An analytical strategy for the identification of carbamates, toxic alkaloids, phenobarbital and warfarin in stomach contents from suspected poisoned animals by thin-layer chromatography/ultraviolet detection.

In this study, an analytical strategy to identify brucine, strychnine, methomyl, carbofuran (alkaline compounds), phenobarbital, and warfarin (acid compounds) using thin-layer chromatography (TLC) screening with ultraviolet (UV) detection at 254 nm in stomach content is shown. The optimum mobile phase was found to be a chloroform: ethyl acetate: diethylamine (0.5:8.5:1) mixture for alkaline substances while a mixture of chloroform: acetone (9:1) has given better results for acidic substances. As for extraction, an equal proportion between distillated water and crude material (1:1) is required. For alkaline compounds, a filtration system was created in order to avoid any interferences from the biological matrix while for acidic compounds only centrifugation (4000 rpm/10 minutes) was required to obtain an appropriate sample. After the respective pretreatments, a one-step liquid-liquid extraction (LLE) has been employed for alkaline substances using a 3 mL of chloroform: ethyl ether (2:1) mixture for 2 min while acidic analytes used 3 mL of chloroform only during 5 min. For both methodologies described, the respective organic layers were dried down and re-suspended with 50 µL of methanol for further TLC plate application. The methodologies have been developed, successfully validated and applied to gastric contents from real case samples of suspected animal poisoning. Positive results from TLC/UV screening were confronted with HPLC-UV and confirmed by GC-MS.

In vitro effects of Blepharocalyx salicifolius (H.B.K.) O. Berg on the viability of Echinococcus ortleppi protoscoleces.

Scolicidal agents are important in the treatment of cystic echinococcosis. This study evaluated the scolicidal activity of the plant Blepharocalyx salicifolius (H.B.K.) Berg against Echinococcus ortleppi protoscoleces. The parasite species was identified by amplifying a fragment of the gene cytochrome c oxidase subunit 1 (COX 1). B. salicifolius crude extract at concentrations of 100, 200, 300 and 400 mg/mL was analyzed at different times (5, 10, 15, 30, 45 and 60 min). N-butanol and ethyl acetate fractions (100 and 200 mg/ mL) were also analyzed at 5, 10, 15 and 30 min. Both fractions showed 100% scolicidal activity at the concentration of 200 mg/mL at 5 min. Gallic acid, identified as the major compound of the ethyl acetate fraction- was responsible for the observed scolicidal activity. The results showed that crude extract and fractions of B. salicifolius have scolicidal effect against E. ortleppi protoscoleces.

Analysis of oral mucosa of users of crack through micronucleus technique.

BACKGROUND: The high consumption of crack has become a serious public health problem. The properties of this drug as well as the effects caused in the body have been approached in different research studies. However, there is no knowledge about the carcinogen level this substance can cause to the user. The presence of micronuclei as biomarkers of genotoxic action reflects the degree of cellular exposure to carcinogens. Considering this fact, the following research aimed to assess the frequency of micronuclei in the oral mucosa of those chemically dependent on crack. METHODS: The sample consisted of buccal mucosa cells from 10 controls, non-smokers, and non-users of drugs and 10 individuals chemically dependent on crack admitted to a hospital. For cell staining, Feulgen technique was applied. RESULTS: Of the 1000 cells analyzed for each sample, the exposed group had an average of 4.3 micronuclei, presenting significant difference (p < 0.01) when compared with the control group, which averaged 0.1 micronuclei. Regarding pictotic cells, the exposed group is also significantly different from the control group (p < 0.01). The karyorrhexis is the nuclear change with the greatest difference between the two groups. It has an average of 347.9 cells undergoing apoptosis for the exposed group, while the control group presented 34.4 cells, obtaining a significant difference, p < 0.001. CONCLUSIONS: The results assessed by micronucleus technique suggested that crack together with other factors associated with the drug might be linked to an increased incidence of micronuclei.