DIFFERENCE SPECTROSCOPIC METHOD FOR THE ESTIMATION OF ACEBUTOLOL HYDROCHLORIDE IN BULK AND IN ITS FORMULATION

{ DOWNLOAD AS PDF }

ABOUT AUTHORS:
Jadhav Santosh*, Mali Audumbar, Pawar Seemarani, Kharat Rekha, Tamboli Ashpak
Department of Pharmaceutics, Sahyadri College of Pharmacy, Methwade,
Sangola-413307, Solapur, Maharashtra, India.
*jadhavsan88@gmail.com

ABSTRACT:
A simple, precise and accurate difference spectroscopic method has been developed for the estimation of Acebutolol Hydrochloride in bulk drug form by difference spectrophotometric method. Acebutolol Hydrochloride has exhibited maximum absorbance at about 233nm and 234nm in acidic and basic solution respectively. Beer’s law was obeyed in the concentration range of 2-10 µg/ml in both the cases. The regression of coefficient was found to be r²=0.9992. The LOD and LOQ value were found to be 0.2670ppm and 0.8091ppm respectively. The proposed method was successfully applied for the determination of Acebutolol Hydrochloride in bulk drug. As per ICH guidelines the result of the analysis were validated statistically and were found to be satisfactory.

REFERENCE ID: PHARMATUTOR-ART-2308

INTRODUCTION:
Chemically acebutolol hydrochloride is a(N-[3-Acetyl-4-[2-hydroxy-3[(1-methylethyl) amino] propoxy] phenyl] butanamide) hydrochloride is a cardio-selective betablocker used in the management of hypertension, angina pectoris and cardiac arrhythmias. Acebutolol hydrochloride (Fig.1.) is a cardioselective, hydrophilic β-adrenoreceptor blocking agent with mild intrinsic sympathomimetric activity (ISA) for use in treating patients with hypertension and ventricular arrhythmias^[1].

Molecular Formula: C18H29ClN2O4.

Molecular weight: 372.9 g/mole

Objective:
Acebutolol hydrochloride shows improved absorbing interference by the technique of different spectrophotometry. Thus the objective of the present study was to develop new analytical difference spectrophotometry method and its validation parameters for the proposed method according to ICH guidelines for the estimation of Acebutolol hydrochloride bulk drug ^{[2, 3, 4]}.

MATERIALS AND METHODS:

Chemical and reagents:
Acebutolol hydrochloride [bulk drug] used were of analytical reagent grade purchased from research lab fine chem. industries Mumbai, India, NaOH and  HCL were purchased from Poona chemical laboratory and double distilled water was used throughout the analysis.

Instrumentation:
A shimadzu 1800 UV/VIS double beam spectrophotometer with 1cm matched quartz cells was used for all spectral measurements.

Selection of common solvents:
1N HCL and 1N NaOH were selected as a common solvent for developing spectral characteristics of drug.

Preparation of solution:
Standard stock solution containing Acebutolol hydrochloride was prepared by dissolving 10mg in 100ml of methanol and then diluted with 1N NaOH and 1N HCL separately to get series of dilution ranging from 2-10 µg/ml and then absorbance recorded at 233 nm and 234 nm respectively against reagent blank. Calibration curve was prepared by plotting concentration versus difference in absorbance and found to be linear in the concentration range of 2-10 µg/ml ^{[2, 3 , 4]}.

VALIDATION^{[2, 5, 6, 7, 8, 9]}.
The proposed method was validated according to ICH (Q2) R1 guidelines for validation of analytical procedures. As per the ICH guidelinesthe method validation parameters checked were Selectivity, linearity, precision and accuracy.

Selectivity:
The selectivity of the method was assessed by analyzing standard drug, and pharmaceutical product, comparing the maxima and minima of the standard with that of the sample to determine whether the pharmaceutical product and excipient lead to interfere in the estimation.

Limit of Detection and Limit of Quantification:
The Limit of Detection (LOD) is the smallest concentration of the analyte that gives the measurable response. LOD was calculated using the following formula

LOD = 3.3 σ /S

The Limit of Quantification (LOQ) is the smallest concentration of the analyte, which gives response that can be accurately quantified. LOQ was calculated using the following formula

LOQ = 10 σ/S

Where, σ is standard deviation of the response and
S is the slope of the calibration curve.

LOD& LOQ of Acebutolol hydrochloride was found to be 0.2670µg/ml & 0.8091µg/ml respectively.

Linearity:
Different volumes of stock solutions were suitably diluted with corresponding medium ( 2,4,6,8, and 10 µg/ml) to get the desired concentrations. Each solution was analyzed in triplicate. The amplitude values were plotted against the corresponding concentrations to obtain the linear calibration curve^{[10, 11, 12, 13]}.

Table 1: Linearity of Acebutolol hydrochloride by difference spectrophotometry

Figure-3-Showing linearity of Acebutolol

Range:
2-10 µg/ml.

Precision:
Precision of analytical methods were expressed in relative standard deviation (RSD) of a series of measurements. The intra-day and inter-day precisions of the proposed methods were determined by estimating the corresponding responses (i.e. three concentrations / three replicates each) of the sample solution on the same day and on three different days respectively.Precision was calculated as inter-day and intra-day coefficient of variation ^[14].

1N HCL

1N NaoH

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT editor-in-chief@pharmatutor.org

Subscribe to Pharmatutor Alerts by Email

FIND OUT MORE ARTICLES AT OUR DATABASE

Accuracy:
The accuracy of the method was determined by recovery experiments. A known amount of standard Acebutolol hydrochloride corresponding to 2, 4, 6 and 8, 10% of the label claim(standard addition method)was added to preanalysed sample of tablet. The recovery studies were carried out in triplicate at each level ^[15].

RESULT AND DISCUSSION:
The optical characteristics such as Beer’s law limits, percent relative standard deviation and percent range of error were found to be within the limit and satisfactory. All of the analytical validation parameter for the proposed method was determined according to ICH guidelines.The method was found to provide high degree of precision and reproducibility.

The recovery studies showed that the result were within the limit indicating no interference. The proposed method is simple, sensitive, accurate and precise and can be successfully employed for the routine analysis of the Acebutolol hydrochloride in bulk drug.

CONCLUSION:
The proposed method is simple, accurate, precise and selective for the estimation of Acebutolol hydrochloride in bulk drug. The method is economical, rapid and do not require any sophisticated instruments contrast to chromatographic method. it can be effectively applied for the routine analysis of Acebutolol hydrochloride in bulk drug.

ACKNOWLEDGEMENT:
The authors sincerely thanks to Sahyadri College of Pharmacy, Methwade, Sangola, Solapur, Maharashtra, India for providing experimental facilities to carry out this work.

REFERENCES:
1. Topale PR, Gaikwad NJ and Tajane MR. Indian Drugs. 2003; 40:19-121.
2. Jin Y, Chen H, Gu S, Zeng F.; Determination of aceclofenacin human plasma by reversed-phase high performance liquid chromatography. Chinese J. Chromatography. 2004; 22(3), 252 – 254.
3. Srinivasan KK, Shirwaikar A., Joseph A., Jacob S., Prabu SL. Simultaneous estimation of aceclofenac and paracetamol in solid dosage form by ultraviolet spectrophotometry. Indian Drugs. 2006; 43(2): 141 – 145.
4. Shanmugam S., Cednil Kumar A., Vetrichelvan T., Manavalan R., Venkappyya D., Pandey VP. Spectrophotometric method for estimation of aceclofenac in tablets. Indian Drugs. 2005; 42(2): 106 – 107.
5. Validation of analytical procedure: text and methodology, ICH Harmonized Tripartite Guideline, Q2 (R1), 2005; 1-3
6. N. sultana, M.S. Arayne, S.Shamim, M.Akhtar and S.Gul.,J.BrajChem Soc., 2011, 22(5),987.
7. Practical pharmaceutical chemistry by A.H. Beckett, J.B.Stenlake. Fourth edition Part-second 293-296.
8. International Conference on Harmonisation. Draft Guideline on Validation of Analytical Procedures: Definitions and Terminology, Federal Register, 1995; 60, 11260.
9. Raja RK, Sankar GG, Rao AL and SeshagiriRao JVLN. Development and Validation of RP HPLC method for the estimation of Aceclofenac in Tablet Dosage form. Indian Drugs. 2005; 42(10): 693 – 695.
10. F.W. Fifield and D. Kealey, 5th Edition, Black Well Science Ltd. Principles and Practice of Analytical Chemistry, 2000, 270 – 276.
11. G.C. Hokanson, A Life Cycle Approach to the Validation of Analytical Methods During Pharmaceutical Prod-uct Development, Part – II: Changes and the Need for Additional Validation, Pharm. Tech., 1994, 92 -100.
12. J.M. Green. A Practical Guide to Analytical Method Validation, Anal. Chem. News and Features, 305A–309A, 1996
13. J. Vessman, Selectivity or Specificity? Validation of Ana-lytical Methods from the Perspective of an Analytical Chemist in the Pharmaceutical Industry, J. Pharm and Biomed. Anal., 1996, 14, 867 – 869.
14. BaokarShrikrishna, PawarVinod, Sonawane S.H., High Performance Liquid Chromatographic Method Development and Validation of Cholesterol Inhibitor Drug.Journal of Pharmacy Research, 2011, 4(7), 2313-2316
15. Shrikrishna B. Baokar, B. Shirke, V. Sivanand, G.K. Pratheesh, Analytical method development and validation for esteemation of sildenafil citrate from tablet dosage form by RP-HPLC, Int. J. Res. Phar.Sci. 2011, 2(2), 130-136.

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT editor-in-chief@pharmatutor.org

Subscribe to Pharmatutor Alerts by Email

FIND OUT MORE ARTICLES AT OUR DATABASE