Optimization and intralaboratorial validation of an analytical method by HPLC/UV for identification and quantification of p-phenylenediamine in henna dye hair and eyebrows
DOI:
https://doi.org/10.22239/2317-269x.01265Keywords:
p-fenilenodiamina, Henna, Method ValidationAbstract
Introduction: p-Phenylenediamine (PPD), aromatic sensitizing amine, has been added to henna powder to modify its natural color to black, increasing its fixation time, a practice that is prohibited in eyelash and eyebrows dyes. Objective: The objective of this study was to optimize and validate, at an intra-laboratory level, an analytical method by HPLC/UV for identification and quantification of PPD in hen dyes for hair and eyebrows. Method: In the method, C8 reverse phase column, mobile phase 1% triethanolamine (pH 8.4) and
acetonitrile (99: 1, v/v), detection 280nm, injection volume: 10uL, flow 1.0 mL/min, column temperature 32°C, run time 10 min, linearity 5-45 μg/mL (n = 5) with correlation coefficient 0.9982, were used. For Cochran test (homoscedasticity): 0.3350 and Critical (0.3934) with 99% confidence. Limits of detection 1.17 μg/mL and quantification 3.54 μg/mL. The coefficient of variation of repeatability was 0.12% and the intermediate precision by F-test yielded p value of 0.283 with 95% of confidence. Accuracy results comprised acceptance criteria of 90%-107%. Results: Of the 19 analyzed samples, 14 presented PPD content between 1.74 and 3.65% w/w, in disagreement with Legislation. Conclusions: The proposed method can contribute to monitoring of quality and safety of use of these products.
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