Development and validation of a method for the determination of regulated fragrance allergens by High-Performance Liquid Chromatography and Parallel Factor Analysis 2.

This work presents the development and validation of a multivariate method for quantitation of 6 potentially allergenic substances (PAS) related to fragrances by ultrasound-assisted emulsification microextraction coupled with HPLC-DAD and PARAFAC2 in the presence of other 18 PAS. The objective is the extension of a previously proposed univariate method to be able to determine the 24 PAS currently considered as allergens. The suitability of the multivariate approach for the qualitative and quantitative analysis of the analytes is discussed through datasets of increasing complexity, comprising the assessment and validation of the method performance. PARAFAC2 showed to adequately model the data facing up different instrumental and chemical issues, such as co-elution profiles, overlapping spectra, unknown interfering compounds, retention time shifts and baseline drifts. Satisfactory quality parameters of the model performance were obtained (R2≥0.94), as well as meaningful chromatographic and spectral profiles (r≥0.97). Moreover, low errors of prediction in external validation standards (below 15% in most cases) as well as acceptable quantification errors in real spiked samples (recoveries from 82 to 119%) confirmed the suitability of PARAFAC2 for resolution and quantification of the PAS. The combination of the previously proposed univariate approach, for the well-resolved peaks, with the developed multivariate method allows the determination of the 24 regulated PAS.

Ultrasound-assisted emulsification microextraction coupled with high-performance liquid chromatography for the simultaneous determination of fragrance allergens in cosmetics and water.

A simple, inexpensive, and environmentally friendly method based on ultrasound-assisted emulsification microextraction followed by solidification of floating organic drop and high-performance liquid chromatography coupled to diode array detection was developed for the simultaneous determination of 18 potentially allergenic fragrance substances. Several parameters affecting the microextraction process were investigated in detail by the "one-variable-at-a-time" approach. Optimal conditions were the following: 50 µL of 2-dodecanol as extraction solvent, 10 mL of sample containing 150 g/L of salt, and 5 min of sonication at 35°C. Under the optimized conditions, method showed good linearity in the selected ranges, with squared correlation coefficients ranging from 0.948 to 0.999. Limits of detection ranged from 0.001 to 0.154 µg/mL and enrichment factors from 9 to 237. Precision of the method, expressed as relative standard deviation, was checked at two levels obtaining good results (3.3-14.4%). Recovery studies were made in baby bath water and in eau de cologne showing acceptable accuracy. Finally, the developed method was successfully applied to different commercial cosmetic and water samples. The most commonly found analyte was linalool followed by cinnamal and lilial. Most of the analyzed samples contained at least one of the target compounds.