The current studies entail systematic quality by design (QbD)-based development of simple, precise, cost-effective and stability-indicating high-performance liquid chromatography method for estimation of ketoprofen. Analytical target profile was defined and critical analytical attributes (CAAs) were selected. Chromatographic separation was accomplished with an isocratic, reversed-phase chromatography using C-18 column, pH 6.8, phosphate buffer-methanol (50 : 50v/v) as a mobile phase at a flow rate of 1.0 mL/min and UV detection at 258 nm. Systematic optimization of chromatographic method was performed using central composite design by evaluating theoretical plates and peak tailing as the CAAs. The method was validated as per International Conference on Harmonization guidelines with parameters such as high sensitivity, specificity of the method with linearity ranging between 0.05 and 250 µg/mL, detection limit of 0.025 µg/mL and quantification limit of 0.05 µg/mL. Precision was demonstrated using relative standard deviation of 1.21%. Stress degradation studies performed using acid, base, peroxide, thermal and photolytic methods helped in identifying the degradation products in the proniosome delivery systems. The results successfully demonstrated the utility of QbD for optimizing the chromatographic conditions for developing highly sensitive liquid chromatographic method for ketoprofen.
Environmental stress and advancing age is considered as the main cause of skin aging. However, environmental stress (especially UV radiations) accelerates the process of skin aging by manifolds. Coenzyme Q10 (CoQ10), an essential compound of cellular bioenergetics also acts as a strong antioxidant and protects the body against aging. High molecular weight and structure specific lipophilic nature of this molecule is a bottle neck in effective delivery through topical route. Preparation of a novel proniosomal (PN) gel formulation of CoQ10 employing systematic design of experiment (DoE) approach is a step ahead in transcending the constraints of the topical delivery. I-optimal mixture design was employed for systematic optimization of proniosomal formulation and evaluation of experimental data was performed for entrapment efficiency and in vitro release. Hydration of PN gel formulation with phosphate buffer (pH 7.5) results in submicron niosomes vesicles of spherical shape, which appeared dark against bright surroundings in TEM study. Animal skin was treated with UV radiations followed by treatment of PN gel CoQ10 and conventional CoQ10 present in a gel base. The effectiveness of the treatment was evaluated on the basis of biochemical estimation and histopathological studies. By using CoQ10 PN gel formulation, levels of superoxide dismutase (SOD), catalase (CA), glutathione (GSH) and total proteins were restored by 81.3%, 72.1%, 74.8 and 77.1%, respectively to that of control group. Histopathological studies revealed better protection of skin treated with CoQ10 PN gel compared to free CoQ10. Prepared PN gel was found undisturbing with the normal histology hence, tolerated by animal skin compare to conventional gel.
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