[Development of 2,4,6-trinitrophenol examination methods to evaluate the features of its distribution in warm-blooded animals].

Abstract

The objective: Is to study a 2, 4, 6-trinitrophenol (2.4,6-TNP) distribution in warm-blooded animals using physical chemical analysis methods after intragastric injection of toxicant. The methods of thin-layer chromatography, spectrophotometry and high-efficient liquid chromatography were used in the study process. Four-months-old rats of the Wistar line (males) were considered as a model of warm-blooded organisms. The investigated substance in amount of three-times LD50 was intragastrically injected in the aqueous-suspension state. 2.4,6-TNP was isolated by a mixture of acetone-acetonitrile (1:1) in a double (by 0.5 of hour) infusing mode from the bioactive matrix of experimental animals, sustaining the mass ratio of isolated agent to bioactive matrix equaled to 2:1. Purification and preliminary identification of analyte were conducted on «Sorbfil» plates (mobile phase - acetone - chloroform (7:3)), confirming identification - by absorption in dimethylformamide medium and by retention time in column (64×2 mm) of «Separon C-18» sorbent during elution by acetonitrile-water (2:8) mixture. The evaluation of 2, 4, 6-trinitrophenol quantitative content by optical density of dimethylformamide solution of analyte at 379 nm was carried out. The analyte in unchanged form was found in blood, parenchymatous and hollow organs, their contents and blood of experimental animals. The highest content of 2.4,6-TNP (mg/100 gr) was revealed in gastric content (149.88±22.70), gastric tissue (97.89±4.86), blood (15.91±0.90) and muscles (10.87±1.91).