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Spectrophotometric method for simultaneous estimation of amlodipine besylate in pharmaceutical formulation

Adv J Pharm Life sci Res, 2013 1;1:16-21 Spectrophotometric Method for Simultaneous Estimation of Amlodipine Besylate in
Pharmaceutical Formulation

Naresh Kalra*, Suresh Choudhary

Department of Pharmaceutical sciences, Alwar Pharmacy College, IET, North Extension M.I.A, Alwar, India

ABSTRACT
The present study deals with a rapid, simple, accurate, and rugged UV spectrophotometric methods have been
developed for the estimation of some water insoluble calcium channel blockers using hydrotropic solubilization phenomenon in pure and as well as in dosage forms. The calibration graphs were linear in the 5–40 µg/ml concentration range (r>0.997) for Amlodipine. All the methods were validated as per USP guidelines. The detection was made on 350- 200 nm in methanol and 0.1M sodium Benzoate observed maximum absorption at 237nm. Hence this method is simple, rapid, and accurate and can be successfully applied for routine analysis. Keywords: Spectrometric analysis; Amlodipine; Method validation; Hydrotrophy
INTRODUCTION:
Amlodipine (a dihydropyridine) is a calcium- The decrease in intracellular calcium inhibits the channel blocker. It is given as besylate and used in the contractile processes of the myocardial smooth muscle management of hypertension and angina pectoris. It is cells, causing dilation of the coronary and systemic chemically 3-ethyl-5-methyl-2-(2aminoethoxymethyl)- arteries, increased oxygen delivery to the myocardial 4-(2-chlorophenyl)-1,4-dihydro-6-methyl pyridine-3,5- tissue, decreased total peripheral resistance, decreased dicarboxylate mono benzene sulphonate1. It is official systemic blood pressure, and decreased after load. in BP2.Various methods have been employed to Another possible mechanism is that amlodipine enhance the aqueous solubility and hydrotrophy is one inhibits vascular smooth muscle carbonic anhydrase I of them. Maheshwari et al. have developed various activity with consecutive pH increase which may be involved in intracellular calcium influx through solubilization phenomenon to analyze poorly water- soluble drugs like hydrochlorothiazide3, aceclofenac4 Objective
Hydrotropic solution may be a proper choice To develop an accurate, selective precise and to preclude the use of organic solvents because organic specific method for estimation of water insoluble solvent having a problems related like toxicity, cost calcium channel blockers in bulk and as well as in etc. Several analytical methods that have been reported dosage form. There is a broad scope for hydrotropic
for the estimation of amlodipine besylate in biological agent in quantitative estimation of poorly water soluble fluids and/or pharmaceutical formulations include UV drugs. The present work is an attempt to study and spectrophotometric6-10, and high-performance liquid develop spectroscopic methods employing hydrotropic chromatographic estimation (HPLC)11, 12. Amlodipine solubilization phenomenon for quantitative estimation is a calcium channel blocking agent. It inhibits the and validation of the method according to ICH influx of extracellular calcium across the myocardial and vascular smooth muscle cell membranes. ____________________________________________ Estimation of Amlodipine in dosage form using
*Corresponding Author: Naresh Kalra
hydrotropic solubilization technique
Asst. Prof, Alwar Pharmacy College, North extension, IET MIA Alwar (Raj) India, M 09413787016 E-mail: [email protected] MATERIALS AND METHODS:
____________________________________________ Instrument:
spectrophotometer (Shimadzu UV-1800) with 1 cm Advanced Journal of Pharmacie and Life science Research Adv J Pharm Life sci Res, 2013 1;1:16-21 Matched quartz cells, band pass 1.8 nm, Shimadzu AX Estimation of amlodipine from dosage form in
-200 electronic weighing balance and Ultrasonicator, methanol
Twenty tablets containing each of 10mg of Drug Sample: Amlodipine were obtained as gift
amlodipine were accurately weighed and finally now sample from Sun Pharma limited, Bangalore. dried in a glass mortar. A weigh equivalent to 10mg of Chemicals and Reagent: Methanol A.R. Grade (Loba
amlodipine was accurately transferred to a 10ml of Chemie, Mumbai). & Sodium benzoate From SD fine standard flask 4ml of methanol was added and swirled Chemicals. Commercially available pharmaceutical gently a period of 10min.The clear supernatant dosage form Amdepin manufactured by Cadila solution was then transferred to 10ml standard flask pharmaceuticals limited, Cadila corporate campus, through Whatmann No.1 filter paper .The residue was Sarkhej- Dholaka road, Bhatt, Ahmedabad-22500 were further extracted twice with 2ml each of methanol and procured from local market for estimation. passed the same filter paper and the volume was made up to 10ml with methanol. The resulting solution had a Estimation of amlodipine in pure and in dosage
form in methanol:
Accurately pipette 1ml of the above solution into a 10ml standard flask and made up to volume with Preparation of standard stock solution of Amlodipine
methanol. The final solution had a concentration of in Methanol
Weighed accurately about 50mg of amlodipine Accurately pipette out 1ml of solution B into 10ml and transferred into a 50ml standard flask dissolved standard flask and the volume was made up using and made unto the volume using methanol. This methanol to obtain concentration of 10mg/ml of solution had a concentration of 1mg per ml of amlodipine. The solution was measured at 237 nm and the concentration of amlodipine was calculated from Accurately pipette out 5ml of solution A into a 50ml standard flask and the volume was made up to 50ml using methanol. The resulting solution had a Estimation of amlodipine in dosage form using
concentration 100mg/ml of amlodipine (solution 0.1M sodium Benzoate as hydrotropic agent:
B).Accurately pipette out 4ml of solution B separately into 10ml of standard flask and the volume was made Preparation of 0.1M Sodium Benzoate
up to 10ml using methanol. The resulting solution had a concentration of 40mg/ml of amlodipine (solution Sodium Benzoate and transferred into a 100ml standard flask dissolved and made up the volume using Study of spectral characteristic of amlodipine in
methanol
Preparation of standard stock solution of amlodipine
After enabling the initial adjustment and blank in 0.1M Sodium Benzoate
correction, using methanol the solution C was scanned Weighed accurately about 50mg of amlodipine in the entire UV range from 350 to 200 nm. A broad and transferred into a 50ml standard flask dissolved band of absorption spectrum was observed with and made up the volume using 0.1M Sodium Benzoate. This solution had a concentration of 1mg Preparation of calibration curve of amlodipine in
Accurately pipette out 5ml of solution A into a 50ml methanol
standard flask and the volume was made up to 50ml using 0.1M Sodium Benzoate. The resulting solution solution B into eight 10ml standard flask. Volume was had a concentration 100mg/ml of amlodipine (solution made up to 10ml using methanol. The absorption of each solution was measured at 237nm against reagent Accurately pipette out 4ml of solution B separately blank. The readings were plotted by taking absorbance into 10ml of standard flask and the volume was made up to 10ml using 0.1M Sodium Benzoate. The Advanced Journal of Pharmacie and Life science Research Adv J Pharm Life sci Res, 2013 1;1:16-21 resulting solution had a concentration of 40mg/ml of The Weight of 20 tablets in methanol & 0.1M Sodium Benzoate is the 0.9844gm & 0.9844gm & Each tablet contains (label claim) amlodipine is 10mg. Study of spectral characteristic of amlodipine in
So that the Average weight of tablet is 0.0992 gm & 0.1M Sodium Benzoate
0.0992 in methanol & 0.1M Sodium Benzoate & the After enabling the initial adjustment and blank Weight of powder taken is 0.0992gm. So that the correction, using methanol the solution C was scanned Average content of amlodipine /tablet determined by in the entire UV range from 400 to 200 nm. A broad the proposed method is 10.082mg & 10.042 mg. & band of absorption spectrum was observed with The Percentage of label claim for methanol and 0.1M Sodium Benzoate is 100.2% & 100.02%. Preparation of calibration curve of amlodipine in
Method validation14,15
0.1M sodium benzoate
Linearity and range
Calibration curve was prepared for amlodipine solution B into eight 10ml standard flask. Volume was is methanol at 364nm and entire calibration data at the made up to 10ml using 0.1M Sodium Benzoate .The selected wave length as summarized in table absorption of each solution was measured at 364nm No.2.These shows that amlodipine. The obeyed Beer’s law in the concentration range 5 to 40mg/ml at 364nm Estimation of amlodipine from dosage form in 0.1M
Repeatability /precision
sodium benzoate
Repeatability expresses the precision under the Twenty tablets containing each of 10mg of same operating condition area short interval of time. It amlodipine were accurately weighed and finally is also termed as intraday precision. The precision of powdered in a glass mortar. A weigh equivalent to the analytical procedure is usually expressed as the 10mg of amlodipine was accurately transferred to a standard deviation of a series of measurements. The 10ml of standard flask 4ml of 0.1M Sodium Benzoate results obtained from nine observations of the same was added and swirled gently for a period of concentration of sample solution the main volume 10min.The clear supernatant solution was then transferred to 10ml standard flask through Whatmann No.1 filter paper .The residue was further extracted Robustness
twice with 2ml each of 0.1M Sodium Benzoate and The robustness of the method was determined passed the same filter paper and the volume was made by using sodium benzoate from three different up to 10ml with 0.1M Sodium Benzoate. The resulting suppliers. SD fine chemicals, Mumbai. E Merck. Ltd, solution had a concentration of 1mg/ml (solution A). Mumbai, Thomas Baker chemicals Ltd.mumbai. Accurately pipette 1ml of the above solution into a 10ml standard flask and made up volume with 0.1M Recovery studies
Sodium Benzoate. The final solution had a concentration of 100mg/ml of amlodipine (solution B). determined by performing recovery studies. A fixed Accurately pipette out 1ml of solution B into 10ml amount of drug from dosage form was taken and pure standard flask and the volume was made up using standard drug at three different concentrations within methanol to obtain concentration of 10mg/ml of beer’s range was added. The total concentration was amlodipine. The solution was measured at 364 nm and found by the proposed method. The determination with the concentration of amlodipine was calculated from each concentration was repeated three times and average percent recovery of the added standard was calculated and results are tabulated in Table No. 5. RESULT AND STATISTICAL EVALUATION:
Advanced Journal of Pharmacie and Life science Research Adv J Pharm Life sci Res, 2013 1;1:16-21 Table 1: Results of analysis of amlodipine in dosage form
Formulatio
Amount of drug % label claim
concentration
in found in µg*
Table 2: Calibration data of amlodipine at 237 nm and 364 nm
Parameter
Amlodipine at 237 nm
Amlodipine at 364 nm

Table 3: Precision data of Amlodipine in Methanol and in 0.1 M Sodium benzoate

Concentration obtained by the
Mean Value
concentration
in µg /ml
In Methanol
In 0.1M Sodium
In Methanol
In 0.1M Sodium
benzoate
benzoate
Table 4: Robustness with Methanol and 0.1M Sodium benzoate from three different suppliers
Methanol
Drug taken in mg
Drug obtained in mg*
% of recovery
0.1 M sodium
Drug taken in mg
Drug obtained in mg
% of recovery
benzoate
Advanced Journal of Pharmacie and Life science Research Adv J Pharm Life sci Res, 2013 1;1:16-21 Table 5: Recovery Studies of Amlodipine in Methanol and in 0.1M Sodium benzoate
Formulation
Label claim
Concentration of
Concentration
% Recovery
pure drug in mg
found in mg*
Amlodipine
in methanol
Amlodipine
benzoate
*Average of 5 determinations
Table 6: Calibration data and Molar absoptivity values of amlodipine
Concentration in
In Methanol at 237nm
In 0.1M Sodium benzoate at
Absorbance*
Absorbance*
*Average of 5 determinations
ACKNOWLEDGEMENT:
3. Maheshwari R K, Chaturvedi S C, Jain N K, The authors are thankful to Sun Pharma limited, Application of hydrotropic solubilization phenomenon in spectrophotometric analysis of Bangalore for gift Sample of amalodipine & hydrochlorothiazide tablets. Indian Drugs Management of alwar Pharmacy College for providing other necessary requirement for this work. 4. Maheshwari R K. Application of hydrotropic solubilization in the analysis of aceclofenac. REFERENCES:
5. Maheshwari R K, Chaturvedi S C, Jain N K. 1. RB Kakd, Spectrophotometric method for pharmaceutical dosage forms. Indian Drugs pharmaceutical preparations. Research J. 2. British Pharmacopoeia Vol. 1, Department of Hydrotropic solubilization of nifedipine. Health Social Services and Public Safety, H.M. Stationary Office London, 2004;126. 7. Kasture AV and Ramteke M. Simultaneous Advanced Journal of Pharmacie and Life science Research Adv J Pharm Life sci Res, 2013 1;1:16-21 estimation of atenolol and amlodipine besylate estimation of amlodipine besylate in bulk drug in combined dosage form. Indian J Pharm Sci and their dosage forms by using hydrotropic agent. Eurasian J. Anal. Chem. 2010; 5(3): 8. Saleh A M, Daabis N A. Study of interaction Determination of Simvastatin and Ezetimibe in Tablets by HPLC. E-Journal of Chemistry. spectrophotometric estimation of amlodipine 14. M.E. Zorn. R.D.Gibbons and W.C Sonzongi. 10. Poochikian G D, Cradock J C. Enhanced Evaluation of approximate methods for the chartreusin solubility by hydroxyl benzoate calculating the limit of detection and limit of quantitation. Enviorn Sci., Technol. 1999; 11. Rao JR, Kadam SS, Mahadik K R. RP-HPLC determination of amlodipine and benzepril 15. P.D Sethi. Quantitative analysis of drug in hydrochloride in tablets. Indian Drugs 2000; pharmaceutical formulation third edition CBS publishers and distributors, New Delhi, Page development and validation for quantitative Advanced Journal of Pharmacie and Life science Research

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