Rapid RP-HPLC method optimization for vitamin D3 quantification in soft-gelatin capsules via Response Surface Methodology (RSM)
Year: 2026
Type: article
Abstract: Reliable quantification of fat-soluble vitamins such as vitamin D3 (VD3) in soft-gelatin dosage forms is challenging because of co-formulated antioxidants and variable excipients. A rapid and robust RP-HPLC method was optimized and validated for VD3 quantification in soft-gelatin capsules containing Vitamin E as an antioxidant using response-surface methodology (RSM). A Box–Behnken design (BBD) within a Design of Experiments (DoE) framework was used to optimize column temperature, flow rate, and mobile phase composition. The final 5-minute method employed an Acclaim RSLC 120 C18 column with ACN:MeOH:Milli-Q water (87.5:9:3.5) as the mobile phase, a flow rate 0.8 mL min -1, and column temperature of 20.8°C. Sample clean-up evaluation identified the SOLA HRP SPE cartridge as providing the highest recovery (98.83 ± 0.65%). Validation demonstrated specificity, linearity (0.02 – 6.00 mg L-1, R2 = 0.999), accuracy (recoveries 98.2 – 99.5%), precision (%RSD 0.66%), and sensitivity (LOD 2.5 µg L-1, LOQ 6.7 µg L-1). This study highlights the utility of RSM - BBD for efficient HPLC method optimization, providing a generalizable strategy for other fat-soluble vitamins and complex formulations.
Source: Results in Chemistry
Language: English
Subfield: Analytical Chemistry
Field: Chemistry
Domain: Physical Sciences
Open Access status: gold
Estimated APC paid: $1,870
Funder: Thermo Fisher Scientific
OpenAlex ID: W7127635022