Simultaneous Determination of Paracetamol and Domperidone in Pharmaceutical Dosage Form by First Order Derivative

UV Spectrophotometry

Audumbar Mali¹*, Sujata Kolekar², Jeeja Panachery³, Ashpak Tamboli⁴

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, Mumbai, Maharashtra, India.

³Department of Pharmacognosy and Phytochemistry, Shri D.D. Vispute college of Pharmacy and Research Centre, Devad-Vichumbe, New Panvel, Mumbai, Maharashtra, India.

⁴Department of Pharmaceutical Chemistry, Sahyadri College of Pharmacy, Methwade,

Sangola-413307, Solapur, Maharashtra, India.

*Corresponding Author E-mail: maliaudu442@gmail.com

ABSTRACT:

Derivative spectrophotometry offers a useful approach for the analysis of drugs in multi-component formulation. In this study a first order derivative spectrophotometric method is applied for the simultaneous determination of Paracetamol and Domperidone in Tablet dosage form. The measurements were carried out at wavelengths of 262 and 297 nm for Paracetamol and Domperidone respectively. The method was found to be linear (r²=0.999) in the range of 5-25 μg/ml for Paracetamol in the presence of 20μg/ml of Domperidone at 262 nm. The linear correlation (r²=0.999) was obtained in the range of 5-25μg/ml for Domperidone in the presence of 20μg/ml of Paracetamol at 297 nm. The method was successfully used for simultaneous determination of Paracetamol and Domperidone in tablet dosage form without any interference from excipients and prior separation.

KEY WORDS: Paracetamol, Domperidone, UV visible spectrophotometry, Method Validation, First order derivative method.

1. INTRODUCTION:

Paracetamol (PR) or acetaminophen (the name of the drug in US, Scheme 1) is the most famous drug in treatment of pain and fever. It is used as antipyretic, analgesic and anti-inflammatory drug, due to inhibiting prostaglandin synthesis cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2). However, Paracetamol does not cause cancer like phenacetin. Also, it has no effect on respiration.

Although there are lots of drugs that work like paracetamol, it is still the most important, because it is cheap, effective, has no side effects and most important, safe. Can be used alone to treat little to moderate pain, but if we combine it with anti-inflammatory steroid drugs or opioid it can treat intense pain [3]. Even then it is safe, but the overuse of it can lead to hepatic toxem. It can also lead to serious condition if it is taken it with alcohol. It is considered as the primary reason for toxemia in USA, UK and the New Zealand.[1-3] Literature survey revealed the estimation methods of Paracetamol or with other drugs by UV spectrophotometry [4, 5], RP-HPLC[6, 7] calorimetric method, flow injection analysis, and HPTLC. [8, 9]

Domperidone is an antiemetic and antinauseant and acts on dopamine receptor system as an antagonist. Chemically it is 5-chloro-1-[1-(2, 3-dihydro-2-oxo-1H-benzimidazole-1-yl) propyl)-4-piperidyl]-2,3-dihydro-1H-benzimidazol-2-one.[10, 11] Literature survey reveals UV spectrophotometric method [12, 13], atomic absorption spectrometry, spectroflurometry, HPLC[14] and HPTLC [15] methods for its determination.

Application of derivative technique of spectrophotometry offers a powerful tool for quantitative analysis of multi-component mixtures. When derivatised, the maxima and minima of the original function take zero values, and the inflections are converted into maxima or minima, respectively. The derivative curves are more structured than the original spectra, thus enabling very tiny differences between the original spectra to be identified. Derivative spectrophotometry provides selectivity and offers a solution in resolving the overlapping spectra in multi-component analysis without previous chemical separation. In the last decades, this technique has rapidly gained application in the field of pharmaceutical analysis to overcome the problem of interference, due to substances other than analytes, commonly present in pharmaceutical formulations or for combination of two or more drug substances.[16, 17] Lack of any published method for simultaneous spectrophotometric determination of Paracetamol and Domperidone, therefore, provoked us to investigate the application of derivative spectrophotometry for simultaneous determination of these compounds in pharmaceutical dosage forms using zero-crossing method.

Fig. 1: Chemical structure of Paracetamol

Fig. 2: Chemical structure 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 Paracetamol and Domperidone API was supplied as gift sample by Cipla Pharmaceutical LTD, Pune, Maharashtra, India. The commercial formulation Cetadomas purchased from the local market Solapur, Maharashtra, India.

2.3 Method development: [18-21]

2.3.1Preparation 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 100 μg/ml for each drug. Dilutions from stock solution were prepared in the range of 5-25μg/ml for Paracetamol and 5-25μg/ml for Domperidone. Methanol was used as a blank solution.

2.3.2Spectrophotometric Measurements:-

Zero-order spectra of standard solutions of Paracetamol (20μg/ml) and Domperidone (20μg/ml) versus their solvent blank were recorded in the range of 200-400 nm (Figure 3). The first order derivative spectra of these solutions were obtained in the same range of wavelength against their blanks (Figure 4). The values of first order derivative amplitudes for Paracetamol in the presence of Domperidone and vice versa were measured at 262nm (zero-crossing of Paracetamol) and 297 nm (zero-crossing of Domperidone), respectively. The calibration curves for derivative spectrophotometry were constructed by plotting the drug concentration versus the absorbance values of the first order derivative spectrum, at 262nm for Paracetamol and at 297 nm for Domperidone.

Figure 3: Zero order spectra (overlain) of Paracetamol 20 μg/ml and Domperidone 20 μg/ml

Figure 4: First order derivative spectra (overlain) of Paracetamol 20 μg/ml and Domperidone 20 μg/ml

2.3.3 Analysis of commercial tablet formulation:-

Contents of 20 tablets were weighed and their average weight was determined and powdered. Accurately weighed powder equivalent to fill weight of one tablet was transferred to 100 ml calibrated flask containing 50 ml of methanol and sonicated for 30 minutes. The volume was then made up to the mark with methanol. The resulting solution was then filtered through whatmann filter paper (#41). From this solution, 1 ml was transferred to another 10 ml calibrated flask and diluted up to 10 ml which gives 200μg/ml concentration of solution. Then 1 ml of this solution was further diluted to 10 ml to get approximate concentration 20μg/ml of Paracetamol and 20μg/ml of Domperidone.

Table 1: Assay of tablet dosage form.

Sr.No.	Sample Solution Concentration (µg/ml)	Amount found (%)*	Mean % found	%RSD
1	20	100.81
2	20	98.13	100.16	0.5486
3	20	101.56

*n=3, % RSD = % Relative Standard Deviation.

3. RESULTS AND DISCUSSION:-

3.1 Linearity and Range:-

3.2 Linearity:-

Calibration curves were constructed using six replicates of Paracetamol solutions between 5-25 μg/ml in the presence of 5-25 μg/ml of Domperidone. The same procedure was used for solutions containing Domperidone 5-25 μg/ml in the presence of 5-25 μg/ml of Paracetamol. The calibration curves were constructed (Fig. 5 and Fig. 6) and statistical analysis was performed. The regression equations of calibration curves were y=0.0162x-0.0042 (r²=0.999) at 262 nm for Paracetamoland y=0.0194x-0.0015 (r²=0.999) at 297 nm for Domperidone for first order derivative spectrophotometry methods. The range was found to be 5-25μg/ml for both drugs for first order spectrophotometry methods.

Table 2: Stastical data for the calibration graphs for determination of Paracetamol and Domperidone by Proposed methods.

Parameters	Paracetamol	Domperidone
Linearity range (µg/ml)*	5-25	5-25
r²± S.D*	0.999	0.999

3.3 Accuracy:-

For accuracy determination, the analysed samples were spiked with extra 80%, 100% and 120% of the standard solution of both drugs and the mixtures were reanalysed by the proposed method. The experiment was conducted in triplicate. This was done to check for the recovery of the drug at different levels in the commercial tablet formulations. The mean recoveries and %RSD are illustrated in Table 3. The data indicates that the proposed derivative spectrophotometric method is highly reproducible during one run and between different runs. [22, 23]

Fig.5: Calibration curve for Paracetamol at 262 nm

Fig.6: Calibration curve for Domperidone at 297 nm

Table 3: Results of drug content and analytical recovery of Paracetamol and Domperidone

Parameters	Paracetamol	% R.S.D	Domperidone	% R.S.D
Labelled claim	10 mg	-	10 mg	-
% Drug content ± S.D	99.23 ± 0.2590	0.57	101.35 ± 0.2158	0.64
Analytical recovery at 80 % ± S.D	102.06 ± 0.4259	0.38	98.18 ± 0.3217	0.31
Analytical recovery at 100 % ± S.D	100.21 ± 0.4587	0.34	101.02 ± 0.2589	0.47
Analytical recovery at 120% ± S.D	98.78 ± 0.1234	0.20	99.20 ± 0.4581	0.28

Fig.7: First order derivative overlay spectra of Paracetamol and Domperidone at 5,10,15,20 and 25 μg/ml Concentrations

3.4 Precision:-

To determine the precision of the method, Paracetamol and Domperidone solutions at a concentration of 20μg/ml were analysed each three times for first order spectrophotometric method. Solutions for the standard curves were prepared fresh every day. [22, 23]

Table 4: Results of Intra and Inter Day Precision:-

Parameters	Intra Day Precision		Inter Day Precision
Parameters	S.D*	% RSD*	S.D*	% RSD*
Paracetamol	0.0063	0.6589	0.0053	0.6982
Domperidone	0.0082	0.7590	0.0041	0.7891

3.5 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 0.3547 μg/ml and 1.0645 μg/ml respectively of Paracetamol for first order derivative and 0.3862µg/ml and 1.1584µg/ml for area under the curve methods respectively.[24, 25]

3.6 Analysis of the Marketed Formulation:-

There was no interference from the excipients commonly present in the tablets. The drug content was found to be 100.16% first order spectrophotometric methods. It may therefore be inferred that degradation of Paracetamol and 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 Paracetamol and Domperidone in pharmaceutical dosage form.[24,25]

Table 5: Summary of validation parameters:-

Parameter	Paracetamol	Domperidone
λ range	200-400 nm	200-400nm
Regression Equation (y=mx+c)	Y=0.0162x-0.0042	Y=0.0194x-0.0015
Measured wavelength	262nm	297nm
Linearity range	5-25µg/ml	5-25µg/ml
Slope	0.0162	0.0194
Intercept	0.0042	0.0015
Correlation coefficient (R²)	0.999	0.999
Limit of Detection (LOD) µg/ml	0.3547	0.3862
Limit of Quantitation (LOQ) µg/ml	1.0645	1.1584
Accuracy (Mean % Recovery)	99.23	101.35
Precision (%RSD)	0.57	0.64

4. CONCLUSION:

From the results of this study it can be concluded that the proposed first order derivative spectrophotometric method can be used for simultaneous determination of Paracetamol and Domperidone. This method is simple, rapid, practical, reliable and inexpensive and can be used for routine analysis of simultaneous determination of these compounds without any prior separation in quality control laboratories.

5. 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.

6. REFERENCES:-

1. Mali Audumbar Digambar, Hake Gorakhnath and Patil Manoj Kumar. Zero order and area under curve spectrophotometric methods for determination of Paracetamol in pharmaceutical formulation. Bulletin of Advanced Scientific Research. 2015; 1(3): 102-105.

2. Pawar P.Y., Hapse Sandip, Salve Megha And Agarkar Arti. Spectrophotometric Estimation And Validation of Paracetamol And Domperidone By Different Method From Pure And Tablet Dosage Form. Int. J. Drug Res. Tech. 2013; 3(2): 37-44.

3. Ahmad M. El-Zinati and Monzir S. Abdel-Latif. Simultaneous Determination of Paracetamol and Tramadol in Pharmaceutical Tablets by Derivative UV-Vis Absorption Spectrophotometry. The Open Analytical Chemistry Journal. 2015; 8:1-5.

4. Rupali S. Joshi, Nilima S. Pawar, Sameer S. Katiyar, Devendra B. Zope and Amol T. Shinde. Development and validation of UV spectrophotometric methods for simultaneous estimation of Paracetamol and Ibuprofen in pure and tablet dosage form. Pelagia Research Library Der Pharmacia Sinica. 2011; 2(3): 164-171.

5. Dr.Pratap Y.Pawar, Lalasaheb S Zanje, Satish S Tambe, Aditya U Nandgaonkar, Pradip V Funde, Aniket A Vyavhare. Simultaneous Estimation of Pregabalin and Paracetamol By UV Spectroscopic Method In Bulk and Tablet Formulation. World Journal of Pharmacy And Pharmaceutical Sciences. 2014; 3(4); 743-750.

6. K Karunakaran, G Navaneethan and KP Elango. Development and Validation of a Stability-Indicating RP-HPLC Method for Simultaneous Determination of Paracetamol, Tramadol HCl and Domperidone in a Combined Dosage Form. Tropical Journal of Pharmaceutical Research. 2012; 11(1): 99-106.

7. Jagdish Kakadiya, Naiya Parmar, Nehal Shah. Development And Validation Of RP-HPLC Methodfor Simultaneous Estimation Of Promethazine Hydrochloride And Paracetamol InCombined Liquid Formulation. Asian Journal of Research in Biological and Pharmaceutical Sciences. 2014;2(1): 11 - 26.

8. K. G. Baheti, Shamim Shaikh, Nishil Shah and M.H. Dehghan. Validated Simultaneous Estimation of Paracetamol and Etoricoxib in Bulk and Tablet by HPTLC Method. International Journal of Research in Pharmaceutical and Biomedical Sciences. 2011; 2(2): 672-675.

9. Kajal S. Mahajan, Renu S. Chauhan, Shailesh A. Shah, Dinesh R. Shah. Development of HPTLC Method for Simultaneous Estimation of Paracetamol and Flupirtine Maleate in their Combined Tablet Dosage Form. Journal of Pharmacy and Applied Sciences. 2015; 2(1): 1-7.

10. 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.

11. Sunil Singh, Surabhi Sharma, Ajit Kumar Yadav and Hemendra Gautam. Simultaneous Estimation of Naproxen and Domperidone Using UV Spectrophotometry In Tablet Dosage Form. Bulletin of Pharmaceutical Research 2013; 3(2):66-70.

12. Kaushik KN, Banerjee SK. UV Spectrophotometric method development for the determination of domperidone in tablet formulation. Asian J. Res. Chem. 2009; 2(4): 432-3.

13. Surekha J. Babar, Varsha B. Mane, Dr. S. B. Bhise. Development And Validation Of UV-Spectrophotometric Methods For Simultaneous Estimation Of Paracetamol And Domperidone In Bulk And Tablet Dosage Form. International Journal of Pharmacy and Pharmaceutical Sciences. 2012; 4, Issue 4, 206-209.

14. Sharma Suparna, Sharma Anuj Kumar, Singh Ompal, Chaturvedi Ashwani Kumar, Verma Vikrant, Arya Rajat Kumar, Singh Umesh Kumar. RP-HPLC Method Development and Validation of Domperidone in Solid Dosage Form. The Pharma Innovation. 2012; 1(4): 32-36.

15. Patel B, Patel M, Patel J, Suhagia B. Simultaneous determination of omeprazole and domperidone in capsules by RP‐HPLC and densitometric HPTLC. J. Liq. Chromatogr. Rel. Technol. 2007; 30(12):1749-62.

16. Audumbar Digambar Mali, Ritesh Bathe and Ashpak Tamboli. Simultaneous Determination of Cefixime Trihydrate and Ofloxacin in Pharmaceutical Dosage Formby Second Order Derivative UV Spectrophotometry. International Journal of Advances in Pharmaceutical Analysis. 2015; 5(2): 42-46.

17. Audumbar Digambar Mali. Simultaneous Determination Of Carvedilol and Hydrochlorothiazide In Pharmaceutical Dosage Form By First Order Derivative UVSpectrophotometry. International Journal of Pharmacy and Pharmaceutical Sciences. 2015; 7(9): 371-374.

18. Lakshmi S, Lakshmi KS and Tintu T Simultaneous Spectrophotometric Estimation of Paracetamol and Lornoxicam in tablet dosage form. Int J Pharm Pharm Sci. 2010; 2(4):166‐168.

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, Mane Pandurang, Tamboli Ashpak. Development And Validation of UV Spectrophotometric Estimation of Diclofenac Sodium Bulk and Tablet Dosage Form Using Area Under Curve Method. Pharmatutor Magazine. 2015; 3(4): 21-25.

21. Kapil K, Naik S, Garima J and Mishra N Spectrophotometric Method for Simultaneous Estimation of Paracetamol and Domperidone in Tablet Formulation Asian J. Research Chem. 2009; 2(2): 112‐114.

22. Mali Audumbar Digambar, Hake Gorakhnath, Patil Manoj Kumar, Bathe Ritesh, Tamboli Ashpak. Zero Order and Area under Curve Spectrophotometric Methods for Determination of Ampicillin Trihydrate in Pharmaceutical Formulation. GCC Journal of Science and Technology. 2015; 1(1): 6-12.

23. 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.

24. Audumbar Digambar Mali, Ritesh Bathe and Ashpak Tamboli. Estimation of Ampicillin Trihydrate in Bulk and Formulation by First Order Derivative Area under Curve UV Spectrophotometric Methods. International Journal of Advances in Scientific Research.2015; 1(05): 239-243.

25. 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.

Received on 10.02.2016 Accepted on 28.02.2016

Asian J. Pharm. Res. 6(1): January -March, 2016; Page 22-26

DOI: 10.5958/2231-5691.2016.00004.6