D Kolekar et al. Chemometric assisted QbD method for estimation of bilastine and montelukast sodium Vol 3 | Issue 1 | Jan – Mar 2024 Indian J Pharm Drug Studies | 16 Original Article Chemo metric assisted Spectrophotometric Method Development through Quality by design Approach for the estimation of Bilastine and Montelukast sodium in combined solid dosage form Bhagyashri D Kolekar1, Namrata N Gawade1, G K Dyade1, Nilesh Y Jadhav2 From, 1Dept of Post Graduate Studies in Pharmaceutical Quality Assurance, SVPM’S College of Pharmacy, Malegaon (BKII), Baramati, Pune, 2SSPM’S Dr N. J. Paulbudhe College of Pharmacy, Survey No 45/1B, Vasant Tekadi, Savedi, Ahmednagar, Maharashtra, India ABSTRACT Objective: Quality by design (QbD) is a systematic process for pharmaceutical development recommended by regulatory agencies like USFDA. Development of various pharmaceutical processes including analytical methods by applying Quality by design aids in ensuring the robustness of the method. QbD approached chemo metric assisted UV-VIS spectrophotometric analytical method was developed for the estimation of Bilastine (BSE) and Montelukast sodium (MKS) from their combined dosage forms. Materials and Method: Simultaneous equation method was selected from the nature of spectra, solvent 50 % alcohol was utilised; and for method 274.5 nm and 351.5 nm was the wavelength for measurement of absorbance of bilastine and montelukast sodium respectively. Effect of input variables on spectrum characteristics were studied for selection of critical parameters and developed method was validated as per ICH Q 2 R1 regulatory guidelines. Linearity of the drugs was ascertained over the conc range 1-32 mcg/ml (microgram/ml) for BSE and 1-20 mcg/ml for MKS. Results and Discussion: The percentage purity of assay was found 98.09 % for BSE and 103.62 % for MKS; and the accuracy study data were varied from 0.2523 to 0.5221 for BSE and 0.2512 to 1.2515 for MKS. Precision study was shown acceptable data as SD data varied from 0.1902 to 0.5773 for BSE and from 0.2828 to 0.5458 for MKS. Conclusion: The developed method is rigid, robust and efficient for the estimation of BSE and MKS from the composition of dosage form. QbD was applied to build rigid robust method through risk assessment at early stage and defining the design space at the later stage. Key Words: Bilastine, Montelukast Sodium, Qbd, ICH, Simultaneous Equation Method ilastine (BSE) chemically 4-[2-[4-[1-(2-Ethoxy ethyl)-1H-Benzimidazole-2-yl]-1-piperidinyl] ethyl]- α, α-dimethyl benzene acetic acid [1] is an antihistamine, non-sedative histamine H1 receptor antagonist; by binding and preventing activation of H1 receptor bilastine reduces development of allergic symptoms due to release of histamine from mast cell [2]. Literature survey revealed that various analytical methods have been reported for estimation of BSE such as UV spectrophotometric methods lonely [3-6], UV spectrophotometric method with MKS or other drug [7], for estimation of BSE with MKS or other by RP-HPLC [8-14], stability indicating RP-HPLC [15-17], green QbD HPLC [18], LC-MS/MS [19], stability indicating UPLC [20] and RP- UFLC [21] alone or in combination with other drugs. Access this article online Received – 21th Dec 2023 Initial Review – 30th Dec 2023 Accepted – 08th Jan 2024 Quick Response Code Montelukast sodium (MKS) chemically Sodium [1-[[[(1 R)-1-(3-[(E) 2-(7-chloroquinoline-2-yl) ethenyl] phenyl)-3-[2- (1-hydroxy-1-methylethyl) phenyl] propyl] sulfanyl] methyl] cyclopropyl] acetate [1] is an anti-asthmatic, leukotriene receptor antagonist. It can completely block the binding of CYSLT’S to receptor that they can inhibit the binding of inflammation mediator LTD4 [2]. Literature survey revealed that various analytical methods have been reported for estimation of MKS includes UV spectrophotometric method lonely [22-28], UV spectrophotometric method with BSE or other drug [29-33], for estimation of MKS Qbd technique [34], RP-HPLC methods [35-40], stability indicating HPLC with other [41-43] and HPTLC [44] alone or in combination with other drugs. Montelukast sodium is official in Indian and British Pharmacopoeia [45, 46]. Chemical structure of both these drugs is shown in (Figure 1). _________________________________________ Correspondence to: G K Dyade, Department of PG Studies in Pharm Quality Assurance, SVPM’S College of Pharmacy, Malegaon (BKII), Baramati, Pune, Maharashtra, India. Email: pharmacyresearchsvpmcop@gmail.com B mailto:pharmacyresearchsvpmcop@gmail.com D Kolekar et al. Chemometric assisted QbD method for estimation of bilastine and montelukast sodium Vol 3 | Issue 1 | Jan – Mar 2024 Indian J Pharm Drug Studies | 17 Figure 1: Chemical structure of Drug molecule Quality by design concept is applied for the development of pharmaceutical processes to assure a predefined product quality. QBD concepts are mentioned in ICH guidelines Q8 (R2) (Pharmaceutical development), Q9 (Quality risk management), and Q10 (Pharmaceutical quality system) [47- 49] shown in (Figure 2). ICH guidelines Q8 (R2) defines QBD as a “a systematic approach to development that begins with predefined objectives and emphasizes product and process understanding and process control, based on sound science and quality risk management”[50]. QBD approach in analytical method summarizes a complete understanding of how the analytical technique attributes and operating conditions affect the analytical performance. Factors to study in analytical quality by design (AQbD) approach may include the type of analytical technique chosen, reagents used and instrument parameters. Fig No 2: Analytical QbD approach There are similar advantages of applying QbD principles to analytical methods as to manufacturing processes and product [51]. A QbD approach can be beneficial in the development of suitable, robust, low cost and eco-friendly (eco-friendly solvent, chemicals) method which is applicable at any stage of the lifecycle of the product. Also some regulatory guidelines have mentioned flexibility of changing analytical method without revalidation if the AQbD approach has been implemented during analytical method development. The first stage of AQbD approach is to fix an analytical target profile (ATP) for the method. ATP defines the goal of the analytical method development process and it is the sign of method performance [52, 53]. For analytical method validation ICH Q2 (R1) has given various method performance characteristics for an analytical method. In development of UV-VIS spectrophotometric method, QbD approach was implemented with the study of the effect of method input variables on spectral shape, intensity of absorbance, and absorbance maxima λmax and critical parameters were selected for the proposed method and method was validated as per ICH guidelines Q2 (R1). MATERIALS AND METHODS Instrumentation: Analysis was performed with a Shimadzu Double beam UV-Visible spectrophotometer (Shimadzu, Kyoto, Japan) with spectral bandwidth of 2 nm and wavelength accuracy of ± 1 nm with 10 mm matched Quartz cells was used. Electronic balance Afcoset balance (The Bombay Burmah Trading corpo Ltd) with accuracy ±0.1 mg Model No. ER 200A was utilised for weighing and for degassing the solution Digital Ultrasonic cleaner 1.8 Ltr (Labman scientific Instruments Chennai) was used. Reagents and Chemicals: Pharmaceutically pure samples of BSE and MKS from Glenmark Pharmaceuticals, Nashik, Maharashtra, India were procured as a gift sample and the commercial formulation Bilafav-M Tablet containing bilastine 20 mg and montelukast sodium 10 mg was procured from the local market. AQbD approach application in method development: AQbD approach was applied to study the influence of input variable parameters on spectrophotometric analytical method performance shown in (Figure 3). Figure 3: Diagram showing the relationship between input variable parameters and the spectrophotometric method performance characteristics D Kolekar et al. Chemometric assisted QbD method for estimation of bilastine and montelukast sodium Vol 3 | Issue 1 | Jan – Mar 2024 Indian J Pharm Drug Studies | 18 Solvent selection: BSE is freely soluble in 10% acetic acid glacial, chloroform and in ethanol, slightly soluble in 1 N HCl and soluble in 1N NaOH and sparingly soluble in water; whereas MKS is very soluble in water, methylene chloride and ethanol. Although the solubility of the procured drugs were studied in alcohol 90%, 0.1 N HCl and 0.1 N NaOH separately; and found that BSE is soluble in ethanol, 0.1 N NaOH, slightly soluble in 0.1 HCl however MKS soluble in ethanol and insoluble in NaOH and HCl. Both drugs solubilises in 50% alcohol, hence selected as a common solvent. Each drugs solution with known conc was scanned in UV range of 400 nm to 200 nm. It was found that alcohol 50% is suitable with respect to stable, robust and precise in producing result. Preparation of stock solutions and standard solutions: 10 mg each of drug BSE and MKS were separately and accurately weighed; and transferred into separate 25 ml volumetric flask. Dissolved into solvent alcohol 50% and volume was made to 25 ml with this solvent. Working standard solution was prepared by diluting 5 ml to 10 ml with 50 % alcohol. Subsequent standard solution of each drug with conc 16μg/ml was prepared by diluting aliquot of stock solution to 10 ml with 50% alcohol into 10 ml capacity volumetric flask. Selection of wavelength and conc range: From UV spectra it was found that BSE has measurable absorbance at 274.5 nm and 281.5 nm (Figure 4) and less interference was observed by MKS; similarly MKS has maximum absorbance at 351.5 nm and negligible interference by BSE was accounted. Chemometric method using simultaneous equation was applied and which was reasonable remedy to overcome interference at each other’s absorbance. To study linearity, working conc range 1 to 32μg/ml for BSE and 1 to 20μg/ml for MKS was selected. Also combined drug solution was prepared simulated to marketed formulation. Selected critical parameters based upon above discussion, observations were listed in (Table 1) and by using these; method was validated as per ICH guidelines and by analysing marketed preparations. Experimental Method for estimation: From the overlain spectra simultaneous equation method was applied for estima- tion of both the analytes from their combined dosage form. Figure 4: Overlaid spectra of BSE and MKS Table No 1: Selected critical parameter for UV-VIS analytical method of BSE and MKS Parameter Selected variables for simultaneous equation method BSE MKS Wavelength 274.5 351.5 Solvent 50% alcohol 50% alcohol Scan speed Fast Fast Sampling interval 0.2 nm 0.2 nm Simultaneous Equation Method for estimation of bilastine and montelukast sodium: BSE was shown maximum absorbance i.e. λmax at 274.5 nm where moderate interference by MKS found and MKS has maximum absorbance i.e. λmax at 351.5 nm where negligible interference by BSE observed. At 274.5 nm the λmax of BSE, the MKS was shown consistency in the absorptivity; hence two wavelengths 274.5 and 351.5 nm were considered as 1 and 2 respectively for the said method to estimate BSE and MKS. The equation A= abc was applied for x (BSE) and y (MKS) determination. Working standard solutions of BSE and MKS containing 12μg/ml conc were separately prepared and used for the method. Where CX = Conc of BSE in sample solution A1 and A2 = absorbance of sample solution at 1 and 2 wavelength ay1 and ay2 = absorptivity of MKS at 1 and 2 wavelength of standard solution ax1 and ax2 = absorptivity of BSE at 1 and 2 wavelength of standard solution Cy = Conc of MKS in sample solution As = Absorbance of Sample solution at 2 wavelength VALIDATION of the METHOD Selected critical parameters should meet the performance characteristics of the analytical method so as to attain analytical target profile of the method. An ICH guideline Q2 R1 was applied to study methods performance with critical parameters in order to implement AQbD approach. The method was validated as per ICH guidelines System suitability: System suitability is studied to demonstrate the suitability of the developed procedure under consideration for the analytical method. Six replicates of working standard solutions with conc 20μg/ml and 16μg/ml of BSE and MKS respectively were prepared separately and absorbance was recorded, calculated SD and % RSD of the response. D Kolekar et al. Chemometric assisted QbD method for estimation of bilastine and montelukast sodium Vol 3 | Issue 1 | Jan – Mar 2024 Indian J Pharm Drug Studies | 19 Linearity: The linearity of an analytical method is its ability to obtain response i.e. absorbance which is directly proportional to the conc of analyte. Series of working standard solutions were prepared in conc. range of 1-32 μg/ml for BSE and 1-20 μg/ml for MKS and scanned in 400 to 200 nm range in spectrum mode of the spectrophotometer, absorbance of the standard solutions were recorded at their respective wavelength; i.e. 274.5 for BSE and 351.5 nm for MKS in spectrum order. Microsoft office excel software tool was used to obtain the standard regression curve and its analysis as slope, intercept, and correlation coefficient. Assay of formulation: Assay was carried out by proposed methods and assay was validated by statistical parameters. Estimation of formulations by simultaneous equation method: Tablet powder equivalent to 10 mg BSE and 5 mg MKS was weighed and transferred into 25 ml volumetric flask. Dissolved into 50% alcohol, mixed well for 10 mins and volume was made to 25 ml with the solvent. Solution was filtered through what man filter paper and aliquots of solution were further diluted with the 50% alcohol to obtain tablet sample solution. Solution was scanned in the range of 400 to 200 nm to obtain absorbance of tablet solution at 274.5 nm and 351.5 nm in spectrum order. Obtained absorbance were utilised to estimate unknown conc of formulation; and results were statistically validated to obtain % of nominal conc, standard deviation and % of RSD. Accuracy and Precision: The accuracy of an analytical method expresses the closeness of an agreement between test result and true result. Accuracy study was performed by recovery study i.e. standard addition method; diluted standard solutions of BSE and MKS were prepared and standard solutions added in 80,100 and 120% proportionate to the tablet solution. Three replicates at each of these three levels were prepared, measured and % of conc, SD and RSD were calculated. The precision study was carried out by performing assay of tablet six times; also the reproducibility in result was studied by inter day and intraday precision. Limit of Detection (LOD) and Limit of Quantitation (LOQ): The LOD and LOQ of BSE and MKS by the proposed method were determined using calibration graph method and calculated as 3.3σ/s and 10 σ/s for LOD and LOQ respectively; σ is the standard deviation of calibration curve and s is the slope of regression line. Robustness and Ruggedness: It is measure of capacity of analytical procedure to remain unaffected by small but deliberate variations in method parameter. RESULTS AND DISCUSSION Method development comprises numerous steps, and of which solvent selection, selection of method for measurement are significant one. Uses of aqueous solvents, eco-friendly solvents like hydrotropic have got remarkable weightage due to low cost, readily available and environmentally sound. Drugs underlying analysis must have appreciable solubility in the selected solvent. Chemical structure of the drug and physico-chemical properties available in the literature guides about use of appropriate solvent in the method. From UV spectra two wavelengths were selected as 274.5 nm (λmax of BSE) and 351.5 nm (λmax of MKS) for calculation of both drugs in combined solution shown in (Fig No 4). System Suitability: The absorbances of six replicates of standard solutions of respective drugs conc are reported in (Table No 2). The SD and % RSD was found for BSE and MKS and meets the system suitability requirements indicate method was suitable for analysis. Table No 2: System suitability study of BSE and MKS Conc in μg /ml Absorbance of BSE Conc in μg /ml Absorbance of MKS 20 μg /ml 0.3124 16 μg /ml 0.5269 20 μg /ml 0.3110 16 μg /ml 0.5443 20 μg /ml 0.3281 16 μg /ml 0.5146 20 μg /ml 0.3115 16 μg /ml 0.5156 20 μg /ml 0.3395 16 μg /ml 0.5332 20 μg /ml 0.3292 16 μg /ml 0.5581 SD RSD 0.006662 0.41972 SD RSD 0.01281 0.29142 Linearity: The calibration curve of both drugs was found to be linear shown in (Figure 5) in the conc range of 1-32 μg/ml for BSE and 1-20 μg/ml for MKS as shown in (Figure 6). The regression equation of line and parameters slope, r2 value and intercept (Figure 7) are tabulated in (Table 3), which proved the linear relationship between conc and obtained response. Assay: The assay was carried out by the proposed method. The spectrum of formulation by method was shown in (Fig No 8). The assay of formulation was carried out by proposed method and calculated % of nominal conc and RSD was found within acceptable limits are summarized in (Table No 4). The results indicated applicability of the method for estimation of formulation. Accuracy and Precision: The accuracy study was carried out at 3 levels; and the results of accuracy are summarised in (Table 5), the obtained results were within acceptable limit; and methods accuracy was justified by calculating % drug content. The precision study was carried out by performing assay of solutions; further the reproducibility in result was studied by interday and intraday precision. The values obtained SD and % RSD was shown methods precision and are summarised in (Table 5). D Kolekar et al. Chemometric assisted QbD method for estimation of bilastine and montelukast sodium Vol 3 | Issue 1 | Jan – Mar 2024 Indian J Pharm Drug Studies | 20 Limit of Detection (LOD) and Limit of Quantitation (LOQ): The LOD and LOQ of BSE and MKS by the proposed method were calculated and shown in (Table 6). Robustness and Ruggedness: Robustness was studied and capacity of analytical procedure to measure analyte was remain unaffected by small but deliberate variations in method parameter. The analytical method was found rugged during development; similarly the result was produced shown in (Table 6) by performing the analysis by different analyst. Figure 5: Overlaid spectra of Bilastine obtained in linearity study Figure 6: Overlaid spectra of Montelukast obtained in linearity study Fig No 7: Calibration curve of BSE and MKS Table 3: Parameters of regression equation obtained in Microsoft excel Parameters BSE MKS Detection wavelength 274.5 351.5 Beer’s law limit (μg/ml) 1–32 μg/ml 1–20 μg/ml Correlation coefficient (r2) 0.9967 0.9998 Regression equation (y = mx + c) Y = 0.0203X - 0.0037 Y = 0.0302X - 0.0013 Fig No 8: Spectra of formulation obtained in the assay Table 4: Results of assay of formulation by proposed method Name of the Formulation - BILAFAV M (Bilastine 20 mg, Montelukast sodium 10 mg) B No -23S2GTB843, MFG Date- Jul 2023, EXP Date–Jun 2025 Formulation Drug Label Claim (mg/Tablet; n=6) Amount found/mg Drug Content % Std Deviation % RSD Method BSE 20 19.618 98.092 0.15702 0.16092 MKS 10 10.362 103.62 0.21932 0.24942 Table No 5: Results of accuracy and precision Parameter Level of study Drug Name S.D. % RSD Precision Intraday Precision BSE 0.19025 3.9635 MKS 0.28281 0.2828 Inter day precision BSE 0.57732 0.83165 MKS 0.54582 0.64584 Accuracy study of BSE and MKS 80% BSE 0.25236 0.21612 100% 0.28842 0.24982 120% 0.52218 0.44192 80% MKS 0.25123 0.34451 D Kolekar et al. Chemometric assisted QbD method for estimation of bilastine and montelukast sodium Vol 3 | Issue 1 | Jan – Mar 2024 Indian J Pharm Drug Studies | 21 100% 0.68602 0.88492 120% 1.25152 1.81023 Table No.6: Results of LOD and LOQ, robustness Parameters BSE MKS LOD μg/ml 0.7253 0.6726 LOQ μg/ml 1.1761 0.8458 Robustness (conc 20 BSE and 16 μg/ml MKS) 0.3724 -to- 0.3973 (± 2 nm) 0.5308 -to- 0.5229 (± 2 nm) Ruggedness Analyst 1 SD ± 0.20681 RSD ± 3.53719 SD ± 0.64246 RSD ± 2.9893 Analyst 2 SD ± 0.20167 RSD ± 5.16971 SD ± 0.062412 RSD ± 2.55771 CONCLUSION Both the drugs were estimated from their combined formulation by simultaneous equation method. Results were found within acceptable limits, statistical data obtained were shown rigidity of the method. The validated method was employed 50% alcohol as solvent thus become economical. The proposed method is precise, accurate, robust and reproducible hence can be routinely used for simultaneous estimation of bilastine and montelukast sodium from combined dosage form ACKNOWLEDGEMENT Authors are thankful to Glenmark Pharmaceuticals, Nashik, Maharashtra. India for providing drugs as gift sample and Management, Principal of SVPM’S College of Pharmacy Malegaon (BKII),Baramati Dist. Pune, Maharashtra, India for providing necessary facilities, chemicals, instruments etc. for research. REFERENCES 1. The Merck Index. An Encyclopaedia of chemicals, drugs and Biological. 15th ed. the Royal Society of Chemistry Cambridge UK. 2013. p. 214, 1164. 2. Alison Brayfield. Martindale (The complete drug reference). 39th ed. Pharmaceutical press London UK. 2017(A): 617-1214. 3. T Anju, SK Hakeema, G Divya Jyothi, et al. Quantitative Determination and Validation of Bilastine in Bulk and Pharmaceutical Dosage Form by Using UV-Spectroscopy. Inter J Pharm and Pharma Res. 2022;23(2):136-146. 4. Kuldip Makawana, Divya Patel, Dhruv Thakor, et al. UV Spectrophotometric method development and validation for the estimation of bilastine in pharmaceutical dosage form. Inter J Res Pub and Rev. 2022;3(3):1286-1295. 5. OS Supe, MM Maste, SS Suryawanshi, et al. Development and standardization of stability indicating UV spectrophotometric method for assessment of bilastine in bulk and pharmaceutical dosage formulation. Inter J Pharma Sci and Res. 2021;13(2):962- 968. 6. Andressa Tassinari DaSilva, Gabriela Rossi Brabo, Isadora Dias Marques, et al. UV Spectrophotometric method for quantitative determination of Bilastine using experimental design for robustness. Drug Analytical Research. 2017;1(2):38-43. 7. R Mohan Raj, ASK Sankar, T Vetrichelvan. Analytical method development and validation for simultaneous estimation of bilastine and montelukast sodium by UV spectrophotometry. World J pharm pharma sci. 2010;10(1):680-687. 8. Riya Mistry, Rajashree Mashru. Analytical Method Development and Validation for Simultaneous Estimation of Bilastine and Montelukast Sodium in their Combined Dosage form by Derivative UV-Spectroscopy and RP- HPLC Method. Inter J Pharma Res Health Sci, 2021;9(3):3313-3318. 9. Syed Nizamuddin, S Appala Raju. Development and Validation RP-HPLC Method for Simultaneous Estimation of Bilastine and Montelukast in Bulk and Pharmaceutical Dosage. Asian Pacific J Health Sci.2022;9(3):242-247. 10. Narmada Vallakeerthi, Rachala Swetha, T Tejaswi, et al. Development and Validation of RP HPLC Method for the Simultaneous Estimation of Bilastine and Montelukast in Tablet Dosage Form. Chem Sci Inter J. 2023;32(3):52-61. 11. Bagul Rashmi Arun, Gosavi Seema. A Research Article on Analytical Method Development and Validation of Antihistamine Drugs Bilastine and Montelukast Sodium by RP-HPLC and UV Spectrophotometric Method. Inter J Pharma Res Applications. 2022;7(6):539-550. 12. Swati M Andhale A, Anna Pratima G Nikalje B. Simultaneous Estimation of Bilastine and Montelukast in Bulk by Rp-Hplc and Assessment of Its Applicability in Marketed Tablet Dosage Form. J Pharma Res Inter. 2022;34(3B):8-25. 13. Aejaz Ahmed, et al. Development of RP-HPLC Method for Simultaneous Determination of Bilastine and Montelukast by Qbd Approach and Its Validation. Inter J Life Sci Pharma Res. 2023;13(5):199-220. 14. Chandra Umesh, Kumar Manish, Sharma Shrestha, et al. New RP-HPLC Assay Method Development and Validation for Simultaneous Quantitation of Bilastine and Montelukast in Bulk and Tablet Dosage Form. Inter J Curr Res Rev. 2021;13(10): 179-188. 15. Peethala Prathyusha, Raja Sundararajan, Palyam Bhanu, et al. A new stability indicating RP-HPLC method for determination of Bilastine in bulk and pharmaceutical formulation. Res J Pharm Tech. 2020;13(6):2849-2853. 16. Paulo Roberto Rodrigues Martini, et al. Bilastine: stability- indicating a method using environmentally friendly by reversed- phase high-performance liquid chromatography (RP-HPLC). Drug analytical Res. 2022;6(n1):13-20. D Kolekar et al. Chemometric assisted QbD method for estimation of bilastine and montelukast sodium Vol 3 | Issue 1 | Jan – Mar 2024 Indian J Pharm Drug Studies | 22 17. Chetan Vilasrao Patil, Shabnam Khan, Ramakant Sharma, et al. Stability indicating rp-hplc method development and validation for simultaneous estimation of bilastine and montelukast in bulk and tablet dosage form. Inter J Pharma Sci Med. 2023;8(3):132- 149. 18. Aya Roshdy, Randa Abdel Salam, Ghada Hadad et al. Green quality by design HPLC approach for the simultaneous determination of Bilastine and Montelukast. BMC Chemistry. 2023;17(43):1-23. 19. Shital Patel, T Yunus Pasha. Stability Indicating Isocratic HPLC Method for Bilastine and Characterization of Forced Degradation Products by LC-MS/MS. Inter J Life Sci Pharma Res. 2022; 13(1):83-93. 20. Shaista Firdous, SH Rizwan. Development and Validation of Stability Indicating UPLC Method for the Estimation of Bilastine in Bulk and Pharmaceutical Dosage Form, Inter J Pharm Sci Rev Res. 2020;65(1):131-135. 21. Lingareddygari SR, Ravikrindhi NR, Beludari IM, et al. Design of Experiments Approach for method development and validation of Bilastine in Pure and Pharmaceutical Dosage Form using RP- UFLC. Orient J Chem. 2023;39(3):736-745. 22. Kuldeep Singh, et al. Validated UV spectroscopic method for estimation of montelukast sodium. Inter J Pharma Sci Res. 2015; 6(11):4728-4732. 23. Shanmukha Kumar JV, Geeta Swarupa P, Vardhan SVM, Ramachandran. D2 Spectrophotometric determination of montelukast sodium in bulk and pharmaceutical formulations Scholars Res Library Der Pharma Chem. 2012;4(2):720-724. 24. K Pallavi, P Srinivasa Babu. Validated UV Spectroscopic Method for Estimation of Montelukast Sodium from Bulk and Tablet Formulations, Inter J Adv Pharm Bio Chem. 2012;1(4): 450-453. 25. Meenu Chaudhary, Praveen Kumar, Divya Thapliy. Analytical method development and validation for determination of montelukast by UV-spectroscopy in API & in pharmaceutical dosage forms. Intern J Pharm Bio Sci. 2018;8(4):482-487. 26. Dlivan Fattah Aziz, Yehia Ismail Khalil. Newly Simple Quantitative Determination of Montelukast Sodium by Ultraviolet-Spectrophotometry. J Sci Tech. 2022;6(2):24-28. 27. Kumar Raja Jayavarapu, et al. Accurate and simple UV-VIS spectroscopic method for the estimation of montelukast sodium in pure and marketed formulations. J Glob Trnd Pharma Sci. 2017;8(2):3994-3997. 28. Selvadurai Muralidharan, et al. Newly Developed and Validated Method of Montelukast Sodium Estimation in Tablet Dosage Form by Ultraviolet Spectroscopy and Reverse Phase-High Performance Liquid Chromatography. PTB Reports. 2016;2(2): 27-30. 29. Nitin Kumar Patel, Pradeep Chouhan, Suresh Kumar Paswan, et al. Development and validation of a UV spectrophotometric method for simultaneous estimation of combination of Montelukast sodium in presence of Levocetirizine Dihydrochloride. Scholars Res Lib Der Pharmacia Lettre. 2014;6 (3):313-321. 30. Pinkal P, Zinal AP, Sweta P, et al. Spectrophotometric Absorption Correction for the Method Simultaneous Estimation of Montelukast Sodium and Bilastine in Tablet Dosage Form, Inter J Educ App Sci. 2022;9(2):20-28. 31. Mandeep Yadav, Ravi Kant, Sonia Yadav, et al. Analytical method development and validation by simultaneous estimation of montelukast sodium and bilastine by UV spectrophotometry. Eur Chem Bull Section A-Res Paper. 2023;12(10):2082 – 208. 32. Detroja K, H Vekaria. Advanced Derivative Spectroscopic Method for Estimation of Montelukast and Bilastine in Their Tablet Dosage Form. Inter J Pharma Sci Drug Res. 2021;13(3): 268-274. 33. Jignesh Maniya, Hasumati Raj, Hasmukh Vaghani, et al. Development and Validation of Spectroscopic Method for Simultaneous Estimation of Acebrophylline and Montelukast Sodium in Combined Dosage Form. Indo American J Pharma Res. 2012;2(10):1027-1036. 34. Pintu Prajapati, Jayesh Tamboli, Ashish Mishra, et al Risk Assessment-Based Enhanced Analytical Quality-by-Design Approach to Eco-Friendly and Economical Multicomponent Spectrophotometric Methods for Simultaneous Estimation of Montelukast Sodium and Bilastine. J AOAC Inter. 2021;104(5): 1453-1463. 35. Akiful Haque M, et al. Stability Indicating Rp-Hplc Method for the Estimation of Montelukast in Pharmaceutical Dosage Form. IOSR J Pharm Bio Sci. 2012;1(6):31-36. 36. K Naga Raju, T Gopala Swamy A Lakshmana Rao. Development and validation of RP-HPLC method for the determination of montelukast sodium in bulk and in pharmaceutical formulation, Intern J pharma, Chem Biological Sci. 2011;1(1):12-16. 37. Singh R M, Saini P K, Mathur S C, et al. Development and Validation of a RP-HPLC Method for Estimation of Montelukast Sodium in Bulk and in Tablet Dosage Form. India J Pharm Sci. 2010;72(2):235-237. 38. Mandeep Yadav, Vikas Jogpal, Jyoti Gupta, et al. Analytical method development and validation of Montelukast sodium and Bilastine by HPLC. Eur Chem Bull Sec A-Res Paper. 2023; 22(3):517-526. 39. Varun Dasari, et al. Development and validation of anti-allergic drugs: RP-HPLC. Dogo Rangsang Res J. 2023;13(3):97-107. 40. N. Rashmitha, et al. A Validated RP-HPLC Method for the Determination of Impurities in Montelukast Sodium. E-J Chem. 2010;7(2):555-563. 41. Akshaya G. Sinhe, Neelam Khan. Stability Indicating RP-HPLC Method Development and Validation of Few Bulk Drug Combinations and Their Formulation. Bulletin of Environment, Pharm Life Sci. 2021;10(6):80-88. 42. V Padhiyar, P Patani, N Tiwari. Development and validation of stability indicating rp-hplc method for simultaneous estimation of montelukast sodium and bilastine from its pharmaceutical dosage form, J Eme Tech Innov Res. 2021; 8(6): 865-877. 43. Rajeswari Sunkara, M Ajitha. Stabilty Indicating RP-HPLC Method Development and Validation for Simultaneous Estimation of Montelukast and Bilastine in Bulk and Pharmaceutical Dosage Form. World J Pharma Sci. 2022; 10(01):90–97. 44. Rathore A S, Sathiyanarayanan L, Mahadik K R. Development of Validated HPLC and HPTLC Methods for Simultaneous Determination of Levocetirizine Dihydrochloride and Montelukast Sodium in Bulk Drug and Pharmaceutical Dosage Form. Pharm Anal Acta. 2010;1(1):1-6. 45. Indian Pharmacopoeia, 8th Ed. Govt. of India, ministry of Health and family welfare, The Indian pharmacopoeia commission Ghaziabad. 2018;2;2628. 46. British Pharmacopoeia, Medicines and Healthcare products regulatory agency, London. 2019;3:331. 47. International Conference on Harmonization. ICH Harmonized tripartite Guideline-Pharmaceutical development, Q8 R2. 4 version. 2009;1-28. D Kolekar et al. Chemometric assisted QbD method for estimation of bilastine and montelukast sodium Vol 3 | Issue 1 | Jan – Mar 2024 Indian J Pharm Drug Studies | 23 48. ICH Expert working group. ICH harmonized tripartite Guideline- Quality Risk Management Q9. In current step 4 version. 2005;1- 23. 49. ICH Expert working group. ICH Harmonized tripartite Guideline-Pharmaceutical Quality system Q 10. In current step 4 version. 2008;1-21. 50. ICH Expert working group. ICH Harmonized tripartite Guideline-Validation of analytical procedures: Text and methodology Q 2 R1. In current step 4 version.2005;1-17. 51. Schweitzer M, et al. Implications and opportunities of applying QbD principles to analytical measurements. Pharm Tech. 2010; 34:12-29. 52. Vogt F G, Kord A S. Development of quality-by-design analytical methods. J Pharm Sci. 2011;100:797-812. 53. Bhatt D A, Rane S I. QbD approach to analytical RP-HPLC method development and its validation, Int Journal of Pharma Science. 2011; (3):179-187. How to cite this article: Bhagyashri D Kolekar, Namrata N Gawade, G K Dyade, Nilesh Y Jadhav. Chemo metric assisted Spectrophotometric Method Development through QBD Approach for the estimation of Bilastine and Montelukast sodium in combined solid dosage form. Indian J Pharm Drug Studies. 2024; 3(1):16-23. Funding: None Conflict of Interest: None Stated Key Words: Bilastine, Montelukast Sodium, Qbd, ICH, Simultaneous Equation Method