Determination of arsenic species in fish, crustacean and sediment samples from Thailand using high performance liquid chromatography (HPLC) coupled with inductively coupled plasma mass spectrometry (ICP-MS)

Suitable techniques have been developed for the extraction of arsenic species in a variety of biological and environmental samples from the Pak Pa-Nang Estuary and catchment, located in Southern Thailand, and for their determination using HPLC directly coupled with ICP-MS. The estuary catchment comprises a tin mining area and inhabitants of the region can suffer from various stages of arsenic poisoning. The important arsenic species, AsB, DMA, MMA, and inorganic arsenic (As III and V) have been determined in fish and crustacean samples to provide toxicological information on those fauna which contribute to the local diet. A Hamilton PRP-X100 anion-exchange HPLC system employing a step elution has been used successfully to achieve separation of the arsenic species. A nitric acid microwave digestion procedure, followed by carrier gas nitrogen addition- (N2)-ICP-MS analysis was used to measure total arsenic in sample digests and extracts. The arsenic speciation of the biological samples was preserved using a Trypsin enzymatic extraction procedure. Extraction efficiencies were high, with values of 82-102%(As) for fish and crustacean samples. Validation for these procedures was carried out using certified reference materials. Fish and crustacean samples from the Pak Pa-Nang Estuary showed a range for total arsenic concentration, up to 17 microg g(-1) dry mass. The major species of arsenic in all fauna samples taken was AsB, together with smaller quantities of DMA and, more importantly, inorganic As. For sediment samples, arsenic species were determined following phosphoric acid (1 M H3PO4) extraction in an open focused microwave system. A phosphate-based eluant, pH 6-7.5, with anion exchange HPLC coupled with ICP-MS was used for separation and detection of AsIII, AsV, MMA and DMA. The optimum conditions, identified using an estuarine sediment reference material (LGC), were achieved using 45 W power and a 20 minute heating period for extraction of 0.5 g sediment. The stability and recovery of arsenic species under the extraction conditions were also determined by a spiking procedure which included the estuarine sediment reference material. The results show good stability for all species after extraction with a variability of less than 10%. Total concentrations of arsenic in the sediments from the Pak Pa-Nang river catchment and the estuary covered the ranges 7-269 microg g(-1)and 4-20 [micro sign]g g(-1)(dry weight), respectively. AsV was the major species found in all the sediment samples with smaller quantities of AsIII. The presence of the more toxic inorganic forms of arsenic in both sediments and biota samples has implications for human health, particularly as they are readily 'available'.     

The critical level of cadmium in renal cortex: the concept and its limitations

Because renal cortex is a primary target organ of Cd the seriousness of human exposure is commonly assessed in terms of the extent to which it permits renal cortical Cd concentrations to approach a critical value. The critical value has been defined in terms of PCC-10, the population critical concentration at which 10% of exposed individuals may be expected to show abnormal renal function. In chronically exposed human populations, PCC-10 appears to lie in the range of 180 to 220 μg Cd/gram fresh weight of cortex. However, Cd accumulated in the kidney under these conditions is distributed between more than one compartment, in all likelihood differing greatly in their toxicological characteristics. For instance, a major fraction of this Cd is bound to metallothionein, and may as such be relatively inert. Compartmentation of tissue Cd, in turn, is strongly influenced by the nature of exposure, and the PCC-10 value of 200 μg/g may therefore have limited general significance. The problem of risk assessment is further complicated by the high variability of Cd absorption. Nevertheless, human renal Cd levels commonly reach values as high as 1/3 to 1/6 of the chronic PCC-10, and further increases in Cd intake should therefore be avoided.     

Bioleaching of zinc and copper from anaerobically digested swine manure: effect of sulfur levels and solids contents.

Anaerobically digested swine manure (ADSM) generally contains high concentrations of zinc (Zn) and copper (Cu). These metals levels exceed the land application regulations of municipal biosolids of many European countries and are on the borderline of exceptional quality in the U.S. Environmental Protection Agency (U.S. EPA) 40 CFR part 503 standards. From this perspective, a series of batch tests were conducted to evaluate the feasibility of bioleaching of Zn and Cu from ADSM so that the produced biosolids could safely be applied to land. The effect of different substrate levels (sulfur) and total solids content (as high as 9%) on metal solubilization was investigated. The results showed that metal solubilization efficiency for both Zn and Cu declined significantly with an increase in total solids from 3 to 6% and then to 9% at the same substrate level. Metal solubilization increased proportionately with increases in substrate concentration up to 6% of total solids content. However, at 9% total solids content, metal solubilization was insignificant at all substrate levels tested. At a 6% total solids level and 10 000 mg/L of substrate, 94% of Zn and 72% of Cu were solubilized. Bioleaching was also found to be effective in destruction of pathogens with approximately 2.5 log-scale reduction. The residual biosolid was found to meet the Class A biosolids standands of U.S. EPA 40 CFR part 503.     

Isolation and characterization of melanopsin and pinopsin expression within photoreceptive sites of reptiles

Non-mammalian vertebrates have multiple extraocular photoreceptors, mainly localised in the pineal complex and the brain, to mediate irradiance detection. In this study, we report the full-length cDNA cloning of ruin lizard melanopsin and pinopsin. The high level of identity with opsins in both the transmembrane regions, where the chromophore binding site is located, and the intracellular loops, where the G-proteins interact, suggests that both melanopsin and pinopsin should be able to generate a stable photopigment, capable of triggering a transduction cascade mediated by G-proteins. Phylogenetic analysis showed that both opsins are located on the expected branches of the corresponding sequences of ortholog proteins. Subsequently, using RT-PCR and RPA analysis, we verified the expression of ruin lizard melanopsin and pinopsin in directly photosensitive organs, such as the lateral eye, brain, pineal gland and parietal eye. Melanopsin expression was detected in the lateral eye and all major regions of the brain. However, different from the situation in Xenopus and chicken, melanopsin is not expressed in the ruin lizard pineal. Pinopsin mRNA expression was only detected in the pineal complex. As a result of their phylogenetic position and ecology, reptiles provide the circadian field with some of the most interesting models for understanding the evolution of the vertebrate circadian timing system and its response to light. This characterization of melanopsin and pinopsin expression in the ruin lizard will be important for future studies aimed at understanding the molecular basis of circadian light detection in reptiles.