The present study is aimed at evaluation of phytosomal gel of the petroleum ether extract of root bark of Onosma echiodes for wound healing activity in rats. Extract of root bark of O. echioides was standardized by isolated naphthoquinone dimer using HPTLC. Phytosomes (equivalent to 2% w/w of naphthoquinones) of the standardized extract were prepared by thin film hydration technique. The wound healing efficacy of the formulation was evaluated in rats by inflicting excision and incision wounds followed by treatment of the wounds topically. The parameters evaluated for healing included determination of breaking strength and tensile strength of healed skin for incision model and percentage wound contraction, hydroxyproline content, granulation tissue free radicals and catalase in excision wound model. The formulation treated group showed a significant healing (p<0.005) of both the excision and incision wounds with respect to wound contraction and tensile strength respectively, as compared to vehicle treated group. The oxidative stress of the granulation tissue was also found to be reduced as indicated by reduced lipid peroxidation and increase in catalase activity. The phytosomal gel of O. echioides effectively exhibited wound healing effect.
Aim: The aim of work is to develop and validate simple, specific, accurate, precise HPLC method for the estimation of Curcumin (CUR) and Thymoquinone (THQ) simultaneously in bulk and its formulation as per ICH guidelines for analytical method development and validation.
Study Design: Developing RP-HPLC method using C-18 Inertsil column and optimization of variables for estimation of Thymoquinone and Curcumin in bulk and formulation.
Place and Duration of Study: The present work was carried out at Shri D. D. Vispute College of Pharmacy and Research Center, Panvel in year 2021.
Methodology: The RP-HPLC method was developed with an isocratic condition of mobile phase comprising acetonitrile and water in a ratio of (82:18) v/v, at a flow rate of 0.9 mL/minute over Inertsil ODS, 250× 4.6 mm, 5 µm column, at 30°C column oven temperature. Photodiode array at 256 nm was used for detection
Results: Retention time for curcumin and thymoquinone was found to be 3.5 and 4.3 mins respectively. The method showed excellent linear response in concentration range of 4-18 µg/mland 10-45 µg/mlfor thymoquinone and curcuminrespectively with correlation coefficient (R2) values of0.999 for both. System precision and method precision studies were less than the maximum allowable limit percentage of relative standard deviation ≤ 2.0 i.e., 1.61 % and 1.62 % for curcumin and 0.47 % and 0.42 % for thymoquinone respectively. Mean % Recovery for both the drugs were within acceptance limits. The developed and validated HPLC method is simple, accurate, precise and suitable for analysis as all the results were within acceptance criteria.
Conclusion: The developed RP-HPLC method at single wavelength was validated according to ICH guideline with respect to system suitability, specificity, linearity, accuracy, precision and robustness and can be used for routine quality monitoring of drug in pharmaceutical dosage form.
A rapid, specific UV spectrophotometric method has been developed using a solvent methanol to determine Tofisopam content in bulk and pharmaceutical formulations. At a pre-determined wavelength at 310 nm, it was proved linear in the range of 4-24 μg/ml and exhibited good correlation coefficient (R2=0.9996) and excellent mean recovery (98-102%). The method was validated statistically and parameters like linearity, precision, accuracy, specificity, and assay were studied according to International Conference on Harmonization guidelines. The obtained results proved that the method can be employed for the routine analysis of Tofisopam in bulk as well as in the commercial formulations.
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