Validation of a simple chromatographic method for naringenin quantification in skin permeation experiments

Naringenin is a flavonoid that can be found in citrus fruits (e.g., Citrus sp). This natural compound is known for its antioxidant activity, antitumor, and neuroprotective potential, also acting directly in controlling the inflammatory response. Topical and transdermal routes are attractive alternatives that could circumvent the low oral bioavailability. However, a simple analytical method capable of determining naringenin in skin layers is still demanded. Thus, this work aimed to validate a selective and straightforward chromatographic method for naringenin determination in skin permeation studies. The developed method uses a reversed-phase C₁₈ column as stationary phase and a mobile phase composed of methanol/phosphoric acid 0.01 M (65:35, v/v), eluted at a flow rate of 0.6 mL/min with detection at 290 nm. The method was linear (r² > 0.99) in a broad concentration range of 0.5-10.0 μg/mL, precise with an overall variation coefficient lower than 2%, and accurate with naringenin recovery from the skin layers higher than 85%. Additionally, the method was sensitive (LD = 0.10 μg/mL, LQ = 0.20 μg/mL), selective against skin matrices as well as naringenin degradation products, and robust regarding methodology parameters. Therefore, the method was suitable to be used in skin permeation studies employing naringenin.