Validation of a chromatographic method for α-phellandrene quantification  in skin permeation studies

Danillo Santos Negreiros; Camila Oliveira Cardoso; Manuel Alves Falcão; Ana Clara Nogueira Brathwaite; Geisa Nascimento Barbalho; Marcílio Cunha-Filho; Joyce Kelly Rosário Silva; Guilherme Martins Gelfuso; Taís Gratieri

doi:10.1590/s2175-97902025e24754

Authors

Danillo Santos Negreiros Laboratory of Food, Drugs, and Cosmetics (LTMAC), School of Health Sciences, University of Brasília, Brasília, DF, Brazil , University of Brasília
Camila Oliveira Cardoso Laboratory of Food, Drugs, and Cosmetics (LTMAC), School of Health Sciences, University of Brasília, Brasília, DF, Brazil , University of Brasília
Manuel Alves Falcão University of Brasília , Laboratory of Food, Drugs, and Cosmetics (LTMAC), School of Health Sciences, University of Brasília, Brasília, DF, Brazil
Ana Clara Nogueira Brathwaite Laboratory of Food, Drugs, and Cosmetics (LTMAC), School of Health Sciences, University of Brasília, Brasília, DF, Brazil , University of Brasília
Geisa Nascimento Barbalho Laboratory of Food, Drugs, and Cosmetics (LTMAC), School of Health Sciences, University of Brasília, Brasília, DF, Brazil , University of Brasília
Marcílio Cunha-Filho Laboratory of Food, Drugs, and Cosmetics (LTMAC), School of Health Sciences, University of Brasília, Brasília, DF, Brazil , University of Brasília
Joyce Kelly Rosário Silva Laboratory of Biotechnology of Enzymes and Biotransformation (LABEB), Federal University of Pará, Belém, PA, Brazil , Federal University of Para
Guilherme Martins Gelfuso Laboratory of Food, Drugs, and Cosmetics (LTMAC), School of Health Sciences, University of Brasília, Brasília, DF, Brazil , University of Brasília
Taís Gratieri Laboratory of Food, Drugs, and Cosmetics (LTMAC), School of Health Sciences, University of Brasília, Brasília, DF, Brazil , University of Brasília

DOI:

https://doi.org/10.1590/s2175-97902025e24754

Keywords:

Essential oil, Alpha phellandrene, Validation, Analytical method, HPLC-UV, Topical application

Abstract

Alpha-phellandrene (α-PHE), a cyclic monoterpene abundant in volatile oils, has anti-inflammatory and healing properties. A cutaneous formulation using this bioactive could provide benefits in wound healing. However, a simple method that quantifies α-PHE in the skin is still required. Thus, the study aims to develop and validate a high-performance liquid chromatographic (HPLC-UV) method to quantify α-PHE extracted from the skin. The method uses a reversed-phase C₁₈ column (15 cm × 4.6 mm, 5 μm) and a mobile phase consisting of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B) flowing at 0.5 mL/min. The phase concentration (A/B %) along the 40-min run time varied as described: 15/85 (0.01–10 min), 0/100 (10.1–20 min), and 15/85 (20.1–40 min). α-PHE was detected at 263 nm. The method was selective against skin and formulation interferents, linear (r = 0.9995) in a concentration range of 0.5–15.0 μg/mL, precise with an overall variation coefficient lower than 5.5%, accurately achieving recovery from the skin within 92–102%, and sensitive for the proposed application (detection limit = 0.05 µg/mL, quantification limit = 0.14 μg/mL). Finally, the validated method was tested in a skin penetration study, attesting to its success in developing topical formulations containing α-PHE.

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Validation of a chromatographic method for α-phellandrene quantification in skin permeation studies

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