Mutagenic activity of the Ganges water with special reference to the pesticide pollution in the river between Kachla to Kannauj (U.P.), India

Water samples from Ganga river were collected from 3 different locations viz. Kachla, Fatehgarh and Kannauj (U.P.). High performance liquid chromatography analysis of samples by liquid-liquid extraction procedure showed the presence of some pesticides like DDT, alpha-BHC, aldrin, dieldrin etc. DDT, alpha-BHC, DDD, aldrin and dieldrin were present at concentration ranges of 3.33-5.33 ppb, 1.73-3.01 ppb, 0.88-2.41 ppb, 1.17-2.81 ppb and 0.49-4.11 ppb, respectively. The organophosphorus pesticides like dimethoate and methyl parathion were also detected at the concentration levels of 0.41-0.56 ppb and 0.16-0.50 ppb, respectively. The organic substances in the test samples were extracted by XAD-resin and liquid-liquid extraction procedures, and the extracts were assayed for mutagenic potential by the Ames Salmonella/microsome test. The test samples exhibited a remarkable degree of mutagenicity with TA98, TA100 and TA97a strains with the probable role of contaminating pesticides in the river water.     

Mercury levels in freshwater fish of the state of South Carolina

Samples of fish from freshwater sources of rivers, lakes and ponds all over the state of South Carolina were collected during the Summer of 1974 and 1975. The fish collected were Bass, Bluegill, Redbreast, Catfish, Shad, Carp, Crappie, Mudfish and Pike. Samples were analyzed using the flameless atomic absorption procedure outlined by Hatch and Ott, and Uthe et al as modified for use with Perkin-Elmer, Coleman MAS-50 mercury analyzer. Triplicate samples of fish tissue were analyzed by wet digestion method. The mean mercury levels in ppb were determined for baseline mercury levels. A significant finding of this report is that those species for which fish of widely differing weights were analyzed, larger fish had higher mercury levels. Mercury levels exceeding the U.S. Food and Drug Administration guideline of 500 ppb for fish tissues have been found in the Mudfish from Edisto River and Pike fish from Lake Murray. Higher levels of mercury occurred in the highly vascularized blood tissues of liver and kidney than in muscle. Carnivorous and bottom-feeding fishes are the most reliable indicators of mercury pollution.     

Determination of organochlorine pesticide and polychlorinated biphenyl residues in fatty fish by tandem solid-phase extraction cleanup

A rapid, multiresidue solid-phase extraction (SPE) technique for determination of organochlorine pesticide and polychlorinated biphenyl (PCB) residues in nonfatty fish was modified for use with fatty fish. In the modified procedures, samples are extracted with acetonitrile, and the extract is cleaned up with both C18 and Florisil SPE columns. Residues are determined by gas chromatography with electron capture detection. The original method was modified for use with fatty fish by reducing the amount of tissue extracted and by using an improved Florisil SPE cleanup. Recovery data are presented for 24 fortified organochlorine pesticide residues (0.12 ppm) and 3 fortified PCB residues (0.80 ppm) from flounder, bluefish, and shad samples, which contained 0.8, 5.4, and 22.6% fat, respectively. For the 3 types of fish, recoveries of 23 of 24 fortified organochlorine pesticide residues ranged from 55 to 129%, and recoveries of 3 fortified PCB residues ranged from 55 to 104%. There were no significant differences in recovery based on fish species and/or fat content for the majority of residues studied. This SPE method and the official AOAC method yielded comparable results for fish containing incurred organochlorine residues.     

Confirmation and quantitation of selected sulfonylurea, imidazolinone, and sulfonamide herbicides in surface water using electrospray LC/MS

A multianalyte method has been developed for the confirmation and quantitation of 16 selected sulfonylurea, imidazolinone, and sulfonamide herbicides in surface water. Samples are acidified, the analytes are extracted using RP-102 extraction cartridges, and the extracts are cleaned-up using an anion-exchange cartridge (SAX) stacked on top of an alumina cartridge. The final extracts are evaporated to dryness, redissolved in 10:90 or 20:80 acetonitrile/water, and analyzed using electrospray LC/MS. Confirmation criteria were defined so that identification of the analytes could be made with a reasonable degree of scientific certainty. Confirmation required that (1) LC/MS retention times of the analytes be within 1% of the retention times of the standards, (2) the molecular ion and two characteristic fragment ions per analyte be present, and (3) ion abundance ratios of the fragment ions relative to the molecular ion be within 20% of the ion ratios obtained for the standards. Each of the analytes in all of the samples used to validate this method met these confirmation criteria. Quantitation of the analytes at 0.1 and 1.0 ppb was demonstrated, with average recoveries between 70% and 114% and relative standard deviations that were less than 13%. The limit of quantitation (LOQ) for this method is 0.1 ppb (ng/mL) for all analytes. Six water types were used for the validation of this method.     

Determination of leucogentian violet in chicken fat by liquid chromatography with electrochemical and ultraviolet detection: interlaboratory study

A laboratory trial was completed for a liquid chromatographic method that can quantitate leucogentian violet (LGV) in chicken fat at 10 ppb. With this method, LGV is isolated from the fat matrix by a series of liquid-liquid extractions. This trial evaluated 2 detection systems: electrochemical (EC) and ultraviolet (UV). The participating laboratories determined incurred residues at 2 levels as well as fat samples fortified at 5, 10, and 20 ppb. Using UV detection, the 3 laboratories reported the following range of recoveries: 71.0-89.6% at 5 ppb, 74.7-83.9% at 10 ppb, and 77.2-79.0% at 20 ppb. When these same samples were chromatographed with EC detection, the 2 reporting laboratories obtained the following average recoveries: 79.0 and 92.5% at 5 ppb, 75.9 and 85.4% at 10 ppb, and 77.3 and 79.8% at 20 ppb. The average concentrations found for the first level of incurred sample were 6.3, 6.3, and 5.4 ppb with coefficients of variation (CVs) of 2.4, 7.6, and 33.7%, respectively, when UV detection was used. Samples chromatographed with EC detection averaged 6.3 and 6.4 ppb with CVs of 4.0 and 8.2%, respectively. The second level of incurred sample gave average concentrations of 27.6, 29.0, and 10.9 ppb with CVs of 11.0, 5.0, and 42.8%, respectively, when the UV detection system was used. With the EC detector, the concentrations averaged 27.2 and 30.7 ppb with CVs of 15.7 and 3.5%, respectively.     

混合增效剂存在下停流FIA光度法测定钼—Mo—SCN^_—MG体系

本文研究了在PVA—Triton X-100存在下,用Mo—SCN~-—MG体系测定微量钼的停流流动注射-光度法,克服了体系稳定性差的缺点,成功地用于钢铁样品和钼铁矿附近井水中微量钼的测定。比尔定律范围40～200ppb,RSD<1％,回收率103％～108％,测定频率24样/h。     

Prominent increase of macrophage migration inhibitory factor in the sera of patients with uveitis

Ciguatera is a significant food-borne disease caused by potent polyether toxins (ciguatoxins) which accumulate in the flesh of ciguateric reef fish at risk levels > 0.1 ppb for Pacific ciguatoxins. Research on ciguatera has been severely hindered by the lack of analytical methods that detect and characterize low levels of ciguatoxin in crude extracts of fish. Here we report a new procedure for ciguatoxin analysis based on gradient reversed-phase HPLC/tandem mass spectrometry (HPLC/MS/MS). The method gave a linear response to pure Pacific and Caribbean ciguatoxins (P-CTX-1 and C-CTX-1) and the structurally related brevetoxin (PbTx-2) spiked into crude extracts of fish. Levels equivalent to 40 ppt P-CTX-1, 100 ppt C-CTX-1, and 200 ppt PbTx-2 in fish flesh could be detected by HPLC/MS/MS. Using P-CTX-1 as an internal standard, the analysis of extracts of 30 ciguateric fish from the Caribbean Sea (8 toxic, 12 borderline, and 10 nontoxic by mouse bioassay) confirmed the reliability of the method and allowed an estimated risk level of > 0.25 ppb C-CTX-1 to be established. HPLC/MS/MS provides a sensitive analytical approach, not previously available, for the determination of Pacific and Caribbean ciguatoxins at sub-ppb levels in fish flesh.     

Isolation of priority polycyclic aromatic hydrocarbons from natural sediments and sludge reference materials by an anti-fluorene immunosorbent followed by liquid chromatography and diode array detection

The selective isolation of PAHs from complex environmental mixtures was accomplished by means of a new methodology based on antigen--antibody interactions. This method consists on the extraction of PAHs from water samples onto an anti-fluorene immunosorbent (IS) followed by liquid chromatography with diode array detection. Environmental sediments and a sludge reference material containing PAHs were analyzed using this methodology in order to validate the performance of the IS for the cleanup procedure of such materials. Sediments were extracted by sonication with dichloromethane/methanol (2:1), and the extracts were brought to a volume of 100 mL of water in order to perform the extraction with the anti-fluorene IS. The reliability of the cleanup achieved by the IS was well demonstrated in the analysis of sediment and sludge complex samples containing the priority PAHs established by the U.S. EPA at concentrations varying from 56 micrograms/kg to 26 mg/kg. Results were compared to those obtained with conventional cleanup procedures showing a better selectivity for PAHs. The chromatograms presented a clear baseline allowing the determination and quantification of PAHs at the ppb level. Using immunosorbents, extraction, trace enrichment, and cleanup were accomplished in only one step.     

Growth-regulatory activity of the growth arrest-specific gene, GAS1, in NIH3T3 fibroblasts

The method described detects and confirms presence of pentobarbital residues in dry, extruded feeds at concentrations of 5-20 ppb. Dried feed is ground to a uniform powder and shaken overnight in methanol. A portion of the methanolic extract is evaporated, and the residue is reconstituted in phosphate-buffered saline. The aqueous extract is cleaned with a solid-phase extraction cartridge designed to extract barbiturate residues from biological matrixes. Dimethyl sulfoxide, tetramethylammonium hydroxide, and iodomethane are added to derivatize pentobarbital, 1,3-Dimethyl-pentobarbital is then acidified with dilute hydrochloric acid and extracted with isooctane. The organic layer is transferred and evaporated under a stream of nitrogen. The residue is reconstituted in a small volume of ethyl acetate for analysis by gas chromatography/mass spectrometry. The limit of detection is approximately 0.7 ppb. The method was validated with pentobarbital-fortified feed samples containing high concentrations of meat and bone meal.     

High pressure liquid chromatographic determination of aflatoxins in wines and other liquid products

A previously developed high pressure liquid chromatographic (HPLC) method was evaluated by an extensive recovery study of aflatoxins from wines and other liquid products. The HPLC column and the preliminary cleanup procedures were both modified for this study. Four aflatoxins (B1, B2, and G2) were recovered at 80-116% from 28 samples of various liquid commodities spiked at the 1 microgram/L (1 ppb) level. The detection limit of the proposed method for wines and fruit juices was about 0.02 ppb. A total of 53 samples were analyzed for aflatoxins during this study.     

Analysis of malachite green and metabolites in fish using liquid chromatography atmospheric pressure chemical ionization mass spectrometry

Malachite green (MG), a traditional agent used in aquaculture, is structurally related to other carcinogenic triphenylmethane dyes. Although MG is not approved for use in aquaculture, its low cost and high efficacy make illicit use likely. We developed sensitive and specific methods for determination of MG and its principal metabolite, leucoMG (LMG), in edible fish tissues using isotope dilution liquid chromatography atmosphere pressure chemical ionization mass spectrometry. MG and LMG concentrations were measured in filets from catfish treated with MG under putative use conditions (ca. 250 and 1000 ppb, respectively) and from commercial trout samples (0-3 and 0-96 ppb, respectively). Concentrations of LMG in edible fish tissues always exceeded those of MG. A rapid cone voltage switching acquisition procedure was used to simultaneously produce molecular ions for quantification and diagnostic fragment ions for confirmation of MG and metabolites. The accurate and precise agreement between diagnostic ion intensity ratios produced by LMG in authentic standards and incurred fish samples was used to unambiguously confirm the presence of LMG in edible fish tissue. This suggested the validity of using LMG as a marker residue for regulatory determination of MG misuse. Additional metabolites derived from oxidative metabolism of MG or LMG (demethylation and N-oxygenation) were identified in catfish and trout filets, including a primary arylamine which is structurally related to known carcinogens. The ability to simultaneously quantify residues of MG and LMG, and to confirm the chemical structure of a marker residue by using LC/MS, suggests that this procedure may be useful in monitoring the food supply for the unauthorized use of MG in aquaculture.     

Analysis of penicillin G in milk by liquid chromatography

A liquid chromatographic (LC) method that was previously developed for penicillin G residues in animal tissues has been adapted to milk and milk products. After protein precipitation with sodium tungstate, samples are applied to a C18 solid-phase extraction cartridge, from which penicillin is eluted, derivatized with 1,2,4-triazole-mercuric chloride solution, and analyzed by isocratic liquid chromatography (LC) on a C18 column with UV detection at 325 nm. Quantitation is done with reference to penicillin V as an internal standard. Penicillin G recoveries were determined to be > 70% on standards fortified at 3-60 ppb. Accuracy approached 100% using the penicillin V internal standard. The detection limit for penicillin G residues was 3 ppb in fluid milk. Samples may be confirmed by thermospray/LC at concentrations approaching the detection limit of the UV method.     

Determination of zinc in serum, blood, and ultrafiltrate fluid from patients on hemofiltration by graphite furnace/atomic absorption spectroscopy or flow injection analysis/atomic absorption spectroscopy

Two methods were optimized for the determination of zinc in samples of blood, serum, and ultrafiltrate fluid from patients with chronic renal impairment undergoing hemofiltration. In the first procedure, after acid digestion of the samples, Zn in blood and serum is determined by a system coupled to flow injection analysis and atomic absorption spectroscopy. The method is rapid, automated, simple, needs small amounts of sample, and has acceptable analytical characteristics. The analytical characteristics obtained were as follows: determination range of method, 0.05-2.0 ppm of Zn; precision as coefficient of variation (CV), 5.3%; recovery, 95-105%; and detection limit (DL), 0.02 ppm. The second method is optimized for ultrafiltrate fluid because the sensitivity of the first procedure is not suitable for the levels of Zn (ppb or ng/mL) in these samples. The technique chosen was atomic absorption spectroscopy with electrothermal atomization in a graphite furnace. The analytical characteristics obtained were as follows: determination range of method, 0.3-2.0 ppb Zn; CV, 5.7%; recovery, 93-107%; and DL, 0.12 ppb. The methods were used to determine zinc in samples of blood, serum, and ultrafiltrate fluid from 5 patients with chronic renal impairment undergoing hemofiltration to discover whether there were significant differences in the zinc contents of blood, serum, and ultrafiltrate fluid after the hemofiltration process. An analysis of variance of the experimental data obtained from a randomly selected group of 5 patients showed that zinc concentrations in the ultrafiltrate fluid, venous blood, and venous serum do not vary during hemofiltration (p < 0.05), whereas in arterial blood and serum, the time factor has a significant effect.     

Determination of nanogram amounts of iodine in foods by radiochemical neutron activation analysis

Three different neutron activation analysis methods have been developed for the determination of ppb levels of iodine in food samples. The methods are based on the separation of iodine using (i) toluene extraction followed by bismuth sulfide coprecipitation; (ii) bismuth sulfide coprecipitation followed by radiochemical purification with palladium iodide; and (iii) radiochemical isolation by bismuth sulfide coprecipitation. The accuracy of these methods was evaluated by analysing replicate samples of reference materials. The measured values of iodine in A-11 Milk Powder, H-4 Animal Muscle and H-9 Mixed Human Diet from the International Atomic Energy Agency (IAEA) are statistically indistinguishable from the IAEA recommended values, and those for the Standard Reference Materials 1571 Orchard Leaves and 1577 Bovine Liver from the National Institute of Standards and Technology (NIST) are in good agreement with the NIST information values. The precision, in terms of relative standard deviation, is 5% at 50-100 ppb and 10% at 5-20 ppb levels of iodine. The absolute detection limits of these methods vary between 0.5 and 10 ng of iodine. All three methods were used to measure the iodine content of several food samples. The method involving bismuth sulfide coprecipitation followed by radiochemical purification provides the best detection limit and highest precision.     

Ion selective method for the determination of nitrite in smoked fish

In a direct ion selective electrode method for determining nitrite in smoked fish, the nitrite extracted from the sample is converted to nitrous acid with a measured addition of an acid buffer. The released nitrous acid is measured by a nitrogen oxide electrode. A known addition procedure is used to accurately measure the amount of nitrite in solution and the nitrite concentration is determined directly from Gran's plot paper. Recoveries of nitrite from spiked smoked chub samples ranged from 94.0 to 100.0% with an average of 96.7 and a relative standard deviation of +/-1.9% at the 100 ppm level. Results from the ion selective method were comparable with those from the official first action AOAC method 24.037-24.038.     

Mitochondrial DNA mutations in multiple symmetric lipomatosis

We describe an analytical technique for measuring residues of imidacloprid, a relatively new and highly active insecticide, in water and soil using high-performance liquid chromatography (HPLC). All analyses were performed on reversed-phase HPLC with UV detection at 270 nm using a mobile phase of acetonitrile-water (20:80, v/v). Fortified water samples were extracted with either solid-phase extraction (SPE) or liquid-liquid extraction methods. A detection limit of 0.5 microgram/l was achieved using the SPE method. The imidacloprid residues in soils were extracted with acetonitrile-water (80:20, v/v), and the extract was then evaporated using a rotary evaporator. The concentrated extract was redissolved in 1 ml of acetonitrile-water (20:80, v/v) prior to analysis by reversed-phase HPLC. A detection limit of 5 micrograms/kg was obtained by this method which is suitable for analysis of environmental samples. Accuracy and precision at 10 and 25 micrograms/kg soil samples were 85 +/- 6% and 82 +/- 4%, respectively.     

A radioimmunoassay for the determination of arginine vasotocin (AVT): plasma and pituitary concentrations in fresh- and seawater fish

A specific radioimmunoassay was developed and characterized for the measurement of arginine vasotocin (AVT) in teleost fish. Specificity of the antibody for AVT was demonstrated by parallelism of a series of AVT standards with serially diluted pituitary and plasma extracts. Crossreactivity of the antibody with the other teleost neurohypophysial peptide, isotocin, was less than 1% and the sensitivity of the assay was 0.24 fmol/assay tube. AVT was extracted from plasma by reverse-phase liquid chromatography [efficiency of 87.6 +/- 9.3% (n = 5)] and demonstrated as an effective procedure for plasma volumes ranging from 0.4 to 1.2 ml. Plasma AVT concentrations measured in a range of euryhaline and stenohaline teleost fish were between 10(-12) and 2 x 10(-11) M (1-20 pg/ml). There were no consistent differences between plasma AVT levels in euryhaline fish (flounder, trout, and eel) adapted to fresh water (FW) and sea water (SW). In flounder, pituitary AVT levels in FW- and SW-adapted fish were also similar.     

Blood levels of chlorinated hydrocarbon residues in the population of a continental town in Croatia (Yugoslavia)

Plasma or serum samples of 147 individuals from the general population of a continental town in Croatia (Yugoslavia) were analyzed for chlorinated hydrocarbons in 1975. The compounds were determined by gas chromatography and identified by comparing their retention times with those of known standards. All samples contained p,p'-DDE; mean concentration was 35.3 ppb. Only 20 samples contained p,p'-DDT; mean concentration was 22.7 ppb. Concentrations of alpha-BHC, lindane, and p,p'-TDE were 3.25, 4.09, and 11.6 ppb, respectively.     

Use of solid phase extraction disks for the GC-MS analysis of acidic and neutral herbicides in drinking water

An analytical procedure was developed in which thirteen herbicides (10 acidic and 3 neutral compounds) may be extracted from drinking water samples using solid phase extraction disks and subsequently analyzed by gas chromatography-mass spectrometry. Using 8 replicate samples consisting of reagent water fortified with nanogram quantities of each herbicide, the average percent recoveries and method detection limits were determined for each analyte. The method was found to be suitable for the determination of individual herbicides in drinking water at concentrations of approximately 10 ng/L.     

Analysis of aromatic sulfonates in water by solid-phase extraction and capillary electrophoresis

The separation of 14 different aromatic sulfonates of environmental concern by capillary (zone) electrophoresis (CZE) is presented. A new off-line solid-phase extraction (SPE) enrichment procedure, that is compatible with CE analysis, was developed, using the styrene-divinylbenzene adsorbent LiChrolut EN. The combined method of SPE and CE allows the determination of aromatic sulfonates in water samples in the low microgram/l range. Separations are performed with a simple sodium borate buffer at pH 9.3. Analytes are detected by UV absorbance and fluorescence emission with a Xe-lamp excitation source, and both principles are compared. The recoveries for most of the sulfonates are > 70% for the extraction from spiked tap and river water. The average method precision is < 20% for replicate analyses. Very hydrophilic sulfonates cannot be extracted by this method. The detection limit of the combined method of SPE enrichment and CE analysis is approximately 0.1 microgram/l for 200-ml water samples. The performance of the method was checked with the analysis of river and contaminated seepage water.