IAJPS 2016, 3 (2), 98-103
Prasanna et al
CODEN (USA): IAJPBB
ISSN 2349-7750
ISSN: 2349-7750
INDO AMERICAN JOURNAL OF
PHARMACEUTICAL SCIENCES Available online at: http://www.iajps.com
Research Article
SIMULTANEOUS ESTIMATION OF DICLOFENAC SODIUM AND PANTOPRAZOLE SODIUM UV SPECTROPHOTOMETRIC METHOD Paras R Kathiriya, Sweetu D Patel, Divya Patel, Prasanna K Pradhan*, Umesh M Upadhyay 1 Sigma Institute of Pharmacy, Baroda, Gujarat, India. Abstract: A Simple, precise, accurate and reproducible UV Spectrophotometric method has been developed and validated for simultaneous estimation of Diclofenac Sodium (DICLO) and Pantoprazole Sodium (PANTO) in its combined dosage form using second order derivative. Zero crossing points 272.8nm and 296.2nm were used for estimation of PANTO and DICLO respectively. The linearity range was found to be 5-15 μg/ml for PANTO and 17.5-52.5 μg/ml for DICLO. The % recoveries for both were found to be well within range. The developed method was validated as per ICH guideline and on application to marketed formulation for evaluation found to be useful. Keywords: Diclofenac Sodium (DICLO), Pantoprazole Sodium (PANTO), Second Order Derivative, Development, Validation.
Corresponding author: Pradhan Prasanna K. Sigma Institute of Pharmacy, Vadodara-390018 Gujarat, India. E-Mail: pkpwithu@gmail.com (M): 8141865430
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Please cite this article in press as Sonia Prasanna et al, Simultaneous Estimation of Diclofenac Sodium and Pantoprazole Sodium UV Spectrophotometric Method, Indo Am. J. Pharm. Sci, 2016; 3(2).
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IAJPS 2016, 3 (2), 98-103 INTRODUCTION: [1, 2, 3, 4] Pantoprazole Sodium and Diclofenac sodium are official in Indian Pharmacopoeia (IP), British Pharmacopoeia BP and United State Pharmacopoeia USP. Diclofenac Sodium (DICLO) is Sodium {2-[(2, 6dichlorophenyl) amino] phenyl} acetate having empirical formulaC14H10Cl2NNaO2 with molecular weight 318.13047 gm/mole [1]. DICLO is nonsteroidal anti-inflammatory (NSAIDs) class of drugs that provide analgesic, antipyretic, and antiinflammatory effects that inhibition of both leukocyte migration and the enzyme cyclooxygenase (COX-1 and COX 2), leading to the peripheral inhibition of prostaglandin synthesis. As prostaglandins sensitize pain receptors, inhibition of their synthesis is responsible for the analgesic effects of diclofenac. Antipyretic effects may be due to action on the hypothalamus, resulting in peripheral dilation, increased cutaneous blood flow, and subsequent heat dissipation. Pantoprazole Sodium (PANTO) is Sodium Salt of 5 (Difluoromethoxy) -2-[[3,4-Dimethoxy -2- pyridinyl) methyl] Sulfinyl] -1- H benzimidazole having empirical formula C16H14F2N3NaO4S with molecular weight 405.352 gm/mole. PANTO is an Anti-ulcer, Proton pump inhibitor drug, that suppresses the final step in gastric acid production by covalently binding to the (H+, K+)-ATPase enzyme system at the
Prasanna et al
ISSN 2349-7750
secretory surface of the gastric parietal cell. This effect leads to inhibition of both basal and stimulated gastric acid secretion, irrespective of the stimulus. Besides inhibiting acid secretion, PAN affords protection against NSAID-induced gastric damage, has low potential for drug-drug interactions, and is particularly suitable for administration to elderly patients who often require concomitant treatment with NSAIDs as no dose adjustment is required during concomitant treatment with both drugs. Literature survey reveals that various analytical methods involving Spectrophotometry, HPLC has been reported for the estimation of Pantoprazole Sodium in single form and in combination with other drugs and various analytical methods involving Spectrophotometry, HPLC has been reported for the estimation of Pantoprazole Sodium in single form and in combination with other drugs. Present work describes the development and validation of a simple, precise, accurate and reproducible Spectrophotometric method for simultaneous estimation of Diclofenac Sodium (DICLO) and Pantoprazole Sodium (PANTO) in its combined dosage form. The developed method has been validated as per ICH guideline and successfully applied for estimation of Diclofenac Sodium (DICLO) and Pantoprazole Sodium (PANTO) Sesquihydrate in marketed tablet Formulation.
MATERIALS AND METHODS:
Fig 1: Calibration Curve of PANTO at 296.2nm
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Fig 2: Calibration Curve of DICLO at 272.8nm
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IAJPS 2016, 3 (2), 98-103
Prasanna et al
ISSN 2349-7750
Fig 3: Overlain UV second order spectra of PANTO and DICLO
Table 1: Validation Parameters of PANTO and DICLO Parameters Linearity Range (µg/ml) Correlation Coefficient (r2) Precision:
PANTO 5-15 0.9998
DICLO 15-40 0.9996
Repeatability (%RSD) Intraday (%RSD) Interday (%RSD) Accuracy: LOD (µg/ml) LOQ (µg/ml) Assay (%)
0.97 1.14 1.22 1.04 0.233 0.706 99.44
0.82 0.84 0.99 0.61 0.658 1.993 99.01
Table 2: Intraday and Interday precision data of PANTO and DICLO Drug
PANTO
DICLO
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Conc. Taken µg/mL 5.0 10.0 15.0 15.0 30.0 40.0
Intra_day Conc. observed* µg/mL ± RSD% 5.08 ± 1.32 9.90 ± 0.60 14.73 ±1.17 14.90 ±1.40 29.75 ± 0.96 40.32 ± 1.41
Inter_day Conc. observed* µg/mL ± RSD% 5.28 ± 0.42 10.04 ±1.14 15.02 ± 0.69 14.85 ± 1.30 30.48 ± 1.42 40.42 ± 0.73
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IAJPS 2016, 3 (2), 98-103
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Table 3: Recovery Data of PANTO and DICLO Drug
Spiking (%)
Spike Conc. (µg/ml)
Conc. recovered (µg/ml)
Mean Recovery ±SD(n=3)
%RSD
PANTO
80 100 120
4 5 6
3.97 4.98 5.94
99.18 ± 0.94 99.56 ± 1.52 99.08 ± 0.63
0.96 1.53 0.64
DICLO
80 100 120
12 15 18
12.19 15.37 18.23
100.20 ± 0.46 101.04 ± 0.74 99.84 ± 0.65
0.46 0.74 0.65
Table 4: Assay result of Marketed Formulation Drug PANTO DICLO * (n=3)
Label claim (mg)
Conc. Found (mg) Mean ±SD*
%Assay Mean ±SD
%RSD
10.0 37.5
9.94 ± 0.42 37.13 ± 0.37
99.44 ± 0.67 99.01 ± 0.54
0.673 0.545
Instrumental Conditions Shimadzu UV-1700 a double beam spectrophotometer connected to Computer with UV Probe 2.34 software was used. The measurement carried out using 1 cm quartz cells Reagent and Materials Gift Sample of Pantoprazole Sodium was obtained from Sun Pharma, Vadodara, India and Diclofenac Sodium obtained from Umedica Laboratory Pvt. Ltd., Vapi, India. Marketed formulation Dufex Capsule containing Pantoprazole Sodium Sesquihydrate 20 mg and Diclofenac sodium75mg was procured from local market. Preparation of standard stock solutions [5,6] Accurately weighted 5mg PANTO and 10mg DICLO was taken separately in 100 ml volumetric flask and make volume up to mark with methanol to produce 50 µg/ml of PANTO and 100 µg/ml of DICLO. Both the solution Sonicated for 5 minutes. Calibration curve and wavelength selection [7] The working stock solutions of both diluted with sufficient volume with methanol to get concentration 5µg/ml and 10µg/ml of PANTO and DICLO respectively. They were scanned at range of 200400nm against methanol as solvent blank. Maximum absorbance was obtained at 296.2 nm and 272.8 nm for PANTO and DICLO respectively. These two wavelengths have been used for the determination of PANTO and DICLO in their formulation.
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Preparation of Sample Solution and Formulation Analysis [8,9,10] Take twenty capsules weighed individually made it empty and content mixed in beaker. From this powder weighed accurately 20mg and 75 mg equivalent pantoprazole and Diclofenac sodium respectively. Transferred it in to the 100ml volumetric flask, add about 50ml methanol to dissolved powder. Sonicated for 5 minutes and then volume was made up to mark with methanol. Filter the solution through whatman filter paper and from this taken 10ml and diluted up to the 100 ml with methanol to got concentration 20μg/ml PANTO and 75μg/ml DICLO. From this solution take 5ml and makeup the volume up the 10ml to get concentration 10 μg/ml DICLO and 37.5 μg/ml PANTO, which is used for estimation of PANTO and DICLO. The concentrations of both PANTO and DICLO were determined by measuring the absorbance of derivative spectra at 296.2 nm and 272.8 nm for second order derivative spectroscopic method. Method Validation [11] Methods were validated according to ICH guidelines. Linearity The linearity of method was evaluated by analyzing different concentrations of the standard solutions of PANTO and DICLO. The Beer-Lambert’s concentration range was found to be 5- 15μg/ml for PANTO and 15-40 μg/ml for DICLO for second order derivative Spectrophotometric method. (figure1, figure-2)
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IAJPS 2016, 3 (2), 98-103 Accuracy To ascertain the accuracy of proposed methods, recovery studies were carried out by standard addition method at three different levels (50%, 100% & 150%). (Table-2). Precision The reproducibility of the proposed method was determined by performing tablet assay at different time intervals on same day (Intra-day precision) and on three different days (Inter-day precision) and was expressed in % RSD. (TABLE-2) Limit of detection and Limit of quantification ICH guidelines describe several approaches to determine the LOD and LOQ. These are visual evaluation, signal-to-noise ratio and the use of standard deviation of the response and slope of calibration curve. In present study, the LOD and LOQ were calculated using standard deviation of the response and slope of calibration curve. Calibration curves were repeated 5 times and standard deviation of intercept is calculated. Then LOD and LOQ were measured as per equations given here. LOD= 3.3 (SD/Slope of calibration curve) LOQ= 10 (SD/Slope of calibration curve) SD = Standard deviation of y-intercepts of regression lines. RESULTS AND DISCUSSION: The methods were successfully validated with respect to linearity, accuracy, intra and inter day precision etc. In Derivative spectroscopic method, linearity for detector response was observed in the concentration range of 5-15 μg/ml for PANTO and 15-40 μg/ml for DICLO. (figure-1, figure-2) As shown in Table-2 %RSD values during both intra and inter day precision studies were <2.0% demonstrating repeatability and intermediate precision. the mean percentage recovery values for PANTO and DICLO by second derivative method.(Table-3). LOD for PANTO and DICLO was found0.233 µg/ml and 0.658 µg/ml respectively and LOQ for PANTO and DICLO was found 0.706µg/ml and 1.993 µg/ml respectively. The proposed spectroscopic methods allow for accurate, precise and reliable measurement of PANTO and DICLO simultaneously in combined capsule dosage form. The developed methods were found to be simple, rapid, linear, accurate and precise for the concurrent estimation of drugs in respective two-component oral dosage forms of PANTO and DICLO.
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Prasanna et al
ISSN 2349-7750
The methods were evaluated for validation parameters, linear relation including accuracy, reproducibility and precision. The RSD for all parameters and assay results obtained by this method are in fair agreement. The developed methods can be used for routine quantitative simultaneous estimation of PANTO and DICLO in pharmaceutical preparations. CONCLUSION: From all results it can be conclude that the developed second derivative Spectrophotometric method is simple, accurate, precise, selective, reproducible, and cost effective for simultaneous estimation of Pantoprazole and Diclofenac in marketed formulation. Percentage recovery shows that method is free from interference of excipients used in the tablet formulation. REFERENCES: 1.Taha A “NSAIDs types, uses, contraindications, and their relation to ulcer” http://medicapharm.com/nsaids-types-usescontraindications.html Accessed on 12-02-2015. 2.Pharmacology of Diclofenac and pantoprazole, https://www.medicineindia.org/pharmacology-forgeneric/3036/diclofenacsodiumpantoprazole Accessed on 06-10-2014. 3.Drug bank, drug profile Diclofenac, http://www.drugbank.ca/drugs/DB00586, Accessed on 06-10-2014. 4.Drug bank, drug profile pantoprazole, http://www.drugbank.ca/drugs/DB00213, Accessed on 06-10-2014. 5.Indian Pharmacopoeia , Ministry of Health & Family Welfare; Govt. of India, 6th Edn; Published by the Indian Pharmacopoeial commission, Ghaziabad,2010; pp 1200. 6.Indian Pharmacopoeia , Ministry of Health & Family Welfare; Govt. of India, 6th Edn; Published by the Indian Pharmacopoeial commission, Ghaziabad,2010; 1857. 7.Patel RK, Patel RN, Patel HB, Patel VA, Ganure AL and Patel LJ “Simultaneous Estimation of Pantoprazole and Diclofenac sodium in Combined Pharmaceutical Dosage Form by Derivative Spectrophotometric Method” Asian Journal of Research In Chemistry, 2012; 5, 893-895. 8.Bhatt NM, Chavada VD, Sanyal M, Shrivastav PS, “Manipulating Ratio Spectra for the Spectrophotometric Analysis of Diclofenac Sodium and Pantoprazole Sodium in Laboratory Mixtures and Tablet Formulation” Hindawi Publishing Corporation the Scientific World Journal, 2014. http://dx.doi.org/10.1155/2014/495739.
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IAJPS 2016, 3 (2), 98-103 9.Naik K, Parmar G, Ahiya J, Bodiwala K and Shah S, “Development and Validation of Derivative Spectrophotometric Method for Simultaneous Estimation of Diclofenac and Pantoprazole in Combined Capsule Dosage Form” Asian Journal of Research in Chemical, 2013; 6: 155-157. 10.Shah DA, Patel A, Baldania SL, Chhalotiya UK, and Bhatt KK “Simultaneous Estimation of Pantoprazole Sodium and Levosulpiride in Capsule Dosage Form by Simultaneous Equation Spectrophotometric Method” Hindawi Publishing Corporation the Scientific World Journal, 2013. http://dx.doi.org/10.1155/2013/459820. 11.ICH Topic Q2B, Note for Guidance on Validation of AnalyticalProcedures: Methodology
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GPMP/ICH/281/95, The European Agency for the Evaluation of Medical Products: Geneva, 1996. 12.Patel ZN, Patel PB, Modi JD, Parikh NN, Chaudhari HM, Pradhan PK and Upadhyay UM “RPHPLC Method For Simultaneous Estimation of Levosulpiride and Pantoprazole Sodium In Tablet Dosage Form” Pharma Science Monitor, 2014; 2(5): 0976-7908. 13.Patel PB, Patel ZN, Modi JD, Parikh NN, Lad BN, Pradhan PK and Upadhyay UM “Development And Validation Of RP-HPLC Method For Simultaneous Estimation of Diclofenac Sodium and Eperisone HCl In Tablet, Dosage Form, Pharma Science Monitor, 2014;2(5):0976-7908.
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