Simultaneous UV Spectrophotometric
Methods for Estimation of Ranitidine and Domperidonein
Bulk and Tablet Dosage Form
Audumbar Mali1*, Sujata Kolekar2, Jija
Franklin2, Ritesh Bathe1
1Department of Pharmaceutics, Sahyadri
College of Pharmacy, Methwade, Sangola-413307, Solapur, Maharashtra, India.
2Department
of Quality Assurance, Shri D.D. Vispute
college of Pharmacy and Research Centre,
Devad-Vichumbe, New Panvel,
Raigad-410206, Maharashtra, India.
*Corresponding Author E-mail: maliaudu442@gmail.com
ABSTRACT:
Two
simple, precise, economical, fast and reliable UV methods have been developed
for the simultaneous estimation of Ranitidine and Domperidone
in bulk and pharmaceutical dosage form. Method A is Absorbance maxima method,
which is based on measurement of absorption at maximum wavelength of 314 nm and
286 nm for Ranitidine and Domperidone respectively.
Method B is area under curve (AUC), in the wavelength range of 269-340 nm for Ranitidine
and262-307nm for Domperidone. Linearity for detector
response was observed in the concentration range of 5-25μg/ml for
Ranitidine and 5-25 μg/ml for Domperidone. The accuracy of the methods was assessed by
recovery studies and was found to be 99.18 % and 100.21 % for Ranitidine and
100.19 % and 101.57 % Domperidone by using method A
and B respectively. The developed method was validated with respect to
linearity, accuracy (recovery), precision and specificity. The results were
validated statistically as per ICH Q2 R1guideline and were found to be
satisfactory. The proposed methods were successfully applied for the
determination of for Ranitidine and Domperidone in
commercial pharmaceutical dosage form.
KEY WORDS: Ranitidine, Domperidone,
Simultaneous estimation, Precision, Absorbance maxima method, Area under curve.
1. INTRODUCTION:
Ranitidine is a Ranitidine is chemically Dimethyl
[(5-{[(2-{[(E)-1-(methyl amino)-2 nitro ethenyl]
amino} ethyl) sulfanyl] methyl} furan-2-yl) methyl]
amine. The H2 antagonists are competitive inhibitors of histamine at the
parietal cellH2 receptor. They suppress the normal secretion of acid by
parietal cells and the meal-stimulated secretion of acid.
They accomplish this by two mechanisms: histamine released by cells in
the stomach is blocked from binding on parietal cell H2 receptors which
stimulate acid secretion and other substances that promote acid secretion (such
as gastrin and acetylcholine) have a reduced effect
on parietal cells when the H2 receptors are blocked.
Fig. 1: Chemical structure of
Ranitidine
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. 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. 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. [2]
Fig. 2: chemical structure of Domperidone
A survey of pertinent literature revealed that in estimation of
individual [3] as well as
combination of Ranitidine and Domperidone. Simultaneous determinations of Ranitidine and Domperidone dosage form were also reported like
HPLC [4], RP-HPLC [5, 6, 7, 8],
HPTLC [9, 10, 11] and
UV-Spectroscopy [12-18]. Therefore
an attempt was made to develop a new rapid and sensitive UV Spectrophotometric
method and to validate as per ICH-guidelines. A comprehensive literature
research reveals the lack of a Spectrophotometric analytical method for
simultaneous estimation of Ranitidine and Domperidone in pharmaceutical formulations. A successful attempt was made to
develop accurate, precise and simple method of analysis for estimation of both
the drugs in combined dosage form.
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 Ranitidine
and Domperidone API was supplied as gift sample by Lupin
Laboratory Park Aurangabad, Maharashtra, India. The commercial
formulation Ranidone has purchased from the local market Solapur, Maharashtra, India.
2.3
Method development: -
2.3.1 Preparation of standard stock solution: -
Stock solution was prepared by diluting
10 mg of each drug in sufficient quantity of methanol in separate volumetric
flask and volume was made up to 100 ml to get the concentrations of
100μg/ml for each drug. Dilutions from stock solution were prepared in the
range of 5-25 μg/ml for Ranitidine and 5-25 μg/ml for Domperidone. Methanol was used as a blank solution.
2.3.2 Method A: Absorption Maxima Method: -
For the selection of analytical wavelength, standard solution of Ranitidine and
Domperidonewere
scanned in the spectrum mode from 400 nm to 200 nm separately. From the spectra
of drug λmax of Ranitidine 314 nm [Fig.3], and λmax
of Domperidone, 286 nm [Fig.4], were selected for the analysis.
Aliquots of standard stock solution were made and calibration curve was
plotted. [19-22]
Fig. 3: It shows λmax of
Ranitidine
Fig. 4: It shows λmax of
Domperidone
2.3.3 Simultaneous estimation of Ranitidine and Domperidone:
The wavelength maxima of Ranitidine and Domperidone were determined and found to be 314 nm (λ1) and 286nm (λ2)
respectively where there was no interference among the drugs. The overlain
spectrum is shown in Fig.5.
Fig.5 Isobestic point of
Ranitidine and Domperidone
2.3.4 Method B: Area under Curve Method:
From the spectra of drug obtained after scanning of standard solution
of Ranitidine and Domperidone separately, area under the curve in the range of 269-340 nm and 262-307
nm was selected for the analysis. The calibration curve was prepared in the
concentration range of 5-25 μg/ml for Ranitidine and 5-25 μg/ml for Domperidone at their respective AUC range. Both drugs followed
the Beer-Lambert’s law in the above mentioned concentration range. The
calibration curves were plotted as absorbance against concentration of Ranitidine and Domperidone. The coefficient of correlation (r), slope and intercept values of
this method are given in Table 2.
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. [19-22]
2.3.5 Application of the proposed
methods for the determination of Ranitidine and Domperidone in tablet dosage form:
For the estimation of drugs in the tablet formulation, 20 tablets were
weighed and weight equivalent to 150mg of Ranitidine and 10mg of Domperidone was transferred to 100 ml volumetric flask and ultra sonicated for 20 minutes and volume was made up to the mark
with methanol. The solution was then filtered through a Whatmann
filter paper (No.42). The filtrate was appropriately diluted further.
Relative
Standard Deviation:
In Method-A, the concentration of Ranitidine and Domperidone was determined by measuring the
absorbance of the sample at 314 nm and 286 nm respectively in zero order
spectrum modes. By using the calibration curve, the concentration of the sample
solution was determined.
In Method-B, the concentration of Ranitidine and Domperidone was determined by measuring area under curve in the
range of 269-340 nm and 262-307nm. By using the calibration curve, the
concentration of the sample solution was determined.
Table 1: Table shows Results of
Analysis of Tablet Formulation
|
Method
|
Drug
|
Label
Claim
mg
|
Sample
Solution Concentration (µg/ml)
|
Amount
found (%)*±
|
% Recovery
|
%RSD
|
|
A
|
Ranitidine
|
150 mg
|
20
|
100.19±1.13
|
99.18
|
0.7851
|
|
B
|
Ranitidine
|
150 mg
|
20
|
102.24±0.17
|
100.21
|
|
A
|
Domperidone
|
10 mg
|
20
|
99.15±1.49
|
100.19
|
0.8021
|
|
B
|
Domperidone
|
10 mg
|
20
|
101.58±1.37
|
101.57
|
*n=3, % RSD = %
Fig.6: It shows AUC of Ranitidine
Fig.7: It shows AUC of Domperidone
3. Validation of the developed methods: [23-27]
The methods were validated with respect to accuracy, linearity,
precision and selectivity.
3.1 Accuracy:
Accuracy of an analysis was determined by systemic error involved.
Accuracy may often be expressed as% Recovery by the assay of known, added
amount of analyte. It is measure of the exactness of
the analytical method. Recovery studies carried out for both the methods by
spiking standard drug in the powdered formulations80%, 100%, 120% amount of
each dosage content as per ICH guidelines.
3.2 Linearity:
The linearity of measurement was evaluated by analyzing different
concentration of the standard solution of Ranitidine and Domperidone. Result should be expressed in terms of correlation
co-efficient.
Fig.8:
Calibration curve for Ranitidine
at 314 nm
Fig.9: Calibration curve for Domperidone at 286 nm
3.3 Precision:
The reproducibility of the proposed method was determined by performing
tablet assay at different time intervals (morning, afternoon and evening) on
same day (Intra-day assay precision) and on three different days(Inter-day
precision). Result of intra-day and inter-day precision is expressed in % RSD.
3.4 Sensitivity: -The limit of detection (LOD) and limit of quantitation (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 0.4892 μg/ml and 1.4678μg/ml respectively of Ranitidine and 0.4251
μg/ml and 1.2759 μg/ml
of Domperidone.
Table 2: Optical Characteristics and Precision
|
Sr. No.
|
Parameter
|
Ranitidine
|
Domperidone
|
|
1
|
λ range
|
200-400 nm
|
200-400nm
|
|
2
|
Regression Equation (y=mx+c)
|
Y=0.024x+0.0002
|
Y=0.023x+0.012
|
|
3
|
Measured wavelength
|
314 nm
|
286 nm
|
|
4
|
Linearity range
|
5-25µg/ml
|
5-25µg/ml
|
|
5
|
Slope
|
0.024
|
0.023
|
|
6
|
Intercept
|
0.0002
|
0.012
|
|
7
|
Correlation coefficient (R2)
|
0.999
|
0.998
|
|
8
|
Limit of Detection (LOD) µg/ml
|
0.4892
|
0.4251
|
|
9
|
Limit of Quantitation (LOQ)µg/ml
|
1.4678
|
1.2759
|
Table 3: Results of drug content and
analytical recovery of Ranitidine
and Domperidone
|
Excess drug added to the analyte (%)
|
Drug
|
% Recovery
|
% RSD
|
|
Method A
|
Method B
|
Method A
|
Method B
|
|
80
|
Ranitidine
|
98.46
|
100.24
|
0.5571
|
0.5368
|
|
100
|
100.21
|
102.36
|
0.4840
|
0.8847
|
|
120
|
99.03
|
101.89
|
0.6854
|
0.6713
|
|
80
|
Domperidone
|
98.34
|
101.46
|
0.8681
|
0.5892
|
|
100
|
102.32
|
102.29
|
0.7743
|
0.9851
|
|
120
|
100.65
|
100.21
|
0.6354
|
0.5893
|
|
|
|
|
|
|
|
Table
4: Results of Intra-day and Inter-day Precision
|
Method
|
Drug
|
Intra-day Precision
|
Inter-day Precision
|
|
SD
|
%RSD
|
SD
|
%RSD
|
|
A
|
Ranitidine
|
0.9961
|
0.6694
|
0.8621
|
0.4589
|
|
B
|
0.9624
|
0.5452
|
0.7264
|
0.4530
|
|
A
|
Domperidone
|
0.9476
|
0.8743
|
0.5237
|
0.3843
|
|
B
|
0.6842
|
0.7963
|
0.4439
|
0.4593
|
4. RESULTS AND DISCUSSION:
The methods discussed in the present work provide a convenient and
accurate way for analysis of Ranitidine
and Domperidone in its bulk and pharmaceutical
dosage form. Absorbance maxima of Ranitidine
at 314 nm and Domperidoneat 286 nm were selected for the analysis. Linearity
for detector response was observed in the concentration range of 5-25 μg/ml for Ranitidine
and 5-25 μg/ml for Domperidone. Percent amount found for Ranitidine
and Domperidone in tablet analysis was found in
the range of 100.19 %, 102.24 % and 99.15 %, 101.58 % respectively [Table 1].
Standard deviation and coefficient of variance for three determinations of
tablet formulation was found to be less than ± 2.0 indicating the precision of
the methods. Accuracy of proposed methods was ascertained by recovery studies
and the results are expressed as %recovery. % recovery for Ranitidine and Domperidone was found in the range of 98.46 % and 98.34 % values
of standard deviation and coefficient of variation was satisfactorily low
indicating the accuracy of all the methods.% RSD for Intraday assay precision
for Ranitidine was found to be
0.6694 and 0.5452 for Method A and B, and for Domperidone0.8743 and 0.7963
for Method A and B. Interday assay precision for Ranitidine was found to be 0.4589 and 0.4530for Method A and B and
for Domperidone 0.3843 and 0.4593 for Method A and
B. The LOD and LOQ were found to be 0.4892 μg/ml
and 1.4678μg/ml respectively of Ranitidine
and 0.4251 μg/ml
and 1.2759 μg/ml of Domperidone. Based on the
results obtained, it is found that the proposed methods are accurate, precise,
reproducible and economical and can be employed for routine quality control of Ranitidine and Domperidone in bulk drug and its pharmaceutical dosage form.
5. CONCLUSION:
UV spectrophotometric methods for Ranitidine and Domperidone were developed separately in bulk
and tablet dosage form by, Absorbance maxima method and Area under curve
method. Further, UV Spectrophotometric methods for the simultaneous estimation
of Ranitidine and Domperidone were in bulk and combined dosage form. The methods were validated as
per ICH guidelines. The standard deviation and % RSD calculated for these
methods are <2, indicating high degree of precision of the methods. The
results of the recovery studies showed the high degree of accuracy of these
methods. In conclusion, the developed methods are accurate, precise and
selective and can be employed successfully for the estimation of Ranitidine and Domperidone in bulk and pharmaceutical dosage form.
6. ACKNOWLEDGEMENT:
The authors are highly thankful to the Sahyadri
College of Pharmacy, Methwade, Sangola,
Solapur, Maharashtra, India
for proving all the facilities to carry out the research work successfully.
7. REFERENCES:
1.
Audumbar Digambar Mali, Patil Manoj Kumar. Estimation of Ranitidine in Bulk and
Formulation by First Order Derivative Area under Curve UV-Spectrophotometry
Methods. Asian
J. Pharm. Ana. 2015; 5(2): 41-46.
2.
Pallavi
Salve, Deepali Gharge, Rupali Kirtawade, Pandurang Dhabale, Kishor Burade. Simple Validated
Spectroscopic Method for Estimation of Ranitidine From Tablet Formulation.
International Journal of Pharm Tech Research. 2010;
2(3): 2071-2074.
3.
Amul N. Mishra and A. C. Rana. UV Spectrophotometric Determination of Ranitidine
Hydrochloride in Pure and Pharmaceutical Formulation. Int. J. Chem. Sci. 2009;
7(3): 2208-2210.
4.
Ho c, Huang HM and Hus SY: Simultaneous HPLC analysis of famotidine ranitidine hydrochloride, cimetidine
nizatidine in commercial products. Drug Develop. Ind Pharm1999; 25:379-85.
5.
M. Naresh, T. Ganesh,
A. Biswal and G. Nagarjuna
Reddy. RP-HPLC method development and
validation studies of Ranitidine HCL and Domperidone.
International Journal of Chemical and Natural Science. 2013; 1(1): 25-28.
6.
Md. Ahsanul Haque,
Mohammad Shahriar, Most. Nazma
Parvin. Validated RP-HPLC Method for Estimation of Ranitidine
Hydrochloride, Domperidone and Naproxen in Solid
Dosage Form. Asian J. Pharm. Ana. 2011; 1(3): 59-63.
7.
Kanumula GV, Bhanu R. Simultaneous
determination of ranitidine hydrochloride and domperidone
in pharmaceutical dosage form by RPHPLC. Indian Drugs 2000; 37: 375-378.
8.
Md. Shozan Mondal,
Md. Ahsanul Haque, Mohammad
Safiqul Islam. Development and Validation of RP-HPLC
Method for the Simultaneous Estimation of Domperidone
and Naproxen in Tablet Dosage Form. Journal of Applied Pharmaceutical Science. 2011;
1(7): 145-148
9.
Nanda, R. K. Potawale, S. E.
Bhagwat, V. V. Deshmukh, R.
S. Deshpande, P. B. Development and validation of a
HPTLC method for simultaneous Densitometric analysis of Ranitidine
hydrochloride and Dicyclomine hydrochloride as the
bulk drugs and in the tablet dosage form. Journal of Pharmacy Research. 2010;
3(8): 1997.
10. Abhay R. Shirode,
Amruta M. Jadhav, Vilasrao J. Kadam. HPTLC Method
for Estimation of Domperidone in Bulk and Marketed
Formulation. Asian
Journal of Pharmaceutical Analysis and Medicinal Chemistry. 2015; 3(2): 52- 61.
11. Anita
B. Kadam, Shweta S. Havele, Sunil R. Dhaneshwar.
Validated HPTLC Method for Simultaneous Estimation of Lafutidine
and Domperidone in Bulk Drug and Formulation. Int. J. Pharm. Pharm. Sci. 2012; 4(4):
221-225.
12. Mali Audumbar, Aasha Wagmode, Hake Gorakhnath, Tamboli Ashpak. Zero Order and Area under Curve Spectrophotometric
Methods for Determination of Ranitidine in Pharmaceutical Formulation. Inventi Rapid: Pharm
Analysis and Quality Assurance. 2015; 2: 1-5.
13. S. Pillai
and I. Singhvi. Spectrophotometric Simultaneous Estimation of Ranitidine Hydrochloride
and Ondansetron hydrochloride from Tablet Formulation. Indian Journal of
Pharmaceutical Sciences.2007; 69(4):
601-604.
14. Audumbar Digambar Mali. Estimation
of Ranitidine in Bulk and Formulation by Second Order Derivative Area under
Curve UV-Spectrophotometric Methods. International
Journal of Analytical, Pharmaceutical and Biomedical Sciences. 2015; 4(6): 7-16.
15. Audumbar Digambar Mali, Ritesh Bathe, Ashpak Tamboli. Zero Order and Area under Curve Spectrophotometric
Methods for Determination of Domperidone in
Pharmaceutical Formulation. Asian Journal of Pharmaceutical Technology. 2015;
5(3): 129-134.
16. Ashish K. Jain, Narendra Lariya, Abhinav Agarwal, Anil K. Kharya, Govind P. Agrawal. Development of
Simultaneous Spectrophotometric Method of Amoxicillin Trihydrate
and Ranitidine Bismuth Citrate in Same Dosages Form. Int. J. Chem. Res. 2010; 1(1): 41-45.
17. Audumbar Digambar Mali.
Estimation of Domperidone in Bulk and Formulation by
First Order Derivative Area under Curve UV-Spectrophotometry Methods. International
Journal of Pharmacy and Technology. 2015; 7(1): 8040-8048.
18. Vaibhav R. Mandale, Dr. Subhash S. Palekar, Dr. Prakash T. Sangale, Dr. Tauqueer Kadu, Dr. Atul S. Sayare, Archana M. Karnik. Development
and Validation of UV Spectrophotometric Method for the Simultaneous Estimation
of Ranitidine and Ondansetron in Bulk and Pharmaceutical Formulation. World
Journal of Pharmaceutical Research.2015; 4(2): 1093-1101.
19. Mali
Audumbar, Jadhav Santosh, Tamboli Ashpak, Bathe Ritesh, Patil Manoj. Development and
Validation of UV-Spectrophotometric Method for the Estimation of Quinapril Hydrochloride in Bulk and In Its Formulation. World Journal of Pharmaceutical Research. 2015; 4(1): 1839-1846.
20. Mali
Audumbar Digambar, Jadhav Santosh, Tamboli Ashpak, and Kargane Vijay. Development and Validation of UV Spectrophotometric Estimation of Lisinopril Dihydrate In Bulk And
Tablet Dosage Form Using Area Under Curve Method. World
Journal of Pharmacy and Pharmaceutical Sciences. 2014; 4(2): 589-596.
21. Mali Audumbar, Jadhav Santosh, Hake Gorakhnath, Tamboli Ashpak. Zero Order and Area under Curve Spectrophotometric
Methods for Determination of Fluoxetine Hydrochloride
in Pharmaceutical Formulation. Pharmatutor
Magazine. 2015; 3(6): 49-54.
22. Mali
Audumbar Digambar, Hake Gorakhnath,
Bathe Ritesh. Zero
Order and Area under Curve Spectrophotometric Methods for Determination of
Aspirin in Pharmaceutical Formulation. Indian E-Journal of Pharmaceutical
Sciences.2015; 1(1): 40-44.
23. Audumbar Digambar Mali, Ritesh
Bathe and Ashpak Tamboli. Simultaneous Determination of Cefixime
Trihydrate and Ofloxacin in
Pharmaceutical Dosage Form by Second Order Derivative UV Spectrophotometry. International Journal of Advances in Pharmaceutical
Analysis. 2015; 5(2):
42-46.
24. Mali Audumbar and Gavade Shivaji. Estimation of carvedilol
in bulk and formulation by second order derivative area under curve UV
spectrophotometric methods. International Journal of Pharmaceutical and
Interdisciplinary Sciences. 2015; 1(2): 1-13.
25. Audumbar Digambar Mali.
Simultaneous Determination Of Carvedilol And
Hydrochlorothiazide In Pharmaceutical Dosage Form By First Order Derivative UV
Spectrophotometry. International
Journal of Pharmacy and Pharmaceutical Sciences.2015;
7(9): 371-374.
26.
Audumbar Digambar Mali, Ritesh Bathe, Manojkumar Patil and Ashpak Tamboli. Zero order and area under curve
spectrophotometric methods for determination of Levocetirizine
in pharmaceutical formulation. International
Journal of Advances in Scientific Research.2015; 1(06): 270-275.
27. International Conference on Harmonization (ICH) of Technical
Requirements for the registration of Pharmaceuticals for Human use, Validation
of Analytical Procedures Methodology.
ICH-Q2 (R1), Geneva.1996, 1-8.