.2No.2/ChemTech_Vol_2No.2_pdf/CT=19%20(876-879).pdf]
14) Vrushali T, Vijya V, Ujjwala K, Sashikant D. Novel UV spectrophotometric method for estimation for ramipril and hydrochlorothiazide by simultaneous equation and area under curve method. Int J Applied Pharmaceutics. 2010; 4(2)
[Available from http://www.ijaponline.org/Vol2Issue4/127.pdf]
15) Toral M, Stefnaia P, Silvia Q, Pablo R. Simultaneous determination of amiloride and furosamide by first digital derivative spectroscopy method.
1. [Available from
2. captura.uchile.cl/jspui/bitstream/.../1/Amiloride%20TORAL,%202002.pdf]
16) Sethi PD. High Performance liquid Chromatography. Quantitative analysis of pharmaceutical formulations. 1st edition(reprint 2008);CBS publishers and distributors, New Delhi; volume 1: p. 213-7
9. / Signature of the candidate / (PATEL RONAK VASANTBHAI)
10. / Remarks of the Guide / RECOMMENDED FOR THE DISSERTATION WORK.
11. / Name and Designation of
(in block letters)
11.1 Guide
11.2 Signature / Mrs. Ritu VIVEK Kimbahune,
ASSISTANT PROFESSOR,
DEPARTMENT OF QUALITY ASSURANCE,
NARGUND COLLEGE OF PHARMACY,
BANGALORE.
11.3 Head of the department
11.4 Signature / DR. J. N. NARENDRA SARATH CHANDRA
PROFESSOR, HEAD OF THE DEPARTMENT,
DEPARTMENT OF QUALITY ASSURANCE
NARGUND COLLEGE OF PHARMACY,
BANGALORE.
12. / 12.1 Remarks of Principal
12.2 Signature / FORWARDED AND RECOMMENDED FOR FAVOURABLE CONSIDERATION.