Zero Order and Area under
Curve Spectrophotometric Methods for Determination of Domperidone
in Pharmaceutical Formulation
Audumbar
Digambar Mali1*, Ritesh
Bathe1, Ashpak Tamboli2
1Department of Pharmaceutics, Sahyadri College of Pharmacy, Methwade,
Sangola-413307, Solapur,
Maharashtra,
India.
2Department of Pharmaceutical Chemistry, Sahyadri College of Pharmacy, Methwade,
Sangola-413307,
Solapur, Maharashtra, India.
*Corresponding
Author E-mail: maliaudu442@gmail.com
ABSTRACT:
Simple, fast and reliable spectrophotometry
methods were developed for determination of Domperidonein
bulk and pharmaceutical dosage forms. The solutions of standard and the sample
were prepared in Methanol.The quantitative
determination of the drug was carried out using the zero order derivative
values measured at286nm and the area under the curve method values measured at
279-293 nm. Calibration graphs constructed at their wavelengths of
determination were linear in the concentration range of Domperidone
using 5-25μg/ml (r²=0.999 and r²=0.998) for zero order and area under the
curve spectrophotometric method. All the proposed methods have been extensively
validated as per ICH guidelines. There was no significant difference between
the performance of the proposed methods regarding the mean values and standard
deviations. Developed spectrophotometric methods in this study are simple,
accurate, precise and sensitive to assay of Domperidonein
tablets.
KEY WORDS: Domperidone, UV visible spectrophotometry, AUC, Method Validation, Precission, Accuracy.
1. INTRODUCTION:
Domperidone is chemically known as
5-Chloro-1-[1-[3-(2-oxo-2, 3-dihydro-1H benzimidazol-1-yl)propyl]-4-piperidyl]-1,3-dihydro-2H-benzimidazol-2-one.1,2 Domperidone
is an Antiemetic drug used to treat nausea and vomiting and to stimulate
lactation in women. It is used in the inhibition of receptive relaxation,
causes enhancement of coordinated antral-duodenal
motility and results acceleration of transit in the small intestine.3,4
It stimulates
gastro-intestinal motility and is used as an antiemetic for the short term
treatment of nausea and vomiting of various aetiologies,
including that associated with cancer therapy and with levodopa
or bromocriptine therapy for Parkinsonism.5,6
It is an official drug in IP and BP.
Literature survey revealed several analytical methods UV
spectrophotometry7,8 and HPLC 9
have been reported in bulk, pharmaceutical dosage form for
determination of Domperidone. To our notice, so far
no UV- spectrophotometry method using Zero Order and
Area under Curve (AUC) has been reported for the determination of Domperidonein bulk and tablets. Hence an attempt has been
made to develop new Zero Order and Area under Curve Spectrophotometric methodsmethod for estimation of Domperidonein
bulk and pharmaceutical formulations with good accuracy simplicity, precision
and economy.
Molecular formula: C22H24ClN5O2
Molecular weight: 425.911 g/mol
Fig.1:- The structural formula of Domperidone.
2. MATERIALS AND METHODS:-
2.1. Apparatus and instrumentation
A Shimadzu
1800 UV/VIS double beam spectrophotometer with 1cm matched quartz cells was
used for all spectral measurements. Single Pan Electronic balance (Contech,
CA 223, India) was used for weighing purpose. Sonication of
the solutions was
carried out using
an Ultrasonic Cleaning
Bath (Spectra lab UCB
40, India). Calibrated volumetric glassware (Borosil®) was used for the validation study.
2.2. Materials
Reference
standard of Domperidone API was supplied as gift
sample Zydas Cadila Health Care (Ahmedabad, India).Tablet sample with label claim 10 mg per
tablet were purchased from local market Solapur.
2.3. Method development
2.3.1. Preparation of Standard and Sample Solutions
Stock solution of
10μg/ml of Domperidone was prepared in Methanol for zero
order and area under the curve spectrophotometric analysis. The standard
solutions were prepared by dilution of the stock solution with Methanol in a
concentration range of 5,10,15,20 and 25μg/ml with Methanol. Methanol was
used as a blank solution.
Fig. 2: - Zero order derivative spectrum of Domperidone in Methanol (20µg/ml).
Fig. 3: - UV AUC spectrum of Domperidone
in Methanol (20µg/ml).
2.3.2. Area under curve (Area calculation)
Area under
curve method involves
the calculation of
integrated value of
absorbance with respect to the
wavelength between two selected wavelengths such as λ1 and λ2
representing start and end point of curve region. The area under curve between
λ1 and λ2 was calculated using UV probe software. In this study area
was integrated between wavelength ranges from 279-293nm.
Area calculation: (α+β) =
Where, α is area of
portion bounded by curve data and a straight line connecting the start and end point,
β is the area of portion bounded by a straight line connecting the start and
end point on curve data and horizontal axis, λ1 and λ2 are wavelength
range start and end point of curve region.
10-12
2.3.3. Assay Procedure
Twenty tablets each
containing 10 mg of Domperidone were weighed crushed
to powder and average weight was calculated. Powder equivalent to 10 mg of Domperidone was transferred in 100 ml of volumetric flask.
A 50 ml of Methanol was added and sonicated for 15
minutes. Then solution was further diluted up to the mark with Methanol. The
solution was filtered using Whatmann filter paper no.
41; first 5 ml of filtrate was discarded. This solution was further diluted to
obtain 15µg/ml solution with water subjected for UV analysis using Methanolas blank. Appropriate dilutions were made with
Methanol from stock solution for both zero order and area under the curve
spectrophotometric methods.
Table 1: Assay of tablet dosage form
|
Sr.
No.
|
Sample
Solution Concentration (µg/ml)
|
Amount
found (%)* Zero order derivative
|
Amount
found (%)* AUC
|
Mean
% Found zero order derivative
|
Mean
%
Found
AUC
|
%RSD
zero order derivative
|
%RSD
AUC
|
|
1
|
15
|
99.87
|
100.06
|
|
|
|
|
|
2
|
15
|
102.14
|
100.19
|
100.04
|
100.47
|
0.7813
|
0.9075
|
|
3
|
15
|
98.13
|
101.17
|
|
|
|
|
*n=3,
% RSD = % Relative Standard Deviation.
Fig. 4: - Zero order derivative spectrum of Domperidone
in Methanol (25µg/ml).
Fig. 5: -UV AUC spectrum of Domperidone
in Methanol (25µg/ml).
3. RESULTS AND DISCUSSION
The zero order and area under
the curve spectra for Domperidone were recorded at
the wavelength of 286nm and 279-293 nm respectively.
3.1. Linearity and Range
Under the experimental
conditions described, the graph obtained for zero order and area under the
curve spectra showed linear relationship. Regression analysis was made for the
slope, intercept and correlation coefficient values. The regression equations
of calibration curves were y=0.023x+0.007(r2=0.999) at 286 nm for
zero order derivative spectrophotometry and
y=0.024x+0.014 (r2=0.998) at 279-293 nm for area under the curve spectrophotometry. The range was found to be 5-25μg/ml
for both zero order and area under the curve spectrophotometric methods.
Fig.6: - Linearity of Domperidone by
Absorbance
Fig.7: - Linearity of Domperidone by
AUC.
Table 2: Statistical data for the calibration graphs for
determination of Domperidone by Proposed methods.
|
Parameters
|
Zero
order derivative
|
Area
Under the Curve
|
|
Linearity range (µg/ml)*
|
5-25
|
5-25
|
|
r2
|
0.999
|
0.998
|
3.2. Accuracy
To study the accuracy of the proposed
methods and to check the interference from excipients
used in the dosage forms, recovery experiments were carried out by the standard
addition method. The accuracy for the analytical method was evaluated at 80%,
100% and 120% levels of 15µg/ml standard solution. For Area under curve (AUC)
was measured in wavelength range 279-293 nm and for zero order derivative at
286nm and results were obtained in terms of percent recovery. Three
determinations at each level were performed and % RSD was calculated for each
level.13-15
Fig. 8: - Zero order derivative overlay
of Domperidone at diff. Concentration.
Table 3: Accuracy results for Domperidone
|
Accuracy level
|
Sample conc (µg/)
|
Std. conc
|
Total amnt. Added (µg/m)
|
%Recovery zero derivatie
|
% Recovery AUC*
|
Mean of Zero derivative*
|
Mean of AUC derivative*
|
%
RSD Zero derivative
|
%
RSD AUC
|
|
80
|
15
|
12
|
27
|
102.19
|
99.52
|
|
|
|
|
|
100
|
15
|
15
|
30
|
99.87
|
102.28
|
100.19
|
100.92
|
0.972
|
1.672
|
|
120
|
15
|
18
|
33
|
98.53
|
100.98
|
|
|
|
|
*n=3,
% RSD = % Relative Standard Deviation.
Table 4: Results of Intra and Inter Day Precision
|
Parameters
|
Intra
Day Precision
|
Inter
Day Precision
|
|
S.D*
|
%
RSD*
|
S.D*
|
%
RSD*
|
|
Zero derivative
|
0.72
|
0.8097
|
0.0587
|
0.7148
|
|
Area under the curve
|
0.8219
|
0.7388
|
0.9814
|
1.6780
|
Table 5: Summary of validation parameters
|
Parameter
|
Zero
derivative
|
AUC
|
|
λ range
|
200-400 nm
|
200-400 nm
|
|
Regression Equation (y=mx+c)
|
Y=0.023x+0.007
|
Y=0.024x+0.014
|
|
Measured wavelength
|
286nm
|
279-293nm
|
|
Linearity range
|
5-25µg/ml
|
5-25µg/ml
|
|
Slope
|
0.023
|
0.024
|
|
Intercept
|
0.007
|
0.014
|
|
Correlation coefficient (R2)
|
0.999
|
0.998
|
|
Limit of Detection (LOD) µg/ml
|
0.6479
|
0.9349
|
|
Limit of Quantitation (LOQ) µg/ml
|
1.9442
|
2.8051
|
|
Accuracy (Mean % Recovery)
|
100.19
|
100.92
|
|
Precission (%RSD)
|
0.972
|
1.672
|
3.3. Precision
To determine the precision of
the method, Domperidone solutions at a concentration
of 10μg/ml were analysed each three times for
both zero order and area under the curve spectrophotometric methods. Solutions
for the standard curves were prepared fresh everyday.13-15 (Table-4).
3.4. Sensitivity
The limit of detection (LOD)
and limit of quantification (LOQ) were calculated by using the equations LOD =
3xσ/ S and LOQ = 10xσ/S, where σ is the standard deviation of
intercept, S is the slope. The LOD and LOQ were found to be 0.6479μg/ml
and 1.9442μg/ml respectively for zero order derivative and The LOD and LOQ
were found to be 0.9349µg/ml and 2.8051µg/ml for area under the curve methods
respectively. 13, 14, 15
3.4.5. Analysis of the Marketed Formulation
There was no interference
from the excipients commonly present in the tablets.
The drug content was found to be 99.07% and 101.28%zero order and area under
the curve spectrophotometric methods respectively. It may therefore be inferred
that degradation of Domperidone had not occurred in
the marketed formulations that were analysed by this method.
The low % R.S.D. value indicated the suitability of this method for routine
analysis of Domperidonein pharmaceutical dosage form.
4. CONCLUSION:
No UV or Area under Curve
spectrophotometric methods have been described for the determination of Domperidone. Therefore simple, fast and reliable derivative
spectrophotometric methods were developed for the routine determination of Domperidone. The developed methods can be concluded as
accurate, sensitive and precise and can be easily applied to the pharmaceutical
formulation.
5. ACKNOWLEDGEMENT:
Authors were
thankful to by Zydas Cadila Health
Care (Ahmedabad, India) for providing gift
sample of Domperidone. We would also like to thanks Principal and Management of Sahyadri College of Pharmacy, Methwade,
Sangola, Solapur,
Maharashtra, India for providing all the
facilities to complete this work successfully.
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