Application of gas chromatography for determination of 2-amino-4,6-dinitrophenol and evaluation of its stability in biological material

2-Amino-4,6-dinitrophenol (2-A-4,6-DNP), or picramic acid, is widely used as an intermediate in organic synthesis, a dye in hair coloring products, an agricultural fungicide, and an explosive. Similar to other related compounds (2,4,6-trinitrophenol (2,4,6-TNF) and N-methyl-N,2,4,6-tetranitroaniline), of which it is a metabolite, 2-A-4,6-DNP is toxic to humans and animals. However, certain aspects of its detection in toxicological screening remain poorly understood. In this study, a mixture of acetone and acetonitrile in equal amounts was identified as the most effective extractant for 2-A-4,6-DNP. The application of gas chromatography–mass spectrometry (GC–MS) for detecting 2-A-4,6-DNP in the biological material extracts was analyzed and justified. The proposed method, based on the indirect determination of 2-A-4,6-DNP through O-acetyl derivative quantification, was employed to assess its stability in the biolo- gical material. 2-A-4,6-DNP was found to be detectable in the biological medium within 25 (storage temperature 18–22 °C) to 45 (storage temperature 1–3 °C) days after the contamination.