RESEARCH ARTICLE

Application of Absorbance Ratio Method for Simultaneous Spectroscopic Estimation of Rabeprazole Sodium and Levosulpiride in Combined Pharmaceutical Formulation
Hardik A Bhalodiya1*, Janvika M Boda1, Parula B Patel1
Abstracts: The present research work describes not only simple and rapid but also sensitive, accurate, precise, and absorbance ratio (Q ratio) method for the simultaneous estimation of Rabeprazole sodium and Levosulpiride in reliable pharmaceutical formulation. Absorbance ratio method uses the ratio of absorbances at two selected wavelengths, one combined an isoabsorptive point and other being the max of one of the two components. Rabeprazole sodium and which is isoabsorptive point at Levosulpiride show an 260.57 nm in methanol. The second wavelength used is 284 nm, which is the max of in methanol. The linearity was obtained in the concentration range of 5 - 25 g/ml for Rabeprazole sodium and 19 rabeprazole sodium Levosulpiride. The concentrations of both the drugs were determined by using ratio of absorbances at isoabsorptive 95 g/ml point and at the max of Rabeprazole sodium. The method was successfully applied to pharmaceutical dosage form because no from the tablet excipients was found. The results of analysis have been validated statistically for linearity, LOD, LOQ, interference precision, accuracy, robustness and ruggedness of the proposed method. Present method is economical in the means of both time & cost.

INTRODUCTION Colorimetric Method (28), UV - Spectroscopic Method (29), Rabeprazole sodium, 2-({[4-(3-methoxypropoxy)-3-methyl- High Performance Liquid Chromatographic Methods (30-34) & 2 pyridinyl] methyl}sulphenyl)-1H-benzimidazole sodium, Methods for Estimation from Biological Fluids (35-38) for a is Proton Pump Inhibitor that suppresses gastric acid Levosulpiride in bulk drug &/or individual dosage form secretion by specific inhibition of the gastric H+, K+ &/or combined dosage form (with other drugs) are existing. enzyme Extensive literature survey revealed that, still no ATPase system at the secretory surface of the gastric parietal cell. It is widely used in the treatment of acid- Spectroscopic method has been developed for peptic diseases, such as duodenal, gastric and esophageal simultaneous estimation of Rabeprazole sodium (RAB) & ulceration. It is official in Indian Pharmacopoeia - 2010.Levosulpiride (LEV) in combined dosage form. (1-5) Levosulpiride, N-[{(2S)-Ethylpyrrolidin-2-yl} methyl]-2- The absorbance ratio method (Q Ratio method) is a methoxy-5-sulphamoyl benzamide, is an anti - pshychotic modification of simultaneous equation procedure. It gastro - prokinetic agent. It exerts its anti - pshychotic depends on the property that for a substance which & effect via blocking D2 - receptor in CNS. As gastro-prokinetic obeys law at all wavelengths, the ratio of absorbance at Beers increases two agent, it the release of acetylcholine (Ach) through any wavelengths is a constant value independent of dopamine antagonistic action and inhibits decomposition D2 receptor concentration or path length. (39) of through its acetylcholinesterase inhibitory action, Ach Aim of present work was to develop rapid, simple, resulting in enhancement of gastrointestinal motility. Itreliable, economical, precise and accurate method for is used in treatment of pshychotic & gastro intestinal track simultaneous estimation of binary drug formulation. diseases. It is official in European Pharmacopoeia, Chinese Pharmacopoeia and Japanese Pharmacopoeia. (1, INSTRUMENTS, REAGENTS & MATERIALS 3-7) Combination of Rabeprazole sodium and Levosulpiride Instruments is used to reduce the amount of acid produced by the The instruments used in present study were, stomach and improve gastrointestinal motility, to treat Double beam UV/Visible spectrophotometer conditions such as Heartburn, Gastro Esophageal Reflux 1800 Shimadzu Disease (GERD), Irritable Bowel Syndrome (IBS) and Electronic weighing balance - Shimadzu AUX220 Gastritis. Combined formulation available in the market in the strength of Rabeprazole sodium 20 mg + Levosulpiride Chemical Used 75mg in Tablet dosage form. Methanol GR Grade, Merck Ltd. - Mumbai, India The Review of Literature has revealed that, Colorimetric Method (8), UV - Spectroscopic Methods (9-13), High Glass Wares Performance Liquid Chromatographic Methods (2, 14-23), Glass wares used were calibrated. All High Performance Thin Layer Chromatographic Methods (24, Methods for Estimation from Biological Fluids(26, 27) for 25) & Standard Drugs Rabeprazole sodium in bulk drug &/or individual dosage Analytically pure Standard Drugs of Rabeprazole sodium form &/or combined dosage form (with other drugs) are and Levosulpiride were obtained from Torrent Research existing. Non-aqueous Titration (Potentiometric) Method Centre, Gandhinagar, Gujarat, India as gift Samples and (6), used as such without further purification.
1Department of Quality Assurance, S. J. Thakkar Pharmacy College, Rajkot,

Gujarat, India. E-mail: hardy_pharma@yahoo.com *Corresponding author

Pharmaceutical Formulation The pharmaceutical dosage form used in this study was Rekool - L tablets (Alembic Pharma Pvt. Ltd.) were

Inventi Rapid: Pharm Ana & Qual Assur Vol. 2012, Issue 3 [ISSN 0976-3813]

2012 ppaqa 320, CCC: $10 Inventi Journals (P) Ltd Published on Web 17/04/2012, www.inventi.in

RESEARCH ARTICLE RESEARCH ARTICLE

Table 1: Absorptivity values at both the wavelengths Drug Absorptivity 1 = 260.57 nm 2 = 284 nm RAB aX1 = 0.0237 aX2 = 0.0382 LEV aY1= 0.0062 aY2 = 0.0071 Table 2: Assay result for tablet formulation (n=6) Parameters Tablet Formulation RAB LEV Concentration Taken 15 g/ml 56.25 g/ml Mean Concentration Obtained 14.87 g/ml 55.96 g/ml % Labeled Claim 99.13 % 99.49 % % RSD 0.8034 0.7305 Table 3: Validation Parameters Parameters Value Obtained RAB LEV Range 5 - 25 g/ml 19 - 95 g/ml Figure 1: Overlain Spectra of Rabeprazole Sodium & Levosulpiride in Equation 1 Y = 0.0382 X - 0.0024 Y = 0.0062 X - 0.014 2 Y = 0.0237 X - 0.0014 Y = 0.0071 Methanol X - 0.0069 Linearity R2 1 0.9998 0.9998 2 0.9998 0.9997 %RSD 1 0.1722 - 1.1120 0.5625 - 1.7283 2 0.4533 - 1.7282 0.5034 - 1.7525 LOD 0.2296 g/ml 1.6560 g/ml LOQ 0.6958 g/ml 5.0182 g/ml L1 100.45 % 100.33 % %Recovery L2 100.01 % 99.94% Accuracy (mean, n=3) L3 100.15 % 99.62 % (3 Level, 3 DeterL1 0.8076 0.1538 0.7 mination) %RSD L2 0.4619 0.4530 y = 0.023x -0.014 L3 0.3434 0.8929 0.6 R = 0.999 Repeatability (%RSD) 0.5328 0.4718 Precision Intraday Precision (%RSD) (n = 3) 0.2196 0.3369 0.5 ce = 6, 100%) (n Interday Precision an 0.4 (%RSD) (n=3) 0.6224 0.3838 Figure 2: Calibration curve of Rabeprazole sodium at 260.57 Figure 3: Calibration curve of Rabeprazole sodium at 284 rb nm 0.3 nm Robustness Scanning Speed (n=3) 0.4094 0.4114 Solvent from Different Manufacturers (n=2) 0.5683 0.4409 so Ab Ruggedness (Different Analyst, n=2) 0.6163 0.5176 0.2

dissolved & diluted up to mark with Methanol to 1.2 final concentration of 1000 g/ml of each component. obtain y = 0.038x -0.002 Both the solutions were further diluted with methanol 1 R = 0.999 obtain working standard solutions of 100 g/ml of each. to Concentration in g/ml 0.8 The standard solutions were prepared individually by ce Methodology dilution of the working standard solutions with methanol an 0.6 Selection of Suitable Common Solvent for Solubility to reach concentration range of 5 - 25 g/ml for RAB and rb of Both Drugs: From the Review of Literature, it is 19 so95 g/ml for LEV respectively. Absorbances of 0.4 concluded that Both Rabeprazole Sodium (RAB) & prepared standard solutions having concentration 5, 10, Ab 15, 0.2 and 25 for RAB and 19, 38, 57, 76, and 95 g/ml 20, Levosulpiride (LEV) are soluble in Methanol. So, for were measured at 260.57 nm (isoabsorptive point) LEV Methanol is suitable common solvent for solubility of and 284 nm (max of RAB) and calibration curves both drugs for UV Spectroscopic determination. 0 (Absorbance Vs Concentration) were plotted. Figure 4: Calibration curve Wavelengths for Absorption Figure 5: Calibration curve of Levosulpiride at 284 nm Selection of Analytical of Levosulpiride at 260.57 nm 0102030 Absorptivity were determined using calibration curves at coefficients Concentation in g/ml Ratio Method: From the overlain spectra of two drugs ml (Figure - 1),flask. To this that RAB ml methanol was iso A2 were recorded at 260.57 nm (isoabsorptive volumetric it is evident flask, 15 and LEV show an both the wavelengths. and added and the flaskat 260.57 nm. The 5 min. The volume was sonicated for second wavelength point) and 284 nm (max of RAB) respectively (Figure absorptive point is adjusted up to the mark with methanol. The solution -6), ratioPharmaceutical(QM) was calculated, i.e. A2/A1. and of of absorbance Formulation Assay used is 284 nm, which is the max of RAB. was then filtered through Whatman & Calibration 41.Relativetablets were weighed drugs in the sample was filter paper no. Twenty concentration of two accurately and finely Preparation of Standard Solutions The solution was suitably diluted with methanol to get calculated using equation (1) and (2). The analysis of RAB a Curves: Accurately weighed 50 mg of each RAB and powdered. The tablet powder equivalent to 25 mg LEV final concentration of 15 g/ml of RAB and 56.25 g/mlprocedure mg of LEV were weighed andtablet were transferred to 50 ml volumetric flasks separately, 93.75 was repeated six times with transferred to 25 and of LEV. The absorbances of the sample solution i.e. A1 formulation. procured from local pharmacy. Rekool - L labeled to 0.1 contain 20 mg of Rabeprazole sodium and 75 mg of 0 Levosulpiride in entric coated bilayer tablet as sustained release 051015202530 form.
Inventi Rapid: Pharm Ana & Qual Assur Vol. 2012, Issue 3 [ISSN 0976-3813] Inventi Rapid: Pharm Ana & Qual Assur Vol. 2012, Issue 3 [ISSN 0976-3813] 2012 ppaqa 320, CCC: $10 Inventi Journals (P) Ltd Published on320, CCC: $10 Inventi Journals (P) Ltd 2012 ppaqa Web 17/04/2012, www.inventi.in Published on Web 17/04/2012, www.inventi.in

ce an rb so Ab0 020406080100 -0.014 in g/ml 0.7 0.6 0.5 0.4 0.3 0.2 0.1 y =R Concentration 0.006x = 0.999

ce an rb so Ab 0 020406080100 -0.006 in g/ml 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 y = 0.007x R = 0.999 Concentation