A new, simple, precise, accurate and reproducible RP-HPLC method for Simultaneous estimation of Desloratadine and Montelukast in bulk and pharmaceutical formulations. Separation of Desloratadine and Montelukast was successfully achieved on a ECLEPSE XDB C8 (4.6 x 150mm, 5 mm, Make: Waters) or equivalent in an isocratic mode utilizing K2HPO4 buffer (pH: 8.6) Methanol (60:40%v/v) at a flow rate of 0.8 mL/min and elute was monitored at 261 nm, with a retention time of 2.485 and 3.800 minutes for Desloratadine and Montelukast. The method was validated and the response was found to be linear in the drug concentration range of 50 µg/mL to 150 µg/mL for Desloratadine and 50 µg/mL to 150 µg/mL for Montelukast. The LOD and LOQ for Desloratadine were found to be 2.759, 9.195 respectivly. The LOD and LOQ for Montelukast were found to be 2.9091, 9.6970 respectively. This method was found to be good percentage recovery for Desloratadine and Montelukast were found to be 100.00% and 100.00% respectively indicates that the proposed method is highly accurate. The specificity of the method shows good correlation between retention times of standard with the sample so, the method specifically determines the analyte in the sample without interference from excipients of tablet dosage forms. The method was extensively validated according to ICH guidelines for Linearity, Range, Accuacy, Precesion, Specificity and Robustness

Geetha, K.

Mallesham, B.

Ramarao, Nadendla

Uma Maheswar Rao, Dr. V.

International Journal of Innovative Technology and Research (IJITR)

   B.Mallesham* et al.   (IJITR) INTERNATIONAL JOURNAL OF INNOVATIVE TECHNOLOGY AND RESEARCH Volume No.2, Issue No.5, August – September 2014, 1181 – 1186.  2320 –5547  @ 2013 http://www.ijitr.com All rights Reserved.       Page | 1181  Simultaneous Estimation of Desloratadine and Montelukast in Bulk and Pharmaceutical Formulations by RP-HPLC B.MALLESHAM M.Pharm Department of Pharmaceutical Analysis &QA C M R College of Pharmacy JNTUH, Hyderabad, INDIA. K.GEETHA Department of Pharmaceutical Biotechnology C M R College of Pharmacy JNTUH, Hyderabad, INDIA Dr. V. UMA MAHESWAR RAO M.Pharm.PH.D. Department of Pharmacognosy C M R College of Pharmacy JNTUH, Hyderabad, INDIA. NADENDLA RAMARAO M.Pharm.PH.D. Department of pharmaceutics Chalapathi institute of pharmaceutical sciences Guntur, A.P. INDIA.  Abstract: A new, simple, precise, accurate and reproducible RP-HPLC method for Simultaneous estimation of Desloratadine and Montelukast in bulk and pharmaceutical formulations. Separation of Desloratadine and Montelukast was successfully achieved on a ECLEPSE XDB C8 (4.6 x 150mm, 5 m, Make: Waters) or equivalent in an isocratic mode utilizing K2HPO4 buffer (pH: 8.6) Methanol (60:40%v/v) at a flow rate of 0.8 mL/min and elute was monitored at 261 nm, with a retention time of 2.485 and 3.800 minutes for Desloratadine and Montelukast. The method was validated and the response was found to be linear in the drug concentration range of 50 µg/mL to 150 µg/mL for Desloratadine and 50 µg/mL to 150 µg/mL for Montelukast. The LOD and LOQ for Desloratadine were found to be 2.759, 9.195 respectivly. The LOD and LOQ for Montelukast were found to be 2.9091, 9.6970 respectively. This method was found to be good percentage recovery for Desloratadine and Montelukast were found to be 100.00% and 100.00% respectively indicates that the proposed method is highly accurate. The specificity of the method shows good correlation between retention times of standard with the sample so, the method specifically determines the analyte in the sample without interference from excipients of tablet dosage forms. The method was extensively validated according to ICH guidelines for Linearity, Range, Accuacy, Precesion, Specificity and Robustness. Keywords: RP-HPLC; Desloratadine, Montelukast. I. INTRODUCTION Desloratadine: 13-chloro-2-(piperidin-4-ylidene)-4 azatricyclo [9.4.0.0^{3,8}] pentadeca 1(11),3,5,7,12,14-hexaene. It belongs to the benzocycloheptapyridines. These are aromatic compounds containing a benzene ring and a pyridine ring fused to a seven membered carbocycle. Desloratadine is a second generation, tricyclic antihistamine that which has a selective and peripheral H1-antagonist action.  Figure1: Chemical structure Of Desloratadine Montelukast: 2-[1-({[(1R)-1-{3-[(E)-2-(7-chloroquinolin-2-yl) ethenyl] phenyl}-3-[2-(2-hydroxypropan-2-yl) phenyl] propyl] sulfanyl} methyl) cyclopropyl] acetic acid. Montelukast inhibits the actions of LTD4 at the CysLT1 receptor, preventing airway edema, smooth muscle contraction, and enhanced secretion of thick, viscous mucus. It is an Antilipidemic Agents Figure2: Chemical structure of Montelukast II. MATERIALS AND METHOD A. Instrumentation  The separation was carried out on HPLC system with Waters 2695 alliance with binary HPLC pump, Waters 2998 PDA detector, Waters Empower2 software with Eclipse XBD-C8, (150mm × 4.6 ; 5µm) column.      B.Mallesham* et al.   (IJITR) INTERNATIONAL JOURNAL OF INNOVATIVE TECHNOLOGY AND RESEARCH Volume No.2, Issue No.5, August – September 2014, 1181 – 1186.  2320 –5547  @ 2013 http://www.ijitr.com All rights Reserved.       Page | 1182  B. Chemicals and Reagents  Desloratadine and Montelukast was a gift sample by Dr. Reddy’s Laboratories Ltd., Hyderabad. K2HPO4, Methanol of HPLC grade was purchased from E. Merck (India) Ltd., Mumbai. Ortho phosphoric acid of AR grade was obtained from S.D. Fine Chemicals Ltd., Mumbai and mille Q water. C. HPLC Conditions The mobile phase consisting of K2HPO4 buffer (pH: 8.6) Methanol was degassed and were pumped from the solvent reservoir in the ratio of 60:40v/v was pumped into the column at a flow rate of 0.8 ml/min. The column temperature was 40°C. The detection was monitored at 261nm and the run time was 6min. The volume of injection loop was 10µl prior to injection of the drug solution the column was equilibrated for at least 15 min. with the mobile phase flowing through the system. D. Preparation of standard solution Accurately weigh and transfer 5mg of Desloratadine and 10mg of Montelukast into 50ml of volumetric flask and add 10ml of methanol to each and sonicate 10min (or) shake 5min and makeup the volume with methanol. Pipette out 5.0ml standard stock standard stock into 25ml volumetric flask dilute to volume with methanol and inject into HPLC. E. Preparation Of Sample Solution Accurately weighed 797.6mg of sample. Transfer the sample powder into 50ml of volumetric flask add 10ml methanol, sonicate for 20mins. Then make up the volume with methanol and filter through the 0.45µm filter paper. Transfer 5ml of above solution 25 ml volumetric flask and make up the volume with methanol.  Figure 3: Standard chromatogram for Desloratadine and Montelukast   Figure 4: Formulation chromatogram for Desloratadine and Montelukast F. Method validation 1. System Suitability Studies  The column efficiency, resolution and peak asymmetry were calculated for the standard solutions (Table I).The values obtained demonstrated the suitability of the system for the analysis of this drug combinations, system suitability parameters may fall within ± 3 % standard deviation range during routine performance of the method. TABLE I: SYSTEM SUITABILITY PARAMETERS Parameter  Desloratadine Montelukast Retention time 2.485 3.800 Theoretical plates 5123 5986 Tailing factor 1.39 1.52 % RSD 0.4 0.3 2.  Specificity Specificity is the ability to assess unequivocally the analyte in the presence of components which may expect to be present. Typically these might include impurities, degradants, matrix, etc. 3. Accuracy and precision The accuracy of the method was determined by recovery experiments. The recovery studies were carried out six times. The percentage recovery and standard deviation of the percentage recovery were calculated. From the data obtained, added recoveries of standard drugs were found to be accurate (Table-II&III).      B.Mallesham* et al.   (IJITR) INTERNATIONAL JOURNAL OF INNOVATIVE TECHNOLOGY AND RESEARCH Volume No.2, Issue No.5, August – September 2014, 1181 – 1186.  2320 –5547  @ 2013 http://www.ijitr.com All rights Reserved.       Page | 1183  TABLE II: ACCURACY FOR DESLORATADINE S.No Accuracy level Sample weight μg/ml added μg/ml found % Recovery % Mean    1    50% 398.80 39.640 39.78 100    100 398.80 39.640 39.79 100 398.80 39.640 39.76 100 398.80 39.640 39.77 100 398.80 39.640 39.80 100 398.80 39.640 39.75 100   2   100% 797.60 79.280 79.36 100  100 797.60 79.280 79.43 100 797.60 79.280 79.36 100    3    150% 1196.40 118.920 119.03 100   100 1196.40 118.920 118.90 100 1196.40 118.920 119.01 100 1196.40 118.920 118.95 100 1196.40 118.920 118.98 100 1196.40 118.920 118.98 100 TABLE III: ACCURACY FOR MONTELUKAST S.No Accuracy level Sample weight μg/ml added μg/ml found % Recovery % Mean    1    50% 398.80 79.760 79.83 100   100 398.80 79.760 79.79 100 398.80 79.760 79.89 100 398.80 79.760 79.77 100 398.80 79.760 79.85 100 398.80 79.760 79.84 100   2  100% 797.60 159.20 159.34 100  100 797.60 159.20 159.47 100 797.60 159.20 159.56 100    3    150% 1196.40 239.280 239.28 100   100 1196.40 239.280 239.24 100 1196.40 239.280 239.47 100 1196.40 239.280 239.49 100 1196.40 239.280 239.37 100 1196.40 239.280 239.29 100  Figure 5: AccuracyChromatograms-50%of Desloratadine and Montelukast Figure 6: AccuracyChromatograms-100%of Desloratadine and Montelukast  Figure 7: AccuracyChromatograms-150%of Desloratadine and Montelukast    B.Mallesham* et al.   (IJITR) INTERNATIONAL JOURNAL OF INNOVATIVE TECHNOLOGY AND RESEARCH Volume No.2, Issue No.5, August – September 2014, 1181 – 1186.  2320 –5547  @ 2013 http://www.ijitr.com All rights Reserved.       Page | 1184  The precision of the method was demonstrated by inter-day and intra-day variation studies. In the intraday studies, six repeated injections of standard and sample solutions were made and the response factor of drug peaks and percentage RSD were calculated. In the inter-day variation studies. six repeated injections of standard and sample solutions were made for three consecutive days and response factor of drugs peaks and percentage RSD were calculated. The chromatograms of three different levels shown in Figure 5, 6 &7. From the data obtained, the developed RP-HPLC method was found to be precise. (Table-IV) TABLE IV: PRECISION STUDIES  4. Linearity and range The linearity of the method was determined at five concentration levels. The calibration curve was constructed by plotting response factor against concentration of drugs. The slope and intercept value for calibration curve was Y=43363X (R2=0.999) for desloratidine and Y=49489X (R2=1) for montelukast. The results shows that an excellent correlation exists between areas and concentration of drugs within the concentration range indicated above.  The overlay chromatograms of Linearity for Desloratadine and Montelukast shows in Figure 10 and the results for calibration curves are given in Figure 8&9.          Figure 8: Linearity Curve for Desloratidine Figure 9: Linearity Curve for Montelukast  Figure 10:  Overlay chromatograms of Linearity for Desloratidine and Montelukast 5. Robustness Robustness of the method was determined by making slight changes in the chromatographic conditions.    It was observed that there were no marked changes in the chromatograms which demonstrated that the RPHPLC method developed, are robust (Table-V&VI). TABLE V: ROBUSTNESS FOR DESLORATIDINE S.No Area Area %Assay (Des ) %Assay (Des) (mon) (mon ) 1 4678289 4945121 99 100 2 4677549 4941492 99 100 3 4676508 4945109 99 100 4 4675202 4943465 99 100 5 4677862 4942694 99 100 6 4678822 4949517 99 100 S.No Sample Name RT Area USP Tailing 1 Temp-1 4.955 4960210 1.39 2 Temp-2 2.978 4927514 1.35 3 Flow-1 3.725 4511175 1.32    4 Flow-2 3.722 4512813 1.32    B.Mallesham* et al.   (IJITR) INTERNATIONAL JOURNAL OF INNOVATIVE TECHNOLOGY AND RESEARCH Volume No.2, Issue No.5, August – September 2014, 1181 – 1186.  2320 –5547  @ 2013 http://www.ijitr.com All rights Reserved.       Page | 1185  TABLE VI: ROBUSTNESS FOR MONTELUKAST 6. LOD&LOQ Limit of quantification and detection were predicted by plotting linearity curve for different nominal concentrations of Desloratadine and Montelukast. RSD (σ) method was applied; the LOQ and LOD values were predicted using following formulas (a) and (b). Precision was established at these predicted level (a) LOQ = 10 σ / S (b) LOD = 3.3 σ / S Where  σ = residual standard deviation of response  S = slope of the calibration curve. Figure 11: Chromatograms for LOQ Figure 12: Chromatograms for LOD TABLE VII: LOD and LOQ for Desloratidine and Montelukast   III. RESULTS AND DISCUSSION System suitability results were given by table1 and system suitability parameters are retention time, resolution, tailing and plate count were shown uniformity and %RSD was less than 1. So we can say system is suitable for analysis method specificity was concluded by figure:3 and figure:4 those figures are Desloratadine and  montelukast standard chromatogram and other one is formulation they were not observed placebo and excipients peaks interference with standard and analytic peak so it proves method is selective. The result given in table I say that the method precision passed for both Desloratadine and montelukast studies. The method accuracy was evaluated by recovery studies. Desloratadine and  montelukast recovery was founded 99%&100% as per ICH 97%- 103% and also percentage RSD was very low so method is accurate shown in table II&III. Linearity calibration curve was given below fig: 8&9 and plot the graph three different concentrations versus areas to construct the linear regression equation and to calculate the value of correlation co-efficient. Linear correlation was found to be Y=43363X for Desloratidine and y = 49488X for Montelukast Method robustness results were given by table V&VI, LOQ and LOD Results were given by table VII. IV. CONCLUSION The proposed HPLC method was found to be simple, precise, accurate and sensitive for the simultaneous estimation of    Desloratidine and Montelukast pharmaceutical dosage forms. Hence, this method can easily and conveniently adopt for rountine quality control analysis of    Desloratidine and Montelukast pure and its pharmaceutical dosage forms. V. ACKNOWLEDGEMENT I am thankful to department of Pharmaceutical Analysis and Quality Assurance of CMR College of pharmacy, Jawaharlal Nehru Technological University, Hyderabad, for providing instruments and analytical support. VI. REFERENCES [1]  Desloratadine Drug profile:   www.drugbank.ca/drugs/DB00967. [2] Montelukast Drug profile.   www.drugbank.ca/drugs/DB00471. [3]  Lakshmana Rao, K.Naga Raju and T.Gopala Swamy. “Development and Validation of RP-HPLC method for the determination of Montelukast Sodium in bulk and Pharmaceutical formulation", IJPCBS, vol. 1(1), pp.12-16, 2011. S.No Sample Name RT Area USP Tailing 1 Temp-1 3.267 4511175 1.52 2 Temp-2 1.961 4512813 1.54 3 Flow-1 2.451 6117979 1.51 4 Flow-2 2.459 3585343 1.48 S.No Sample Name Name RT Area 1 LOD Desl 2.451 881757 2 LOQ Desl 2.452 1898664 1 LOD Mont  3.719 1025560 2 LOQ Mont  3.724 2071910    B.Mallesham* et al.   (IJITR) INTERNATIONAL JOURNAL OF INNOVATIVE TECHNOLOGY AND RESEARCH Volume No.2, Issue No.5, August – September 2014, 1181 – 1186.  2320 –5547  @ 2013 http://www.ijitr.com All rights Reserved.       Page | 1186  [4]  Singh RM, Saini PK, Mathur SC, Singh GN, Lal B. “Development and Validation of a RP-HPLC Method for estimation of Montelukast Sodium in bulk and in tablet dosage form" Indian J Pharm Sci., vol. 72(2), pp.235-7, Mar 2010. [5]   R. Vibhuthi,Chhatrala, jitendra patel, “simultaneous estimation of Montelukast Sodium and Desloratadine by RP HPLC in their marketed formulation"International jornal of Chem Tech Research, vol 4,No.4,pp.1402-1407,2012. [6]  SV Patel1,GF Patel1,SG Pipaliya, “Development and “Validation of Derivative Spectroscopic Method for Simultaneous Estimation of Montelukast Sodium and Desloratadine in Bulk and Combined Dosage Form", Inventi Journals (P) Ltd, Pharm Ana & Qual Assur ,vol. 2012, Issue 2 [E- ISSN 0976–755X, P- ISSN 2229–4198],pp.82-85,2012. [7]  Rima M. Bankar, Dipti B. Patel, “RP-HPLC method for simultaneous estimation of montelukast sodium and desloratadine in combined dosage form", pharmatutor.org [8]  Kalyankar tukaram M,kokate ranjeet H,kakaderajndra B, “RP-HPLC Method for Simultaneous Estimation of Montelukast Sodium and Desloratadine from bulk and tablets formulation", International research journal of pharmacy, vol.3(7),ISSN 2230-8407, pp.343-347, 2012.  

SIMULTANEOUS ESTIMATION OF DESLORATADINE AND MONTELUKAST IN BULK AND PHARMACEUTICAL FORMULATIONS BY RP-HPLC

http://www.ijitr.com/index.php/ojs/article/download/370/pdf

SIMULTANEOUS ESTIMATION OF DESLORATADINE AND MONTELUKAST IN BULK AND PHARMACEUTICAL FORMULATIONS BY RP-HPLC

Abstract

Similar works

Full text

Available Versions

International Journal of Innovative Technology and Research (IJITR)