Abstract
Extracts made from the skin of dead Lithodytes lineatus frog individuals with the application of the benzocaine-based anesthetic gel, introduced into the oral cavity, were analyzed by 1H Nuclear Magnetic Resonance to investigate whether the application of this product (oral) can make studies that use extracts from the skins of these animals unfeasible. For comparison, we used skins of another species of anuran following the same death protocol. No trace of the benzocaine substance was found in the 1H-NMR spectra of the skin extracts from any of the tested anuran species. Still, using the hierarchical clustering model, it was possible to observe the formation of well-defined groups between the skin extracts of anurans and the anesthetic used to kill these animals. Our results suggest that the lethal dose of benzocaine in gel used inside the mouth of frogs may have no influence on potential results regarding the chemical composition or even bioassays using extracts made from the skin of these animals killed under this protocol since there was no detection of this substance for the analyzed samples.
Highlights
Studies involving analysis of the chemical composition of the cutaneous secretion of several species of anurans have been widely carried out, because it is possible to find a large number of substances with bioactive properties in the skin of these animals that present, mainly, great antimicrobial potentials [1,2,3,4,5,6]
The methanolic (MET) and dichloromethanic (DCM) extracts made with the skins of individuals of Lithodytes lineatus were compared with the dichloromethanic extract of the Benzotop© (BEN) product to verify the presence or absence of characteristic signs of the benzocaine substance (Fig 3)
It was found that the benzocaine substance was not incorporated into the general composition of the skin extracts of L. lineatus individuals (Fig 4) killed with BEN, in neither of the two extractions, since the signs and the characteristic chemical shifts of the substance in question were not detected
Summary
Studies involving analysis of the chemical composition of the cutaneous secretion of several species of anurans have been widely carried out, because it is possible to find a large number of substances with bioactive properties in the skin of these animals that present, mainly, great antimicrobial potentials [1,2,3,4,5,6]. There are several methods to extract the substances present in the body of frogs. The method using skin extracts requires greater care during treatment in order to avoid indirect contamination of the samples during handling.
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