Development of an Enzyme Immunoassay for the Analysis of the Atrazine Metabolite Hydroxyatrazine Entwicklung eines Enzymimmunoassays zur Analyse des Atrazin-Metaboliten Hydroxyatrazin

A highly sensitive and specific enzyme immunoassay (EIA) is described for the detection of the atrazine metabolite hydroxyatrazine. Polyclonal antibodies were raised in rabbits by immunization with a hapten-bovine serum albumin (BSA) conjugate containing 8 hapten residues per molecule of BSA. An EIA with a horseradish peroxidase (HRP) hapten tracer was optimized in microtitre plates. A concentration of 50% B/B₀ was found at 0.10 μg/L for hydroxyatrazine. A limit of determination for hydroxyatrazine was reached at approximately 0.01 μg/L, i.e. well below the maximum concentration permitted by the EU guidelines for drinking water and the drinking water ordinance of the FRG. The assay did not require concentration or clean-up steps for drinking water or ground water samples. Validation experiments confirmed a good accuracy and precision. Hydroxyatrazine is reported to be the main atrazine metabolite found in soil samples. As organic solvents are usually employed for soil extraction, the influence of methanol as representative organic solvent on the assay was examined. Up to a concentration of 5% (v/v) methanol, the organic solvent did not affect the assay.     

Einsatz eines Immunoassays als Screening-Verfahren für die Untersuchung von Grundwasserproben auf Mecoprop

Application of an Enzyme Immunoassay for the Detection of Mecoprop in a Ground water Screening Procedure A competitive solid-phase enzyme immunoassay based on rabbit polyclonal antibodies was tested for its reliability. This assay, used for the detection of the phenoxycarboxylic acid mecoprop [2-(4-chloro-2-methylphenoxy-)propionic acid], has already been characterized in terms of middle of the test (0.36 μg/L at 50% B/B₀) and detection limit (0.06 μg/L at 80% B/B₀) by Weber (Acta hydrochim. hydrobiol. 22, 53–59 (1994)). For the reliability test, a small number from a batch of one hundred groundwater samples were spiked. The analyst was neither aware of which nor of how many samples were spiked. The spiked samples were all detected as positive. There were no false negative results. From these findings, values of 1 were obtained for the diagnostic sensitivity and for the diagnostic specificity. Based on these results, the mecoprop contents of 228 groundwater samples from various waterlevel gauges of a test field were determined. The test site had previously been treated repeatedly with this herbicide. Confirmation of the enzyme immunoassay results was carried out by GC/MS. This showed that the enzyme immunoassay gave one false positive result only. All other results from immunoassay and GC/MS showed good correlation.     

Entwicklung von Enzymimmunoassays zum Nachweis von Phenoxycarbonsäure-Herbiziden in Trink- und Grundwässern Development of Enzyme Immunoassays for the Detection of Phenoxycarboxylic Acid Herbicides in Drinking Water and Groundwater

Competitive solid phase enzyme immunoassays using polyclonal antibodies were developed for the detection of the phenoxycaboxylic acids MCPB [4-(4-chloro-2-methylphenoxy)butyric acid], Mecoprop [2-(4-chloro-2-methylphenoxy)propionic acid], and 2,4-D [(2,4-dichloro-phenoxy)acetic acid] in drinking water and ground water. The carrier protein for the immunization was bovine serum albumin, horseradish peroxidase conjugates were employed as enzyme tracer. For the three antisera, the optimization of detection limits and test sensitivities was our first consideration. For the mecoprop and 2,4-D antisera, the strongest influences were the pH value and the ionic strength, as much as the use of enzyme tracers with lower affinities. The MCPB antiserum reacted with 2,4-DB[4-(2,4-dichlorophenoxy)butyric acid] with equal specificity, either could be detected at 0.02 μg/L (80% B/B₀), middle of the test (50 B/B₀) lying at 0.1 μg/L. The detection limit with mecoprop antiserum was optimized to 0.02 μg/L, the prescribed limit for drinking water of 0.1 μg/L ling at 60 % B/B₀. The strongest cross-reactivity was found for mecoprop methyl ester. MCPB. 2,4-DB, and dichloroprop have crossreactivities of 50%, 6.7%, and 6.3%, respectively. The 2,4-D antiserum reacts less sensitively with 2,4-D, the detection limit being 0.4 μg/L. The 2,4-D isooctyl ester and 2,4-D methyl ester demonstrate as cross-reacting compounds high cross-reactivites of 3630% and 2230%. The cross-reactions of the compounds 2,4-DB, MCPB, and MCPA [(4-chloro-2-methylphenoxy)acetic acid] lie at 52%, 69%, and 41%. 100 ground water tests were spiked within laboratory. All positive samples were correctly identified. Falsely negative results did not appear.     

Entwicklung eines Enzymimmunoassays zum Nachweis des Herbizids Mecoprop in Trink- und Grundwasser auf der Basis von Antikörpern aus den Eiern von Legehennen

Development of an Enzyme Immunoassay for the Detection of Mecoprop in Drinking Water and Groundwater Based on Antibodies Raised in Chicken Egg Yolk Antibodies against mecoprop were isolated from egg yolk of immunized hens and were used for the development of an enzyme-linked immunosorbent assay. We tested assay parameters (pH and concentration of buffer, incubation temperature, kind of enzyme tracer) to optimize the standard curve for mecoprop. By decreasing the pH, the detection limit was reached at a concentration of 0.35 μg/L mecoprop. The concentration for 50% inhibition (50% B/B₀) was 2.8 μg/L. Dichlorprop and the methyl esters of both mecoprop and dichlorprop showed high crossreactivity (165%, 400% and 233%). Antibodies against mecoprop separated from egg yolk were compared to antibodies raised in rabbit for both sensitivity and specificity. Chicken immunoglobulins were found to be less sensitive and specific than the mammalian IgG's.     

Ein kompetitiver Immunoassay zur Bestimmung des Herbizids Fluazifop in Trink- und Grundwasser

A Competitive Immunoassay for the Determination of the Herbicide Fluazifop in Drinking Water and Groundwater A competitive solid-phase enzyme immunoassay using rabbit polyclonal antibodies was developed for the detection of the herbicide fluazifop [(RS)-2-[4-(5-trifluoromethyl-2-pyridyloxy)phenoxy]propionic acid] in drinking water and groundwater. Present regulatory limits for drinking water in Germany were taken as the critical level. The carrier protein was bovine serum albumin; horseradish peroxidase was used as marker enzyme with 3,3′,5,5′-tetramethylbenzidine as substrate. A high concentration of high-affinity antibodies in the serum, optimization of test conditions (antibody and enzyme tracer concentration, incubation time etc.), and very low cross reactivities to substances of similar structures led to a highly sensitive and specific ELISA with a detection limit below 0.1 μg/L for fluazifop as free acid. On testing the suitability of the assay's use as a screening test with one hundred drinking-water samples, the three samples which had been spiked in the laboratory were recognized as positive with respect to their fluazifop content. Confirmation by gas chromatography-mass spectrometry showed the test results of two other samples to be false positive. False negative results did not appear. The concentration was in the detection limit region of 0.1 μg/L.     

Determination of the Maximum Acceptable Toxicant Concentration (MATC) and the safe Concentration for Certain Aquatic Pollutants

— The 96 h TL₅₀ values, the maximum acceptable toxicant concentration (MATC) and the safe concentrations were determined for various aquatic pollutants, viz. thiotox, chlordane, metasystox, dichlorvos, sevin and carbofuran (pesticides), phenol, pentachlorophenol and dinitrophenol (phenolic compounds), cadmium chloride and copper sulphate (heavy metal), swascofix P₁₄, swanic 6 L and swascol 1P (synthetic detergent) using the larvae (the most sensitive stage of the life cycle) of Cyprinus carpio, a fresh water exotic carp. — The estimated MATC ranged between 0.009 and 400 μg/1 for all the pollutants; — the calculated value of the application factor (AF) ranged between 0.0096 and 0.0978. — The safe concentrations were determined for each pollutant by the lowest value of application factor worked out in the present investigation; — safe concentrations were also worked out by different formulae given by different workers. The values obtained were compared with the safe concentrations value worked out by the authors of the present study.     

菹草(Potamogeton crispus)附着物对水体氮、磷负荷的响应

通过实验模拟了10组氮、磷负荷对菹草(Potamogeton crispus)生长期和衰亡期茎叶附着物的影响.结果显示:随着水体氮、磷浓度的升高,菹草附着物的叶绿素a(Chl.a)含量、附着有机物量、附着无机物量和附着物总量均增加,在氮、磷浓度最高的T10组(总氮12.0 mg/L,总磷1.0 mg/L),附着物的总量达到高峰,附着物的Chl.a含量为2.005~4.765mg/g(DW),附着有机物的量为29.027~94.886 mg/g(DW),附着无机物的量为176.881~397.750 mg/g(DW),附着物总量为205.909~492.636 mg/g(DW).在菹草的快速生长期、稳定期和衰亡期,附着物的Chl.a含量、附着有机物量、附着无机物量和附着物总量均存在显著差异,均表现为衰亡期 >稳定期 >快速生长期,且在各营养盐浓度下均存在这一趋势.菹草衰亡期附着物的Chl.a含量、附着有机物量、附着无机物量和附着物总量分别为稳定期的1.046~1.826、1.046~1.638、1.029~1.858和1.106~1.717倍,为快速生长期的2.324~4.059、2.323~3.640、2.101~3.792和2.280~3.584倍.结果表明水体氮、磷负荷的增加促进了菹草茎叶附着物的生长和积累,加速了沉水植物衰亡.     

Field Study of Enhanced TCE Reductive Dechlorination by a Full‐Scale Whey PRB

This study evaluates the efficiency of a full‐scale, 81 m‐wide permeable reactive barrier (PRB) configured by injection of dairy whey in the downgradient region of a contaminant source zone to enhance the in situ biodegradation of high concentrations (10² to 10³μg/L) of chlorinated ethenes (CEs). Ten biannual whey injections were completed in a 3.5‐year pilot phase and 1.5‐year operational phase. Improved and sustained dechlorination was observed at extraction/injection and downgradient wells in the fully‐operational phase, when dried whey masses were increased from 13.6 kg to 230–360 kg, whey slurry volumes were increased from 2300 L to 307,000–480,000 L, and extraction/injection well loops were employed for the application of whey. At extraction/injection wells, CEs decreased to low (≤10 μg/L) or undetectable levels. At downgradient wells, average trichloroethene concentrations decreased, by as much as 100% (from ≤384.2 during the pilot phase to ≤102.6 μg/L during the operational phase), while average cis‐dichloroethene concentrations decreased by as much as 57.5% (from ≤6466.1 to ≤4912.2 μg/L). Downgradient vinyl chloride averages either increased by as much as 63.8% (from ≤859.6 to ≤1407.9 μg/L) or decreased by 64.0% (from 1375.4 to 880 μg/L). Downgradient ethene + ethane averages increased by as much as 73.2% (from ≤1145.3 to ≤1347.1 μg/L). On the basis of the 2008 average market price, the estimated material cost of whey is $1.96/kg organic carbon or, for the configuration of an 81 m PRB by biannual application of 300 kg whey, $325/year. Carbon substrate cost comparisons and implications for efficient in situ treatment design are discussed.     

Surveying Upstate NY Well Water for Pesticide Contamination: Cayuga and Orange Counties

Private wells in Cayuga and Orange counties in New York were sampled to determine the occurrence of pesticide contamination of groundwater in areas where significant pesticide use coincides with shallow or otherwise vulnerable groundwater. Well selection was based on local groundwater knowledge, risk modeling, aerial photo assessments, and pesticide application database mapping. Single timepoint samples from 40 wells in each county were subjected to 93‐compound chromatographic scans. All samples were nondetects (reporting limits ≤1 μg/L), thus no wells from either county exceeded any of 15 state groundwater standards or guidance values. More sensitive enzyme‐linked immunosorbent assays (ELISA) found two wells with quantifiable atrazine in each county (0.1–0.3 μg/L), one well with quantifiable diazinon (0.1 μg/L) in Orange County, and one well with quantifiable alachlor (0.2 μg/L) in Cayuga County. Trace detections (<0.1 μg/L) in Cayuga County included atrazine (five wells), metolachlor (six wells), and alachlor (one well), including three wells with multiple detections. All 12 Cayuga County wells with ELISA detections had either corn/grain or corn/forage rotations as primary surrounding land uses (although 20 other wells with the same land uses had no detections) and all quantified detections and most trace detections occurred in wells up to 9‐m deep. Orange County trace (<0.1 μg/L) ELISA detections (atrazine three wells, diazinon one well, and metolachlor five wells) and quantified detections were only generally associated with agricultural land uses. Finding acceptable drinking water quality in areas of vulnerable groundwater suggests that water quality in less vulnerable areas will also be good.     

Terbuthylazine and Deethylterbuthylazine in Rain and Surface Water – Determination by Enzyme Immunoassay and Gas Chromatography/Mass Spectrometry

Rain and surface water samples from Southern Germany were investigated from 1991 to 1995 for terbuthylazine and one of its major metabolites, deethylterbuthylazine. The concentrations observed were compared to the concentrations found for atrazine and deethylatrazine in the same water samples. Concentrations ranged from < 0.02 μg/L to 0.7 μg/L for terbuthylazine and from < 0.02 μg/L to 0.6 μg/L for deethylterbuthylazine, compared to concentrations of < 0.02 μg/L to 3 μg/L and < 0.02 μg/L to 0.5 μg/L for atrazine and deethylatrazine, respectively. The ratios of metabolite concentrations to parent compound concentrations were calculated for deethylterbuthylazine to terbuthylazine (DTR) and deethylatrazine to atrazine (DAR). In rain water, DTR of 0.8…3.0 and DAR of 0.3… 1.9 were determined with mean values of 0.9… 1.7 for DTR and 0.6…0.9 for DAR in the different years. The ratios increased during summer periods. The highest ratios were observed in samples from forest stands, showing that degradation of the herbicide has occurred during transport between the source and the sampling site. The DTR in rain water were about 50… 100% higher than the DAR. This indicates a higher degradation rate of terbuthylazine during atmospheric transport. In surface water, DTR of 0.3… 1.2 with mean values of 0.5…0.8 and DAR of 0.2…2.2 with mean values of 0.2… 1.3 were observed. The ratios increased from June to September.     

Studies on Adsorptive Desulfurization by Activated Carbon

Sulfur removal using adsorption requires a proper process parametric study to determine its optimal performance characteristics. In this study, response surface methodology was employed for sulfur removal from model oil (dibenzothiophene; DBT dissolved in iso‐octane) using commercial activated carbon (CAC) as an adsorbent. Experiments were carried out as per central composite design with four input parameters such as initial concentration (C₀: 100–900 mg/L), adsorbent dosage (m: 2–22 g/L), time of adsorption (t: 15–735 min), and temperature (T: 10–50°C). Regression analysis showed good fit of the experimental data to the second‐order polynomial model with coefficient of determination R²‐value of 0.9390 and Fisher F‐value of 16.5. The highest removal of sulfur by CAC was obtained with m = 20 g/L, t = 6 h, and T = 30°C.     

Concentrations and Seasonal Trends of Extracellular Microcystins in Freshwaters of the Czech Republic – Results of the National Monitoring Program

The first outcomes of the national monitoring program on cyanobacteria and their toxins in the Czech Republic are reported. The concentrations of extracellular microcystins (MCs, analyzed by ELISA) in 206 water samples collected from 94 localities have been investigated. Observed MC concentrations were relatively high in comparison with the literature data, although weather conditions during the covered season (year 2004) were less favorable for the cyanobacterial water blooms. Detectable MCs (>0.125 μg/L) were found in 145 waters (70% of all samples) with a median concentration of 0.67 μg/L. The total median, including samples with no detectable MCs, was 0.26 μg/L. The seasonal trend showed an increase from July to September at most localities (maximum concentrations of about 8 μg/L and extremes up to 37 μg/L). However, a peak at the beginning of the season or continuously elevated MC concentrations were also observed at some localities. Positive correlations between increasing concentrations of MCs and the dominancy of Microcystis spp. have been observed. The survey provides the first large-scale study of the occurrence and seasonal variability of extracellular microcystins that are of particular interest for drinking water management.     

Performance of A Combination Process of UV/H2O2/Micro‐Aeration for Oxidation of Dichloroacetic Acid in Drinking Water

The decomposition of dichloroacetic acid (DCAA) in water using a UV/H₂O₂/micro‐aeration process was investigated in this paper. DCAA cannot be removed by UV radiation, H₂O₂ oxidation or micro‐aeration alone, while UV/H₂O₂/micro‐aeration combination processes have proved effective and can degrade this compound completely. With initial concentrations of about 110 μg/L, more than 95.1% of DCAA can be removed in 180 min under UV intensity of 1048.7 μW/cm², H₂O₂ dosage of 30 mg/L and micro‐aeration flow rate of 2 L/min. However, more than 30 μg/L of DCAA was left after 180 min by UV/H₂O₂ combination process without micro‐aeration with the same UV intensity and H₂O₂ dosage. The effects of applied UV radiation intensity, H₂O₂ dose, initial DCAA concentration and pH on the degradation of DCAA have been examined in this study. Degradation mechanisms of DCAA with hydroxyl radical oxidation have been discussed. The removal rate of DCAA was sensitive to operational parameters. There was a linear relationship between rate constant k and UV intensity and initial H₂O₂ concentration, which indicated that a higher removal capacity can be achieved by improvement of both factors. A newly found nitrogenous disinfection by‐product (N‐DBP)‐DCAcAm, which has the potential to form DCAA, was easier to remove than DCAA by UV/H₂O₂ and UV/H₂O₂/micro‐aeration processes. Finally, a preliminary cost comparison revealed that the UV/H₂O₂/micro‐aeration process was more cost‐effective than the UV/H₂O₂ process in the removal of DCAA from drinking water.     

Fine Particles (PM2.5) in Residential Areas of Lucknow City and Factors Influencing the Concentration

Urban populations are exposed to a high level of fine and ultrafine particles from motor vehicle emissions which affect human health. To assess the hourly variation of fine particle (PM_2.5) concentration and the influence of temperature and relative humidity (RH) on the ambient air of Lucknow city, monitoring of PM_2.5 along with temperature and RH was carried out at two residential locations, namely Vikas Nagar and Alambagh, during November 2005. The 24 h mean PM_2.5 concentration at Alambagh was 131.74 μg/m³ and showed an increase of 13.74%, which was significantly higher (p < 0.05) than the Vikas Nagar level. The 24 h mean PM_2.5 on weekdays for both locations was found to be 142.74 μg/m³ (an increase of 66.23%) which was significantly higher (p < 0.01) than the weekend value, indicating that vehicular pollution is one of the important sources of PM_2.5. The mean PM_2.5 at night for all the monitoring days was 157.69 μg/m³ and was significantly higher (p < 0.01) than the daytime concentration (89.87 μg/m³). Correlation and multiple regressions showed that the independent variables, i. e., time, temperature, and RH together accounted for 54%, whereas RH alone accounted for 53% of total variations of PM_2.5, suggesting that RH is the best influencing variable to predict the PM_2.5 concentration in the urban area of Lucknow city. The 24 h mean PM_2.5 for all the monitoring days was found to be higher than the NAAQS recommended by the US‐EPA (65 μg/m³) and can be considered to be an alarming indicator of adverse health effects for city dwellers.     

Oxidation of Benzo(a) Pyrene Catalyzed by Phenol Oxidases of Alga Nitella sp. and Potato Tubers

The kinetics of oxidation of carcinogenic benzo(a)pyrene (0.1 nM solution in phosphate buffer) catalyzed by enzyme protein of soil and water plants (potato tubers and alga Nitella sp.) has been studied. The benzo(a)pyrene oxidation rate is determined by the phenoloxidase activity of the enzyme protein from both sources to an equal degree and expressed by the Michaelis-Menten equation. The values for k_cat and apparent K_m were (0.97 ± 0.14) Ms^?1 and 33 ± 6 μM, respectively. The presence of other compounds (including protein substances) in acetone preparations of potato tubers considerably suppresses its catalytic activity in relation to benzo(a)pyrene. Consequently, in the polluted biosphere some water plants, such as alga Nitella sp., and potato tuber can transform carcinogenic benzo(a)pyrene (BP). As oxidation products, all three BP-quinones, i.e. 1,6-, 3,6-and 6,12-diones, were identified.     

The acute toxicity of silver to some freshwater fishes

In the batch experiment with daily exchange of medium the acute toxicity of Ag from AgNO₃ to Channa punctatus, Lebistes reticulatus and Puntius sophore is determined over 12 to 96 h exposure. In general, the sensitivity of species increases in the order mentioned. The LC_{50, 96h} are 18.89 μg/l Ag for Channa punctatus, 7.55 μg/l Ag for Puntius sophore and 6.44 μg/l for Lebistes reticulatus. For 12 and 48 h exposure the values in the same order of species are: 62 and 30, 44 and 14 as well as 57 and 16 μg/l Ag.     

A Practical Measurement Strategy to Estimate Nonlinear Chlorinated Solvent Sorption in Low foc Sediments

In this study, we tested a practical strategy useful for accurate chlorinated volatile organic compound (cVOC) sorption prediction. Corresponding to the feature of the superposition of adsorption due to thermally altered carbonaceous matter (TACM) with organic carbon‐water partitioning, a nonlinear Freundlich sorption isotherm covering a wide range of aqueous concentrations was defined by equilibrium sorption measurement at one or a few low concentration points with extrapolation to the empirical organic carbon‐water partition coefficient (K_oc,e) near compound solubility. We applied this approach to obtain perchloroethene equilibrium sorption isotherm parameters for TACM‐containing glacial sand and gravel subsoil samples from a field site in New York. Sorption and associated K_oc,c applicable to low (5–500 µg/L) and high (>100,000 µg/L) aqueous concentrations were determined in batch experiments. (The K_oc,c is the organic carbon‐normalized sorption partition coefficient corresponding to aqueous concentration C_w.) The K_oc,c measurements at low concentration (~5 µg/L) were 6 to 34 times greater than the K_oc,e. The importance of this type of data is illustrated through presentation of its substantial impact on the site remedy. In so doing, we provide an approach that is broadly applicable to cVOC field sites with similar circumstances (low carbon content glacial sand and gravel with TACM).     

Atmospheric Particulate Concentration Measured in an Urban Area Bandung

— Atmospheric particulate concentration for total suspended particles (TSP) and for PM₁₀ (particulate matter under 10 micron) was measured in Jalan Braga and ITB campus, Bandung. Six samples were collected over one- or two-day time periods using High Volume Sampler (HVS) for TSP and Low Volume Sampler (LVS) or Anderson Cascade Impactor for PM₁₀. Samples were further analyzed to determine concentrations of metals, sulfate and nitrate. Concentration of NO_x (NO and NO₂) was also measured hourly and simultaneously during the sampling period. The results from this study show that the atmospheric particulate concentration in Jalan Braga for TSP ranged from 304.04 to 363.17, and for PM₁₀ concentration ranged from 277.02 to 336.44 μg/m³. The lead concentrations were 1.42–2.37 μg/m³ in the TSP and 0.81–1.57 μg/m³ in the PM₁₀. The nitrate concentrations were 5.89–6.51 μg/m³ and 2.27–3.45 μg/m³ for the TSP and PM₁₀, respectively. The hourly NO_x concentration varied between 0.14–0.35 ppm. The total elements (metals, sulfate and nitrate) found in the samples contribute from 20 to 25% of the total particulate concentration.     

Effect of equatorial ionization anomaly on the occurrence of spread-F

The unique geometry of the geomagnetic field lines over the equatorial ionosphere coupled with the E–W electric field causes the equatorial ionization anomaly (EIA) and equatorial spread-F (ESF). lonosonde data obtained at a chain of four stations covering equator to anomaly crest region (0.3 to 33 °N dip) in the Indian sector are used to study the role of EIA and the associated processes on the occurrence of ESF. The study period pertains to the equinoctial months (March, April, September and October) of 1991. The ratios of critical frequency of F-layer (f₀F₂) and electron densities at an altitude of 270 km between Ahmedabad (33 °N dip) and Waltair (20 °N dip) are found to shoot up in the afternoon hours on spread-F days showing strengthening of the EIA in the afternoon hours. The study confirms the earlier conclusions made by Raghava Rao et al. and Alex et al. that a well-developed EIA is one of the conditions conducive for the generation of ESF. This study also shows that the location of the crest is also important in addition to the strength of the anomaly.     

Phosphorus Forms and Their Changes in Water of Three Subalpine Lakes in Italy

Total phosphorus and its main forms: dissolved mineral, dissolved organic, particulate organic and particulate mineral in the vertical water column of three subalpine lakes of various types in Italy, has been estimated during the winter-vernal season. The range of variation in the phosphorus content in these waters was as follows: total phosphorus 16 ± 2860 μg/1 PO₄, dissolved mineral phosphorus 4 ± 1040 μg/l PO₄, dissolved organic phosphorus 1 ± 160μg/l PO₄, particulate organic phosphorus 0 ± 290 μg/l PO₄ and particulate mineral phosphorus 1 ± 100 μg/l PO₄, Generally the content of total phosphorus and dissolved mineral phosphorus (phosphates) increased with the degree of eutrophy with the depth and with the progress of the vernal season towards the summer stagnation time. The amount of phosphates increased in water with the depletion of oxygen, both in the verical water column and with the progress of stagnation time. The amounts of dissolved organic phosphorus decressed with the depth of the vertical water column whereas the dissolved mineral phosphorus increased. The development of the particulate organic phosphorus stratification in the vertical water column was clearly visible in the eutrophic lake. The quantities of total phosphorus and its main component, dissolved mineral phosphorus, decreased evidently from January to May in all three lakes, mostly in the eutrophic lake. The reason of this decrease is sorption by lake sediments and to a certain degree sedimentation of phosphorus sorbed by ferric hydroxide. The increase of dissolved mineral phosphorus and that of total phosphorus in the vertical water column and with the progress of summer stagnation had as a reason the liberation of phosphorus from sediments, and not so much decomposition of sedimentating plankton or dissolved organic phosphorus. The share of single (mean) values of phosphorus forms in the total phosphorus was as follows: In the oligotrophic lake the share of particulate mineral phosphorus was extremely high in March (21% of the total), probably because of the inflow of the melting waters from the drainage area. The development of vertical stratification in waters of three subalpine Italian lakes at the end of the vernal season (May) indicates the quantitative prevailing of dissolved mineral phosphorus with its increase with the depth and domination of dissolved organic phosphorus in the trophogenic zone.