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Phenylenediamine (PPD) is an aromatic amine used as a precursor in oxidative permanent hair dyes, with a maximum allowed concentration of 2% and a prohibition on its use in cosmetic products for direct skin application in the EU. Besides hair dyes, PPD and its derivatives are used in the synthesis of azo dyes, antioxidants, tattoo pigments, and pharmaceuticals The determination of PPD in various matrices, such as permanent hair colorants, windshield washer fluid, blood, and urine is typically performed using RP-HPLC. The aim of this paper was to apply an experimental design to optimize the HPLC method for the determination of PPD in biological samples (urine, serum) and cosmetic products ensuring sensitivity, selectivity, and reproducibility of the analytical results. Four crucial factors: acetonitrile content in the mobile phase (%), pH value of the aqueous phase, temperature and injection volume were selected and Response Surface Methodology (RSM) was performed to optimize the chromatographic conditions. The optimal conditions obtained are: acetonitrile content in the mobile phase 70%, pH value of the aqueous phase 3.07, column temperature 45 ºC, and injection volume 1µL.

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Application of Experimental Design for Optimizing the HPLC Method to Determine p – Phenylenediamine (PPD)

  • 会议论文

  • 发表日期：

    2026年

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  • Emina Bečić,
  • Hanna Helać,
  • Irmela Ivazović,
  • Ajka Čamdzić,
  • Mirza Dedić,
  • Elma Omeragić

摘要

Phenylenediamine (PPD) is an aromatic amine used as a precursor in oxidative permanent hair dyes, with a maximum allowed concentration of 2% and a prohibition on its use in cosmetic products for direct skin application in the EU. Besides hair dyes, PPD and its derivatives are used in the synthesis of azo dyes, antioxidants, tattoo pigments, and pharmaceuticals The determination of PPD in various matrices, such as permanent hair colorants, windshield washer fluid, blood, and urine is typically performed using RP-HPLC. The aim of this paper was to apply an experimental design to optimize the HPLC method for the determination of PPD in biological samples (urine, serum) and cosmetic products ensuring sensitivity, selectivity, and reproducibility of the analytical results. Four crucial factors: acetonitrile content in the mobile phase (%), pH value of the aqueous phase, temperature and injection volume were selected and Response Surface Methodology (RSM) was performed to optimize the chromatographic conditions. The optimal conditions obtained are: acetonitrile content in the mobile phase 70%, pH value of the aqueous phase 3.07, column temperature 45 ºC, and injection volume 1µL.