Development and validation of an LC-MS/MS method for determination of p-phenylenediamine and its metabolites in blood samples

作者：Mohamed Khaled M^*; Cromarty Duncan; Steenkamp Vanessa

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2015, 997: 1-6.

DOI：10.1016/j.jchromb.2015.05.030

• p-Phenylenediamine
• Metabolites
• Blood
• LC-MS/MS
• Validation
• Stability

In some developing countries, p-phenylenediamine (PPD) is used in combination with Henna as hair dye or skin decoration. A sensitive LC-MS/MS method was developed and validated for the simultaneous determination of p-phenylenediamine (PPD) and its metabolites N-acetyl-p-phenylenediamine (MAPPD) and N,N-diacetyl-p-phenylenediamine (DAPPD) in human blood. Acetanilide was used as an internal standard (IS). The LC-MS/MS was operated under multiple reaction-monitoring mode using the electrospray positive ionization technique. The transition ions m/z 109 --> 92, m/z 151 --> 92, m/z 193 --> 92, and m/z 136 --> 77 were selected for the quantification of PPD, MAPPD, DAPPD, and IS, respectively. The linear range was 10-2000 ng/mL for all the compounds. The absolute recoveries were 51.94, 56.20 and 54.88% for PPD, MAPPD and DAPPD, respectively. Intra- and inter-assay imprecision were lower than 14% (RSD), and the bias of the assay was lower than 15% for all the compounds. The stability studies demonstrated that critical degradation for PPD in blood samples and autosampler occurred after 6 h, while MAPPD and DAPPD were stable in blood samples and the autosampler up to 48 h and 24 h, respectively. This newly developed method allows for the detection of PPD and its metabolites in blood samples in the clinical and forensic setting.

• 出版日期2015-8-1