SUMMARY, CONCLUSION & RECOMMENDATIONS

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1 196 Chapter-5 SUMMARY, CONCLUSION & RECOMMENDATIONS

2 197 CHAPTER Summary, Conclusion and Recommendations Summary and Conclusion are drawn based on the work carried out by the author on development and Validation of various methods for individual medicinally important organic drugs. In this perspective, analytical chemistry of medicinally important compounds had been taken up by using high performance liquid chromatography which is most popular chromatographic technique in confiding for Related Substances, Assay and Dissolution profile and results were incorporated in the current thesis. Whereas the selected drugs rare and required accurate analytical separation technique to quantify its drug substance, the current research program fulfills the need of accurate quantification of drug substance. The first chapter illuminates the general introductory about the basic principles liquid chromatography, general introduction for development and validation of analytical procedures and also enlightened the prominence of medicinally important organic compounds in the society for day to day life. Second chapter is confined for the literature survey related to the current research program. An eextensive literature survey was done on selected medicinal compounds related to the current research work to have better clarity and vision before going to initiate the research program. The overall outcome of the literature survey described that no methods were reported for two medicinal compounds and few methods were reported for the rest of three medicinal compounds.

3 198 Nonetheless those reported methods were shown less separation between impurities and with the main peak, and are troublesome gradient methods, lower analytical techniques, tedious procedures, different and difficult detection techniques and were not meeting the current regulatory requirements. Hence the research program was devoted on Levofloxacin, Cinacalcet Hydrochloride, Plerixafor, Dexibuprofen and Milnacipran Hydrochloride. The chapter-3.1 & 4.1 reveals that to develop a validated specific stability indicating isocratic reversed-phase liquid chromatographic method for the quantitative determination of Levofloxacin as well as its related substances in pharmaceutical dosage forms in presence of degradation products and its process related impurities. Forced degradation studies were performed on Levofloxacin as per International Conference on Harmonisation (ICH) prescribed stress conditions using acid, base, oxidative, water hydrolysis, thermal stress and photolytic degradation to show the stability indicating power of the method. Significant degradation was observed during oxidative stress, minor degradation in acidic stress and no degradation was observed in other stress conditions. The chromatographic method was optimized using the samples generated from forced degradation studies and the spiked impurity solution. The analysis was carried out using a 50x4.6 mm; 3.0 µm YMC Pack Pro- C18 column using the eluent containing the mixture of 1.0% (v/v) triethylamine (ph 6.30), methanol and acetonitrile (7.7:1.3:1.0) was

4 199 pumped at a flow rate of 0.8mLmin -1 with Ultraviolet (UV)-detection at 235nm. The chapter-3.2 & 4.2 investigated to develop a validated specific stability indicating isocratic reversed-phase liquid chromatographic method using UPLC for the quantitation of Cinacalcet HCl in bulk drug and its tablet dosage form in a shorter chromatographic run time. The analysis was carried out using a 100mm length with 2.1mm internal diameter, 1.7µm particle size Kinetex XB-C18 column using the eluent containing the mixture of 0.1% (v/v) acetic acid in water with ph adjusted to 5.40 using triethylamine and acetonitrile (2:3) was pumped at a flow rate of 0.5mLmin -1 with Ultraviolet detection at the wavelength of 220nm and the injection volume was 1.0µL. The chapter-3.3 & 4.3 devoted to develop a validated selective & specific stability indicating simple isocratic RP-HPLC method for the quantitation of Plerixafor and its related substances quantitatively in API and its pharmaceutical dosage form. The stability indicating nature of the method was proven by performing the forced stress/degradation studies on Plerixafor such as acid, alkali, oxidative hydrolysis etc. The samples of stress studies and impurity spiked sample were used to develop and optimize the current HPLC method. An isocratic reverse phase HPLC analytical method with Zorbax Bonus RP, 150x4.6mm; 3.5µm column was employed for the optimum chromatographic separations. The eluent contains a mixture of buffer and acetonitrile in the ratio of 3:1. The flow rate was kept at 0.8mL.min -1, the injection volume was 10µL, the column temperature

5 200 was maintained at 40 C and the column eluent was monitored at 210 nm. The total chromatographic runtime was 40 minutes. The chapter-3.4 & 4.4 confined that Dexibuprofen is commonly used as NSAID in place of ibuprofen to achieve higher pharmacological activity. The main aim and focus of present study is to develop RP-HPLC method and validation in stability indicating manner as per ICH guidelines for the quantification of Dexibuprofen in API and pharmaceutical formulations. The optimized chromatographic conditions includes a C-18 RP-HPLC column as stationary phase (i.e. Thermo Hypersil ODS, 150x4.6mm; 5µm) and the mobile phase pumped with the flow rate of 1.0mL.min -1 and the eluent was monitored at 220nm were established. The eluent contains a combination of buffer (acetic acid & triethylamine buffer) and acetonitrile in the ratio of 40:60% (v/v). The developed LC method is a simple, rapid, precise, specific and accurate for the determination of Dexibuprofen in the range of 20% to 200% of working concentrations with 0.2 µg ml -1 value of LOQ and 0.05 µg ml -1 value of LOD. The chapter-3.5 & 4.5 demonstrated that a simple and precise isocratic reverse phase High Performance Liquid Chromatography analytical method was developed and validated for the analysis of dissolution samples of Milnacipran drug product for In-vitro bioavailability studies in different dissolution mediums to proceed for bio-waiver claim. The analysis was carried out with the optimized chromatographic conditions which includes Hypersil BDS C8 (150x4.6 mm; 5µm) column as stationary phase with 30 C as column

6 201 temperature, 215nm as detection wavelength and the mobile phase pumped in the flow rate of 1.0mL.min -1. The eluent consist a disodium phosphate buffer of ph 5.5 and Acetonitrile in the ratio of 70:30. The current LC method was developed against qualified working standard and validated as per ICH requirements. As a final point, based on the above accomplished extensive research program, the recommendation were drawn and concluded in accordance to the current requirements of regulatory bodies by bearing in mind about all the parameters which backbones the pharmaceutical industry and research development centers The current thesis put forward the simple, precise, accurate, and cost effective, less time consuming, reproducible, reliable and rugged methods for selective medicinally important organic compounds which were playing vital role in the treatment of human diseases. All the above reported methods can be easily adopted for the purity quantification of the respective medicinal compounds in as such form as well as pharmaceutical dosage forms. The proposed methods will also accommodate and fulfills the current need in regular quality control testing laboratories of pharmaceutical industry. The introduced methods can be used abundantly in the abbreviated new drug development, Novel drug delivery system development etc.

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