Optimization, validation and application of headspace solid-phase microextraction gas chromatography for the determination of 1-nitro-2-phenylethane and methyleugenol from Aniba canelilla (H.B.K.) Mez essential oil in skin permeation samples.

Aniba canelilla (H.B.K.) Mez is an aromatic plant from the Amazon region whose essential oil has 1-nitro-2-phenylethane (NP) and methyleugenol (ME) as major compounds. Despite of the scientifically proven antifungal and anti-inflammatory activities for these compounds, there is no report up to date about the topical permeation or quantification of NP and ME on skin samples. The aim of this study was the validation of an optimized bioanalytical method by solid-phase microextraction in headspace mode in gas chromatograph with flame ionization detector (HS-SPME-GC-FID) for the determination of NP and ME from the oil in different samples from permeation study, such as porcine ear skin (PES) layers (stratum corneum, epidermis and dermis) and receptor fluid (RF). For this propose polydimethylsiloxane fibers (100 μm) were used and HS-SPME extraction condition consisted of 53 °C, 21 min, and 5% w.v NaCl addition. The wide range of the calibration curve (2.08-207.87 μg mL for NP and 0.40-40.41 μg mL for ME), the presence of matrix interferences and the intrinsic characteristics of HS-SPME required a data linearization using Log. Thereby, data and the gained results presented homoscedasticity, normalization of residues and adequate linearity (r > 0.99) and accuracy for both compounds. In order to verify the applicability of the validated method, the HS-SPME-GC-FID procedure was performed to determine the amount of NP and ME permeated and retained in samples after Franz diffusion cell study from different dosages (20, 100 and 200 μL) of A. canelilla oil. Compounds permeation showed a progressive increase and penetration dependence based on the dosage applied. Furthermore, retention was in order receptor fluid >> dermis >> epidermis >> stratum corneum for both compounds, suggesting NP and ME could penetrate deep tissue, probably due to the partition coefficient, mass, size, and solubility of these compounds. In conclusion, the proposed method by HS-SPME-GC-FID to quantify 1-nitro-2-phenylethane and methyleugenol from Aniba canelilla essential oil was able to determine selectively, precisely and accurately these main compounds in skin permeation samples.