Determination of the threshold odor concentration of main odorants in essential oils using gas chromatography-olfactometry incremental dilution technique.

An essential oil, obtained by steam distillation of Clinopodium tomentosum (Kunth) Govaerts (Lamiaceae), collected in Ecuador, was analyzed by gas chromatography-olfactometry (GC-O) and GC-MS techniques. To our knowledge, the composition of this essential oil is described here for the first time, both from the chemical and olfactometric viewpoints. A preliminary analysis by GC-MS and using Kovats' retention indexes, lead to characterize and quantify the oil constituents, while GC-O was then applied for the identification of the main odorants. By the incremental dilution method (AEDA, CHARM Analysis), applied to the GC-O technique, the flavor dilution (FD) chromatogram was obtained. In order to calculate the TOC values of the main odorants, the relationship between the odorant concentration at the sniffing port and that one in the injected solution was established. This relationship was calculated by comparing the injected amount with the TOC value of a reference compound (limonene), obtained by dynamic dilution olfactometry. A good agreement was found between calculated and measured TOC values of few odorants.

Simultaneous determination of unmodified sevoflurane and of its metabolite hexafluoroisopropanol in urine by headspace sorptive extraction-thermal desorption-capillary gas chromatography-mass spectrometry.

Unmodified sevoflurane and its metabolite, hexafluoroisopropanol (HFIP), have both been proposed as biomarkers of exposure in post-shift urine for operating room personnel exposed to inhalation anaesthetic sevoflurane. We used headspace sorptive extraction (HSSE) and thermal desorption-capillary GC-MS to assess sensitively both compounds in the urine matrix (after a HFIP deconjugation step). In GC-MS splitless mode, calibration plots (approximately 15-650 microg/L) were linear (r2 > 0.9910) and the limits of detection (1 microg/L for both biomarkers) showed increased sensitivity for HFIP with respect to the previously described headspace GC-MS method. The method was suitable for biological monitoring of both biomarkers of exposure to sevoflurane.

Exposure to heptachlor: evaluation of the effects on the larval and adult epidermis of Rana kl. esculenta.

Widely used in the past against termites and soil insects, the chlorinated insecticide heptachlor (H) is a toxic contaminant which represents a risk for both terrestrial and aquatic organisms. Like many organochlorine pesticides, heptachlor and heptachlor epoxide (HE), with oxidation products synthesized by many plant and animal species, degrade slowly since many of the derived compounds are persistent. This increases the status of heptachlor as a hazardous pollutant. In the present experimental study we exposed specimens of Rana kl. esculenta, from the tadpole stage through to their complete metamorphosis, to three different concentrations of heptachlor (4, 40 and 400 ppb). Mortality and HE bioaccumulation were evaluated on all the experimental groups. Since amphibian integument directly interacts with the environmental constituents (water, air and soil), we investigated the toxic effects on the ventral epidermis of both tadpole and adult samples by employing such histo-cytopathological biomarkers as ultrastructural morphology, certain enzyme activities (acid and alkaline phosphatases, AcPase, and AlkPase; succinic dehydrogenase, SDH; alpha-naphtyl butyrate esterase, ANBE; nitric oxide synthase/NADPH diaphorase, NOS/NADPHd). Also, the levels of reactive oxygen species (ROS) in the different conditions were evaluated. The results obtained were of ecological relevance, in particular as regards the effects of this environmental toxicant on the samples of tadpole epidermis. Severe morphological alterations were observed in the larval epidermal cells (apical and skein cells), whereas the cell epidermis (keratinocytes and mitochondria-rich cells) of the adult survivors showed changes in enzyme activities, particularly those involved in the protective response to xenobiotic injury. In general, morpho-histochemical studies, analysis of HE bioaccumulation and mortality showed a relation to the H doses employed.

Differential retention of PCB congeners in cockroaches Blattella germanica.

Organisms differ widely in their ability to metabolize and eliminate polychlorinated biphenyls (PCBs). We evaluated the retention of PCB congeners, reference standard mixture, experimentally injected into cockroach Blattella germanica, in relation to sex and time, and its elimination through the feces, exuviae and oothecae. The nymphs that died prematurely had a higher average PCB retention than those with a longer lifetime, 88% of the injected dose for those that died within 1 to 10 days, and 76% for those within 11-20 days. Diverse PCB congeners showed differential retention, and particularly the more volatile, low weight congeners were recovered in lower concentrations, the medium-weight ones were intermediate, while the high-weight congeners attained the highest concentration. PCBs were also detected in the excrements and in exuviae, which may therefore act as detoxification paths, and in the oothecae, thus showing that PCBs can be transferred from mother to progeny.

Effects of environmental pollution on the liver parenchymal cells and Kupffer-melanomacrophagic cells of the frog Rana esculenta.

In vertebrates, the biotransformation processes of xenobiotics are performed mainly by the liver which involves both hepatocytes and Kupffer-melanomacrophagic cells through enzymatic and nonenzymatic mechanisms. In this study, we investigated the liver of Rana esculenta adult frogs collected at two sample rice fields, one heavily polluted and one relatively unpolluted. Water pollution was determined by chemical analysis on tadpoles. The specific activities of some enzymes (glucose-6-phosphate dehydrogenase (G6PDH), acid and alkaline phosphatases (AcPase and AlkPase), succinic dehydrogenase (SDH), and catalase) were studied in the liver of adult frogs to identify the possible changes induced by contamination in the metabolic processes which depend on the function of the liver. The production of reactive oxygen species (ROS) were also evaluated through histochemical techniques. In the polluted samples, hepatocytes showed variations in the activity of G6PDH, AlkPase, and SDH and a moderate to intense ROS expression. Prominent changes were observed in Kupffer cells (KCs) and melanomacrophages (MMPs), both showing intense reactivity for AcPase and catalase and variations in melanin content and distribution. Results thus indicate a general adaptive response of liver parenchyma to environmental pollution. The possible role of both KCs and MMPs as scavengers of foreign substances is discussed.