RP-HPLC Determination of Atenolol in Its Pharmaceutical Dosage Forms
Amrish Sharma*, Mukul Tailang, Sonali Khare and Tripti Shukla
People’s Institute of Pharmacy and Research Centre, Bhanpur, Bhopal-462 037 (M.P.)
*Corresponding Author E-mail: amrish_sharma21@rediffmail.com
ABSTRACT
KEYWORDS: RP HPLC, Validation, Atenolol.
INTRODUCTION:
Atenolol (C14H22N2O3 and Molecular weight is 266.34) is 4-[2-Hydroxy-3-[(1-methyl ethyl) amino]propoxy] benzeneacetamide1 is a Cardio vascular drug possessing β-adrenergic blockerand antihypertensive activity2. All the b adrenoceptor blocking agents are synthetic compounds and competitively inhibit the actions of adrenergic agonist on b-receptors. It is indicated for the treatment of hypertension, angina pectoris, cardiac arrhythmias, myocardial infraction and CHF. It is official in USP3, BP4and IP5. A survey of literature reveals that it is estimated by HPLC6-7, HPTLC8, Colorimetry9, Spectrophotometry10 and Flourimetry11. It is also studied with some other drugs with HPLC12, GLC13and Spectrophotometrically14.
Chemical structure:
Figure-1: Chemical structure of Atenolol
MATERIAL AND METHODS:
Reagents And Chemicals:
Acetonitrile HPLC grade, Methanol HPLC grade, KH2PO4, Triethylamine.
Instruments:
SHIMADZU HPLC (V.P SERIES), SPD-20 ATVP WITH UV-visible detector.
Hypersil C-18 column, Phenomenex Gemini, 250´4.6mm,5m particle size, Injector- Rheodyne.
Pump LC- 20 AT, Mode of operation -Isocratic, Temperature-Ambient, Flow rate-1ml / min and UV detection at 238nm.STANDARDIZATION OF METHOD
Acetonitrile, Methanol, KH2PO4 were mixed in the ratio of (250 : 250 : 500) and was used as a mobile phase.
Buffer solution was prepared by dissolving 0.05M KH2PO4+0.1% H3PO4 adjunct with Triethylamine.
An accurately weighed 500mg of Atenolol was dissolved in100 ml mobile phase. This gave a concentration of 5000mg/ml of Atenolol. From this solution 4, 4.5, 5, 5.5, 6, ml of the solutions were taken and the volume was made up to 25ml with the mobile phase to get concentration ranges of 800mg/ml, 900mg/ml, 1000mg/ml, 1100mg/ml, 1200mg/ml which represents the range of 80%, 90%, 100%, 110%, 120%. These concentration ranges were used for further work of the analysis.
About 10 tablets were weighed (each tablet contains 50mg Atenolol powder). Powder equivalent to average weight of 10 tablets was taken and dissolved in 100ml of the mobile phase and solution was filtered through what’s man filter paper No. 40. This gave a concentration of 5000mg/ml of Atenolol from this solution 4, 4.5, 5, 5.5, 6, ml of the solutions were taken and the volume was made up to 25ml with the mobile phase to get concentration ranges from 800mg/ml to 1200mg/ml which represents the range of 80%, 90%, 100%, 110%, 120%. 20ml of the standard and sample solutions were injected respectively and the scans were recorded. Each solution was run five times at an interval of 15 min, to ensure elution of earlier injection.
The method is found to be specific because it doesn’t shows any interference with placebo solutions.
Varying quantities of the standard stock solution was diluted with the mobile phase to give concentration of 80, 90, 100, 110, and 120 % of the Atenolol. The injections were made at an interval of 15 min and peak area was determined. A calibration curve was determined by plotting the peak areas obtained against concentrations. There exists a linear relationship showing concentrations ranging from for Atenolol 800mg/ml to 1200mg/ml. From the data obtained, correlation coefficient for the Atenolol was found 0.99.
Precision:
Repeatability expresses the precision under the same operating conditions. Responses to the repeated injection of the same standard solution should have R.S.D< 1.0%. It is also termed as intra-assay precision. Injection of sample from each solution should have R.S.D< 2.0%. The R.S.D. in the present experimentation was found to be 0.17% and for system precision it was found to be 0.3% which indicates the precision and accuracy of the method.
The number of theoretical plates was 3153 and tailing factor was 1.5. The retention time was 4 min for the developed RP HPLC. The number of the theoritical plates was high indicating the efficient performance of the column.
Recovery Studies:
Determination of accuracy by direct comparison to reference standard is a preferred technique. Recovery studies were performed by spiking the blank matrix of the sample at different levels (80%, 100%, 120%) of the known level in the sample. Average recovery of the analyte was found to be in the range of 99.0-102% at different levels of spiking.
Table 1 : RESULTS WITH ACCEPTANCE CRITERIA
|
S. No |
Parameter |
Acceptance criteria |
Results obtained |
|
|
1 |
Specificity |
Should not interfere with the placebo |
Passes |
|
|
2 |
Linearity |
Correlation cofficient not less than 0.99 |
0.99 |
|
|
3 |
Precision |
R.S.D <1.0% for system precision |
0.3 % |
|
|
R.S.D<2.0% for method precision |
0.17 %
|
|||
|
4 |
Accuracy |
R.S.D for % recovery at each accuracy level not more than 2.0% |
Accuracy with concentration |
R.S.D |
|
80% |
1.13% |
|||
|
100% |
0.2% |
|||
|
120%
|
0.02%
|
|||
|
Recovery of the spiked drug (98-102%) |
concentration |
% Recovery |
||
|
80% |
100.45% |
|||
|
100% |
99.89% |
|||
|
120% |
100.88% |
|||
RESULTS AND DISCUSSION:
The developed reverse phase HPLC method utilizes acetonitrile, methanol and KH2PO4 in ratio of (250:250:500) adjunct with triethylamine as a mobile phase and column-phenomenex Gemini C18 (5µ-250×4.6mm) as a stationary phase. The method precision and system precision were performed and found to be within the limits. The recovery study reveals the accuracy and precision of the method employed for the present studies.
CONCLUSION:
It is clear from the present study that the prescribed method of analysis is simple, accurate, Specific and precise in operation and can be employed for routine batch analysis of Atenolol tablets.
Authors are thankful to Cadila Pharmaceuticals ltd. Ahmedabad for providing gift samples of the pure drug Atenolol.
REFERENCES:
1. Gov. of India ministry of Health and Family Welfare “Indian Pharmacopoeia” 1996 Vol-I, PP-145.
2. Barar, F.S.K. Essentials of Pharmacotherapeutics, Third edition, 2000, PP: 211-212.
3. United States Pharmacopoeia,22 nd Edition 1996.
4. British pharmacopoeia, , volume-1, 1998 PP.122.
5. Gov. of India ministry of Health and Family Welfare “Indian Pharmacopoeia” volume-I, 1996, PP: 72.
6. Avadhanulu, A.B; srinivas, K.S; Anjaneyulu; Indian Drugs 1996, PP-33-40.
7. Patki, R.V; Tambankar, C.D; Tipnis, H.P; Indian Drugs, 1994, PP-31(12), 33-40.
8. Chandrashekar, T.G; Rao, P.S.N; Smrita, K; Vyas, SK. Dutt, C; Analytical abstract, vol. 57, 1995, PP- 4972.
9. Zakhari, N.A; Hassan, S.M; Shabrawy, Y; Analytical abstract, vol-54, 1992, PP-6G78.
10. Wang, S; Analytical abstract, , vol. 53, 1991, PP-4040.
11. Zhao, H.C; Yan, N.H; Guo, J.X; Analytical abstract, , vol.57, 1995, PP-9G67.
12. Jain, R; Jain, C.L; Analytical Abstract, , vol-55, 1993, PP-5G38.
13. Erram, S.V; Tipnis, H.P; Indian Drugs, 1992, PP-436-438.
14. Rao,G.R; Avadhanulum A.B; Giridhar,R; Pantulu, A.r Kokate, C.K; Analytical abstract, vol, 53, 1991, PP-8G85.
Received on 30.07.2009 Modified on 27.09.2009
Accepted on 24.10.2009 © AJRC All right reserved
Asian J. Research Chem. 2(4):Oct.-Dec. 2009 page 485-487