RESUMO
An isocratic reverse-phase High performance liquid chromatography (HPLC) method using a chemometric modelwas developed and validated for the simultaneous determination of ambroxol hydrochloride, terbutaline sulfate,and guaiphenesin in their combined dosage form. Central composite design which is a subset of response surfacemethodology was used as the chemometric model. The separation of three drugs was carried out by using a PhenomenexC18 column and detection at 220 nm using a UV detector. Based on initial trials, the three factors selected for thedesign were methanol (MN) concentration, mobile phase pH, and flow rate in the range of 55%–65% (v/v), 4–5 units,and 0.6–1 ml/min, respectively. The impact of the selected factors on the responses like retention time of first peak(tR1), resolution of 2nd and 3rd peak (Rs2,3), and theoretical plates of the first peak (TP1) were evaluated. Derringers’desirability function was used to optimize the responses. The conditions which were optimized for the assay of drugswere MN, ACN, and 50 mM KH2PO4 (pH 5) in the ratio of 55:10:35 at a flow rate of 0.8 ml/minute. The developedand optimized method was validated as per the International conference on harmonization (ICH) guidelines and canbe used for routine analysis in quality control laboratories.
RESUMO
Objective: To evaluate the anti-obesity activity of ethanolic extract of cashew apple using various in vitro and in vivo models. Methods: Phytochemical screening was carried out in ethanolic extract of cashew apple, followed by quantification of phenol and flavonoid. Antioxidant potential was evaluated using 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) scavenging assays. The inhibitory effect of ethanolic extract of cashew apple on α-amylase and pancreatic lipase was also studied. In addition, anti-obesity activity was determined in two in vivo models, lipid emulsion model and atherogenic diet-induced obese rat model. Levels of postprandial plasma triglycerides were assessed in lipid emulsion model, whereas serum lipid profile, in vivo antioxidants and histopathological studies of the carotid artery and liver were performed in an atherogenic diet-induced obese model. Results: Phytochemical screening revealed the presence of carbohydrates, alkaloids, polyphenols, terpenoids, and steroids. The in vitro assays showed inhibition of α-amylase and pancreatic lipase and strong antioxidant potential. Ethanolic extract of cashew apple showed significant and time-dependent inhibitory activity on postprandial triglycerides after administration of lipid emulsion for 5 h. Ethanolic extract of cashew apple at 200 and 400 mg/kg on day 60 showed a significant reduction in body weight, body mass index and atherogenic index, whereas lipid profile and liver function marker levels in the serum were decreased in a dose-dependent manner at time intervals (day 0, 20, 40, and 60) compared to the atherogenic diet-induced obese rats. Histological observations showed reduced non-alcoholic fatty liver deposits and decreased atherosclerotic fatty streak plaques (carotid artery) after treatment with ethanolic extract of cashew apple. Conclusions: Ethanolic extract of cashew apple ameliorates obesity, which may be partly mediated by its delayed absorption of cholesterol and carbohydrates.
RESUMO
A simple, sensitive, precise, accurate and robust high performance liquid chromatographic method has been developed for simultaneous estimation of saxagliptin (SAXA) and dapagliflozin (DAPA) in pharmaceutical formulation. Design of experiments (DoE) was applied for multivariate optimization of the experimental conditions of RP-HPLC method. Risk assessment was performed to identify the critical method parameters. Three independent factors; mobile phase composition, flow rate and column temperature were used to design mathematical models. Central composite design (CCD) was used to study the response surface methodology and to study in depth the effects of these independent factors. Desirability function was used to simultaneously optimize the retention time and resolution of SAXA and DAPA. The optimized and predicted data from contour diagram consisted of acetonitrile and ortho phosphoric acid (0.1%) in the ratio of 50:50 respectively, at a flow rate of 0.98 ml/min and column temperature 31.4 °C. Using these optimum conditions baseline separation of both drugs with good resolution and run time of less than 6 min were achieved. The optimized assay conditions were validated according to ICH guidelines. Hence, the results clearly showed that Quality by design approach could be successfully applied to optimize RP-HPLC method for simultaneous estimation of SAXA and DAPA.