Teratologic assessment of maleic hydrazide and daminozide, and formulations of ethoxyquin, thiabendazole and naled in rats

Teratogenicity studies were conducted in rats treated orally from days 6-15 of gestation with single daily doses of 400-1600 mg/kg of maleic hydrazide, 300-1000 mg/kg daminozide, 125-500 mg/kg ethoxyquin or thiabendazole, or 25-100 mg/kg naled. Dams were killed on the 22nd day of gestation, and fetuses were evaluated by routine teratologic methods. No adverse effect was related to any treatment other than an increased incidence of anomalous fetuses at the highest dose (500 mg/kg) of thiabendazole.     

Teratogenic evaluation of the pesticides baygon, carbofuran, dimethoate and EPN

Baygon was administered IG once daily to CD rats (5 to 50 mg/kg), on the 7th-19th day of gestation or to CD-1 mice (5 to 60 mg/kg) on days 6-16 of gestation. Baygon, at dose levels which were not maternally lethal, did not produce fetotoxicity, fetal lethality or malformations in the fetuses. Baygon was not teratogenic in the CD rat or CD-1 mouse at maternally nontoxic dose levels. Carbofuran was administered IG once daily to CD rats (0.05 to 5.0 mg/kg), on the 7th-19th day of gestation or to CD-1 mice (0.1 to 20 mg/kg) on days 6-16 of gestation. At dose levels which were not maternally lethal, carbofuran did not produce fetotoxicity, fetal lethality or malformations in the fetuses. Carbofuran was not teratogenic in the CD rat or CD-1 mouse at maternally nontoxic dose levels. Dimethoate was administered IG once daily to CD-1 mice (10 to 80 mg/kg), on the 6th-16th day of gestation. At dose levels which were not maternally lethal, dimethoate did not produce fetotoxicity, fetal lethality or malformations in the fetuses. Dimethoate was not teratogenic in the CD-1 mouse at maternally nontoxic dose levels. EPN was administered IG once daily to CD-1 mice (1.0 to 12.0 mg/kg) on the 6th-16th day of gestation. EPN, at dose levels up to those which were maternally lethal, did not produce fetotoxicity, fetal lethality or an increase in malformations. EPN was not teratogenic in the CD-1 mouse at maternally nontoxic dose levels.     

A teratological assessment of four trihalomethanes in the rat

Abstract

Four trihalomethanes were administered by gavage to Sprague‐Dawley rats from day 6 to day 15 of gestation. Chloroform (Ch) was administered at levels of 100, 200 and 400 mg/kg and bromoform (Br), bromodichloromethane (BDCM) and chlorodibromomethane (CDBM) were administered at levels of 50, 100 or 200 mg/kg/day. A separate control was used for each compound. Maternal weight gain was depressed in all groups receiving Ch and at the highest dose levels of BDCM and CDBM. Ch administration caused decreased maternal hemoglobin and hematocrit values at all dose levels and also produced increased serum inorganic phosphorus and cholesterol at the highest dose. Liver enlagement was observed at all dose levels of Ch but in no other treatment groups. Evidence of a fetotoxic response was observed with Ch, CDBM and Br but not BDCM. No dose‐related histopathological changes were observed in either mothers or fetuses as a result of treatment. None of the chemicals tested produced any teratogenic effects.     

Photomirex: A teratogenicity and tissue distribution study in the rabbit

Abstract

Adult New Zealand white does were intubated orally with single daily doses of 0, 5, or 10 mg of photomirex (8‐monohydromirex) per kg body weight from the 6th through to the 18th day of gestation. Pregnancies were interrupted at term by cesarian section and fetuses removed and evaluated by following routine teratologic methods. Both maternal and fetal tissues were analyzed for residues of photomirex. None of the treated does showed any sign of toxicity. Except for a significant reduction in the mean fetal weight of the 10 mg/kg group all other parameters which evaluated fetal survival and fetal development were within the control range. Photomirex was found in all tissues examined. In the doe, the highest levels were found in fat followed by liver, kidney, spleen, heart, brain and blood. Photomirex was readily transferred across the placenta and accumulated in the fetus. However, in the fetus the highest levels were found in the heart, followed by liver, brain and blood. There were no teratogenic effects at the doses used in this study.     

Detection and occurrence of pentachlorophenol residues in chicken liver and fat

Abstract

A method for the detection of pentachlorophenol (PCP) residues in chicken liver and fat is presented. A detection limit of 0.002 mg/kg was achieved. Recoveries from liver and fat were in the range 82–88% and 95–97%, respectively.

Low level residues of PCP were found in all 1072 liver and 723 fat samples. These levels were <0.010 mg/kg in 92.7% of the fat and 75.6% of the livers. Only 0.75% of the liver samples had PCP levels>0.1 mg/kg. None of the more toxic impurities of PCP were detected in the chicken tissues.     

Teratogenic evaluation and fetal deposition of hexabromobenzene (HBB) and hexafluorobenzene (HFB) in CD-1 mice

HBB (hexabromobenzene) and HFB (hexafluorobenzene) were tested for their teratogenic potential in CD-1 mice. HBB and HFB were administered to pregnant mice from the 6th to the 16th day of gestation by gastric intubation. Neither HBB nor HFB were teratogenic or fetotoxic at doses up to 98.6 mg HBB/kg and 65.3 mg HFB/kg. No maternal toxicity was noted. HBB concentration in the fetuses indicated little, if any accumulation. No HFB was detected in the fetal or maternal tissues 24 hours after the last dose.     

Study on 14C-BHA absorption and distribution in rat

BHA was administered to rats at doses of 5 or 500 mg/kg for seven days. ¹⁴C-BHA absorption was investigated on the eighth day and compared with animals receiving a single dose of 5 or 500 mg/kg BHA. Absorption of ¹⁴C-BHA was proportionately greater at the high dose rats. ¹⁴C distribution was 3–6 times higher in the forestomach than in the pyloric region of the stomach. The urinary excretion of BHA conjugates was not dependent on pretreatment with BHA or on the dose.     

Impact of molybdenum nanoparticles on survival,activity of enzymes,and chemical elements in <Emphasis Type="Italic">Eisenia fetida</Emphasis> using test on artificial substrata

The influence of molybdenum oxide nanoparticles (MoO₃) on the growth and survival of Eisenia fetida was established. The activity of antioxidant enzymes and changes in concentration of molybdenum in the body of E. fetida were determined. The degree of bacterial bioluminescence inhibition in extracts of substrates and worm was studied using luminescent strain Escherichia coli K12 TG1. The enzymatic activity of substrates before and after exposure with nanoparticles and worms was assessed. Nanoparticles have concentrations of 10, 40, and 500 mg/kg of dry matter, and substrata are made of artificial soil (substrate A) and microcrystalline cellulose (substrate B). Spherical nanoparticles MoO₃, yellow in color, with size 92?±?0.3 nm, Z-potential 42?±?0.52 mV, molybdenum content 99.8 mass/%, and specific area 12 m²/g were used in the study. A significant decrease by 23.3 % in weight was registered (for MoO₃ NPs at 500 mg/kg) on substrate A (p?≤?0.05). On substrate B, the maximum decrease in weight by 20.5, 33.3, and 16.9 % (p?≤?0.05) was registered at a dose of 10, 40, and 500 mg/kg, respectively; mortality was from 6.6 to 73 %. After the assessment of bacterial bioluminescence inhibition in substrates A and B (extracts) and before worms were put, the toxicity of substrates was established at doses of 40 and 500 mg/kg, expressed in inhibitory concentration (IC) 30 and IC 50 values. Comparatively, on days 7 and 14, after exposure in the presence of E. fetida, no inhibition of bioluminescence was registered in extracts of substrates A and B, indicating the reduction in toxicity of substrates. The initial content of molybdenum in E. fetida was 0.9?±?0.018 mg/kg of dry matter. The degree of molybdenum accumulation in worm tissue was dependent on the dose and substrate quality. In particular, 2–7 mg/kg of molybdenum accumulated from substrate A, while up to 15 kg/kg of molybdenum accumulated from substrate B (day 7). Molybdenum concentration decreased by 64.8 and 57.4 % at doses 40 and 500 mg/kg, respectively, on day 14. The reaction of antioxidant enzymes was shown in an insignificant increase of glutathione reductase (GSR) and catalase (CAT) at concentrations of 10 and 40 mg/kg in substrate A, followed by the subsequent reduction of their activity at the dose of 500 mg/kg MoO₃. The activity of GSR in substrate B against the presence of MoO₃ nanoparticles decreased, with significant difference of 33.5 % (p?≤?0.05) at the dose of 500 mg/kg compared with untreated soil. In experiments with substrate A, an increase of catalase activity was registered for the control sample. The presence of MoO₃ nanoparticles at the concentration of 10 mg/kg in the environment promoted enzymatic activity on days 7 and 14, respectively. A further increase of nanoparticle concentration resulted in the decrease of catalase activity with a minimum value at the concentration of MoO₃ of 500 mg/kg. In the experiment with substrate B at the concentration of MoO₃ nanoparticles of 40 mg/kg, enzymatic activity increases on day 7 of exposure. However, the stimulating effect of nanoparticles stops by day 14 of the experiment and further catalase activity is dose dependent with the smallest value in the experiment with MoO₃ having the concentration of 500 mg/kg.     

Effect of quinalphos on blood enzymes and its acute toxicity in bubalus bubalis

Abstract

The acute toxic effects of quinalphos (0,0‐diethyl 0–2‐quinoxalyl phosphorothioata) uere investigated in male buffalo calves. Quinalphos was administered in single oral doses of 5, 7.5, 8.5 and 10 mg/kg body wt. and its effects on erythrocyte and plasma cholinesterases, serum aspartate aminotransferase and blood glucose were studied at various time intervals. The lowest dose (5 mg/kg) produced no apparent toxic symptoms. All the animals given highest dose (10 mg/kg) died within 60–82 hours after dosing. Quinalphos at all the dose levels markedly inhibited the erythrocyte and plasma cholinesterases (68–100%) and significantly elevated the levels of serum aspartate aminotransferase and blood glucose. Seven days after the administration of quinalphos, the blood cholinesterases in survivors remained inhibited to the extent of 41–77% whereas the levels of serum aspartate aminotransferase and blood glucose were comparable to control values.     

Effect of Isoflavones on Reproductive Performance,Testosterone Levels,Lipid Peroxidation,and Seminal Plasma Biochemistry of Male Rabbits

Abstract

The objective of this study was to determine the effect of either 2.5 mg/kg Body Weight or 5 mg/kg Body Weight (BW) doses of isoflavones on semen quality, testosterone levels, lipid peroxidation and semen biochemistry of male New Zealand White rabbits. Animals were given both 2.5 mg/kg BW and 5 mg/kg BW doses of isoflavones. The tested doses were given to rabbits orally every other day for 13 weeks. Treatment with isoflavones caused an increase (p < 0.05) in libido (by decreasing the reaction time), sperm concentration, sperm motility (%), total motile sperm per ejaculate (TMS), packed sperm volume (PSV), total functional sperm fraction (TFSF), total sperm output, initial fructose concentration and normal sperm, while dead sperm was reduced compared to control animals. On the other hand, ejaculate volume, initial hydrogen ion concentration (pH) and plasma testosterone levels did not change in treated animals with both doses of isoflavones as compared to control. Concentrations of thiobarbituric acid-reactive substances (TBARS), total lipids, and low density lipoprotein were significantly (p < 0.05) reduced in seminal plasma of rabbits treated with either 2.5 mg/kg BW or 5 mg/kg BW doses of isoflavones. While, the activities of glutathione S-transferase (GST), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), acid phosphatase (AcP), and alkaline phosphatase (AlP) were significantly (p < 0.05) increased in seminal plasma of treated animals. Also, total cholesterol, percentage cholesterol (out of total lipids), and high density lipoprotein were significantly (p < 0.05) increased, while triglyceride did not change in seminal plasma of treated animals. Supplementation at either level of isoflavones did not cause changes in live body weight (LBW), dry matter intake (DMI), and relative weights of testes and epididymis. The present results showed that either 2.5 mg/kg BW or 5 mg/kg BW doses of isoflavones caused an improvement of some semen characteristics and did not have negative effects on male fertility.     

Diminution of migration of phthalic acid esters in tequila beverage by the year of production

Abstract

The presence of diethyl-phthalate (DEP), dibutyl-phthalate (DBP), butylbenzyl-phthalate (BBP), diethylhexyl-phthalate (DEHP) and diisononyl-phthalate (DINP) was determined in 295 tequila samples. They were grouped by age of maturation (white, aged, extra aged or ultra aged) and year of production (between 2013 and 2018). Gas Chromatography coupled with Mass Spectrometry was used for identification and quantification. The results showed that 65 samples (22% of the total) were phthalate free. DEP (0.13-0.27?mg/kg), BBP (0.05–2.91?mg/kg) and DINP (1.64–3.43?mg/kg) were detected in 11 (3.73%), 37 (12.54%) and 5 (1.69%) samples, respectively. But, these concentrations did not exceed the maximum permitted limits (MPL) of phthalates for alcoholic beverages. DBP (0.01–2.20?mg/kg) and DEHP (0.03–4.64?mg/kg) were detected in 96 (32.54%) and 224 (75.93%) samples, from them only 10 (3.39%) and 15 (5.08%) samples, respectively, exceeded the MPL for alcoholic beverages and they were few tequilas produced in the year 2014 or before. DEHP was the most frequent phthalate found in tequila and observed DEHP concentrations were 2-times higher in ultra aged tequilas compared to those in white tequilas. We concluded that all tequilas produced in 2015 and after, satisfied the international standards for these compounds.     

Determination of Acaricide Residues in Saudi Arabian Honey and Beeswax Using Solid Phase Extraction and Gas Chromatography

Determination of acaricide residues of flumethrin, tau-fluvalinate, coumaphos, and amitraz in honey and beeswax was carried out using a rapid extraction method utilizing C-18 SPE cartridges and an analytical method utilizing GC with ECD, NPD, and MSD detectors for the four acaricides. Recovery percentages from the extraction method ranged from 90–102%, while the minimum detection levels ranged from 0.01–0.05 mg/kg for the acaricides. Nine of the 21 analyzed samples were found to be contaminated with the acaricides tau-fluvalinate and coumaphos. Neither flumethrin nor amitraz was detected in any of the honey or wax samples. Coumaphos was found only in honey samples in which two samples exceeded the tolerance levels set by EPA and EC regulations. It has not been detected in beeswax. Five honey samples and eight beeswax samples were found to be contaminated with tau-fluvalinate. One of the wax samples was contaminated with a relatively high residue of tau-fluvalinate and contained above 10 mg/kg.     

Determination of fumonisins in milk

Abstract

Fumonisin B₁ (FB₁) and fumonisin B₂ (FB₂) were determined in milk by liquid chromatography (LC) following immunoaffinity column cleanup. Recoveries from milk spiked with 5–50 ng each fumonisin/ml averaged 79–109%. The aminopentol hydrolysis product of FB₁ (AP₁) was determined by LC after cleanup on a C₁₈solid phase phase extraction column; mean recoveries were 69–83% at spiking levels of 50–100 ng AP₁/ml milk. Detection limits were of the order 3–7 ng/ml for FB₁ and FB₂, and 20–25 ng/ml for AP₁. A stability study showed no losses of FB₁ and FB₂ in milk under conditions of freezing, refrigeration and boiling. A transmission study using four cows dosed with pure FB₁, either orally (1.0 and 5.0 mg FB₁/kg b.w.) or by i.v. injection (0.05 and 0.20 mg FB₁/kg b.w.) showed no detectable residues of FB, or AP₁ in the milk, with or without hydroiytic treatment with β‐glucuronidase/sulfatase to liberate any conjugates.     

Pentachlorotoluene and pentabromotoluene: Results of a subacute and a subchronic toxicity study in the rat

Abstract

Pentachlorotoluene (PCT) and pentabromotoiuene (PBT) are environmental contaminants detected in the Great Lakes ecosystem. In view of the paucity of toxicity data and the potential for human exposure, a subacute (28 day) and a subchronic (91 day) study were conducted in the rat. In the subacute study, groups of 10 male and 10 female rats were fed the diet containing PCT or PBT at 0, 0.5, 5.0, 50 or 500 ppm for 28‐days. In the subchronic study, the group size was increased to 15, the dose levels were 0, 0.05, 0.5, 5.0, 50 and 500 ppm in the diet and the exposure period was 91 days. Growth rate and food consumption were not affected by exposure to either chemical in the subacute and subchronic study. Clinical observations revealed no abnormalities. Decreased hemoglobin was observed in female rats fed 5.0 ppm PCT and higher levels in the subacute (28 day) study. In the same study the hematocrit value and erythrocyte numbers of females fed 5.0 or 500 ppm PCT diets were significantly lower than the control. In both subacute and subchronic studies mild dose‐dependent histopathological changes were observed in the thyroid, liver and kidney of rats fed PCT and PBT diets. In general male rats were more susceptible than females to the treatment of PCT and PBT. Based on these data, it was concluded that the no observable adverse effect level for PCT was 50 ppm in the diet (3.5 mg/kg b.w./day) and that of PBT was 5.0 ppm (0.35 mg/kg b.w./day).     

Simultaneous analysis of thiabendazole,carbendazim and 2-aminobenzimidazole in concentrated fruit juices by liquid chromatography after a single mix-mode solid-phase extraction cleanup

A method using liquid chromatography and a single mix-mode solid-phase extraction cleanup for the simultaneous analysis of thiabendazole [2-(1,3-thiazol-4-yl)-1H-benzoimidazole], carbendazim [(methyl N-(1H-benzoimidazol-2-yl)-carbamate)] and 2-aminobenzimidazole (1H-benzimidazol-2-amine) in concentrated fruit juices is described. The three fungicides were isolated from the samples and concentrated by solid-phase extraction on Oasis MCX cartridges. The determination was performed by liquid chromatography with a diode array, detecting at 288 nm. In this method, the average recoveries from blank control juice samples spiked in a concentration range of 0.01–0.10 mg/kg were in the range of 81–95%, with relative standard deviations below 8%. Detection limits and limits of quantification were 4 μ g/kg and 10 μ g/kg, respectively, for each fungicide. Real juice samples were analyzed by the proposed method. The results were compared to those from liquid chromatography-tandem mass spectrometry.     

Chlorinated Pesticides (2,4-D and DDT) Biodegradation at High Concentrations Using Immobilized Pseudomonas Fluorescens

Abstract

Degradation of two chlorinated pesticides (2,4-D and DDT) using a 54-mL glass column packed with tezontle (a low-cost basaltic scoria) was tested. Bacteria were cultured in YPG (yeast, peptone, and glucose) liquid medium at 32°C. The rich medium was pumped during 24 h through the column to inoculate it. Later, the wasted medium was discharged and the pesticide added. Optical densities, TOC, and pesticide concentration were determined. Pesticide removals for 2,4-D (with initial concentration between 100 and 500 mg/L) were about 99%. DDT removal (at initial concentration of up to 150 mg/L) was as high as 55–99%. TOC removals for 2,4-D was in the 36-87% interval, whereas for DDT they were as high as 36–78%.     

Determination of organochlorine and organophosphate pesticide residues in fruits,vegetables and sediments

Abstract

A method is described for the determination of organochlorine and organophosphate pesticide residues in fruits, vegetables and sediments. The concentrated solvent extract was sealed in a polymeric membrane tube, dialysed in cyclohexane and the solvent replaced with hexane. The organophosphates were analysed on a specific thermionic detector without further clean‐up. For the organochlorine pesticides the extract was eluted through 3 g of alumina and analysed on GC/ECD. The clean‐up for sediment extract was carried out on a10 g alumina column with 100 mL hexane containing 5% acetone and the eluate was concentrated to 5 mL.

The detection limit for organophosphates on a 40 g sample and a final volume of 10 mL was on the average 0.01 mg/kg. The detection limit for organochlorine pesticides, with the final volume of 25 mL, was 0.005 mg/kg for all pesticides except for p,p'‐DDT and endosulfan sulphate, which was 0.01 mg/kg.

The detection limit for oganochlorine pesticides in sediment, with the final volume of 2 mL, was less than 1 μg/kg and for organophosphate pesticides less than 10 μg/kg when the final volume was made to 0.5 mL. At the detection limits the method produced a very high coefficient of variation for both organochlorine and organophosphate pesticides.     

Persistence of fensulfothion in a sandy‐loam soil and uptake by rutabagas,carrots and radishes using microplots

Abstract

Field microplots were treated with 141 and 282 ppm fensulfothion and 37.1 and 74.2 ppm fensulfothion sulfone. These concentrations are equivalent to field treatment rates of 8.48 and 16.96 kg Al/ha, fensulfothion, and 2.23 and 4.47 kg Al/ha, fensulfothion sulfone, respectively, for banded application (10 cm wide, rows 80 cm apart). The half‐lives in a sandy loam soil were 30–39 and 14–23 days, respectively. Fensulfothion sulfone and sulfide were the main derivatives found in fensulfothion treated soil.

The maximum levels of these derivatives were 21.22 and 22.95 ppm, respectively for the 8.48 kg/ha treatment and 33.90 and 42.45 ppm, respectively, for the higher treatment, which occurred between 30–60 days.

Carrots appeared to take up more fensulfothion from soil than rutabagas or radishes. The residue levels at harvest decreased in the order carrot peel > pulp > rutabagas root > peel > pulp. Residue levels of fensulfothion and sulfone in radishes were similar to those found in rutabagas. The ratio sulfoxide/sulfone in rutabagas ranged from 0.4–1.5 and in carrots from 1.7–7.6. This phenomenon is thought to be due to oxidative enzyme systems present in rutabagas. Dimethyl phosphorothioic acid, but not dimethyl phosphoric acid was detected (max. 1.33 ppm) in some rutabagas samples but not in carrots.     

Comparative tolerance of Pinus radiata and microbial activity to copper and zinc in a soil treated with metal-amended biosolids

A study was conducted to evaluate the effects of elevated concentrations of copper (Cu) and zinc (Zn) in a soil treated with biosolids previously spiked with these metals on Pinus radiata during a 312-day glasshouse pot trial. The total soil metal concentrations in the treatments were 16, 48, 146 and 232 mg Cu/kg or 36, 141, 430 and 668 mg Zn/kg. Increased total soil Cu concentration increased the soil solution Cu concentration (0.03–0.54 mg/L) but had no effect on leaf and root dry matter production. Increased total soil Zn concentration also increased the soil solution Zn concentration (0.9–362 mg/L). Decreased leaf and root dry matter were recorded above the total soil Zn concentration of 141 mg/kg (soil solution Zn concentration, >4.4 mg/L). A lower percentage of Cu in the soil soluble?+?exchangeable fraction (5–12 %) and lower Cu²⁺ concentration in soil solution (0.001–0.06 μM) relative to Zn (soil soluble?+?exchangeable fraction, 12–66 %; soil solution Zn²⁺ concentration, 4.5–4,419 μM) indicated lower bioavailability of Cu. Soil dehydrogenase activity decreased with every successive level of Cu and Zn applied, but the reduction was higher for Zn than for Cu addition. Dehydrogenase activity was reduced by 40 % (EC₄₀) at the total solution-phase and solid-phase soluble?+?exchangeable Cu concentrations of 0.5 mg/L and 14.5 mg/kg, respectively. For Zn the corresponding EC₅₀ were 9 mg/L and 55 mg/kg, respectively. Based on our findings, we propose that current New Zealand soil guidelines values for Cu and Zn (100 mg/kg for Cu; 300 mg/kg for Zn) should be revised downwards based on apparent toxicity to soil biological activity (Cu and Zn) and radiata pine (Zn only) at the threshold concentration.     

Contents and Leachability of Heavy Metals (Pb,Cu, Sb,Zn, As) in Soil at the Pantex Firing Range,Amarillo, Texas

ABSTRACT

Pantex firing range soil samples were analyzed for Pb, Cu, Sb, Zn, and As. One hundred ninety-seven samples were collected from the firing range and vicinity area. There was a lack of knowledge about the distribution of Pb in the firing range, so a random sampling with proportional allocation was chosen. Concentration levels of Pb and Cu in the firing range were found to be in the range of 11-4675 and 13-359 mg/kg, respectively. Concentration levels of Sb were found to be in the range of 1-517 mg/kg. However, the Zn and As concentration levels were close to average soil background levels. The Sn concentration level was expected to be higher in the Pantex firing range soil samples. However, it was found to be below the neutron activation analysis (NAA) detection limit of 75 mg/kg.

Enrichment factor analysis showed that Pb and Sb were highly enriched in the firing range with average magnitudes of 55 and 90, respectively. Cu was enriched ~6 times more than the usual soil concentration levels. Tox-icity characteristic leaching procedure (TCLP) was carried out on size-fractionated homogeneous soil samples. The concentration levels of Pb in leachates were found to be ~12 times higher than the U.S. Environmental Protection Agency (EPA) regulatory concentration level of 5 mg/L. Sequential extraction (SE) was also performed to characterize Pb and other trace elements into five different fractions. The highest Pb fraction was found with organic matter in the soil.