In vivo distribution and fractionation of indium in rats after subcutaneous and oral administration of [(114m)In]InAs

Two in vivo experiments were carried out in this study. In the first experiment five rats were given two subcutaneous injections of [(114m)In]InAs. Major sites of accumulation were spleen, liver and kidney. The intracellular distribution of indium was examined by differential centrifugation. The cytoplasmic fraction contained most of the indium activity followed by the mitochondrial fraction. Both outcomes are in close agreement with the results obtained in previous studies. Chromatographic separations on a preparative size exclusion column were carried out. It was shown that indium was mostly bound to high molecular mass compounds in serum and in the cytoplasmic fraction of spleen, liver and kidney. In a second experiment five rats were given four oral doses of [(114m)In]InAs over a short period. Prior to this experiment the in vitro solubility of cold InAs in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) was determined using graphite furnace atomic absorption spectroscopy. In the case of the SGF only 1.3% of an InAs suspension dissolved after 48 hours incubation at 37 degrees C. InAs was not soluble in SIF. Uptake of InAs after oral administration was minimal (<1%). Due to incomplete removal of traces of [(114m)In]InAs from the gastrointestinal tract, it was impossible to calculate accurately the in vivo distribution over the different organs. As the uptake and consequently the activity in the organs were very low, no further chromatographic separations could be carried out. Considering this very low uptake, it can be concluded that InAs will not accumulate in the body after oral exposure.     

Association of metals (mercury, cadmium and zinc) with metallothionein-like proteins in storage organs of stranded dolphins from the Mediterranean sea (Southern Italy)

Selected toxic and essential metals (mercury, Hg; cadmium, Cd; and zinc, Zn) were determined in the liver and in the kidney collected from 13 dolphins (Stenella coeruleoalba and Tursiops truncatus) stranded along the southern coasts of Italy from 1991 to 1999. Liver samples were also analysed for their methyl mercury (MeHg) and selenium (Se) contents. For subcellular fractionation, liver and kidney samples were homogenized in Tris-HCl buffer; after centrifugation, the supernatant (cytosol) was separated from pellets (insoluble fraction), heated at 80 degrees C for 10 min and centrifuged in order to separate the precipitate containing the thermolabile high molecular weight proteins. The cytosol heat-stable fraction, including metallothionein-like proteins (MTLPs), was then purified by gel filtration chromatography on Sephadex G-75 column. The three subcellular fractions collected (insoluble fraction, thermolabile fraction and purified heat-stable fraction) were analysed for their Hg, Cd and Zn contents. The analytical results confirm previous similar studies on toothed whales in showing that: (1) in the liver, as well as in the kidney, Hg was mainly found in the insoluble fraction, therefore, metallothioneins seem to have no role in the Hg detoxification; (2) in the liver, the molar ratio between Se and inorganic Hg was very close to 1; this suggests that the final compound of MeHg detoxification could be HgSe (tiemannite); (3) in almost all the samples, Cd and Zn were detected in the purified heat-stable fraction (including MTLPs). The mechanism of Cd detoxification and Zn homeostasis is also discussed.     

Metal Levels in Raccoon Tissues: Differences On and Off the Department of Energy's Savannah River Site in South Carolina

Levels of arsenic, cadmium, chromium, copper, lead, manganese, selenium, and strontium⁸⁸ were examined in heart, kidney, muscle, spleen and liver of raccoons (Procyon lotor) from four areas on the Department of Energy's Savannah River Site (SRS), including near a former reactor cooling reservoir and a coal ash basin, and from public hunting areas within 15 km of the site. Mercury is mentioned briefly because it is discussed more fully in another paper. We test the hypotheses that there are no differences in metal levels between raccoons on SRS and off the SRS (off-site), and among different locations on the SRS. Although raccoons collected off-site had significantly lower levels of mercury and selenium in both the liver and kidney, there were few consistencies otherwise. There were significantly higher levels of cadmium in liver of on-site compared to off-site raccoons, and significantly higher levels of chromium and strontium⁸⁸ in kidney of on-site compared to off-site raccoons. Copper and manganese were highest in the liver; cadmium, lead, mercury and selenium were highest in the liver and kidney; chromium was highest in the spleen and muscle; arsenic was highest in the heart, and strontium⁸⁸ was slightly higher in the kidney than other organs. Where there were significant differences on site, chromium, manganese were highest in raccoon tissues from Steel Creek; arsenic, lead and selenium were highest in the Ash Basin; cadmium was highest at Upper Three Runs; and strontium⁸⁸ was highest at Upper Three Runs and Steel Creek. The patterns were far from consistent.     

Biomonitoring of heavy metals (Cd,Hg, and Pb) and metalloid (As) with the Portuguese common buzzard (Buteo buteo)

The accumulation of heavy metals in the environment may have a wide range of health effects on animals and humans. Thus, in this study, the concentrations of arsenic (As), cadmium (Cd), lead (Pb), and mercury (Hg) in the blood and tissues (liver and kidney) of Portuguese common buzzards (Buteo buteo) were determined by inductively coupled plasma-mass spectrometer (ICP-MS) in order to monitor environmental pollution to these elements. In general, Hg and As were the elements which appeared in the highest and lowest concentrations, respectively. A highest percentage of non-detected concentration was found for blood Cd (94.6 %) but, in turn, it was the only metal that was detected in all kidney samples. The kidney was the analyzed sample which showed the highest concentrations of each element evaluated. Statistically, significant differences among blood, liver, and kidney samples were observed for As and Cd (P?相似文献   

Historical pollution of seldom monitored trace elements in Sweden--Part B: sediment analysis of silver, antimony, thallium and indium

Sediment cores from four boreal and remote lakes in a south to north transect in central Sweden were analysed for acid leachable silver, antimony, thallium and indium in the solid sediment phase and the corresponding porewater. Dating of the cores was made by their content of acid leachable lead and the (206)Pb/(207)Pb ratio, in one lake also by (210)Pb. The impact of diagenesis on element redistribution in the sediments was included and found to be minor except for thallium. The results show lowered concentrations towards the north and most intense accumulation after the Second World War, which is taken as evidence for atmospheric deposition being the primary source. Indium has declining concentrations in recent strata while silver and antimony increase. Thallium has lowered acid-leachable concentrations in recent strata. For all metals the impact of domestic industrialisation as well as the early industrialisation of central Europe is discernible. Only thallium appears to reach a geological background at depths that correspond to the late 18th century. For the other metals elevated levels are concluded.     

Impact of Hazardous Exposure to Metals in the Nile and Delta Lakes on the Catfish, Clarias lazera

The impact of metal pollution in the Nile and Delta lakes was studied in Clarias lazera. Lake Edku is grouped as the site highest in metal concentrations. Compared to Lakes Maryût and Edku, the Nile stations displayed lower levels of metal contamination signifying their selection as reference sites. Metals were accumulated most by the liver, less by muscles and least by serum. Iron showed the highest bioconcentration factors (BCF) in liver, muscles or serum compared to other metals. Even after the refinement of the surrounding water, fish seem to uphold the metals that have been previously ingested. Mercury, in particular, seems to eventually reside in muscles rather than in liver. The positive relationship between total serum protein and metal contents may indicate the formation of specific metal-binding proteins. Meanwhile, serum hyperalbuminemia in polluted fish is probably a shock response towards the high stress index induced by metals. The hypothesis that most of serum copper is bound to ceruloplasmin is documented also in C. lazera. Moderate, but not extennsive, exposure to metals was quite stimulatory to liver alkaline phosphatase (AP) biosynthesis. AP could, thus, be considered as a promising bioindicator for assessing sublethal water intoxicosis.     

Multielement analysis in the fish hepatic cytosol as a screening tool in the monitoring of natural waters

The possibility of direct measurement of trace elements in hepatic cytosol of European chub (Squalius cephalus) by high-resolution inductively coupled plasma mass spectrometry (HR ICP-MS) after cytosol dilution with Milli-Q water and subsequent acidification was investigated. Due to low detection limits of this procedure, determination of 13 elements (As, Cd, Co, Cu, Fe, Mn, Mo, Pb, Sb, Sn, Sr, V and Zn) was possible in the chub hepatic cytosol, exhibiting excellent measurement repeatability in duplicates. Some of these elements were also measured by HR ICP-MS in acid digested cytosols (Cd, Co, Cu, Fe, Mn, Mo, Sr, V and Zn). Good agreement of the results obtained after sample dilution and sample digestion indicated that complex organic matrix of hepatic cytosol did not affect measurement reliability. Cytosolic concentrations of 13 trace elements in the chub liver were quantified in the following order: Fe, Zn>Cu>Mn>Mo>Sr, V, Cd>Co>As, Pb>Sn>Sb. Unlike Cd, Cu, Fe, Mn and Zn for which the cytosolic concentrations were previously reported after measurement by AAS, cytosolic concentrations of eight additional trace elements characteristic for the liver of chubs inhabiting the low contaminated river water were reported here for the first time (in nanogrammes per gramme)—Mo, 136.8–183.6; Sr, 32.7–63.0; V, 17.5–69.0; Co, 24.3–30.7; As, 9.9–29.5; Pb, 5.8–35.6; Sn, 5.5–12.4; and Sb, 0.9–2.6. The simultaneous measurement of large number of trace elements in the cytosolic fractions of fish tissues, which comprise potentially metal-sensitive sub-cellular pools, could be beneficial as a screening tool in the monitoring of natural waters, because it would enable timely recognition of increased fish exposure to metals.     

Oxidative stress in Cyprinus carpio to analyze microcystin impact in eutrophic shallow lakes: a preliminary study

Microcystins (MCYSTs) are toxins produced by cyanobacteria in aquatic environment and are of high potential risk to aquatic organisms. The physiological responses and pathobiological developments that they elicit in fish have been extensively studied, mainly through acute toxicity experiments. This study was designed to examine the seasonal fluctuation of biochemical markers of oxidative stress in different tissues of a natural population of Cyprinus carpio inhabiting a shallow Mediterranean lake, along with the respective MCYSTs concentrations in blood and tissues at environmentally relevant MCYSTs values. MCYSTs content was assessed in liver, kidney, intestine, brain and muscle along with the MCYSTs in lake water and scum applying ELISA technique. Catalase activity, GSH/GSSH relative concentrations and lipid peroxidation were used as biochemical markers. Our results suggest that common carp of Lake Pamvotis exposed to naturally fluctuating concentrations of MCYST in water and scum contained stably high MCYST concentrations in all tissues that might pose a threat to public health. Liver and kidney were the primary target organs. Tissue concentrations did not correlate with the response of any of the elements of the antioxidant defence system. Hepatic catalase, GSH content and TBARS in all tissues tested followed the fluctuations of major limnological parameters, i.e. water temperature and oxygen concentration, chlorophyll-a, MCYST in water and scum, suggesting that they should be cautiously used to monitor exposure to MCYSTS in natural freshwater ecosystems.     

Distribution of mercury in the tissues of five species of freshwater fish from Lake Mead, USA

Total mercury (Hg) concentrations were determined in seven tissues (skeletal muscle, liver, blood, gonad, brain, gill, and heart) of 59 striped bass and four tissues (muscle, liver, blood, and gonad) of 69 largemouth bass, 76 channel catfish, 12 bluegill, and 22 blue tilapia collected from Lake Mead, USA. Mercury levels generally increased according to trophic level and fish length. For striped bass, mean Hg levels (ng g(-1), wet mass) were highest in the liver (531), followed by muscle (309), heart (186), gonad (136), brain (77), gill (52), and blood (36). Similarly, Hg levels in the catfish and tilapia were liver > muscle > gonad > blood. In contrast, largemouth bass and bluegill had the highest levels in muscle, followed by liver, gonad, and blood. Generally, Hg levels were strongly correlated among the tissues, especially for blood/muscle and blood/liver. As the body burden of Hg increased, the concentration in blood and organs increased relative to the concentration in muscle. The trend was most pronounced for the liver. These relationships could form the basis of a predictive model and suggest that blood and muscle (plugs) could be useful for a non-lethal measure of Hg concentration and exposure in fish. For the striped bass, elevated Hg levels in the tissues were also correlated with degree of emaciation. Liver-to-muscle ratios were similar to literature values, except for tilapia with an average ratio of approximately 1.7, which is higher than generally reported for non-piscivores. Finally, this study demonstrates the usefulness of a solid sampling approach in trace element monitoring, especially as pertaining to in vivo analysis, analysis of a large number of samples and reduction of contamination risk.     

饶河入鄱阳湖口处沉积物中重金属分布特征及形态研究

为了解饶河入鄱阳湖湖口重金属污染状况,在鄱阳湖饶河入湖段设置了3个采样点,用原子吸收分光光度计对沉积物进行了重金属Cu、Pb、Zn的测定,分析了这3种元素在各采样点的形态分布特征.结果表明:饶河人鄱阳湖口处沉积物中Cu、Pb、Zn的污染较为严重,呈现出复合污染的趋势.该区域沉积物中3种元素的分布受水流的影响较大,其形态...     

Organochlorine Pesticides and PCBs in Wild Boars from Calabria (Italy)

At present, there are no specific studies on the evaluation of environmental toxicological risks in Calabria (south of Italy) and on the presence of contaminants in the fauna of this region. The aim of the present research was to investigate the levels of contamination by OC pesticides and PCBs in some organs and tissues of wild boars (utilized as 'biological indicator') from various areas of Calabria. Quantitative determinations of organochlorines were carried out using GC-ECD and confirmed with GC-MS in 154 samples from wild boars (heart, liver, lung, kidney, muscle tissue and spleen) during the hunting season from 2000 to 2002. The results indicate the low residual levels of DDE in 8 samples and DDT in 4 samples and PCBs residues (Aroclor 1232) below the detection limits were found. Therefore, these results show that the Calabria region is not at contamination risk from organochlorines and moreover is free from health problems for the consumer of boar meat.     

Seasonal variation of the metal (Zn, Fe, Mn) and metallothionein concentrations in the liver cytosol of the European chub (Squalius cephalus L.)

Seasonal variations of biometric parameters as well as metal and protein concentrations were analyzed in the liver cytosol of indigenous European chub (Squalius cephalus L.). Fish sampling was carried out in the reproductive (spring) and non-reproductive period (autumn) in order to define parameters which are seasonally dependent. The specimens caught in the spring period had higher biometric (liver mass, condition and hepatosomatic indices) and some biochemical parameters (metallothionein (MT) and Mn) that are characteristic for the reproductive period. Contrary, Fe hepatic cytosolic concentrations were significantly lower in specimens from spring than in autumn period. Total cytosolic proteins, Cu and Cd concentrations were not seasonally dependent. Range of constitutive levels was determined for total cytosolic proteins (17.7 to 24.7 mg ml(-1)) from the whole set of data. Constitutive range of seasonally dependent parameters MT (207 to 337 microg ml(-1)) and Mn (0.11 to 0.19 microg ml(-1)) is based on the data from non-reproductive period. In order to avoid the interference of reproductive cycle on the level of selected biochemical parameters, it is recommended to perform chub sampling during non-reproductive period.     

Metal release from serpentine soils in Sri Lanka

Ultramafic rocks and their related soils (i.e., serpentine soils) are non-anthropogenic sources of metal contamination. Elevated concentrations of metals released from these soils into the surrounding areas and groundwater have ecological-, agricultural-, and human health-related consequences. Here we report the geochemistry of four different serpentine soil localities in Sri Lanka by coupling interpretations garnered from physicochemical properties and chemical extractions. Both Ni and Mn demonstrate appreciable release in water from the Ussangoda soils compared to the other three localities, with Ni and Mn metal release increasing with increasing ionic strengths at all sites. Sequential extraction experiments, utilized to identify “elemental pools,” indicate that Mn is mainly associated with oxides/(oxy)hydroxides, whereas Ni and Cr are bound in silicates and spinels. Nickel was the most bioavailable metal compared to Mn and Cr in all four soils, with the highest value observed in the Ussangoda soil at 168?±?6.40 mg kg^?1 via the 0.01-M CaCl₂ extraction. Although Mn is dominantly bound in oxides/(oxy)hydroxides, Mn is widely dispersed with concentrations reaching as high as 391 mg kg^?1 (Yudhaganawa) in the organic fraction and 49 mg kg^?1 (Ussangoda) in the exchangeable fraction. Despite Cr being primarily retained in the residual fraction, the second largest pool of Cr was in the organic matter fraction (693 mg kg^?1 in the Yudhaganawa soil). Overall, our results support that serpentine soils in Sri Lanka offer a highly labile source of metals to the critical zone.     

Metal exposure and effects in voles and small birds near a mining haul road in Cape Krusenstern National Monument, Alaska

Voles and small passerine birds were live-captured near the Delong Mountain Regional Transportation System (DMTS) haul road in Cape Krusenstern National Monument in northwest Alaska to assess metals exposure and sub-lethal biological effects. Similar numbers of animals were captured from a reference site in southern Cape Krusenstern National Monument for comparison. Histopathological examination of selected organs, and analysis of cadmium, lead, and zinc concentrations in liver and blood samples were performed. Voles and small birds captured from near the haul road had about 20 times greater blood and liver lead concentrations and about three times greater cadmium concentrations when compared to those from the reference site, but there were no differences in zinc tissue concentrations. One vole had moderate metastatic mineralization of kidney tissue, otherwise we observed no abnormalities in internal organs or DNA damage in the blood of any of the animals. The affected vole also had the greatest liver and blood Cd concentration, indicating that the lesion might have been caused by Cd exposure. Blood and liver lead concentrations in animals captured near the haul road were below concentrations that have been associated with adverse biological effects in other studies; however, subtle effects resulting from lead exposure, such as the suppression of the activity of certain enzymes, cannot be ruled out for some individual animals. Results from our 2006 reconnaissance-level study indicate that overall, voles and small birds obtained from near the DMTS road in Cape Krusenstern National Monument were not adversely affected by metals exposure; however, because of the small sample size and other uncertainties, continued monitoring of lead and cadmium in terrestrial habitats near the DMTS road is advised.     

Bacterial contamination and health risks of drinking water from the municipal non-government managed water treatment plants

Water quality and bacterial contamination from 18 drinking water municipal plants in three locations at Giza governorate were investigated. The average total count of bacteria detected after four stages of treatments in the investigated plants was 32 CFU/1 mL compared to 2330 cfu/mL for raw water, with a reduction percentage of 98.6. Although there is a relatively high removal percent of bacterial contamination from the water sources, however, several bacterial pathogens were identified in the produced water prepared for drinking including Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, and Shigella spp. After 3 days of water incubation at 30 °C, the amount of bacterial endotoxins ranged from 77 to 137 ng/mL in the water produced from the municipal plants compared to 621–1260 ng/mL for untreated water. The main diseases reported from patients attending different clinics and hospitals during summer 2014 at the surveyed locations and assuredly due to drinking water from these plants indicated that diarrheas and gastroenteritis due to E. coli and Campylobacter jejuni constituted 65.7% of the total patients followed by bacillary dysentery or shigellosis due to Shigella spp. (7.9%) and cholera due to Vibrio cholera (7.2%). There was an increase in serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) as well as urea and creatinine values of guinea pigs consuming water produced from the non-governmental plants for 6 months indicating remarkable liver and kidney damages. Histological sections of liver and kidney from the tested animal revealed liver having ballooning degeneration of hepatocytes and distortion and fragmentation of the nuclei, while the section of the kidney showed irregularly distributed wrinkled cells, degenerated Bowman’s capsule, congested blood vessels, and inflammatory cells.     

Kidney,liver and bone cadmium content in the western sandpiper in relation to migration

Cadmium content was measured in kidney, liver and tarsus bones of western sandpipers (Calidris mauri) at a temperate migratory stopover site (Fraser Delta, British Columbia, Canada) and a wintering site (Playa el Agallito, Chitre, Panama) over a two year period. Cadmium content in liver and kidney was age and sex dependent. Adult females generally had lower kidney and liver cadmium than adult males (P < 0.05), but a sex difference was not detected in juveniles. Cadmium increased with age in kidney, liver and to a lesser extent in bone (P < 0.001) with average "steady-state" kidney and liver content being reached within the sandpipers first year. In general, tissue cadmium residues in adult males and females were independent of sampling location although for bone, site-specific differences did occur (P < 0.001). Bone cadmium was lower in females sampled from their wintering grounds as compared to temperate stopover sites suggesting that bone cadmium may be mobilized during periods of feather molt. Comparison of cadmium residues among sandpipers of increasing age suggest that exposure is occurring along the Pacific Coast, at stopover sites as the birds migrate north to Alaska and south to Panama. This study points to the importance of considering the ecology of the species (e.g., in this case migratory behavior) in interpreting trace metal residues.     

Assessment of prenatal exposure to persistent organohalogen compounds from cord blood serum analysis in two Mediterranean populations (Valencia and Menorca)

Prenatal levels of organohalogen compounds (OHCs), including polybromodiphenyl ethers (PBDEs), polychlorobiphenyls (PCBs), hexachlorobenzene (HCB), hexachlorocyclohexanes and DDTs, have been investigated in two different cohorts from the Spanish Mediterranean area, Valencia and Menorca, by analysis of cord blood serum. Comparison of the observed median concentrations to other cohorts shows low PBDE levels (2.8-6.5 ng g(-1) lipid) but high values of HCB (0.68 ng ml(-1), 260 ng g(-1) lipid) and PCBs (0.65 ng ml(-1), 240 ng g(-1) lipid) in the cohort of Menorca indicating that Mediterranean areas, and not only high latitude regions, may contain population groups highly exposed to some of these pollutants. Significant differences in the concentrations of these compounds have been found between the two cohorts. Differences in maternal population such as age or body mass index cannot explain this variation. One possible cause of variability is the seven year time period elapsed between the two cohort recruitments but the strong differences observed do not allow exclusion of local diet differences. Thus, the different average PCB congener distributions between the two cohorts suggest an influence from materials containing diverse PCB mixtures. The congener mixtures found in Menorca could reflect a diet contribution from some western Mediterranean fish species. These results provide baseline information on prenatal OHC background levels in the Mediterranean area.     

A preconcentration procedure for the determination of cadmium in biological material after on-line cloud point extraction

In this paper, a method involving on-line preconcentration with cloud point extraction for the determination of cadmium in biological samples is presented. The procedure is based on the sorption of micelles containing Cd(II) ions and the reagent 4-(5'-bromo-2'-thiazolylazo)orcinol (Br-TAO) in a minicolumn packed with polyester. The surfactant Triton X-114 was used in the formation of micelles. After sorption, the Cd(II) ions were desorbed from the minicolumn with acid eluent and determined by flame atomic absorption spectrometry. Parameters influencing the cloud point extraction were studied. The method showed a detection limit of 0.5 μg l(-1) and an enhancement factor of 27. The accuracy was tested by determination of cadmium in certified reference materials (spinach leaves 1570a and tomato leaves 1573a) from the National Institute of Standards and Technology.     

Cadmium Contamination in Wild Birds as an Indicator of Environmental Pollution

Cadmium (Cd) is an environmental pollutant that has serioustoxicity in humans and animals and causes Itai-Itai disease.However, there is little available information on itscontamination in wildlife as an indicator of environmentalpollution. The Cd contents in the kidney and liver of 85 wildbirds from 9 different prefectures in Japan were investigated.The ranges of the Cd contents in the kidney and liver in allbirds were ND-174.4 and ND-21.2 g g^-1 dry wt.,respectively. The mean Cd contents were higher in theoil-contaminated birds than those in the non-contaminated ones.Furthermore, a strong correlation was obtained only between theCd contents in the kidney and those in the liver of theoil-contaminated seabirds and not in the other non-contaminatedones. These results suggest that wild birds reflect the levelof environmental contamination which should be monitored.     

A pilot study on the determination of perfluorooctanesulfonate and other perfluorinated compounds in blood of Canadians

A pilot study was conducted to provide preliminary data on the concentrations of perfluorooctanesulfonate (PFOS), perfluorooctanoic acid (PFOA) and perfluorooctanesulfonamide (PFOSA) in the blood of Canadians. A set of 56 human serum samples was collected from non-occupationally exposed Canadians and analyzed by microbore HPLC-negative ion electrospray tandem mass spectrometry. PFOS was the main component of perfluorinated organic compounds (PFCs) and was detected in all 56 blood specimens at an average concentration of 28.8 ng mL(-1) and a range from 3.7 to 65.1 ng mL(-1). The concentration of PFOA was an order of magnitude lower than that of PFOS and was found only in 16 samples (29%) at concentrations above the limit of quantification (LOQ). PFOSA was not detected at levels above the method detection limit (MDL) in any of the samples. The levels of PFCs observed in the sample group of non-occupationally exposed humans in Canada were similar to the levels reported in a previous US study with a similar sample pool size. Two distinct PFOS isomers in human serum were identified by accurate mass determination.