建立柱前在线自动衍生-高效液相色谱法快速测定染发剂中过氧化氢的含量

目的：建立柱前在线自动衍生-高效液相色谱法快速测定染发剂中过氧化氢的含量，并与《化妆品安全技术规范》（2015年版）高效液相色谱柱前衍生法进行方法学比较。方法：对照品和样品在自动进样器中完成自动衍生反应后直接进入高效液相色谱系统进行分析（将被测样品中的过氧化氢与三苯基膦进行衍生化反应，生成氧化三苯基膦），由安捷伦ZORBAX SB-C₁₈（4.6 mm×250 mm，5 μm）色谱柱分离。结果：柱前在线自动衍生-高效液相色谱法和《化妆品安全技术规范》（2015年版）高效液相色谱柱前衍生法的线性范围分别为2.4~192.7 μg·mL^-1和9.6~192.7 μg·mL^-1，线性回归方程分别为Y=39.19X-187.9（r=0.999 6）和Y=43.15X-160.8（r=0.999 7），加样回收率分别为92.7%~99.5%和91.4%~99.0%，精密度RSD分别为0.70%和0.30%（n=6），检测下限分别为18.1 μg·g^-1和23.5 μg·g^-1。柱前在线自动衍生-高效液相色谱法和《化妆品安全技术规范》（2015年版）高效液相色谱柱前衍生法测定3种不同品牌染发剂中过氧化氢的含量分别为5.83%~8.40%和5.68%~8.27%，RSD为1.6%~2.6%。结论：柱前在线自动衍生-高效液相色谱法快速测定化妆品中过氧化氢的重复性好，灵敏度高，且比《化妆品安全技术规范》（2015年版）检测方法操作简单，检出下限更低，适用于染发剂中过氧化氢的常规分析检测。

Objective:To establish a high performance liquid chromatography with on-line automated pre-column derivatization method to measure the content of H₂O₂ in hair dye,and to compare the new established method with high performance liquid chromatography with pre-column derivatization method published in《Safety and Technical Standards for Cosmetics》(2015 version).Methods:After automated derivatization in the autosampler,the standards and samples were directly into the HPLC system for analysis(hydrogen peroxide in the samples were reacted with triphenylphosphine to produce triphenylphosphine oxide).The analysis was carried out on an Agilent SB-C₁₈(4.6 mm×250 mm,5 μm)column.Results:The hydrogen peroxide linear ranges of high performance liquid chromatography with on-line automated pre-column derivatization method and high performance liquid chromatographic with pre-column derivatization method were 2.4-192.7 μg·mL^-1(Y=39.19X-187.9)(r=0.999 6)and 9.6-192.7 μg·mL^-1(Y=43.15X-160.8)(r=0.999 7),respectively.The average recovery rates were 92.7%-99.5% and 91.4%-99.0%,with RSD of 0.7% and 0.3%(n=6),respectively.The detection limits were 18.1 μg·g^-1 and 23.5 μg·g^-1,respectively.The content of hydrogen peroxide in three different batches of hair dye products were 5.83%~8.40% by on-line automated pre-column derivatization method and 5.68%~8.27% by high performance liquid chromatographicwith RSD of 1.6%~2.6%.Conclusion:High performance liquid chromatography with on-line automated pre-column derivatization method has good reproducibility and high sensitivity.It is more simple and its detection limit is lower than that of 《Safety and Technical Standards for Cosmetics》(2015 version).It is suitable for routine analysis and detection of hydrogen peroxide in hair dye products.

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