RP-HPLC METHOD DEVELOPMENT AND VALIDATION OF TETRAHYDROCURCUMIN USING MULTILEVEL FULL-FACTORIAL DESIGN IN BULK, NANOEMULSION AND LIPOSOMES
*Neha Chhabra, Anita Solanki, Rajani Athawale and Supriya Mahajan
In this paper, an application of Design of Experiment (DoE) concept for the development of a simple yet precise reverse phase HPLC method for analysis of tetrahydrocurcumin in bulk drug and dermal formulations including nanoemulsion and liposomes is described. The method optimization was accomplished by applying multi-level full-factorial design model using Design ExpertÂ® (Version 10) software. 3 fators (F1, F2 and F3) and 2 responses (R1 and R2) were used viz mobile phase ratio of acetonitrile and methanol, flow rate, % v/v acetic acid, Retenton time and number of theoretical plates respectively to obtain the right combination that gave single sharp peak. The optimized HPLC conditions (acetonitrile and methanol in the ratio of 53:47, 0.26 % v/v with glacial acetic acid, at a flow rate of 0.6 ml/min and temperature maintained at 25ËšC Â± 0.5ËšC gave single sharp peak at 4.5 minutes. The developed method was validated using ICH guidelines (for linearity, precision, robustness, Limit of quantification and limit of detection) and then analysed using ANOVA and 3-dimensional plots. Values of adequate precision 21.614 indicated that the model could be used to navigate the design space. â€œProb>5â€ value less than 0.0500 indicated that the model terms were significant. It was concluded that the proposed method was simple, sensitive, and highly robust for routine analysis of bulk drug and other dermal formulations of tetrahydrocurcumin.
Keywords: Tetrahydrocurcumin, RP-HPLC, Method development, Validation, Full factorial design.
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