Application of Box-Behnken Design for Ultrasound- Assisted Extraction and Recycling Preparative HPLC for Isolation of Anthraquinones from Cassia singueana

Introduction – Cassia singueana Del. (Fabaceae) is a rare medicinal plant used in the traditional medicine preparations to treat various ailments. The root of C. singueana is a rich source of anthraquinones that possess anticancer, antibacterial and antifungal properties. Objective – The objective of this study was to develop ultrasound-assisted extraction (UAE) method for achieving a high extraction yield of anthraquinones using the response surface methodology (RSM), Box-Behnken Design (BBD), and a recycling preparative HPLC protocol for isolation of anthraquinones from C. singueana. Methodology – Optimisation of UAE was performed using the Box-Behnken experimental design. Recycling preparative HPLC was employed to isolate anthraquinones from the root extract of C. singueana. Results - The BBD was well-described by a quadratic polynomial model (R2 = 0.9751). The predicted optimal UAE conditions for a high extraction yield were obtained at: extraction time 25.00 min, temperature 50C and solvent-sample ratio of 10 mL/g. Under the predicted conditions, the experimental value (1.65 ± 0.07%) closely agreed to the predicted yield (1.64%). The obtained crude extract of C. singueana root was subsequently purified to afford eight anthraquinones. Conclusion - The extraction protocol described here is suitable for large-scale extraction of anthraquinones from plant extracts

Entomotoxicology in burnt bodies: a case of maternal filicide-suicide by fire

One of the most common methods of maternal filicide is by fire. In this case study, a 40-year-old female and her children were found completely burned in a burnt out car. All bodies showed a degree of destruction by fire consisting to a level 3 of the Crow-Glassman Scale (CGS) and early stage of insect activity. Toxicological analyses were performed on soft tissues and body fluids still available. The results were positive for diazepam and its metabolites only for children with blood concentrations consistent with therapeutic doses of benzodiazepines. Home video surveillance cameras confirmed sedation prior to death recording the mother while administering some drops of sedative drugs in a soft drink to the children just a couple of hours before setting fire to the car. Based on autopsy findings, all victims were still alive at the time of fire. The cause of death was determined as carbon monoxide poisoning and fatal thermal injuries by fire. This case study has a special focus on the entomotoxicology and the potential role of insects in death investigations of burnt bodies, supposed to be an inadequate substratum for insect colonization. It demonstrates that in burnt bodies, arthropod colonization can be quite immediate after fire is extinguished. Toxicological analyses performed on larvae actively feeding on the children’s bodies were positive for diazepam and its metabolites in small amount compared with blood concentrations, whereas the larvae collected from the mother’s body were totally negative. These data, according to the autopsy findings and the toxicological results from the victim’s blood and tissues, supported the suspect of a non-lethal sedation prior to death, which is a common behaviour in maternal filicide

Intraguild predation influences oviposition behavior of blow flies (Diptera: Calliphoridae)

The objective of the present study was to determine whether blow flies (Diptera: Calliphoridae) are able to identify larvae of an intraguild predator species in the substrate and avoid laying eggs there. Blow flies oviposited in traps with different treatments: substrate only and substrate with larvae of Chrysomya albiceps (Wiedemann, 1819), Chrysomya megacephala (Fabricius, 1794), or Chrysomya putoria (Wiedemann, 1830). Ch. megacephala, Ch. putoria, and Lucilia eximia (Wiedemann, 1819) avoided laying eggs in the trap containing Ch. albiceps larvae. Cochliomyia macellaria (Fabricius, 1775) did not oviposit differently in each substrate but had overall low abundance. The prevalence of species on corpses may be influenced by the ability of the species to detect the presence of other species, mainly predators. In this sense, intraguild predation may result in misinterpretations of a crime scene and should be considered when assessing the minimum postmortem interval