¹Department of Pharmaceutics, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia
²Department of Pharmacy, Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia
³Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia
⁴Department of Dermatology and Venereology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia DOI : 10.29228/jrp.410 High Performance Liquid Chromatography (HPLC) was used to determine vitamin D3 content from the topical dosage form. Self-nanoemulsifying drug delivery systems (SNEDDS) based hydrogel was aimed to optimize the delivery of vitamin D3 through the skin due to its high lipophilicity. Quantifying vitamin D3 was performed using the internal standard, vitamin D2, on LiChrocart C18 125-4 reverse phase column. Several characteristics of validation were referred including specificity, linearity, range, accuracy, precision, limits of detection, and quantification. The mobile phase was optimized as a mixture of acetonitrile-methanol-water for injection 94:3:3 v/v, and the flow rate was optimized at 0.8 mL/min at 25°C. Vitamin D3 detection and quantification were observed at a wavelength of 266 nm using an ultra-violet (UV) detector. The analysis method performed no excipient interference, including separating vitamin D3 from its matrix. Extraction of vitamin D3 from its matrix was observed using the dialysis membrane, with agitation (vortex) and no agitation (immersion). The drug content determination was found to be 108.22 % ± 14.77% for the immersion method and 72.04% ± 33.37% for the vortex method. Further, the high variability of the results indicated the dialysis membrane was not a suitable method for determining lipophilic drug content from the colloidal polymer matrix. The other techniques with negligible variation, such as reverse dialysis or subsequent solvent addition, are preferred. Keywords : Vitamin D3; HPLC; drug content; SNEDDS; hydrogel