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1.
An analytical method has been developed for the quantification of two herbicides (ethidimuron and methabenzthiazuron) and their two main soil derivatives. This method involves fluidized-bed extraction (FBE) prior to cleanup and analysis by reverse-phase liquid chromatography with UV detection at 282 nm. FBE conditions were established to provide efficient extraction without degradation of the four analytes. (14)C-labeled compounds were used for the optimization of extraction and purification steps and for the determination of related efficiencies. Extraction was optimal using a fexIKA extractor operating at 110 degrees C for three cycles (total time = 95 min) with 75 g of soil and 150 mL of a 60:40 v/v acetone/water mixture. Extracts were further purified on a 500 mg silica SPE cartridge. Separation was performed on a C18 Purosphere column (250 mm x 4 mm i.d.), at 0.8 mL min(-1) and 30 degrees C with an elution gradient made up of phosphoric acid aqueous solution (pH 2.2) and acetonitrile. Calibration curves were found to be linear in the 0.5-50 mg L(-1) concentration range. Besides freshly spiked soil samples, method validation included the analysis of samples with aged residues. Recovery values, determined from spiked samples, were close to 100%. Limits of detection ranged between 2 and 3 microg kg(-1) of dry soil and limits of quantification between 8 and 10 microg kg(-1) of dry soil. An attempt to improve these performances by using fluorescence detection following postcolumn derivatization by orthophthalaldehyde-mercaptoethanol reagent was unsuccessful.  相似文献   

2.
Enzyme-linked immunosorbent assay for microcystins in blue-green algal blooms   总被引：18，自引：0，他引：18  
A direct competitive enzyme-linked immunosorbent assay (ELISA) for the freshwater blue-green algal toxin microcystin (MCYST) in algae and water was developed. The assay involves coating anti-MCYST-variant leucine-arginine (LR) antibody to the ELISA plate and the use of MCYST-LR-peroxidase as the enzyme marker. The linear portion of the standard curve for MCYST in phosphate buffer containing saline (PBS) was 0.5-10.0 ng/mL (25-500 pg/assay). The minimum detection level for MCYST-LR was 0.20 ng/mL (10 pg/assay). Contaminated water could be directly used in the ELISA. The overall analytical recoveries for MCYST-LR added to water at levels of 1-20 ng/mL was 83.4%. For analysis of cellular MCYST, the toxin was first extracted from the algae with 0.1M ammonium bicarbonate, diluted with PBS to less than 0.5 mg dried algae/mL (less than 5.0 mg wet weight/mL) and directly used in the ELISA. C-18 reverse-phase Sep-Pak cartridges effectively adsorbed MCYST from the toxin-containing solutions. The toxin could be recovered from the cartridge by eluting with 60% methanol. Using this approach, an algae extract that was relatively free of MCYST was prepared and was used in a recovery study. The overall analytical recovery of MCYST added to the algae extract in the range of 0.25-20 ppm was 83% with a coefficient of variation of 11.9%. The detection limit for MCYST in dried algae was about 0.25-0.5 microgram/g (0.25-0.5 ppm) lyophilized algae sample. This method was applied for the analysis of several naturally occurring algal blooms. Limited samples were also analyzed for MYCST by liquid chromatography.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

3.
Organic acids in fresh leaves, fruits, and dried rinds of Garcinia cowa (G. cowa) were determined by high-performance liquid chromatography. Fresh leaves, fruits, and dried rinds were extracted with water at 120 degrees C for 20-30 min under 15 lbs/in(2) pressure. Also, dried rinds were extracted with solvents (acetone and methanol) using a Soxhlet extractor at 60 degrees C for 8 h each. The samples were injected to HPLC under gradient elution with 0.01 M phosphoric acid and methanol with a flow rate of 0.7 mL/min using UV detection at 210 nm. The major organic acid was found to be (-)-hydroxycitric acid present in leaves, fruits, and rinds to the extent of 1.7, 2.3, and 12.7%, respectively. (-)-Hydroxycitric acid lactone, and oxalic and citric acids are present in leaves, fruits, and rinds in minor quantities. This is the first report on the composition of organic acids from G. cowa.  相似文献   

4.
Precolumn derivatization applying o-phthaldialdehyde (OPA) was used to analyze free lysine, histidine, and ornithine, precursors of the respective biogenic amines cadaverine, histamine, and putrescine, which are considered indicators of fish quality and safety. This method uses 75% methanol to eliminate the use of strong acids as the extraction solution. Each analysis took 35 min, was reproducible, and allowed separation of primary amino acids in fish samples. A binary solvent delivery system coupled with a fluorescence detector and an Ultrasphere ODS column were utilized for HPLC separation. Linearity of the calibration curves was very good (r(2) = 0.99) for the amino acids of interest. Minimum concentrations of detection were 40 pmol/mL for histidine and lysine and 70 pmol/mL for ornithine. Average recoveries were 72% for lysine, 93% for histidine, and 98% for ornithine. This method used solvent gradient elution to study the levels of these analytes in mahi-mahi, bigeye tuna, and flounder.  相似文献   

5.
A rapid solid-phase extraction (SPE) method was developed for the determination of bentazone and the phenoxy acids 2,4-D, dichlorprop, MCPA, and mecoprop in Norwegian environmental water samples. Cartridges with a high-capacity cross-linked polystyrene-based polymer were used for off-line preconcentration. The effects of elution solvent, elution volume, sample volume, sorbent mass, pH, and flow rate on the recoveries of the pesticides were investigated using HPLC. Average recovery of >90% was achieved with 500 mg sorbents using 2 mL of methanol with 5% NH3 as elution solvent. The recoveries were independent of sample pH in the tested range of pH 1-7. Using a sample volume of 200 mL, the limits of determination for the phenoxy acids and bentazone are 0.02 microg/L. Sample volumes up to 2000 mL at a flow rate of 60 mL/min could be handled without any loss of analytes, which makes it possible to lower the limits of determination. The SPE method was compared to a routinely used liquid-liquid extraction method. Three different water matrices spiked at 1.0 and 0.05 microg/L were extracted, and the quantification was performed by GC-MS. Both methods permitted the determination of phenoxy acids and bentazone in distilled water, creek water, and well water down to a level of 0.05 microg/L with recoveries >80% for 200 mL samples. Important advantages of the SPE method compared to the liquid-liquid extraction method were the short extraction times, lack of emulsions, use of disposable equipment, and reduced consumption of organic solvents.  相似文献   

6.
Low molecular mass organic acids (LMMOAs) and hydroxamate siderophores (HS) are molecules secreted by microbes and have previously been found in soil solution and in cultures. Mycorrhizal fungi are suggested to be involved in the nutrient uptake processes of trees and weathering of minerals. In this study soil samples taken from the O and E horizons of a podzol were extracted with 10 mM potassium phosphate buffer at pH 7.2. Variable parameters included addition of methanol to the extraction buffer and the use of ultrasonication or rotary shaking during extraction. LMMOAs and HS content of the soil extracts were determined. Analysis of soil extracts were carried out by liquid chromatography mass spectrometry (LC–MS) and the extraction results compared to results for soil solution samples obtained by centrifugation of the soils sampled. The extraction yields were significantly increased by addition of methanol to the extraction buffer, especially for the O horizon samples. Rotary shaking of the samples for 90 min gave slightly higher yields than ultrasonication for 15 min but the reduction in extraction time makes ultrasonication an attractive option. Of the HSs determined, ferricrocin was found in all samples. Optimal extraction conditions showed citric acid and isocitric acid to be the most abundant organic acids in the O and E horizons, respectively.  相似文献   

7.
《Soil biology & biochemistry》2012,44(12):2417-2422
Low molecular mass organic acids (LMMOAs) and hydroxamate siderophores (HS) are molecules secreted by microbes and have previously been found in soil solution and in cultures. Mycorrhizal fungi are suggested to be involved in the nutrient uptake processes of trees and weathering of minerals. In this study soil samples taken from the O and E horizons of a podzol were extracted with 10 mM potassium phosphate buffer at pH 7.2. Variable parameters included addition of methanol to the extraction buffer and the use of ultrasonication or rotary shaking during extraction. LMMOAs and HS content of the soil extracts were determined. Analysis of soil extracts were carried out by liquid chromatography mass spectrometry (LC–MS) and the extraction results compared to results for soil solution samples obtained by centrifugation of the soils sampled. The extraction yields were significantly increased by addition of methanol to the extraction buffer, especially for the O horizon samples. Rotary shaking of the samples for 90 min gave slightly higher yields than ultrasonication for 15 min but the reduction in extraction time makes ultrasonication an attractive option. Of the HSs determined, ferricrocin was found in all samples. Optimal extraction conditions showed citric acid and isocitric acid to be the most abundant organic acids in the O and E horizons, respectively.  相似文献   

8.
A simple and highly sensitive reversed-phase high-performance liquid chromatographic method (RP-HPLC) has been developed for the determination of steviol (SV) using dihydroisosteviol (DHISV) as an internal standard (IS). SV and DHISV were derivatized by reaction of the acids with 4-(bromomethyl)-7-methoxycoumarin in an aprotic solvent (DMF or acetone). The resulting ester derivatives were separated on an ODS column (250 x 4.6 mm i.d., 5 microm particle size) using fluorescence detection with excitation at 321 nm and emission at 391 nm. The mobile phase consisted of acetonitrile/water (80:20 v/v) with a flow rate of 1 mL min(-)(1). A linear relationship was observed for concentrations between 0.5 and 50 microg/mL of SV, and the detection limit was 100 pg. For application of this method to samples of beer fortified with stevioside, a simple procedure for extraction of the beer with diethyl ether and derivatization in DMF was applied. Whereas beer samples spiked with SV gave a linear response over the range 0.1-15 microg/mL beer, no SV could be detected in beer samples enriched in stevioside that had been stored for over 3 years. The application of the method to plant samples involved preparation of an acid fraction containing the SV analyte, derivatization, and sample cleanup using small silica columns and thin-layer chromatography. A sensitive determination of 594 ng of steviol present in 100 mg of dry plant material was performed with high precision and accuracy.  相似文献   

9.
The invasion and subsequent spread of the mosquito-borne West Nile virus in the United States has resulted in increased use of methoprene. With the increased need for sensitive detection and monitoring of methoprene in the environment, an analytical LC/ESI-MS/MS method has been developed for the analysis of methoprene and two analogues, kinoprene and hydroprene, in water. To improve the ionization efficiency of the nonpolar analytes, a derivatization step with the Cookson-type reagent 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD) was used. Derivatization improved the limit of detection 100-fold. For tandem MS analyses, limits of detection in environmental water samples (S/N = 3) are about 6 pg/mL for methoprene and 20 pg/mL for kinoprene and hydroprene, resulting in limits of quantification (S/N = 10) of 20 pg/mL for methoprene and 60 pg/mL for hydroprene and kinoprene extracted from 10 mL of water. This method was applied to measure methoprene concentrations in water samples from a treated site.  相似文献   

10.
The performance of a commercially available microtiter plate ELISA kit for the determination of the neonicotinoid insecticide imidacloprid was evaluated for sensitivity, selectivity, influence of organic solvent used for extraction procedure, matrix interference originated from agricultural sample, accuracy, and method comparison with conventional HPLC analysis. The limit of detection for the kit (0.1 or 0.5 ng/mL) was determined. The working range (1-39 ng/mL) experimentally calculated on the basis of a criterion, which is determined as the range from I(20) to I(80), was comparable to that established by the manufacturer (1-50 ng/mL). The linearity of the standard curve based on the kit-assembled standard solutions agreed with the one based on the self-made standard solutions. Specificity studies indicate that the imidacloprid monoclonal antibody can readily distinguish the target compound from other structurally related neonicotinoid analogues and some metabolites, with the exception of clothianidin, the cross-reactivity of which was approximately 12%. To extract imidacloprid from an agricultural sample (apple) as simply and rapidly as possible, some extraction methods were examined. Consequently, the extraction method with hand-shaking for 5 min was the best among the examined methods. For the analysis of imidacloprid in apple samples, it was extracted directly with methanol and the extracts were diluted 10-fold (100-fold in the well) with water prior to ELISA analysis. No significant matrix interference was observed with the dilution factor. Recoveries of imidacloprid from fortified apple samples ranged from 87.7 to 112.0%. The results obtained with the ELISA kit correlated well with those by the reference method (conventional HPLC analysis) for apple samples (r > 0.998). These findings strongly indicate that the ELISA kit may be employed routinely for an on-site imidacloprid residue analysis of apple samples.  相似文献   

11.
Soil organic N accounts for 95-98% of the total soil N content with amino acids (AAs) and amino sugars (ASs) identified as the major soil organic N compounds, but traditional 6 M HCl with reflux or sealed digestions for 24 h and various detection systems have accounted for only 30-40% of soil total N content as AA-N. This study compared traditional HCl extraction methodology with methanesulfonic acid (MSA) hydrolysis and nonderivatized AA and AS quantification by ion chromatography with pulsed amperometric detection for determination of the AA composition of plant litter and soils. MSA (4 M) gave AA-N recovery comparable to or better than 6 M HCl for plant AA digestions (16 h, 121 degrees C, 104 kPa). Use of 4 M MSA (0.5-1.5 h, 136 degrees C, 112 kPa) increased the total recovery of organic N as AAs, ASs, and NH(4)(+) by 46% from soils (n = 22) compared with 6 M HCl (12 h, 110 degrees C, reflux) with a MSA recovery rate of 85.6% of the total N content (n = 22 soils). The shorter MSA soil digestions (0.5-1.5 h) suggested that the majority of soil organic N was not present as protein forms found in plant litter analysis (16 h of digestion). MSA ion chromatographic analysis for soil AA/AS composition is a robust nonderivatization method requiring little sample preparation that can distinguish between small changes in soil AA composition during one growing season due to vegetation and tillage managements.  相似文献   

12.
Abstract

Extraction efficiency for metribuzin [4‐amino‐6‐(1,1‐dimethyl‐ethy 1)‐3‐(methylthio)‐l,2,4‐triazin‐5(4H)‐one] from soil was evaluated using four solutions of methanol:water at ratios of 1:1, 7:3, 4:1, and 1:0 v/v. Two concentrations of metribuzin (O.216 and 2.44 ug/ g soil, unlabeled and I4C‐metribuzin, respectively) were added to surface and subsurface samples of a Dundee silt loam soil. Although the differences in extraction efficiencies were slight due to differences in methanol:water ratios, the metribuzin recovery varied with soil depth. Less metribuzin was recovered from surface soil extracted with 1:1 and 1:0 methanol:water solutions when compared to 7:3 and 4:1 solutions (80 and 81% vs 89 and 88% recoveries, respectively). About equal quantities, 81 to 85%, were recovered from the subsurface soil. Three extraction shaking times (0.5 h followed by another 0.5 h; 4 h:4 h; and 24 h:24 h) were also evaluated using the 4:1 extractant. No recovery differences were observed between the 0.5 h and 4 h shaking times. However, significantly higher recoveries occurred in both the surface and subsurface soil with 24 h shaking. The efficiency of this method was also determined on Eustis loamy sand, Sharkey clay, and Dorovan muck soils of diverse physical and chemical properties. The 4:1 extracting solution consistently yielded among the highest metribuzin recovery (74 to 97%) from the four soil types, and two soil depths of a Dundee soil.  相似文献   

13.
Methiocarb was extracted from surface water samples collected at experimental rice field sites in Louisiana and Texas. The sampling system consisted of a single-stage 90-mm Empore extraction disk unit equipped with a battery-powered vacuum pump. After extraction, the C-18 extraction disks were stored in an inert atmosphere at -10 degrees C and shipped overnight to the laboratory. The disks were extracted with methanol and the extracts analyzed by reversed-phase high-performance liquid chromatography with a methanol/water mobile phase. Methiocarb was detected by ultraviolet absorption at 223 nm and quantified with the use of calibration standards. Recoveries from control surface water samples fortified at 5.0, 10, 50, and 100 ng/mL methiocarb averaged 92 +/- 7%. A method limit of detection for methiocarb in rice field surface water was estimated to be 0.23 ng/mL at 223 nm.  相似文献   

14.
Soil extracts are routinely used to quantify dissolved organic nutrient concentrations in soil. Here we studied the loss and transformation of low molecular weight (LMW) components of DOC (14C-glucose, 1 and 100 μM) and DON (14C-amino acid mixture, 1 and 100 μM) during extraction of soil (0-6 h) with either distilled water or 0.5 M K2SO4. The extractions were performed at 20 °C, at 4 °C, or in the presence of an inhibitor of microbial activity (HgCl2 and Na-azide). We showed that both glucose and amino acids became progressively lost from solution with increasing shaking time. The greatest loss was observed in H2O extracts at 1 μM for both substances (>90% loss after 15 min). Lower temperature (4 °C) and presence of K2SO4 both resulted in reduced loss rates. The presence of microbial inhibitors effectively eliminated the loss of glucose and amino acids. We conclude that microbial transformation of LMW-DOC and DON during H2O or K2SO4 extraction of soil may affect the estimation of their concentrations in soil. This finding has significant implications for methods that rely on chemical extractions to estimate LMW-C components of DOC and DON.  相似文献   

15.
A high-performance liquid chromatography (HPLC) method was developed for measuring the concentrations of clovamide-type phenylpropenoic acid amides (N-caffeoyldopamine and N-caffeoyltyramine) in cell and plasma samples. The separation was performed on a Nova-Pak C18 column using an isocratic buffer with a coulometric electrochemical detector with four electrode channels. Using the HPLC method, N-caffeoyldopamine and N-caffeoyltyramine could be detected with good peak resolutions at respective retention times (4 and 6.4 min). The calibration curves were linear over the ranges (0.1 and 100 microM), and their lower limit of detection was as little as 100 fmol. For quantifying N-caffeoyldopamine and N-caffeoyltyramine in cell and plasma samples, the samples were extracted by extraction methods with more than 95% recoveries. After extraction, the amides were detected with the same sensitivity, peak resolutions, and retention times. Using this method, plasma concentrations of N-caffeoyltyramine were determined in blood samples collected at 12, 24, 30, 36, 48, 60, and 75 min after the oral administrations of N-caffeoyltyramine (0.5 mg and 2 mg/30 g body weight). This HPLC method with an electrochemical detector is the first reported method able to quantify N-caffeoyldopamine and N-caffeoyltyramine in biological samples with excellent detection limits, peak resolutions, discrete retention times, and consistent reproducibility.  相似文献   

16.
Pressurized low-polarity water (PLPW) extraction of phenolic compounds from flax shive was investigated using statistically based optimization and the "one-factor-at-a-time" method. Extraction variables examined using central composite design (CCD) included temperature, flow rate, and NaOH concentration of the extracting water. Extraction of phenolic compounds including p-hydroxybenzaldehyde, vanillic acid, syringic acid, vanillin, acetovanillone, and feruric acid was affected by temperature and NaOH concentration; and extraction of all phenolic compounds, except ferulic acid, increased with temperature and NaOH concentration of the extracting water. Flow rate had little effect on concentration of phenolic compounds at equilibrium, but the extraction rate at the early phase was higher for higher flow rates. The mechanism of PLPW extraction of flax shive phenolics was also investigated using a two-site kinetic model and a thermodynamic model. To determine the extraction mechanism, flow rate was varied from 0.3 to 4.0 mL/min while temperature and NaOH concentration were fixed at 180 degrees C and 0.47 M, respectively. The flow rate tests showed the extraction rates of total phenolic (TP) compounds increased with flow rate and can be described by a thermodynamic model. The results from the thermodynamic model demonstrated that a K(D) value of 30 agreed with the experimental data in the flow rate range of 0.3-4.0 mL/min. When the effect of the three independent variables was evaluated simultaneously using CCD, a maximum TP concentration of 5.8 g/kg of dry flax shive (DFS) was predicted from the combination of a high temperature (230.5 degrees C), a high initial concentration of NaOH (0.63 M), and a low flow rate (0.7 mL/min). Maximum TP concentration of 5.7 g/kg of DFS was obtained from extraction conditions of 180 degrees C, 0.3 or 0.5 mL/min, and 0.47 M NaOH at equilibrium. A second-order regression model generated by CCD predicted a maximum TP concentration of 5.8 g/kg of DFS under the same extraction conditions, which is well matched with the results from experimental data.  相似文献   

17.
通过比较不同的提取溶剂和使用量,就水体中毒死蜱和TCP残留提取的效果及不同的流动相组成和比例对毒死蜱和TCP测定的影响,建立了水体中毒死蜱及TCP的HPLC残留分析方法。结果表明,水体中毒死蜱和TCP最佳提取溶剂为乙酸乙酯,提取次数为2次,用量分别为50和30mL。色谱条件为：流动相为甲醇：水=90：10或乙腈：水=90：10,流速1mL·min^-1;紫外检测波长300nm。当流动相为甲醇：水=90：10时,毒死蜱和TCP的保留时间分别为6．4和3．6min;当流动相为乙腈：水=90：10时,其保留时间分别为5．6和2．5min。毒死蜱和TCP的检出限分别为0．5和0．15ng。当毒死蜱和TCP在水中的添加浓度为0．01～5mg·L^-1时,标准添加回收率分别为91．4％-105．1％和90．6％～105．4％,变异系数分别为0．99％～4．12％和0．29％～9．33％。水样中毒死蜱和TCP的最小检出浓度分别为2和0．6ng·mL^-1。  相似文献   

18.
A method is described to separate and quantitate free amino acids from soil solutions extracted rapidly by centrifugation at 10,000 g for 5 min. Centrifugation of soil samples with forces up to 20,000 g did not affect the content of ammonium and water soluble organic carbon in solution, therefore it was concluded that the integrity of microbial cells was unaffected.The method of amino acid analysis involved treatment of the soil solution with tritiated 1-Fluoro-2,4-dinitrobenzene under alkaline conditions after addition of a known concentration of a standard mixture of amino acids. After extraction, the dinitrophenyl (DNP)-amino acids were separated by two-dimensional thin layer chromatography. Quantification was effected by counting the 3H-activity of each spot and comparing it to that of spots of a standard amino acid mixture of known concentration. The method has been tested and found satisfactory on a variety of cultivated soils, except for an Andept soil with a high content of metallic ions in solution.The total free amino acid content in cultivated and virgin soil samples ranged between 0.32 and 4.72 μg g?1 soil.Alanine, phenylalanine and threonine were the most abundant followed by valine and the aspartic-glutamic acid complex. Basic amino acids, those containing S (methionine and cystine), and the isoleucine-leucine pair were the least abundant and frequently not detected. With the Chernozemic order the total amino acid content decreased in the following order: Brown > Dark Brown > Black. Virgin soil samples contained more but not always a wider range of free amino acids than did cultivated soils. A Luvisolic soil cropped for 50 yr to a 5-yr rotation contained fewer and a narrower range of free amino acids than did the same soil cropped to a wheat-fallow rotation.  相似文献   

19.
A method for the simultaneous determination of 10 pesticides (organochlorines, organophosphorus compounds, and pyrethrins) in soils using capillary gas chromatography with atomic emission detection (GC-AED) is reported. Soil samples are first "cleaned-up" with 25 mL of an ascorbic acid solution (pH 2.15). The aqueous phase is extracted with ethyl acetate, and the solid residue is then extracted twice with 10 mL of ethyl acetate. The three resultant organic extracts are combined, concentrated to dryness, and reconstituted in 1 mL of acetone. The pesticides are selectively detected by monitoring chlorine and bromine in the first run and sulfur emission line wavelength in the second run. Each chromatographic run takes 19 min. Detection limits are between 25 and 75 pg, depending on the compound, which corresponds to 1.7 and 5.0 ng/g in the soil samples, respectively. Recoveries of the pesticides from spiked preparations result in an overall mean recovery of 95.3% (n = 120) at fortification levels ranging from 10 to 60 ng/g, depending on the compound. The method is reliable and can be useful for routine monitoring in soils.  相似文献   

20.
In the present work, an ultrasoundic procedure for the extraction of phosphorus (P) available in soil is described. The proposed method is based on extraction by 0.5 M sodium bicarbonate (NaHCO3), following with sonication under different conditions. Phosphorus was determined by the Murphy and Riley method. Sonication time and soil–extractant relative quantities were optimized. A statistical analysis approach was used to find suitable conditions for the ultrasoundic extraction. The main advantages of the sonication method are the reduced times of extractions, which take 10 min in contrast to the 30 min required by a shaking method, and the possibility to reduce soil and extractant quantities from 5 g–100 mL to 2 g–40 mL. Performance of the method was evaluated, and the procedure was utilized to analyze soils from Santiago del Estero, Argentina.  相似文献   

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