Validated Spectrophotometric Methods for the Simultaneous ...

Validated Spectrophotometric Methods for the Simultaneous ... International Journal of PharmTech Research CODEN (USA): IJPRIF ISSN : 0974-4304 Vol.4, No.2, pp 828-834, April-June 2012 Validated Spectrophotometric Methods for the Simultaneous Estimation of Domperidone and Lansoprazole in Bulk Drug and Pharmaceutical Formulation Anil Kumar S.M, Nagraj Javar Gowda*, Siddalinga Swamy M.S. Department of pharmaceutical Analysis, National College of Pharmacy, Balraj Urs Road, Shimog (District), Karnataka, India-500201. *Corres. author: anil_manchi@rediffmail.com Cell: 9964651920 Abstract: Two simple spectrophotometric methods have been developed for simultaneous determination of Domperidone and Lansoprazole in bulk and combined pharmaceutical dosage forms. methanolic sodium hydroxide (70:30) was used as solvent. Method I simultaneous equation method involves measurement of absorbances at two wavelengths 287 nm (λ max of Domperidone) and 294 nm (λ max of Lansoprazole) and Method II first derivative Zero-crossing involves measurement of amplitudes of first derivative spectra of the binary mixtures containing Domperidone and Lansoprazole at 294 nm (zero crossing of Lansoprazole) for the determination Domperidone and at 287 nm (zero crossing of Domperidone) for the determination of Lansoprazole respectively. The linearity lies between 4-36 µg/ml for Domperidone and 2-18 µg/ml for Lansoprazole for both the two methods. The accuracy and precision of the methods were determined and validated statistically. Both the methods showed good reproducibility and recovery with %RSD less than 2. Both the methods were found to be rapid, specific, precise and accurate and can be successfully applied for the routine analysis Domperidone and Lansoprazole in bulk and combined pharmaceutical dosage form. Keywords: Domperidone. Lansoprazole, Simultaneous equation, First derivative zero-crossing method, Method validation. Lansoprazole (2-[[3-methyl-4-(2,2,2-trifluoroethoxy) 1,2,3,41. INTRODUCTION pyridin-2-yl]methylsulfinyl]-1H-benzimidazole) is a Domperidone (6-chloro-3-[1-[3-(2-oxo-3H-proton pump inhibitor (Fig.2), which prevents the benzimidazol-1-yl)propyl]piperidin-4-yl]-1H-stomach from producing acid. It is used for the benzimidazol-2-one) is an antidopaminergic drug treatment of ulcers of the stomach, duodenum, and (Fig.1), developed by Janssen Pharmaceutica, and used NSAID-induced ulcers and gastro esophageal reflux orally, rectally or intravenously, generally to suppress disease (GERD). The combination of Lansoprazole nausea and vomiting. It has also been used to stimulate and Domperidone is very useful in gastro-esophagus lactation and found effective in the treatment of dysfunctions like functional dyspepsia. gastroparesis, a stomach motility condition, and for pediatric Gastro esophageal reflux. Nagraj Javar Gowda et al /Int.J. PharmTech Res.2012,4(2) 829 Hsolvents used in analysis were of spectroscopic grade. ClNOLanspro-D Capsules (label claim 30mg LANS and H30mg DOMP). Batch No. LPD-8002 B of Indchemie, NNOHealth Specialties Pvt, Ltd. was used for analysis. NN2.3 Methanolic Sodium Hydroxide To 300ml of 0.1N sodium hydroxide solution, 700ml Domperidoneof methanol was added to get methanolic sodium Figure. 1- Chemical structure of Domperidone hydroxide (70:30). 2.4 Standard solutions of Domperidone and Lansoprazole HCAccurately weighed 25mg of Domperidone and 3OCFO3Lansoprazole separately, and it was transferred into N Sclean, dry 25ml volumetric flasks. This gave the -1. From this 4ml of concentration of 1000µg mlNDomperidone solution was pipetted out in to 50ml NHvolumetric flask and the volume was made up to the mark by methanolic sodium hydroxide solution. This Lansoprazole -1gave 80µg ml of drug concentration. In the same way Figure. 2 -Chemical structure of Lansoprazole 2ml of the Lansoprazole solution was pipetted out into 50ml volumetric flask and the volume was made up to -1the mark with the same solvent. This gave 40µg ml On literature survey it was found that Domperidone drug concentration. and Lansoprazole can be estimated independently or in combination with the other drugs by several methods. 2.5 Calibration sets for simultaneous equation and One HPTLC method has been reported for 5first derivative zero-crossing methods Domperidone with Rabeprazole, and many HPLC A calibration set containing nine dilutions each of methods have been reported for Domperidone with -1-167,89DOMP (4-36 µg ml) and LANS (2-18 µg ml) was Rabeprazole, Pantaprazole, Omeprazole and 10prepared in methanolic sodium hydroxide and UV Paracetamol. spectra were recorded in the wavelength range of 200-Since no spectrophotometric method is reported for the 350 nm versus solvent blank. simultaneous estimation of Domperidone (DOMP) and Lansoprazole (LANS) in combination therefore in the 2.6 Pharmaceutical Sample solution present work a succesful attempt has been made to Twenty Lanspro-D capsules were weighed accurately. estimate both these drugs simultaneously by two An amount of the powder equivalent to content of one simple UVspectrophotometric methods (Simultaneous unit of capsule was triturated and dissolved separately equation method and First derivative zero crossing in 60 ml methanol: 0.1N sodium hydroxide (70:30 v/v). method). The solution was sonicated for 10 min and filtered into a 100ml volumetric flask through 0.45µ nylon 2. MATERIALS AND METHODS membrane filter. The residue was washed 3 times with 10 ml of methanol: 0.1N sodium hydroxide (70:30 v/v) 2.1 Apparatus and then the volume was completed to 100 ml with the Shimadzu UV-1601 double beam spectrophotometer same solvent. This solution was diluted to 1:10 with connected to a computer loaded with Shimadzu UV methanol: 0.1N sodium hydroxide (70:30 v/v) and Probe 2.10 software was used for all the again this solution was further diluted to 2:10 with the spectrophotometric measurements. The spectral band same solvent. The proposed methods were applied and width was 1 nm and the wavelength scanning speed -1the concentration of each component in the was 2800 nm min. The absorbance spectra of the formulation was determined. reference and test solutions were carried out in 1cm quartz cells over the range of 201-350 nm. 2.7 Simultaneous equation method (Method I): In the simultaneous equation method, the absorbance 2.2 Reagents and solutions signals were measured at 287 nm (λ max of Pure drug samples of Domperidone and Lansoprazole Domperidone) and 294 nm (λ max of Lansoprazole). were kindly gifted by Microlabs, Bangalore. Certified Concentration of each drug was obtained by using the to contain 99.6% and 99.8% purity respectively. The absorptivity values calculated for both the drugs at drugs were used without further purification. All the Nagraj Javar Gowda et al /Int.J. PharmTech Res.2012,4(2) 830 -1-1these two wavelengths. A calibration set containing gm 100 ml Absorbance unit cm-1nine dilutions each of DOMP (4-36µg ml) and LANS E = Absorptivity value, A = Absorbance, b = Path -1(2-18µg ml) was prepared in methanol:0.1N sodium length of quartz cell (1 cm), -1 hydroxide (70:30) and UV spectra were recorded in C = concentration in µg ml the wavelength range 200-350 nm versus solvent blank. Binary mixture solutions for prediction of DOMP and The overlay absorption spectra of standard solutions LANS were prepared as shown in Table 1. DOMP and LANS are shown in the Fig.3 Absorbencies of the above standard solutions of Absorbencies of binary mixture solutions were DOMP and LANS were measured at two wavelengths measured at 287 and 294nm. The concentration of 287 and 294nm, to get the absorptivity values at both each component of the binary mixture was calculated wavelengths for both the drugs from the equation: by using simultaneous equation. AE,*10000bC Table 1. Composition of DOMP and LANS in preparation of Binary mixture -1Mixture No. Concentration (µg ml) DOMP LANSO 1 4 2 C = concentration of x, x2 8 2 Am= Absorbance of Sample solution at ‘n’ nm, n3 16 2 E = Absorptivity of x at ‘n’ nm, xn4 24 2 5 36 2 The absorptivity values ? SD at 287 and 294nm for 6 4 4 both the drugs were: 7 8 4 E = 269.027 ? 1.351 DOMP 2878 16 4 E = 239.27 ? 6.4195 DOMP2949 24 4 E = 475.911 ? 1.1535 LANS28710 36 4 E = 551.55 ? 10.252 LANS294 11 4 8 12 8 8 13 16 8 2.8 First derivative zero crossing method (Method II): In this method the amplitudes of first derivative 14 24 8 spectra of the binary mixtures containing DOMP and 15 36 8 LANS were measured at 294nm (zero crossing of 16 4 12 LANS) for the determination DOMP and at 287nm 17 8 12 (zero crossing of DOMP) for the determination of 18 16 12 LANS respectively. The linearity ranges for DOMP 19 24 12 -1 and LANS were found to be 4 – 36 and 2 – 18µg ml20 36 12 respectively. A calibration set of nine dilutions each of 21 4 18 -1-1DOMP (4-36 µg ml) and LANS (2-18µg ml) in 22 8 18 methanol: 0.1N sodium hydroxide (70:30 v/v) was 23 16 18 prepared and UV spectra were recorded and converted 24 24 18 the same to first derivative spectra in a wavelength of 25 36 18 range 200-350nm versus solvent blank. The first derivative overlay spectra of standard solutions of DOMP and LANS are shown in the Fig.4. Nagraj Javar Gowda et al /Int.J. PharmTech Res.2012,4(2) 831 Table 2. Results of Intra and Inter Day Precision % RSD of Intra Day Precision % RSD of Inter Day Precision Parameters DOMP LANS DOMP LANS Method I 0.8379 1.1103 1.1516 1.1066 Method II 0.7159 1.1712 0.4164 0.9952 Fig.3 Zero-order absorption spectra for DOMP and LANS, ‘a’ represents DOMP showing λ max at 287nm and ‘b’ represents LANS showing λ max at 294nm. Fig. 4 Overlay first derivative absorption spectra of standard dilutions of DOMP and LANS, ‘a ‘represents the zero crossing of DOMP at 287nm and ‘b’ represents the zero crossing of LANS at 294nm 1 0.9 0.8 0.7 0.6 0.5abs0.4y = 0.025x + 0.0220.3R? = 0.9980.2 0.1 0 010203040 con Fig. 5 Calibration curve for determination of DOMP at 289nm.(Method I) Nagraj Javar Gowda et al /Int.J. PharmTech Res.2012,4(2) 832 y = 0.052x + 0.0181.2 R? = 0.9981 0.8 0.6 abs 0.4 0.2 0 Fig.6 Calibration curve for determination of LANS at 294nm. (Method I) 05101520 conc y = 0.025x + 0.0211 R? = 0.9980.9 0.8 0.7 0.6 0.5abs 0.4 0.3 0.2 0.1 0 Fig. 7 Calibration curve for determination of DOMP (Method II) 010203040 con Nagraj Javar Gowda et al /Int.J. PharmTech Res.2012,4(2) 833 3.5y = 0.160x + 0.028 R? = 0.9993 2.5 2 abs1.5 1 0.5 0 05101520 con Fig. 8 Calibration curve for determination of LANS (Method II) Table 3. Data of recovery studies Level of % recovery Mean Recovery* Standard Deviation* 80% 99.45% 0.5851 100% 99.91% 0.5204 120% 99.69% 0.6928 80% 99.51% 0.4490 100% 99.95% 0.4821 120% 99.69% 0.6943 calculated as shown in Table 2. Which showed 3. RESULTS AND DISCUSSION determination of DOMP and LANS were more precise 3.1. LINEARITY AND RANGE: by the proposed methods. Under the experimental conditions described, the graph obtained for simultaneous equation method 3.3. ACCURACY: (Method I) and derivative zero crossing method The interference of excipients in capsules for two (Method II) showed linear relationship. Regression drugs was tested for the application of proposed analysis was made for both the methods, in which for methods to commercial formulation. The study was method I correlation co-efficient, slope and intercept performed by increasing standard addition of known was found to be 0.9989, 0.0257, 0.0224 for DOMP amounts of studied drugs to an unknown concentration and 0.9989, 0.0522 , 0.0182 for LANS respectively as (constant volume) of the commercial pharmaceutical shown in Fig. 5 and 6. For method II correlation co-formulation (Standard addition access the effect of a efficient, slope and intercept was found to be 0.9984, sample matrix changes the analytical sensitivity of the 0.0258, 0.0219 for DOMP and 0.999, 0.1606, 0.0285 method). A constant volume of the unknown solution for LANS respectively as shown in Fig.7 and 8. was added to each of six 10ml volumetric flasks. Then a series of increasing volumes of working standard 3.2 PRECISION: solutions were added. Finally, each flask was made up Intra day and inter day precision for determination of to the mark with solvent and mixed well. The DOMP and LANS by proposed methods were concentration of the working standard solutions added evaluated in terms of % RSD. The binary mixtures of should be chosen to increase the concentration of the DOMP and LANS at five levels within their linearity unknown by minimum 30% in each succeeding flask. range were prepared and experiment was repeated The resulting mixtures were analyzed and recoveries three times (repeatability) and in three different days were determined. The results obtained are compared (intermediate precision).The average % RSD values with expected results. The excellent mean recoveries for determination of DOMP and LANS were Nagraj Javar Gowda et al /Int.J. PharmTech Res.2012,4(2) 834 and standard deviation (Table 3 and 4) suggested good their binary mixture formulation. The summary of the accuracy of the proposed method and no interference validation parameters is depicted in Table 7. from formulation excipients. 4. CONCLUSION 3.4 SENSITIVITY: The proposed spectrophotometric methods enable the The limit of detection (LOD) and limit of quantitation of mixture DOMP and LANS even in the quantification (LOQ) were calculated by using the presence of different excipients, with good accuracy equations LOD = 3 x σ / S and LOQ = 10 x σ /S, and precision, either in laboratory prepared samples or where σ is the standard deviation of intercept, S is the in spiked pharmaceutical dosage form in the linearity slope. . The values of LOD and LOQ are given in -1. range of 4-18µgmlFinally it was concluded that the Table 5. short analysis time and low costs were the main advantages of this spectrophotometric methods for the 3.5. ANALYSIS OF THE MARKETED determination of DOMP in combination with LANS in FORMULATION: routine analysis. High percentage recovery showed Applicability of the methods were tested by analyzing that the methods were free from interferences from the the commercially available formulation containing the excipients and additives commonly used in the binary mixture of DOMP and LANS. The values of % formulations of the drugs. recovery from the formulation as shown in the Table 6 were found to be very close to the label value of 5. ACKNOWLEDGEMENT: commercial pharmaceutical formulation, which The authors are highly thankful to the PES College of showed that the methods were applicable for Pharmacy and Principal, PES College of Pharmacy, simultaneous determination of DOMP and LANS from Bangalore for proving all the facilities to carry out the research work. of pantoprazole and domperidone in pure powder REFERENCES and a pharmaceutical formulation by high-1. Indian pharmacopoeia,1996; 2, A-169. perfomance liquid chromatography and high- performance thin-layer chromatography methods.J 2. 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