UV Spectrophotometric Method Development and Validation of Acotiamide in Bulk and Solid Doage Form
Vinod Matole*, Avinash Birajdar, Suyash Ingle, Sagar Adlinge, Gajanand Nangare, Saili Madur, Smeeta Patil, Anita Shegaonkar
D.S.T.S Mandal’s College of Pharmacy, Solapur-413004, Maharashtra, India.
*Corresponding Author E-mail: matole7414@gmail.com
ABSTRACT:
Objective: A new, simple, sensitive, precise and reproducible UV spectroscopic method was developed for the estimation of Acotiamide in bulk and Tablet Formulation. Methods: The UV spectrum of Acotiamide in water showed λ max at 249 nm. Beer’s law is valid in the concentration range of 3-18µg/ml. This method was validated for linearity, accuracy, precision, ruggedness and robustness. Results: The method has demonstrated excellent linearity over the range of 3-18µg/ml with regression equation y = 0.0523x - 0.0429 and regression correlation coefficient r2= 0.9996. Moreover, the method was found to be highly sensitive with LOD (0.38μg/ml) and LOQ (1.17μg/ml). Conclusion: Depending on results the given method can be successfully applied for assay of Acotiamide in Tablet formulation.
KEYWORDS: Acotiamide, UV spectroscopy, method development and validation, Water Tablet Formulation.
INTRODUCTION:
Acotiamide is a drug manufactured and approved in Japan for the treatment of postprandial fullness, upper abdominal bloating, and early satiation due to functional dyspepsia. It acts as an acetylcholinesterase inhibitor. It is pending approval in the United States; ten clinical trials have been successfully completed2-3.
Figure1: Structure of Acotiamide
The Chemical name of Acotiamide is s (N-[2-[bis(1-methylethyl) amino] ethyl]-2-[(2-hydroxy-4,5-dimethoxybenzoyl) amino] thiazole-4-carboxamide. The molecular formula of Acotiamide is C21H30N4O5S and molecular weight is 250.55gm/mol. It is freely soluble in water, ethanol and methanol. The aim of this study is to give a new, simple, sensitive, precise and reproducible UV spectroscopic method was developed for the estimation of Acotiamide in bulk and Tablet formulation4-6.
MATERIALS AND METHOD:
Materials:
Acotiamide was take as gift sample from Dr. Reddy, Hyderabad. Distilled water was taken of analytical grade.
Instruments:
Analytical balance (Shimadzu AY220), Sonicator (Microclean-1103), UV-Visible spectrophotometer (Systronic 2201).
Experimental:
Preparation of standard stock solution:
Accurately weighed 10mg of Acotiamide transferred to 100ml volumetric flask. It was dissolved in water AND sonicated for 5 minutes. The volume was made up to mark with same diluent to make up final strength.
Procedure for plotting calibration curve:
For calibration curve in a series of 10ml volumetric flasks, 0.3-1.8ml of standard solution was pipetted out separately. The volume was completed to the mark using water. The absorbance was measured at wavelength 249 nm against blank solution.
Analysis of Acotiamide in Tablet formulation:
10 mg equivalent Acotiamide Tablet was weighed and transferred to the 100ml volumetric flask and dissolved in water as a solvent. After that sonicated for 5min and vortex for 2min. 0.8ml of above solution was pipetted out and transferred to the 10ml volumetric flask and make up the volume upto the mark with same solvents and analysed at 249nm. Calculate the % purity of Acotiamide.
Figure 2: UV spectrum of Acotiamide
RESULTS AND DISCUSSION:
The absorption spectrum shows λ max of Dapsone at 249nm.
The proposed method was validated according to ICH Q28 R1 guidelines for validation of analytical procedure.7
Linearity:
Six different concentrations of Acotiamide were prepared and analysed at wavelength 249nm. The regression coefficient was found to be 0.9996. The absorbance was found in limit i.e. 0-2. Hence the analysed parameter was found to be validated (table 1).
Table 1: Results of Linearity
|
Sr. No. |
Concentration(µg/ml) |
Absorbance |
|
1 |
3 |
0.106 |
|
2 |
6 |
0.272 |
|
3 |
9 |
0.431 |
|
4 |
12 |
0.592 |
|
5 |
15 |
0.743 |
|
|
18 |
0.995 |
Table 2: Optimization parameters of Acotiamide
|
Parameters |
Method values |
|
Maximum Wavelength |
249nm |
|
Beer’s Law |
3-18µg/ml |
|
Correlation Coefficient (r2) |
0.9996 |
|
Regression Equation |
y = 0.065x + 0.0353 |
|
Slope (m) |
0.0523 |
|
Intercept (c) |
0.0429 |
Figure 3: Calibration curve for Acotiamide
Accuracy:
The concentration 3,6,9 µg/ml was taken as 50,100,150% and % recovery was found to be in range 99%-101%. Hence the parameter was found to be validated.
Table 3: Results of Accuracy
|
Name of Drug |
Recovery Level in % |
Concentration |
Amount Recovered |
% recovery with SD |
|
|
50 |
3µg/ml |
3.01 |
100.01±0.25 |
|
Acotiamide |
100 |
6 µg/ml |
4.02 |
99.02±0.7 |
|
|
150 |
9 µg/ml |
5.01 |
100.01±0.29 |
Range:
Range is an interval between highest and lowest concentration limit of the analyte i.e. 3-18µg/ml.
Precision:
In precision intra-day and inter-day precision were performed at concentration 9(µg/ml). The obtained results were found within limit i.e. less than 2%RSD.
Table 4: Results of Intra-day Precision
|
Sr. No. |
Concentration |
Absorbance |
|
1 |
9 (µg/ml) |
0.431 |
|
2 |
0.430 |
|
|
3 |
0.430 |
|
|
4 |
0.432 |
|
|
5 |
0.431 |
|
|
6 |
|
0.431 |
|
|
SD |
0.000753 |
|
|
%RSD |
0.174725 |
Table 5: Results of Inter-day precision
|
Sr. No. |
Concentration |
Absorbance (Day1) |
Absorbance (Day2) |
|
1 |
9 (µg/ml) |
0.431 |
0.432 |
|
2 |
0.430 |
0.432 |
|
|
3 |
0.430 |
0.431 |
|
|
4 |
0.432 |
0.431 |
|
|
5 |
0.431 |
0.430 |
|
|
|
|
0.431 |
0.431 |
|
|
SD |
0.000753 |
0.000753 |
|
|
%RSD |
0.174725 |
0.17459 |
Limit of Detection (LOD):
The limit of detection was found to be 0.38 µg/ml (table 6).
Limit of Quantification (LOQ):
The limit of quantification was found to be 1.17µg/ml (table 6).
Table 6: Results of LOD and LOQ
|
LOD |
0.38 µg/ml |
|
LOQ |
1.17µg/ml |
Ruggedness:
The change in analyst with same concentration and environmental condition didn’t affect the results.
Table 7: Results of Ruggedness
|
Concentration |
Absorbance (Analyst1) |
Absorbance (Analyst2) |
|
10µg/ml |
0.485 |
0.486 |
|
0.487 |
0.485 |
|
|
0.484 |
0.487 |
|
|
0.488 |
0.486 |
|
|
0.485 |
0.485 |
|
|
0.486 |
0.485 |
|
|
Average |
0.485833 |
0.485667 |
|
SD |
0.001452 |
0.000812 |
Robustness:
The change in wavelength (249nm and 252nm) and concentration (10µg/ml) didn’t affect the results.
Assay:
The assay was performed by using Acotrust100 Tablet at concentration 8µg/ml. The % purity was found to be 99.8%.
Table 8: Results of Robustness
|
Wavelength |
249nm |
252nm |
|
Concentration |
10µg/ml |
10µg/ml |
|
Absorbance |
0.485 |
0.490 |
|
0.487 |
0.491 |
|
|
0.484 |
0.491 |
|
|
0.488 |
0.492 |
|
|
0.485 |
0.492 |
|
|
0.486 |
0.491 |
|
|
Average |
0.485833 |
0.491167 |
|
SD |
0.001452 |
0.000753 |
Table 9: Results of Assay
|
Formulation |
Labeled Amount |
Amount obtained |
% purity |
|
Acotrust100 Tablet |
100mg |
99.8 |
99.8% |
CONCLUSION:
An analytical UV spectrophotometric method was developed and validated thoroughly for quantitative determination of Acotiamide in bulk drug and Tablet Formulation. The presented method was found to be simple, precise, accurate, rugged, reproducible and gives an acceptable recovery of the analyte, which can be directly easily applied to the analysis of pharmaceutical Tablet formulation of Acotiamide.
ACKNOWLEDGEMENT:
Authors are thankful to the Principal, College of Pharmacy Solapur, for providing the necessary facilities.
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Received on 12.04.2020 Modified on 29.04.2020
Accepted on 21.05.2020 ©Asian Pharma Press All Right Reserved
Asian J. Pharm. Ana. 2020; 10(3): 147-149.
DOI: 10.5958/2231-5675.2020.00026.5