Improving plant bioaccumulation science through consistent reporting of experimental data.

Experimental data and models for plant bioaccumulation of organic contaminants play a crucial role for assessing the potential human and ecological risks associated with chemical use. Plants are receptor organisms and direct or indirect vectors for chemical exposures to all other organisms. As new experimental data are generated they are used to improve our understanding of plant-chemical interactions that in turn allows for the development of better scientific knowledge and conceptual and predictive models. The interrelationship between experimental data and model development is an ongoing, never-ending process needed to advance our ability to provide reliable quality information that can be used in various contexts including regulatory risk assessment. However, relatively few standard experimental protocols for generating plant bioaccumulation data are currently available and because of inconsistent data collection and reporting requirements, the information generated is often less useful than it could be for direct applications in chemical assessments and for model development and refinement. We review existing testing guidelines, common data reporting practices, and provide recommendations for revising testing guidelines and reporting requirements to improve bioaccumulation knowledge and models. This analysis provides a list of experimental parameters that will help to develop high quality datasets and support modeling tools for assessing bioaccumulation of organic chemicals in plants and ultimately addressing uncertainty in ecological and human health risk assessments.

Comparison of Accelerated Solvent Extraction (ASE) and Energized Dispersive Guided Extraction (EDGE) for the analysis of pesticides in leaves.

Various techniques have been evaluated for the extraction and cleanup of pesticides from environmental samples. In this work, a Selective Pressurized Liquid Extraction (SPLE) method for pesticides was developed using a Thermo Fisher Scientific Accelerated Solvent Extraction (ASE) system. This instrument was compared to the newly introduced (2017) extraction instrument, the Energized Dispersive Guided Extraction (EDGE) system, which combines Pressurized Liquid Extraction (PLE) and dispersive Solid Phase Extraction (dSPE). We first optimized the SPLE method using the ASE instrument for pesticide extraction from alfalfa leaves using layers of Florisil and graphitized carbon black (GCB) downstream of the leaf homogenate in the extraction cell (Layered ASE method). We then compared results obtained for alfalfa and citrus leaves with the Layered ASE method to those from a method in which the leaf homogenate and sorbents were mixed (Mixed ASE method) and to similar methods modified for use with EDGE (Layered EDGE and Mixed EDGE methods). The ASE and EDGE methods led to clear, colorless extracts with low residual lipid weight. No significant differences in residual lipid masses were observed between the methods. The UV-Vis spectra showed that Florisil removed a significant quantity of the light-absorbing chemicals, but that GCB was required to produce colorless extracts. Recoveries of spiked analytes into leaf homogenates were generally similar among methods, but in several cases, significantly higher recoveries were observed in ASE extracts. Nonetheless, no significant differences were observed among pesticide concentrations in field samples when calculated with the isotope dilution method in which labelled surrogates were added to samples before extraction. The extraction time with the ASE methods was ~45 minutes, which was ~4.5 times longer than with the EDGE methods. The EDGE methods used ~10 mL more solvent than the ASE methods. Based on these results, the EDGE is an acceptable extraction instrument and, for most compounds, the EDGE had a similar extraction efficiency to the ASE methods.

A review of measured bioaccumulation data on terrestrial plants for organic chemicals: Metrics, variability, and the need for standardized measurement protocols.

Quantifying the transfer of organic chemicals from the environment into terrestrial plants is essential for assessing human and ecological risks, using plants as environmental contamination biomonitors, and predicting phytoremediation effectiveness. Experimental data describing chemical uptake by plants are often expressed as ratios of chemical concentrations in the plant compartments of interest (e.g., leaves, shoots, roots, xylem sap) to those in the exposure medium (e.g., soil, soil porewater, hydroponic solution, air). These ratios are generally referred to as "bioconcentration factors" but have also been named for the specific plant compartment sampled, such as "root concentration factors," "leaf concentration factors," or "transpiration stream (xylem sap) concentrations factors." We reviewed over 350 articles to develop a database with 7049 entries of measured bioaccumulation data for 310 organic chemicals and 112 terrestrial plant species. Various experimental approaches have been used; therefore, interstudy comparisons and data-quality evaluations are difficult. Key exposure and plant growth conditions were often missing, and units were often unclear or not reported. The lack of comparable high-confidence data also limits model evaluation and development. Standard test protocols or, at a minimum, standard reporting guidelines for the measurement of plant uptake data are recommended to generate comparable, high-quality data that will improve mechanistic understanding of organic chemical uptake by plants. Environ Toxicol Chem 2018;37:21-33. © 2017 SETAC.

Temperature effect on tert-butyl alcohol (TBA) biodegradation kinetics in hyporheic zone soils.

BACKGROUND: Remediation of tert-butyl alcohol (TBA) in subsurface waters should be taken into consideration at reformulated gasoline contaminated sites since it is a biodegradation intermediate of methyl tert-butyl ether (MTBE), ethyl tert-butyl ether (ETBE), and tert-butyl formate (TBF). The effect of temperature on TBA biodegradation has not been not been published in the literature. METHODS: Biodegradation of [U 14C] TBA was determined using hyporheic zone soil microcosms. RESULTS: First order mineralization rate constants of TBA at 5 degrees C, 15 degrees C and 25 degrees C were 7.84 +/- 0.14 x 10-3, 9.07 +/- 0.09 x 10-3, and 15.3 +/- 0.3 x 10-3 days-1, respectively (or 2.86 +/- 0.05, 3.31 +/- 0.03, 5.60 +/- 0.14 years-1, respectively). Temperature had a statistically significant effect on the mineralization rates and was modelled using the Arrhenius equation with frequency factor (A) and activation energy (Ea) of 154 day-1 and 23,006 mol/J, respectively. CONCLUSION: Results of this study are the first to determine mineralization rates of TBA for different temperatures. The kinetic rates determined in this study can be used in groundwater fate and transport modelling of TBA at the Ronan, MT site and provide an estimate for TBA removal at other similar shallow aquifer sites and hyporheic zones as a function of seasonal change in temperature.

Mineralization and plant uptake of 14C-labeled nonylphenol, nonylphenol tetraethoxylate, and nonylphenol nonylethoxylate in biosolids/soil systems planted with crested wheatgrass.

Microcosm experiments (duration, 150 d) were conducted to evaluate the mineralization and plant uptake of [14C]nonylphenol (NP), [14C]nonylphenol tetraethoxylate (NPE4), and [14C]nonylphenol nonylethoxylate (NPE9) in a soil/biosolids (99.5:0.5 w/w) environment planted with crested wheatgrass (Agropyron cristatum). Three initial nominal concentrations (6, 24, and 47 mg/kg dry wt) each of NP, NPE4, and NPE9 were examined along with unplanted and unplanted poisoned controls. Phenol (22 mg/kg) also was evaluated as a more degradable reference compound. The biosolids were obtained from a municipal treatment plant, and the loamy sand soil was freshly collected. Mineralization ranged from 7% for NP to 53% for phenol, and no enhancement was observed in the planted systems. For NP, NPE4, and NPE9, 14C foliar tissues concentrations were proportional to exposure concentrations but were 10-fold lower than the root concentrations and two- to threefold lower than the soil concentrations. Bioconcentration factors (BCFs) based on 14C measurements ranged from 0.31 (mg compound/kg dry plant/ mg compound/kg dry soil) for systems spiked with NP to 0.52 for systems spiked with NPE9. Results of the NP analysis (initial concentration, 47 mg/ kg) showed a 90% decrease in the soil concentration and an average BCF of 1.0. The lower BCF calculated from the 14C analysis likely resulted from the presence of NP transformation products in the soil that are less available or are translocated by the plants but quantified by the combustion/liquid scintillation counting procedure.

The sky is falling III: The effect of deposition from static solid rocket motor tests on juvenile crops.

A mixture of combustion products (mainly hydrogen chloride, aluminum oxide, and water) and entrained soil, referred to as Test Fire Soil (TFS), can be deposited on crops during static solid rocket motor tests. The impact of a reported worst-case event was previously evaluated by exposing corn and alfalfa to 3200-gTFS/m2 at 54days after emergence. Exposures via soil and leaves were evaluated separately. Reduced growth (soil exposure) and leaf "scorch" (leaf exposure) were attributed mainly to the high chloride concentrations in the TFS (56,000mg/kg). A follow-up study was conducted to evaluate the effect of a typical deposition event (70-gTFS/m2, estimated by radar during several tests) and exposure (soil and leaves simultaneously) on juvenile corn, alfalfa, and winter wheat. Younger crops were used to examine potential age sensitivity differences. Impact was evaluated by comparing the growth, elemental composition, and leaf chlorophyll content of treated and untreated plants. The relationship between deposition exposure and response was also addressed. Growth of corn, alfalfa, and winter wheat exposed to a typical TFS loading was not impacted, although slightly elevated concentrations of aluminum and iron were found in the leaves. At the highest loadings used for the exposure-response experiment, concentrations of chloride and calcium were higher in TFS-exposed corn leaves than in the untreated leaves. Overall results indicate that exposure to a typical deposition event does not adversely impact juvenile crops and that younger plants may be less vulnerable to TFS. However, higher TFS loadings can cause leaf scorch and increase the leaf concentrations of some elements.

Review of existing terrestrial bioaccumulation models and terrestrial bioaccumulation modeling needs for organic chemicals.

Protocols for terrestrial bioaccumulation assessments are far less-developed than for aquatic systems. This article reviews modeling approaches that can be used to assess the terrestrial bioaccumulation potential of commercial organic chemicals. Models exist for plant, invertebrate, mammal, and avian species and for entire terrestrial food webs, including some that consider spatial factors. Limitations and gaps in terrestrial bioaccumulation modeling include the lack of QSARs for biotransformation and dietary assimilation efficiencies for terrestrial species; the lack of models and QSARs for important terrestrial species such as insects, amphibians and reptiles; the lack of standardized testing protocols for plants with limited development of plant models; and the limited chemical domain of existing bioaccumulation models and QSARs (e.g., primarily applicable to nonionic organic chemicals). There is an urgent need for high-quality field data sets for validating models and assessing their performance. There is a need to improve coordination among laboratory, field, and modeling efforts on bioaccumulative substances in order to improve the state of the science for challenging substances.

Uptake of nonylphenol and nonylphenol ethoxylates by crested wheatgrass.

Nonylphenol (NP) and other hydrophobic biodegradation intermediates of nonylphenol ethoxylate (NPE) surfactants have been identified in wastewater treatment biosolids. These biosolids often are land applied, but little is known regarding the potential uptake of biosolid-derived contaminants by plants. Hydroponic experiments, 11 to 14 weeks in duration, were conducted to examine the uptake and translocation of 14C and unlabeled NP, nonylphenol tetraethoxylate (NPE4), and nonylphenol nonylethoxylate (NPE9) by crested wheatgrass (Agropyron cristatum). Phenol also was evaluated for comparison. Plant tissue was analyzed for 14C and for the parent compounds. Volatilization from the hydroponic system and rhizosphere mineralization also were quantified. At the conclusion of the study, most of the plant-associated 14C was found in the roots (NP = 98%, NPE4 = 92%, and NPE9 = 81%). Concentrations of 14C in the foliar tissue ranged from 0.002 to 0.045 mg-equivalent per kg (dry wt), but no parent compounds were detected, implying that the 14C was unextractable or in the form of metabolites. Transpiration stream concentration factors for NP, NPE4, and NPE9, calculated assuming the 14C was parent compound, were 0.012, 0.032, and 0.066, respectively. Little mineralization was observed for NP, NPE4, and NPE9 in the hydroponic system; however, for phenol, 16 to 30% of the added 14C was mineralized.

Plant leaves as indoor air passive samplers for volatile organic compounds (VOCs).

Volatile organic compounds (VOCs) enter indoor environments through internal and external sources. Indoor air concentrations of VOCs vary greatly but are generally higher than outdoors. Plants have been promoted as indoor air purifiers for decades, but reports of their effectiveness differ. However, while air-purifying applications may be questionable, the waxy cuticle coating on leaves may provide a simple, cost-effective approach to sampling indoor air for VOCs. To investigate the potential use of plants as indoor air VOC samplers, a static headspace approach was used to examine the relationship between leaf and air concentrations, leaf lipid contents and octanol-air partition coefficients (Koa) for six VOCs and four plant species. The relationship between leaf and air concentrations was further examined in an actual residence after the introduction of several chlorinated VOC emission sources. Leaf-air concentration factors (LACFs), calculated from linear regressions of the laboratory headspace data, were found to increase as the solvent extractable leaf lipid content and Koa value of the VOC increased. In the studies conducted in the residence, leaf concentrations paralleled the changing air concentrations, indicating a relatively rapid air to leaf VOC exchange. Overall, the data from the laboratory and residential studies illustrate the potential for plant leaves to be used as cost effective, real-time indoor air VOC samplers.

Investigating differences in the root to shoot transfer and xylem sap solubility of organic compounds between zucchini, squash and soybean using a pressure chamber method.

A pressure chamber method was used to examine differences in the root to shoot transfer and xylem sap solubility of caffeine (log Kow=-0.07), triclocarban (log Kow=3.5-4.2) and endosulfan (log Kow=3.8-4.8) for zucchini (cucurbita pepo ssp pepo), squash (cucurbita pepo ssp ovifera), and soybean (glycine max L.). Transpiration stream concentration factors (TSCF) for caffeine (TSCF=0.8) were statistically equivalent for all plant species. However, for the more hydrophobic endosulfan and triclocarban, the TSCF values for zucchini (TSCF=0.6 and 0.4, respectively) were 3 and 10 times greater than the soybean and squash (TSCF=0.2 and 0.05, respectively). The difference in TSCF values was examined by comparing the measured solubilities of caffeine, endosulfan and triclocarban in deionized water to those in soybean and zucchini xylem saps using a modified shake flask method. The measured solubility of organic contaminants in xylem sap has not previously been reported. Caffeine solubilities in the xylem saps of soybean and zucchini were statistically equal to deionized water (21500mgL(-1)) while endosulfan and triclocarban solubilities in the zucchini xylem sap were significantly greater (0.43 and 0.21mgL(-1), respectively) than that of the soybean xylem sap (0.31 and 0.11mgL(-1), respectively) and deionized water (0.34 and 0.11mgL(-1), respectively). This suggests that the enhanced root to shoot transfer of hydrophobic organics reported for zucchini is partly due to increased solubility in the xylem sap. Further xylem sap characterization is needed to determine the mechanism of solubility enhancement.

Quantitative structure-activity relationships for predicting soil-sediment sorption coefficients for organic chemicals.

Sorption coefficients are used to describe the equilibrium distribution of a chemical between a soil or sediment and the aqueous phase that it is in contact with. Although sorption coefficients for a particular organic chemical vary greatly from soil to soil, the observation has been made that sorption generally increases as the organic carbon content of the soil and the hydrophobicity of the chemical increases. This general observation resulted in the acceptance of organic carbon normalized sorption coefficients (KOC) as unique properties or constants of organic chemicals. In turn, KOC values have been estimated by quantitative structure-activity relationships (QSARs) developed by correlation with a variety of physical or chemical properties and structural descriptors related to the hydrophobicity of the chemical such as octanol-water partition coefficients, aqueous solubilities, molecularconnectivity indices, molecular weight, molecular surface area, and reverse-phase high-performance liquid chromatography retention times. The selection and application of the most appropriate QSAR for predicting KOC depend on several factors, including the availability of required input, the appropriateness of model to chemical of interest, and the methodology for calculating the necessary topological or structural information. A review of the existing QSARs for predicting KOC and the limitations of using the KOC approach to estimate sorption coefficients will be presented.

Tracer studies for evaluation of in situ air sparging and in-well aeration system performance at a gasoline-contaminated site.

Field-scale tracer studies were conducted at a gasoline-contaminated site in order to evaluate the effectiveness of in situ air sparging (IAS) and in-well aeration (IWA) in controlling the movement of soil gas and groundwater in the subsurface. The field site was comprised of silty sand (SM) and silty clay (CL), underlain by a clay layer at approximately 7.6 m. Depth to groundwater ranged from 2.4 to 3 m. Soil permeability and the natural hydraulic gradient were both low. Helium was used to trace the movement of soil gas in the unsaturated zone during the IAS field study, and successfully confirmed short-circuit pathways for injected air and demonstrated the limited distribution of injected gases at this site. Fluorescein, bromide, and rhodamine were used to trace the movement of groundwater during the IWA system field study, and successfully documented the inability of the IWA system to recirculate enough groundwater to enhance subsurface dissolved oxygen levels or to remediate groundwater by air stripping at this site. The inability of the systems to remediate the site was likely due to site conditions which consist of low-permeability soils and decreasing permeability with depth. As a result, relatively impermeable layers exist at the depth of the IAS screen and the lower IWA screen. These site conditions are not conducive to successful performance of either remediation system.

The sky is falling II: Impact of deposition produced during the static testing of solid rocket motors on corn and alfalfa.

Tests of horizontally restrained rocket motors at the ATK facility in Promontory, Utah, USA result in the deposition of an estimated 1.5million kg of entrained soil and combustion products (mainly aluminum oxide, gaseous hydrogen chloride and water) on the surrounding area. The deposition is referred to as test fire soil (TFS). Farmers observing TFS deposited on their crops expressed concerns regarding the impact of this material. To address these concerns, we exposed corn and alfalfa to TFS collected during a September 2009 test. The impact was evaluated by comparing the growth and tissue composition of controls relative to the treatments. Exposure to TFS, containing elevated levels of chloride (1000 times) and aluminum (2 times) relative to native soils, affected the germination, growth and tissue concentrations of various elements, depending on the type and level of exposure. Germination was inhibited by high concentrations of TFS in soil, but the impact was reduced if the TFS was pre-leached with water. Biomass production was reduced in the TFS amended soils and corn grown in TFS amended soils did not develop kernels. Chloride concentrations in corn and alfalfa grown in TFS amended soils were two orders of magnitude greater than controls. TFS exposed plants contained higher concentrations of several cations, although the concentrations were well below livestock feed recommendations. Foliar applications of TFS had no impact on biomass, but some differences in the elemental composition of leaves relative to controls were observed. Washing the TFS off the leaves lessened the impact. Results indicate that the TFS deposition could have an effect, depending on the amount and growth stage of the crops, but the impact could be mitigated with rainfall or the application of additional irrigation water. The high level of chloride associated with the TFS is the main cause of the observed impacts.

Root-to-Shoot transfer and distribution of endosulfan in the wetland macrophyte Bidens laevis L.

Endosulfan is genotoxic in somatic cells of Bidens laevis, and reproduction could be affected if translocated from roots to flower buds. Hydroponic experiments were conducted to quantify this transfer. While the root uptake of [(14) C] endosulfan and its transfer to aboveground tissues was relatively low, the resulting average flower bud concentration (1.01 ± 0.76 ng/g) after 30 d of exposure to an aqueous concentration of 5 µg/L could still represent a genotoxic risk for germ cells.

The sky is falling: chemical characterization and corrosion evaluation of deposition produced during the static testing of solid rocket motors.

Static tests of horizontally restrained rocket motors at the ATK facility in Promontory UT, USA result in the deposition of entrained soil and fuel combustion products, referred to as Test Fire Soil (TFS), over areas as large as 30-50 mile (80-130 km) and at distances up to 10-12 miles (16-20 km) from the test site. Chloride is the main combustion product generated from the ammonium perchlorate-aluminum based composite propellant. Deposition sampling/characterization and a 6-month field corrosivity study using mild steel coupons were conducted in conjunction with the February 25th 2010 FSM-17 static test. The TFS deposition rates at the three study sites ranged from 1 to 5 g/min/m. TFS contained significantly more chloride than the surface soil collected from the test site. The TFS collected during two subsequent tests had similarly elevated chloride, suggesting that the results obtained in this study are applicable to other tests assuming that the rocket fuel composition remains similar. The field-deployed coupons exposed to the TFS had higher corrosion rates (3.6-5.0 mpy) than paired non-exposed coupons (1.6-1.8 mpy). Corrosion rates for all coupons decreased over time, but coupons exposed to the TFS always had a higher rate than the non-exposed. Differences in corrosion rates between the three study sites were also observed, with sites receiving more TFS deposition having higher corrosion rates.

Chemical hydrophobicity and uptake by plant roots.

The transpiration stream concentration factor (TSCF), the ratio between a compound's concentration in the xylem to that in the solution adjacent to the roots, is commonly used to describe the relative ability of an organic compound to be passively transported from root to shoot. Widely cited bell-shaped curves relating TSCFto the octanol/water partition coefficient (log Kow) imply that significant root uptake and transfer into shoot tissues occurs only for compounds falling within an intermediate hydrophobicity range. However, recent laboratory and field data for relatively water soluble compounds such as sulfolane, methyl tert-butyl ether (MTBE), and 1,4-dioxane suggest that these relationships are not universally applicable, especiallyfor nonionizable, highly polar, water soluble organics. To re-evaluate the relationship between root uptake and chemical hydrophobicity, TSCFs were measured for 25 organic chemicals ranging in log Kow from -0.8 to 5 using a pressure chamber technique. Using the TSCF values measured in this study, a new empirical relationship between TSCF (0 and 1) and log Kow (-0.8 to 5) is presented that indicates that nonionizable, polar, highly water soluble organic compounds are most likely to be taken up by plant roots and translocated to shoot tissue.

Trichloroethylene uptake into fruits and vegetables: three-year field monitoring study.

Trichloroethylene (TCE) contaminated groundwater migrating into communities surrounding Hill Air Force Base (HAFB) in northern Utah prompted a multiyear monitoring program (2001-2003) to examine the extent of TCE uptake and transfer into edible fruits. During the initial sampling in fall 2001, TCE was detected in a small fraction of the 167 fruit and tree core samples collected from 17 private residences. Samples were analyzed using headspace gas chromatography (GC) with electron capture detection (ECD) with limited confirmation by mass spectrometry (MS) in selected ion monitoring mode. In fall 2002, over 300 samples were collected from the same general locations sampled in 2001. No TCE was found in any of the fruit or vegetable samples above the method detection limit (MDL) for the headspace GC/MS method (approximately 0.1 microg/ kg fresh weight, depending on sample size), but TCE was again detected in several fruit tree trunk core samples. The detection of TCE in fruit in 2001, but not in 2002, may have been due to improvements in the analytical procedure or changes in the environmental conditions impacting transfer to fruit. The 2003 monitoring focused on repeated sampling over several months at five locations that were selected to represent the range of exposure scenarios evaluated during the previous years. No TCE was identified in any of the fruit above the MDL during 2003, however TCE was again found in tree core samples as observed in 2001 and 2002.

Trichloroethylene uptake by apple and peach trees and transfer to fruit.

A greenhouse study was conducted to quantify 14C-trichloroethylene (TCE) uptake and transfer into the edible fruit of apple and peach trees. Trees were subsurface irrigated with solutions of 14C [TCE] that bracketed groundwater concentrations (5 and 500 microg/L) found in residential areas surrounding Hill Air Force Base, UT, where trace amounts of TCE had been found in several fruits during a preliminary field survey. Nondosed control trees were grown within the canopy of the dosed trees and in a separate greenhouse. Tissue samples were analyzed for 14C and TCE using combustion/liquid scintillation counting (LSC) and headspace/gas chromatography/mass spectrometry (HS/GC/MS). Tissue was also extracted and analyzed by GC/MS for dichloroacetic acid (DCAA), trichloroacetic acid (TCAA), and trichloroethanol (TCEt), three specific TCE metabolites that have been previously identified in laboratory and field studies. No 14C was detected in the nonexposed control trees. Exposed trees contained levels of 14C that were proportional to the exposure concentration. 14C concentrations were greatest in leaves followed by branches and fruits. At the end of the study, TCE was detected only in roots implying that the 14C in the leaves, branches, and fruit was associated with unidentified nonvolatile TCE transformation products and/or is nonextractable. However, TCAA and DCAA were positively identified only in leaves collected during the first year from an apple tree exposed to the high dose treatment. Additional data for other chemicals and fruittrees are needed to better understand the potential transfer of organic compounds to edible fruit.

Determination of alkanolamines in cattails (Typha latifolia) utilizing electrospray ionization with selected reaction monitoring and ion-exchange chromatography.

Selected reaction monitoring (SRM) with electrospray ionization was used as a specific detection technique for the analysis of alkanolamines in plant tissue extracts. Ion-exchange chromatography was used as the method of separation. Quantification was based on monitoring the loss of either H2O or 2(H2O) from the protonated molecule [M+H]+. The method provided increased selectivity for all analytes and better detection limits for three of the six analytes investigated compared with an earlier method using selected ion monitoring with liquid chromatography. Instrumental detection limits ranged from 6-300 pg injected for monoethanolamine (MEA), monoisopropanolamine (MIPA), diethanolamine (DEA), methyldiethanolamine (MDEA), diisopropanolamine (DIPA), and triethanolamine (TEA). Method robustness and selectivity were demonstrated by the determination of DIPA and a known transformation product MIPA in over 35 plant extract samples derived from a laboratory study of plant uptake mechanisms.