Eggenreich K, Zach E, Beck H, Wintersteiger R
Institute of Pharmaceutical Chemistry and Pharmaceutical Technology, Karl-Franzens-University, Schubertstrasse 1, A-8010 Graz, Austria.
J Biochem Biophys Methods. 2004 Oct 29;61(1-2):35-46. doi: 10.1016/j.jbbm.2004.04.017.
For the determination of two oxidation hair dyes, 4-amino-m-cresol (4-AC) and 5-amino-o-cresol (5-AC), a sensitive isocratic high performance liquid chromatography (HPLC) method using the reversed phase mode was developed. The hair dyes were pre-column derivatized with fluorescamine prior to injection. Sensitivity could be improved 10-fold for 4-AC and 50-fold for 5-AC by fluorescence detection compared to UV detection. The limit of detection was 1 ng/injection for 4-AC and 100 pg/injection for 5-AC, respectively. For the determination of both compounds in aqueous biological matrices in order to simulate conditions for penetration studies with pig skin, a solid phase extraction procedure using C18 cartridges and acetonitrile (ACN) for elution could be developed. Average recovery was 83.4% with a coefficient of variation (CV) of 2.64% for intra-day assay and 3.20% for inter-day assay for 5-AC and 2.89% and 3.41% for 4-AC, respectively.
为了测定两种氧化型染发剂，即4-氨基间甲酚（4-AC）和5-氨基邻甲酚（5-AC），开发了一种采用反相模式的灵敏等度高效液相色谱（HPLC）方法。染发剂在进样前用荧光胺进行柱前衍生化。与紫外检测相比，通过荧光检测，4-AC的灵敏度可提高10倍，5-AC的灵敏度可提高50倍。4-AC的检测限分别为1 ng/进样，5-AC的检测限为100 pg/进样。为了模拟猪皮渗透研究的条件，以便测定水性生物基质中的两种化合物，可开发一种使用C18柱和乙腈（ACN）进行洗脱的固相萃取方法。5-AC日内测定的平均回收率为83.4%，变异系数（CV）为2.64%，日间测定的变异系数为3.20%；4-AC日内测定的变异系数分别为2.89%，日间测定的变异系数为3.41%。
