Rapid determination of methyl mercury in fish and shellfish: method development

The AOAC official first action method for methyl mercury in fish and shellfish was modified to provide more rapid determination. Methyl mercury is isolated from homogenized, acetone-washed tissue by addition of HCl and extraction by toluene of the methyl mercuric chloride produced. The extract is analyzed by electron capture gas chromatography (GC) on 5% DEGS-PS treated with mercuric chloride solution. The quantitation limit of the method is 0.25 micrograms Hg/g. Swordfish, shark, tuna, shrimp, clams, oysters, and NBS Research Material-50 (tuna) were analyzed for methyl mercury by the AOAC official first action method. All products also were analyzed by the modified method and the AOAC official method for total Hg. In addition, selected extracts obtained with the modified method were analyzed by GC with Hg-selective, microwave-induced helium plasma detection. There was no significant difference between the results for the various methods. Essentially all the Hg present (determined as total Hg) was in the organic form. Coefficients of variation from analyses by the modified method ranged from 1 to 7% for fish and shellfish containing methyl mercury at levels of 0.50-2.30 micrograms Hg/g. The overall average recovery was 100.5%.     

Simple combustion-vapor trapping technique for determination of mercury in fish

A simple combustion train was constructed for analyzing mercury in fish samples. A nitric acid trap was used to capture the mercury vapors which were released later by adding a tin salt. The method is rapid, accurate, and reproducible and permits one person to analyze 40 samples daily. Sample matrix had no apparent effect on the recovery of mercury. Mean recoveries from fish samples spiked with 0.3 and 1.0 microgram mercuric chloride/g ranged from 97 to 105% with an average recovery of 101.5% and a standard deviation of 2.74%.     

Rapid determination of methyl mercury in fish and shellfish: collaborative study

A modification of the official AOAC method for determining methyl mercury in fish and shellfish was studied in 8 laboratories. Methyl mercury is isolated from homogenized, acetone-washed tissue by adding HCl and extracting into toluene the methyl mercuric chloride produced. The extract is analyzed for methyl mercuric chloride by electron capture gas chromatography. Collaborators determined methyl mercury in blind duplicate homogenates at 2 levels in tuna and at 1 level in swordfish and oysters. Collaborators also analyzed single homogenates of swordfish and oysters containing methyl mercury at a second level. Both fortified and unfortified tissues were analyzed. Methyl-bound mercury in the commodities ranged from 0.50 to 2.30 micrograms Hg/g. Reproducibility coefficients of variation ranged from 4 to 15%. Accuracy, measured by comparison to reference values, ranged from 92 to 101%. Recovery from fortified homogenates ranged from 86 to 98%. Reference values and unfortified levels were determined in the author's laboratory by replicate analysis of fortified and unfortified commodities. The method has been approved interim official first action.     

Interlaboratory calibration results of polychlorinated biphenyl analyses in herring

Twenty-three laboratories from 13 countries in continental Europe, Scandinavia, the British Isles, and North America participated in a polychlorinated biphenyl (PCB) Check Sample Program conducted under the auspices of the International Council for Exploration of the Sea (ICES). PCBs were determined in unspiked and spiked (1.00 mg Aroclor 1254/kg oil) herring (Clupea harengus harengus) oil by the participants, each of which used his or her own cleanup and quantitation techniques; a common Aroclor 1254 mixture was used as a standard and a common quantitation technique was used for comparative purposes. Results for the unspiked oil ranged from 0.48 to 3.416 mg PCB/kg oil, while spiked oil results ranged from 0.70 to 3.891 mg/kg. Calculated spike recoveries ranged from 22 to 136%. Serious deficiencies were found in most steps in the procedures. No evidence was found to support the use of a common PCB standard or a common method of calculation using packed column chromatography. The chromatographic stationary phase used appeared to affect the PCB levels obtained. Florisil cleanup adsorbent yielded higher mean results for both unspiked and spiked oils than did alumina. Large coefficients of variation were found (25-50%), the principal source of which was systematic error (interlaboratory).     

Mercury uptake patterns of biota in a seasonally anoxic northern California Reservoir

Biotic uptake of mercury (Hg) in Davis Creek Reservoir, California increased dramatically in conjunction with the entrainment of anoxic hypolimnetic water into the mixed layer. This indicated a seasonal pulse increase of bioavailable Hg associated with thermal destratification. The effect was more pronounced in juvenile bass (70–200% seasonal increases in muscle Hg concentration), as compared to adults (15–25% increases), and was most distinct in zooplankton, which spiked to concentrations of 3–6 mg/kg, dry weight, immediately following fall destratification (130–270% seasonal increases over pre-fall levels). In addition to the general buildup of methyl Hg in the hypolimnion under anoxic conditions, a dense layer of photosynthetic anaerobic bacteria just beneath the thermocline is implicated as a potentially important seasonal source of methyl Hg to reservoir fish. Hg increases in juvenile and adult fish correspond to late summer and fall entrainment of upper hypolimnetic water, while zooplankton spike increases may be partially related to ingestion or adsorption of Hg-scavenging manganese oxides, which precipitate following full turnover. A simple and effective, syringe-based cold vapor atomic absorption method for total Hg is also described.     

Digestion of food samples for total mercury determination

A method of digestion by using a mixture of hydrochloric, nitric, and sulfuric acids has been developed for the determination of total mercury in a wide range of food samples. Good recoveries of mercury were obtained from NBS (National Bureau of Standards) Albacore Tuna and from food samples spiked with inorganic mercury. A detection limit of 0.01 microgram mercury/g can be obtained.     

Development of a Mercury Vapour Air Analyser for Measurement of Hg in Solution

The Tekran 2537A mercury vapour analyser, designed to measure Hg in air by cold vapour atomic fluorescence spectrometry, has been modified to determine Hg in solution. The new ‘front-end’, required to generate Hg° vapour from acidified waters or acid leachates, is described. Using 1% NaBH4 as reducing agent, a 12 mL water sample can be analysed, at a rate of 1 every 6 min, for Hg to a detection limit of 0.8 ppt (ng L-1). Instrumental precision is typically 1% relative standard deviation (RSD) at levels of Hg from 10 to 200 ppt. Results for 10 analyses of the international water standard, NIST 1642b, are 1530±20 ppt Hg, agreeing well with the certified value of 1480±130 ppt. Nineteen geological standard reference materials (soils, sediments and tills) were used to assess accuracy. Results for these samples, digested in aqua regia in triplicate, showed good agreement with recommended values for all but two, SO-3 and TILL-1. However, results by this method for these two standards were confirmed by an independent method, direct atomic absorption spectrometry. Average method precision was shown to be 5% RSD over the range 10 ng g- 1 to 35 μg g-1 Hg.     

Automatic flow-batch system for cold vapor atomic absorption spectroscopy determination of mercury in honey from Argentina using online sample treatment

An automatic flow-batch system that includes two borosilicate glass chambers to perform sample digestion and cold vapor atomic absorption spectroscopy determination of mercury in honey samples was designed. The sample digestion was performed by using a low-cost halogen lamp to obtain the optimum temperature. Optimization of the digestion procedure was done using a Box-Behnken experimental design. A linear response was observed from 2.30 to 11.20 μg Hg L(-1). The relative standard deviation was 3.20% (n = 11, 6.81 μg Hg L(-1)), the sample throughput was 4 sample h(-1), and the detection limit was 0.68 μg Hg L(-1). The obtained results with the flow-batch method are in good agreement with those obtained with the reference method. The flow-batch system is simple, allows the use of both chambers simultaneously, is seen as a promising methodology for achieving green chemistry goals, and is a good proposal to improving the quality control of honey.     

A simplified procedure for the determination of total mercury in soils

Abstract

A saraimplified procedure for the measurement of total mercury in soils and some preliminary results for Saskatchewan are presented. Soil samoles were digested in aqua regia at low temperatures, followed by determination of Hg with a simplified flameless atomic absorption technique. Hg levels in soils analyzed ranged from 0.003 to 0.071 ug/g.     

Speciation analysis of mercury in cereals by liquid chromatography chemical vapor generation inductively coupled plasma-mass spectrometry

A simple and rapid procedure for the separation and determination of inorganic, methyl, and ethyl mercury compounds was described using liquid chromatography (LC) followed by vapor generation inductively coupled plasma-mass spectrometry (VG-ICP-MS). Well resolved chromatograms were obtained within 5 min by reversed-phase liquid chromatography with a C8 column as the stationary phase and a pH 4.7 solution containing 0.5% v/v 2-mercaptoethanol and 5% v/v methanol as the mobile phase. The separated mercury compounds were converted to mercury vapors by an in situ nebulizer/vapor generation system for their introduction into ICP. The concentrations of NaBH4 and HNO3 required for vapor generation were also optimized. The method was applied for the speciation of mercury in reference materials NIST SRM 1568a Rice Flour and NIST SRM 1567a Wheat Flour and also rice flour and wheat flour samples purchased locally. The accuracy of the procedure was verified by analyzing the certified reference material NRCC DOLT-3 Dogfish Liver for methyl mercury. Precision between sample replicates was better than 13% for all the determinations. The detection limits of the mercury compounds studied were in the range 0.003-0.006 ng Hg mL(-1) in the injected solutions, which correspond to 0.02-0.06 ng g(-1) in original flour samples. A microwave-assisted extraction procedure was adopted for the extraction of mercury compounds from rice flour, wheat flour, and fish samples using a mobile phase solution.     

Distribution of mercury species in soil from a mercury-contaminated site

The distribution of mercury species was determined in soil from a site with Hg contamination. Mercury contamination was primarily confined to the top 40 cm of soil, and the concentration of total Hg ranged from 0.5 to 3000 µg Hg g^?1. Of total Hg present, we determined that 91% was inorganic, 0.01% organic (as methyl Hg), and 6% elemental Hg. Furthermore, of total inorganic Hg present, 85% was in the insoluble mercuric sulfide form. Thus, of total Hg present in soil at this contaminated site, 91% was in the relatively insoluble HgS and Hg⁰ forms.     

Gas-liquid chromatographic screening method for determination of methyl mercury in tuna and swordfish

A rapid screening method has been developed for determining methyl mercury in tuna and swordfish. Fish tissue is blended with acidic KBr solution to release methyl mercury, which is then extracted into methylene chloride. After cleanup by partitioning with cysteine, the methyl mercury is extracted into toluene and determined by gas-liquid chromatography. The proposed method compares favorably with the official AOAC atomic absorption method.     

Determination of inorganic tin in biological samples by hydride generation-atomic absorption spectrometry after silica gel cleanup

A method is described for the determination of inorganic tin in biological samples by hydride generation-atomic absorption spectrometry (HG-AAS). A sample is extracted with ethyl acetate after addition of HCl and NaCl. The concentrated extract is passed through a silica gel column. The column is washed with ethanol, water, and 0.2N HCl successively, and then inorganic tin is eluted with 2N HCl and measured by HG-AAS. Recoveries from fish muscle spiked with 0.1 micrograms/g Sn4+ are 78.9 +/- 4.2% (average +/- standard deviation, n = 5). The detection limit is 0.01 micrograms/g as Sn.     

基质固相分散萃取-高效液相色谱串联质谱法测定土壤中残留的磺酰脲类除草剂

建立了基质固相分散萃取-高效液相色谱串联质谱法(MSPD-HPLC-MS/MS)测定土壤中3种磺酰脲类除草剂(氯磺隆、甲磺隆、苯磺隆)残留的分析方法。对基于球磨的基质固相分散萃取条件进行了详细优化,最终确定最佳条件为:0.2 g土壤样品、0.8 g HC-C18粉末状分散剂与直径为8 mm的小钢珠一起球磨10 min后,转移至空的玻璃萃取小柱,用10 m L乙腈洗脱,氮气吹干后用甲醇定容至0.6 m L,再经0.22μm的滤膜抽滤后装入自动进样瓶中。用Syncronis C18反相色谱柱分离,以甲醇(A)~1‰甲酸溶液(B)为流动相进行梯度洗脱,选择反应监测(SRM)模式下进行检测。氯磺隆在20~200μg·kg~(-1),甲磺隆和苯磺隆在10~200μg·kg~(-1)范围内线性良好,相关系数r在0.997 9~0.999 5。土壤样品的平均加标回收率在84.7%~104.6%,相对标准偏差在4.5%~7.9%(n=5)。方法的检出限(S/N=3)0.32~0.68μg·kg~(-1)。该方法简单、效率高、干扰少、回收率高,满足土壤中除草剂的残留分析要求。     

Online YPA4 resin microcolumn separation/preconcentration coupled with inductively coupled plasma optical emission spectrometry (ICP-OES) for the speciation analysis of mercury in seafood

A simple and effective method for the determination of trace amounts of methylmercury (MeHg(+)) and inorganic mercury (Hg(2+)) in seafood was developed by online microcolumn separation/preconcentration combined with inductively coupled plasma optical emission spectrometry (ICP-OES). It was found that Hg(2+) could be quantitatively adsorbed by YPA 4 resin from pH 7.0 to strong acidic medium (6 mol L(-1) HCl) and that MeHg(+) was retained by the YPA 4 microcolumn only at pH 1.0-7.0. Therefore, a strong acidic medium (about 5 mol L(-1) HCl), which could liberate mercury species from biological samples, was used to directly separate inorganic Hg(2+) from total Hg, and MeHg(+) in effluent was retained by YPA 4 column after the effluent was adjusted to pH 1.5. The effects of acidity, sample flow rate and volume, elution solution, and interfering ions on recovery of the two mercury species have been systematically investigated. Under optimal conditions, the limits of detection (LODs) were 72 and 44 ng L(-1) for Hg(2+) and MeHg(+) (as Hg) with online concentration factors of 12.5 and 12.1, respectively. The relative standard deviations (RSDs) for nine replicate determinations at 5 ng mL(-1) levels of mercury species were 2.7 and 2.0% for Hg(2+) and MeHg(+), respectively. The calibration graphs were linear with a correlation coefficient of 0.9902 in the range of 0.5-100 ng mL(-1) for Hg(2+) and 0.9976 in the range of 0.1-100 ng mL(-1) for MeHg(+), respectively. The developed method was successfully applied to the direct determination of MeHg(+) and Hg(2+) in seafood samples, and the recoveries for the spiked samples were in the range of 89.9-102.4% (MeHg(+)) and 87.0-104.6% (Hg(2+)), respectively. The method was validated by analyzing a certified reference material DORM-2 (dogfish muscle), and the determined values were in good agreement with certified values.     

Gas-liquid chromatographic determination of selenium in biological materials, using 4-bromo- and 4-chloro 1,2-diaminobenzene as derivatizing reagents

A method is described for the gas-liquid chromatographic determination of traces of selenium in marine biological materials. The method is based on the reaction of Se(IV) with bromo- and chloro-substituted 1,2-diaminobenzenes. The benzoselenadiazoles so formed are sensitive to electron capture detection. The sample is digested in a nitric-perchloric acid mixture and selenium is reduced to the IV oxidation state. Different aliquots of the digest solution are reacted with either 4-bromo- or 4-chloro-1,2-diaminobenzene to quantitatively form the corresponding 2,1,3-benzoselenadiazole. Recovery of added selenite to a fish meal sample was 95% for the bromo derivative and 101% for the chloro derivative. Different portions of a well mixed fish meal sample were analyzed in independent laboratories by the fluorometric method and by atomic absorption spectrophotometry (hydride generation). The following mean values (microgram/g) were found: present method 1.89, fluorometric method 1.91, atomic absorption method 2.1. The lower limit of detection for the method described was 13 ng, using the bromo derivative, and 27 ng, using the chloro derivative.     

Multi-laboratory study of measurement of chlorobiphenyls and other organochlorines in fish oil

Analysts participating in a multi-laboratory comparative study were asked to identify 4 chlorobiphenyls (CBs) supplied in neat form, to measure the amounts of these present in spiked and unspiked fish oil, to measure other CBs and organochlorines in the unspiked fish oil, and to compare results for their own standard solution and those for standard solutions prepared from the supplied CB compounds. Comparisons were done for a common supplied method and the individual methods used in each laboratory. Participants had no trouble identifying 2,2',5,5'-tetrachlorobiphenyl, 2,2',4,5,5'-pentachlorobiphenyl, and 2,2',4,4',5,5'-hexachlorobiphenyl. Most misidentified 2,2',3,4,5-pentachlorobiphenyl. Approximately one-half of the quantitative comparisons between participants' standards and those prepared from the supplied CBs differed by more than 10%. Two standards prepared from one of the CBs differed by an average of 6.6% (range 0.0-24.5%). Recoveries of added CBs from the fish oil ranged from 24 to 294% for spikes of 63-85 ng/g with no clear distinction between results for the common method vs individual laboratory methods. The common methodology gave a lower coefficient of variation (CV) for most other CBs and organochlorines, but in most cases the CVs for the individual CBs were not smaller than that for Aroclor 1254 equivalents.     

Ion-pair liquid chromatographic determination of some penicillins in canine and equine sera

A sensitive liquid chromatographic method was developed for determining oxacillin, cloxacillin, and dicloxacillin (simultaneously), and penicillin G, amoxicillin, carbenicillin, and ticarcillin in canine and/or equine serum. The method involves filtering diluted serum through a 30,000 molecular weight cut-off filter and separating penicillins from other serum components by ion-pair liquid chromatography using a reverse-phase column eluted with acetonitrile-water solutions. The ultraviolet absorbance of the column effluent was monitored at 230 nm. Recoveries of oxacillin, cloxacillin, dicloxacillin, and penicillin G (spiked at 2.5 micrograms/mL), amoxicillin (spiked at 5 micrograms/mL), and carbenicillin and ticarcillin (spiked at 10 micrograms/mL) from canine and equine serums ranged from 78.3 to 104.4% with coefficients of variation ranging from 3.35 to 5.95%. The limit of detection for these penicillins was 0.02-0.05 microgram/mL.     

Extraction of silver from soils and its determination by atomic absorption spectrometry

A simple, sensitive and reliable method for determining silver in soils by atomic absorption spectrometry with a graphite furnace is described which involves direct analysis of acid digests and avoids pre-concentration of the analyte. Chemical extracts have been investigated as a means of fractionating soil silver and results are reported for a range of soils, from areas of differing geology in Wales, U.K. Background concentrations are compared with areas contaminated by spoil from derelict mines. Uncontaminated soils contained silver in the range 0.01–1 μg Ag/g. The proportion of total silver extracted from a soil spiked and equilibrated with 110m AgNO₃ and unspiked field samples gave good agreement for the extractants 0.1 M and 1.0 M nitric acid, 0.005 M DTPA (diethylenetriaminepentaacetic acid), 0.05 M acetic acid, 1 M ammonium acetate and deionised water, although some extractants did not meet their predicted extraction efficiency. In the field samples soluble plus exchangeable silver was present between 0.4–40 ng Ag/g (< 1% of total), whereas biologically-available silver measured in 0.005 M DTPA was in the range 3–540 ng Ag/g (< 5% of total).     

Flame absorption spectroscopic determination of cadmium, copper, iron, lead, and zinc in mussels

A rapid method is proposed for determination of Cd, Cu, Fe, Pb, and Zn in mussel samples. The elements are extracted with concentrated nitric acid in borosilicate glass tubes at 90 degrees C for 1 h, and determined by flame atomic absorption spectroscopy. Detection limits for a 300 mg sample corresponded to 0.3 microgram Cd/g, 0.7 microgram Cu/g, 33 microgram Fe/g, 0.7 microgram Pb/g, and 6 micrograms Zn/g. The coefficient of variation for 20 independent analyses was 7% for Cd, 7% for Cu, 6% for Fe, 14% for Pb, and 8% for Zn. Recoveries were 107 +/- 3% for Cd, 90 +/- 3% for Cu, 94 +/- 1% for Fe, 90 +/- 5% for Pb, and 96 +/- 3% for Zn. The accuracy of the method was determined by analyzing NBS Oyster Tissues.