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Environ. Sci. Technol.

2006, 40, 45-52

Plant Uptake of Non-Ionic Organic Chemicals

C H R I S C O L L I N S , * , M I K E F R Y E R , A N D
Department Of Environmental Science and Technology, Imperial College, South
Kensington, London, UK SW7 2BP, and Environment Agency of England and
Wales, Howbery Park, Wallingford, Oxfordshire, UK, OX10 8BD

Plant uptake of organic chemicals is an important

process when considering the risks associated with land
contamination, the role of vegetation in the global cycling of
persistent organic pollutants, and the potential for
industrial discharges to contaminate the food chain.
There have been some significant advances in our
understanding of the processes of plant uptake of organic
chemicals in recent years; most notably there is now a
better understanding of the air to plant transfer pathway,
which may be significant for a number of industrial
chemicals. This review identifies the key processes involved
in the plant uptake of organic chemicals including those
for which there is currently little information, e.g., plant lipid
content and plant metabolism. One of the principal
findings is that although a number of predictive models
exist using established relationships, these require further
validation if they are to be considered sufficiently robust
for the purposes of contaminated land risk assessment or
for prediction of the global cycling of persistent organic
pollutants. Finally, a number of processes are identified which
should be the focus of future research.

It is widely recognized that plants can become contaminated
with a range of toxic organic industrial chemicals, e.g., PCBs,
PAHs, explosives, and dioxins (1-7). To this end plant uptake
submodels have been developed within larger exposure
assessment models for the determination of risks posed by
the development of sites which contain toxic organic
chemicals, e.g., CSOIL (RIVM, Netherlands, see (8)), CLEA
(DEFRA and Environment Agency, U.K., see (9)), and CALTOX
(California Department of Toxic Control, U.S., see (10)). Plant
uptake and storage is also a potentially key component in
the global cycling of persistent organic pollutants (11-14).
In recent years our understanding of the uptake of organic
chemicals by vegetation has increased considerably; most
notably the process of plant contamination following aerial
deposition (15-17). However, the most widely quoted review
for the plant uptake of organic chemicals (16) is now over
16 years old. Ryan et al. (18) proposed a screening framework
to identify those chemicals which were most likely to
contaminate vegetation rather than a process driven model.
The current review considers the plant uptake of organic
chemicals and identifies the key processes involved with the
emphasis on those where there have been significant
developments over the last 16 years. An additional aim of the
* Corresponding author e-mail: c.collins@imperial.ac.uk; tel: +44
207 594 7378; fax: +44 207 594 2246.
Imperial College.
Environment Agency of England and Wales.
10.1021/es0508166 CCC: $33.50
Published on Web 11/17/2005

2006 American Chemical Society

review is to provide information to aid the construction of

plant uptake models and improve those that already exist.

Plant Uptake Pathways

The uptake of organic chemicals by plants occurs via a
number of pathways. These are demonstrated in Figure 1.
Passive and Active Uptake from Soil into Plant Roots.
Organic chemicals can be taken up by plant roots via the
vapor or water phases of soil. The uptake of anthropogenic
organic chemicals by plant roots has been shown to be a
passive, diffusive process, with the exception of a few
hormone-like chemicals such as the phenoxy acid herbicides,
for which there is some evidence of active uptake (19).
Experiments involving the uptake of nonionized chemicals
from hydroponic solution into plant roots have demonstrated
that the uptake process consists of two components: (1)
equilibration of the aqueous phase in the plant root with
the concentration in the surrounding solution; and (2)
sorption of the chemical onto lipophilic root solids (20).
Lipophilic root solids include lipids in membranes and cell
walls (21).
Polycyclic aromatic hydrocarbons (PAHs), chlorobenzenes, polychlorinated biphenyls (PCBs), and polychlorinated
dibenzo-p-dioxins/dibenzofurans (PCDD/Fs) have all been
found to accumulate significantly in plant roots (22). Briggs
et al. determined a linear relationship between the octanolwater partition coefficient (KOW) of nonionized chemicals
and the observed root concentration factor (RCF ) chemical
concentration in the root/concentration in external solution)
from their experiments investigating the uptake of Omethylcarbamoyloximes and substituted phenylureas into
barley plants. Lipophilic organic chemicals possess a greater
tendency to partition into plant root lipids than hydrophilic
chemicals. Wild and Jones (23) categorized nonionized
organic chemicals with log KOW > 4 as having a high potential
for retention in plant roots. Schwab et al. (24) reported that
uptake of naphthalene from solution followed a Freundlich
isotherm, the gradient of which was less than one in the
roots of vegetative plants indicating that they had become
saturated with naphthalene, but was not significantly different
from one for older roots indicating that they had not become
saturated. Therefore, looking at root uptake purely as a
partitioning process may be incorrect as this assumes it is
independent of concentration, which it is not if the roots can
become saturated. The plant aqueous phase as distinct from
the lipid phase is only thought to represent an important
storage compartment for organic chemicals with log KOW <
2, and a dimensionless Henrys law constant (KAW) < 100
In addition to the physicochemical properties of the
contaminant, the composition of the plant roots is likely to
influence root uptake behavior. Trapp and Pussemeir (26)
found that the relationship derived by Briggs et al. (20)
overestimated experimentally derived RCFs for carbamates


FIGURE 1. Principal uptake pathways for the uptake of organic chemicals by plants.
in common bean plants. A higher content of solids of a lipidlike nature in barley (Hordeum vulgare) roots compared with
bean (Vicia faba) roots has been advanced as a probable
explanation for this difference (19). Schwab et al. (24) also
found quantitative differences between the uptake of naphthalene in plant roots dependent on their lipid content. There
is, however, very little information readily available for the
lipid contents of plant compartments for different plant
species (see below). The retention in root lipid materials has
been further illustrated by the higher retention of organic
compounds in the peels of carrot and potato compared to
their pulp (27, 28).
Transport from the Root to the Shoot. Water and solutes
are transported upward from the root into other plant parts
through the xylem. This flux is driven by the water potential
gradient, created throughout the plant during transpiration.
For chemicals taken into plant roots to reach the xylem, they
must penetrate a number of layers: the epidermis, cortex,
endodermis, and pericycle. At the endodermis all materials
must pass through at least one cell membrane. It is the
combination of the solubility of chemicals in water and the
solubility within the cell membrane, which is rich in lipids,
that determines the movement of chemicals into roots and
subsequent transport to the plant shoot (29).
Briggs et al. (30) derived a relationship for predicting
concentrations of chemicals in the transpiration stream of
the plant from the concentration in soil solution and the KOW
of the chemical. This relationship was based on experiments
investigating the uptake of a limited number of nonionized
chemicals into barley plants. It was found that the transpiration stream concentration factor (TSCF ) concentration in
xylem/concentration in external solution) was at a maximum
for chemicals with log KOW ca. 1.8. This relationship has been
widely used in both plant modeling (18, 31) and also by some
authors to explain their results (6). Hsu et al. (32) found a
similar relationship for cinmethylin and related compounds
in soybean (Glycine max) plants, as did Burken and Schnoor
(33) using a range of industrial organic pollutants with hybrid
poplar (Populus deltoides nigra, DN34) (Figure 2). It can
be seen from Figure 2. that there are potentially large
differences in the TSCF at high (>4) and low log KOW (<1)
depending on which relationship is used.
The reasons for the KOW optima are not fully understood,
but it appears that more polar chemicals are less able to
cross hydrophobic lipid membranes, while, for reasons


FIGURE 2. Variation in the prediction of the transpiration stream

concentration factor with KOW (17, 26, 27).
uncertain, more lipophilic chemicals also cross the endodermis less efficiently, possibly because they become retained
in the lipid material there (19). Mono- and dichlorophenols
which have a range of log Kow 2.15-3.62 have been demonstrated to possess a high potential for root uptake and
translocation (22); these findings would be predicted by the
relationships for TSCF outlined by Briggs et al. (30), Hsu et
al. (32), and Burken and Schnoor (33). Although a bell-shaped
relationship is reported by all these workers, recent evidence
for highly soluble compounds such as sulfolane and HMX,
log KOW -0.77 and 0.13, respectively, indicates that they may
have TSCF approaching unity (6, 34).
Water and solutes transported in the xylem may diffuse
laterally into adjacent tissues. Chemicals may therefore
become concentrated in plant shoots as a result of the
following: (1) equilibration of the aqueous phase in the plant
shoots with the chemical in the xylem, and (2) sorption of
the chemical onto lipophilic shoot solids (29). Partitioning
to plant shoots has been demonstrated to be linearly related
to KOW for nonionized chemicals (20, 35). Important properties of the plant that may influence chemical concentrations
in the shoots, resulting from root uptake and translocation,
include the lipophilic solids content and the plant transpiration streamflow rate. There is potential for sorption to stem
components as the chemical moves up the stem; this sorption
increases with the increasing lipophilicity, i.e., KOW, of the
chemical (20, 36, 37). This would indicate that the chemical

concentration will be dependent on position in the plant

shoot, and using a single value derived from the TSCF may
possibly overestimate the concentration in plant organs
where transport up the stem is required, e.g., fruits. Additionally the TSCF has been demonstrated to increase over
a short time scale, up to 96 h, with low KOW chemicals reaching
equilibrium more quickly (20).
Hulster et al. (1) reported that courgette and pumpkin
plants accumulate and translocate higher concentrations of
PCDD/Fs from contaminated soil than other fruits and
vegetables and that this is the main contamination pathway
for these species. The authors postulate that this may be due
to crop-specific root exudates which mobilize PCDD/Fs from
the soil and make these compounds available for uptake and
translocation. An additional characteristic of certain plant
species that may influence how they accumulate organic
chemicals from surrounding soil is the presence of oil
channels within roots and shoots. Uptake and translocation
of organic chemicals in these channels has been demonstrated for some oil-containing plants such as carrots (38).
Experimental results have shown that carrots possess a greater
potential for the uptake of nonionized chemicals than other
crop species (39-41). In addition to this inter-species
variation, White et al. (42) also found considerable intraspecies variation in the uptake of DDE in zucchini (Curcubita
Transfer from Soil to Soil Water and Soil Air. The uptake
of organic chemicals by plants is profoundly influenced by
the soil. There are many papers on the partitioning of organic
chemicals from soil to soil solution, a detailed description
of which is beyond the scope of this review. A brief description
is given for information. The concentration in the soil water
of a contaminant is described by the Kd which is the partition
distribution coefficient between the soil and soil water. Nonionized organic pollutants are principally sorbed onto the
organic fraction of the soils solid phase, thus the Kd can be
defined in terms of the soil organic carbon content (Kd ) KOC
fOC, where KOC ) organic carbon to water partition
coefficient and fOC ) fraction organic carbon). The KOC is
readily calculated and linearly related to the KOW. Therefore
soil sorption will increase with KOW, reducing the availability
of high KOW compounds for plant uptake. Correlations
between KOC and KOW have been derived by a number of
workers, but the most widely quoted is Karickhoff (43). It
should be noted that the slope from the fitted linear regression
for this relationship varies with pollutant class, e.g., 1.01
aromatic hydrocarbons, 0.88 chlorinated hydrocarbons, and
0.81 for chlorophenols. The best fit to all compound classes
combined was found to be

log KOC ) (0.989 log KOW) - 0.346fOC

Increases in the fOC therefore reduce both the total amount
of chemical absorbed by vegetation and the optimum KOW
for uptake into plants. More recent work has found that the
situation is complex with compounds sorbing to different
soil fractions of organic carbon (44) as well as other soil
fractions (45).
The partition distribution coefficient between soil and
soil air for organic chemicals is related to the Henrys law
constant. Both Bromilow and Chamberlain (19) and Ryan et
al. (18) indicate that where the dimensionless Henrys law
constant is greater than 10-4 the compound will partition
preferentially to the gas phase from the soil and plant uptake
will be from the soil-air-leaf pathway. The transfer from
the soil to the gas phase is determined by the soil organic
carbon content, porosity, and moisture content of soils.
Gaseous Uptake from Air. Organic chemicals can contaminate plant shoots via the soil-air-plant pathway.
Chemicals may volatilize from the soil into the free atmo-

FIGURE 3. Interpretive framework of McLachlan (13) illustrating

the relationship between log KOA and uptake process for organic
chemicals in the atmosphere.
sphere and subsequently diffuse into the plant leaves via the
cuticle or the stomata. The stomata provide an entry pathway
for carbon dioxide and an exit route for water vapor from the
transpiration stream. Small particle-bound compounds may
be taken up via the stomata, but the stomatal entry route will
generally only be significant for chemicals present in air in
the gaseous phase (17). Chemicals entering the leaves via
the stomata or the cuticle will diffuse into intercellular air
spaces and partition to the aqueous and lipophilic phases of
adjacent plant tissues. Lipophilic leaf tissues include interior
lipids, such as membrane lipids, storage lipids, resins, and
essential oils, and surface lipids, such as cutin and cuticular
waxes (46). Tolls and McLachlan (47) and Hauk et al. (48)
have suggested that, in terms of the storage of organic
chemicals, plant leaves can be divided into two compartments: a relatively small surface compartment with rapid
uptake and clearance kinetics, and a larger reservoir compartment with relatively slow chemical migration. The
composition of the larger reservoir compartment is believed
to influence the retention of organic chemicals, but the
physiological relationship between the two compartments
has not been elucidated.
Gaseous uptake from ambient air has been demonstrated
to be the main uptake pathway into plant foliage for a variety
of organic chemicals, including PAHs (2) PCBs (15) and tetraand hexa-chlorinated PCDD/Fs (49, 50). The bioconcentration of organic chemical vapors into plant leaves has also
been investigated by Bacci and co-workers (51-53). The
results of these experiments were used to derive relationships
between leaf/air bioconcentration factors and KOW and the
dimensionless Henrys law constant (HC), e.g.

Log (BCFHC) ) -1.95 + 1.14 log KOW

Other studies have shown that the partition between plant
foliage and the air is linearly related to the octanol/air
partition coefficient (KOA) for many organic chemicals (21,
47, 54). The interpretative framework of McLachlan (16)
suggests that for semivolatile organic chemicals with log KOA
< 8.5, gaseous uptake will be the primary process of foliar
uptake, with partitioning of the chemical between the leaf
and air approaching equilibrium, but they will accumulate
less than chemicals with a higher KOA (Figure 3). For those
chemicals with log KOA between 8.5 and 11, gaseous uptake
into the foliage will still be the primary process of foliar uptake,
but this uptake will be kinetically limited (i.e., the plant will
not come into contact with enough air for it to become
saturated with the chemical) and will not approach equilibrium. Finally, those compounds with a log KOA > 11 will
be particle-bound when they deposit to plant surfaces, so


desorption from the particle into the leaf would be required

to contaminate the plant shoot. It should be noted that
McLachlans framework derived from an offsite aerial source,
for local sources soil volatilization and subsequent gaseous
deposition may also play a role for those compounds with
log KOA > 11. Both Wang and Jones (55) and Kipopoulou et
al. (5) found a linear correlation between log KOA and shoot
concentration factor (SCF ) shoot concentration of X/soil
concentration of X) when investigating crop uptake of
chlorobenzenes (log KOA 4.5-7) and PAHs (log KOA 7.5-13),
Interspecies variability in organic chemical concentrations
in plant foliage has been demonstrated by Buckley (56).
Schrieber et al. (57) measured cuticular permeabilities for 40
plant species and which ranged over 2 orders of magnitude,
in any specific properties. Bohme et al. (15) found a 20-fold
variation between 5 different grasses and herbs. Since
lipophilic chemicals will tend to partition to leaf lipids,
differences in the lipid content of plant foliage would seem
the most reasonable explanation for this behavior. For foliar
uptake models, such as that of Muller et al. (58), the leaf lipid
content represents the most sensitive plant characteristic
for the uptake of lipophilic chemicals. A study by Simonich
and Hites (59) supports this with interspecies variability in
experimental results found to be reduced when PAH concentrations were normalized to leaf lipid contents. Both
Bohme et al. (15) and Komp et al. (60) found that interspecies
variability in leaf/air bioconcentration factors for a variety
of semivolatile organic chemicals, where gaseous uptake was
the dominant uptake process, could not be explained by
variability in extractable lipid contents. The authors concluded that not just lipid quantity but also lipid quality need
to be considered when evaluating interspecies variability in
leaf/air bioconcentration factors. Unfortunately neither of
these workers specifies what they mean by lipid quality.
Schreiber and Schonherr (61) and McCrady and Maggard
(62) have found that the exposed surface area of the plant
foliage influences the foliar uptake rate for a variety of species.
Simonich and Hites (59) suggested that this plant characteristic will be particularly significant if chemical partitioning
to foliage does not approach equilibrium, because those
plants with a larger exposed area will come into contact with
more chemical.
Particulate Deposition to Plant Surfaces. Organic chemicals can be transferred from soil by the re-suspension of soil
particles, and then subsequently to plant foliage via dry and
wet deposition. Other atmospheric aerosols containing
organic compounds will also contaminate plant foliage
following dry and wet deposition. A study by Kao and
Venkataraman (63) found that re-suspended soil dust
contributed approximately 4% of the total ambient PCDD/F
concentrations measured, but could contribute between 20
and 90% of the deposition to environmental surfaces because
of the high deposition velocity of soil particles.
The dry deposition of particles suspended in the air to
plant surfaces involves diffusion, interception, impaction,
and sedimentation processes (64). Once particles have been
deposited on plant foliage the chemicals bound to them are
subject to removal and degradation. However, during the
period of contact between plant leaves and deposited
particles, particle-bound organic chemicals may diffuse into
the plant cuticle and become adsorbed to lipophilic tissues
or may permeate into the leaf interior. The plant cuticle can
be divided into an outer skin, made up of cuticular waxes
embedded within the cutin matrix, and an inner volume
element, which mainly consists of the cutin matrix. Lipophilic
chemicals that permeate into the inner volume element of
the cuticle can pass through the epidermis into the leaf
interior with relative ease (46). The permeability of plant
cuticles to organic chemicals in solution has been found to


be linearly related to the KOW of the chemical and inversely

related to its molecular volume (65). However, the situation
regarding the uptake of chemicals from ephemeral particulate
deposits is far more complex and the extent to which the
chemical is reversibly or irreversibly bound to the particle
will significantly influence the rate of transfer.
Nakajima et al. (66) have investigated the uptake of PAHs
from the atmosphere into Azalea leaves. The authors suggest
that for benzo[a]pyrene and perylene, dry deposition of
suspended particles with subsequent permeation into the
cuticle represents the major pathway of contamination.
Welsch-Paulsch et al. (67) also determined that the dry
deposition of large particles represented an important uptake
pathway for hepta- and octo-chlorinated PCDDs into grass
leaves. The octanol/air partition coefficient has been suggested as a suitable indicator of the significance of the dry
deposition contamination pathway for semivolatile organic
chemicals (SVOCs). The interpretative framework of
McLachlan (16) identifies particle-bound deposition as the
primary process of plant uptake for SVOCs with log KOA >
11. Cousins and Mackay (25) suggest that for chemicals with
log KOA > 9, particle-bound deposition becomes relatively
more important than gaseous deposition. When plant uptake
models were compared for incorporation into the CSOIL
model (8), the SCF (stem concentration factor ) stem
concentration/soil concentration) between the model and
the data were only in reasonable agreement when soil
resuspension was included in the model using the relationship proposed by Hetrick and McDowell-Boyer (68).
Wet deposition is thought to be the dominant deposition
mechanism for chemicals with log KAW < -6 (25). Relatively
little information is available regarding the wet deposition
of particle-bound and gaseous phase organic chemicals to
plant surfaces and this pathway is seldom considered when
looking at the contamination of vegetation (17). Cousins and
Mackay (25) suggest that the majority of wet deposition is
not intercepted and retained by vegetation, but is washed
directly to soil. However, it should be noted that wet
deposition can nevertheless be an important indirect factor
via rain splash. Dreicer et al. (69) investigated the rain splash
pathway for tomato plants, and they found that only finer
soil particles were retained on plant surfaces. There is some
evidence that PAH burdens in soil accumulate preferentially
in these finer fractions (70). Kaup et al. (71) sequentially
extracted maize leaves and reported that of the particle
associated PAH and PCCD/F approximately 20% could
potentially be eroded and some 30-60% had desorbed from
the particle into the epicuticular waxes. Greater penetration
was achieved by the more volatile compounds.
Interspecies variability in organic chemical concentrations
in plant foliage also occurs for chemicals following particulate
deposition. Significant differences between plant species have
been observed in mass loadings of soil to plant surfaces, as
a result of contact with soil particles (17). Plant characteristics
that affect the rate of particle deposition and retention include
the exposed surface area of the foliage and the presence or
absence of leaf hairs. Little and Wiffen (72) observed that
rough or hairy leaf surfaces were more efficient at collecting
aerosols than smooth surfaces. This observation was explained by increased surface area and the projection of
roughness elements through the leaf-air boundary layer.
Pinder et al. (73) found that the largest soil loadings occurred
on broad-leaved species growing close to the ground, while
lster and Marschner (74) reported that direct contact
between soil particles and plant surfaces was a significant
uptake pathway for PCDD/Fs to hay.
Influence of Plant Composition on Uptake. As previously
stated the composition of the plant is believed to significantly
affect the uptake of organic chemicals. There are a number
of authors who have reported that the lipid content or lipid

TABLE 1. Details of Water, Lipid, Carbohydrate, and Fiber Content of Crops Relevant to Risk Assessment (69)
crop class





leaf crops (lettuce, pak choi, cabbage)

bulbs, stems and tubers (onion, leek, potato)
root crops (carrot, parsnip)
fruit crops (tomato, pepper, pea, bean)





quality is of significance (2, 15, 19, 58). A number of predictive

uptake models use lipid content as an input parameter (31,
75, 76). Hung and Mackay (75) and Chiouet al. (76) also
require the content of fiber and carbohydrate, respectively.
There are, however, very little readily accessible data for these
parameters. The best source was found to be the website of
the U.S. Department of Agriculture where details of the crop
composition of important food staples can be found (77).
From Table 1 it can be seen that there was some variability
within the major crop classes for water, lipid, carbohydrate,
and fiber content. The major differences were the higher
water content of leaf crops, the lower carbohydrate content
of leaf crops, and the higher lipid content of root and fruit
crops. These values are lower than the default values
suggested by the models of Trapp and Mathies (31), Hung
and Mackay (75), and Chiou et al. (76), but are in agreement
with Flindt (78) and Tam et al. (79). Therefore these models
may possibly overestimate the accumulation of organic
chemicals in plants. However, it should also be noted that
other plant components such as waxes, lignin, and suberin
may also absorb organic chemicals and these would not be
extracted using the normal lipid extraction methods, so using
values for lipid alone may underestimate total potential
accumulation. It may be the reservoir capacity of these other
plant components that are responsible for the differences in
uptake reported by Bohme et al. (15) and Komp et al. (59)
that could not be related solely to lipid content.

Other Potentially Significant Processes

Plant Metabolism. Chemicals accumulated in plants may
become metabolized, thus reducing their concentration
within the plants tissues. A fuller review of this process can
be found in Burken (80). The process can be divided into
transport, where the uptake will be driven by those processes
described above; transformation reactions, e.g., oxidation;
conjugation, e.g., with glutathione; and finally sequestration,
e.g., into the cell wall. Plant metabolism has been reported
for a relatively small number of industrial organic chemicals
although more is known of herbicides. Those chemicals where
evidence has been reported include trichloroethylene, benzene, and pyrene (81-83); with most success being reported
for the explosives (84, 85). Metabolic processes and rates will
be specific to particular chemicals and plant species (8587). The toxicity of the metabolites and bound residues from
these processes is an area that requires investigation.
Photolytic Degradation on Plant Surfaces. Photolytic
degradation may reduce chemical concentrations in plants
and has been demonstrated to be a significant loss mechanism for 2,3,7,8-TCCD from spruce (Picea abies) foliage (88).
Trapp and Matthies (89) found it was the most significant
loss process for 2,3,7,8-TCDD in their PLANTX model.
Photolytic degradation on plant leaves is enhanced by the
tendency of leaves to orient themselves to maximize the
interception of sunlight. The extent of photolytic degradation
on leaf surfaces will influence the aerial deposition and direct
contact plant uptake pathways.
Volatilization from Foliage to Air. The plant transpiration
flux will move some chemicals to sub-stomatal tissues from
which they will subsequently be lost via volatilization, thus
reducing plant concentrations (90). Volatilization is likely to
be a significant transport pathway for chemicals with high

water solubility and vapor pressure. This pathway was found

to be significant for benzene (91) and TCE (92). Phytovolatilization is the principal mechanism in the phytoremediation
of MTBE-contaminated groundwaters (93). This process will
occur in parallel with the gaseous uptake processes described
Growth Dilution. The importance of growth dilution for
the contamination of vegetation with organic chemicals has
not been clearly established. Trapp and Matthies (31) and
Hung and Mackay (75) include it in their models, but these
assume that plants are in the middle stage of exponential
growth, they do not describe the biomass accumulation over
a season. There are two scenarios where growth dilution
might be potentially important: (1) where there is an acute
exposure event and growth subsequently results in dilution
of this spike of contamination, and (2) where the uptake
of the chemical per unit mass is slower than the accumulation
of dry matter per unit mass. There is little data regarding the
first and it would be critically dependent on the growth stage
of the crop; the earlier in the season the greater potential for
dilution. The second scenario is highly probable. There is
little known about the kinetics of uptake, but it is known that
plants with KOA > 8.5 may not become saturated with
chemical from aerial deposition gaseous exposure from the
McLachlan framework discussed above, and Schwab et al.
(24) found that older roots of tall fescue (Festuca arundinacea,
Schreber) and alfalfa (Medicago sativa, L.) were not saturated
with naphthalene, but young roots were. It would therefore
appear that growth will have a significant role.

Focus for Future Research

This paper has identified key pathways for the plant uptake
of organic chemicals. Where possible, predictive relationships
have been presented or referenced to enable the construction
of plant uptake models. Future research will need to address
those areas where at present the understanding of a
potentially important pathway is lacking. There is a significant
amount of information on the root uptake and aerial
deposition of organic chemicals to plants that allows for the
understanding of the potential for the transfer of these
chemicals. This is not always at the mechanistic level, but
is probably sufficient for an estimation of the potential risk
from the ingestion of plant material contaminated by these
Where relationships are proposed (for example, those for
the accumulation from the gas phase of Bacci et al. (52) and
McLachlan (16)) these have not been subsequently validated
by a range of independent experiments as is required for a
truly robust model. This absence of validation is also true for
the relationship derived by Briggs et al. (30) which was
developed for pesticides and is used in a number of models
for the uptake of industrial chemicals (18, 94). Such validation
would determine if this is a better relationship than that of
Hsu et al. (32) or Burken and Schnoor (33). Recent work
would also suggest that at low KOW, these relationships do
not hold (6, 34). There is therefore a need for high-quality
laboratory and field studies to quantify key processes so
proposed relationships can be validated and hence be used
with confidence.
At present the contribution to the foliar contamination
from distant aerial sources cannot always be separated from


that arising from local sources. The former has been well
studied, but the information on the latter is lacking.
Particulate deposition from local sources may be a particularly
significant pathway and is a pressing area for investigation.
There are virtually no data to allow for quantification of the
gaseous soil-air-plant pathway, which would also arise from
a local source. Separation of the local from the off-site is
required for a balanced risk assessment for contaminated
A number of researchers have related uptake of organic
chemicals to the lipid content of the plant component to
predict the contaminant accumulation. However, few data
are easily available for a value to be chosen that would be
technically defensible. This data gap needs to be addressed
if these parameters are to be used with confidence in models.
Species and interspecies variability may play a major role
here, but one needs to remember the law of diminishing
returns; it may not be practicable to determine the lipid and
other accumulating components for a wide range of species
particularly if there are significant intra-species differences.
Finally, there is little information on the toxicity of the
plant metabolites and bound residues formed from the
uptake of such chemicals and this may significantly influence
the risk prioritization of those chemicals investigated.

Literature Cited
(1) Hulster, A.; Muller, J. F.; Marschner, H. Soil-Plant Transfer of
Polychlorinated Dibenzo-p-Dioxins and Dibenzofurans to
Vegetables of the Cucumber Family (Cucurbitaceae). Environ.
Sci. Technol. 1994, 28, 1110-1115.
(2) Simonich, S. L.; Hites, R. A. Vegetation-Atmosphere Partitioning
of Polycyclic Aromatic Hydrocarbons. Environ. Sci. Technol.
1994, 28, 939-943.
(3) Lovett, A. A.; Foxall, C. D.; Creaser, C. S.; Chewe, D. PCB and
PCDD/DF congeners in locally grown fruit and vegetable
samples in Wales and England. Chemosphere 1997, 34, 14211436.
(4) Thomas, G.; Sweetman, A. J.; Ockenden, W. A.; Mackay, D.;
Jones, K. C. Air-pasture transfer of PCBs. Environ. Sci. Technol.
1998, 32, 936-942.
(5) Kipopoulou, A. M.; Manoli, E.; Samara, C. Bioconcentration of
polycyclic aromatic hydrocarbons in vegetables grown in an
industrial area. Environ. Pollut. 1999, 106, 369-380.
(6) Groom, C. A.; Halasz, A.; Paquet, L.; Olivier, L.; Dubois, C.;
Hawari, J. Accumulation of HMX (octahydro-1,3,5,7-tetranitri1,3,5,7-tetrazocine) in indigenous and agricultural plants grown
in HMX-contaminated anti-tank firing-range soil. Environ. Sci.
Technol. 2002, 36, 112-118.
(7) Wu, W. Z.; Schramm, K. W.; Xu, Y.; Kettrup, A. Contamination
and distribution of polychlorinated dibenzo-p-dioxins and
dibenzofurans (PCDD/F) in agriculture fields in Ya-Er Lake area,
China. Ecotoxicol. Environ. Saf. 2002, 53, 141-147.
(8) Rikken, M. G. J.; Lijzen, J. P. A.; Cornelese, A. A. Evaluation of
Model Concepts on Human Exposure; RIVM (National Institiute
for Public Health and the Environment): Bilthoven, The
Netherlands, 2001.
(9) DEFRA. The Contaminated Land Exposure Assessment Model
(CLEA): Technical Basis and Algorithms; WRc, Department for
Environment, Food, and Rural Affairs: London, 2002.
(10) McKone, T. E.; Maddalena, R. L.; Bennett, D. H. CALTOX Total
Exposure Model for Hazardous Waste Sites; California Department of Toxic Substances Control: Sacramento, CA, 2003. http://
www.dtsc.ca.gov/ScienceTechnology/caltox.html (last accessed
Jan 2005).
(11) Dalla Valle, M.; Dachs, J.; Sweetman, A. J.; Jones, K. C. Maximum
reservoir capacity of vegetation for persistent organic pollutants: Implications for global cycling. Global Biogeochem. Cycle
2004, 18.
(12) Barber, J. L.; Thomas, G. O.; Kerstiens, G.; Jones, K. C. Current
issues and uncertainties in the measurement and modelling of
air-vegetation exchange and within-plant processing of POPs.
Environ. Pollut. 2004, 128, 99-138.
(13) Wegmann, F.; Scheringer, M.; Moller, M.; Hungerbuhler, K.
Influence of vegetation on the environmental partitioning of
DDT in two global multimedia models. Environ. Sci. Technol.
2004, 38, 1505-1512.


(14) Scheringer, M.; Salzmann, M.; Stroebe, M.; Wegmann, F.; Fenner,
K.; Hungerbuhler, K. Long-range transport and global fractionation of POPs: insights from multimedia modeling studies.
Environ. Pollut. 2004, 128, 177-188.
(15) Bohme, F.; Welsch-Pausch, K.; McLachlan, M. S. Uptake of
airborne semivolatile organic compounds in agricultural
plants: Field measurements of interspecies variability. Environ.
Sci. Technol. 1999, 33, 1805-1813.
(16) McLachlan, M. S. Framework for the interpretation of measurements of SOCs in plants. Environ. Sci. Technol. 1999, 33, 17991804.
(17) Smith, K. E. C.; Jones, K. C. Particles and vegetation: implications
for the transfer of particle-bound organic contaminants to
vegetation. Sci. Total Environ. 2000, 246, 207-236.
(18) Ryan, J. A.; Bell, R. M.; Davidson, J. M.; OConnor, G. A. Plant
Uptake of Non-Ionic Organic-Chemicals from Soils. Chemosphere 1988, 17, 2299-2323.
(19) Bromilow, R. H.; Chamberlain, K. Principles governing uptake
and transport of chemicals. In Plant Contamination: Modelling
and Simulation of Organic Chemical Processes; Trapp, S.,
McFarlane, J. C., Eds.; Lewis Publishers: London, 1995; pp 3864.
(20) Briggs, G. G.; Bromilow, R. H.; Evans, A. A.; Williams, M.
Relationships between Lipophilicity and the Distribution of NonIonized Chemicals in Barley Shoots Following Uptake by the
Roots. Pestic. Sci. 1983, 14, 492-500.
(21) Paterson, S.; Mackay, D.; Bacci, E.; Calamari, D. Correlation of
the Equilibrium and Kinetics of Leaf Air Exchange of Hydrophobic Organic Chemicals. Environ. Sci. Technol. 1991, 25, 866871.
(22) DuarteDavidson, R.; Jones, K. C. Screening the environmental
fate of organic contaminants in sewage sludge applied to
agricultural soils. 2. The potential for transfers to plants and
grazing animals. Sci. Total Environ. 1996, 185, 59-70.
(23) Wild, S. R.; Jones, K. C. Organic Chemicals Entering Agricultural
Soils in Sewage Sludges - Screening for Their Potential to
Transfer to Crop Plants and Livestock. Sci. Total Environ. 1992,
119, 85-119.
(24) Schwab, A. P.; Al-Assi, A. A.; Banks, M. K. Adsorption of
naphthalene onto plant roots. J. Environ. Qual. 1998, 27, 220224.
(25) Cousins, I. T.; Mackay, D. Strategies for including vegetation
compartments in multimedia models. Chemosphere 2001, 44,
(26) Trapp, S.; Pussemeir, L. Model calculations and measurements
of uptake and translocation of carbamates by bean plants.
Chemosphere 1991, 22, 327-345.
(27) Fismes, J.; Perrin-Ganier, C.; Empereur-Bissonnet, P.; Morel, J.
L. Soil-to-root transfer and translocation of polycyclic aromatic
hydrocarbons by vegetables grown on industrial contaminated
soils. J. Environ. Qual. 2002, 31, 1649-1656.
(28) Wild, S. R.; Jones, K. C. Polynuclear Aromatic Hydrocarbon
Uptake by Carrots Grown in Sludge-Amended Soil. J. Environ.
Qual. 1992, 21, 217-225.
(29) McFarlane, J. C. Plant transport of organic chemicals. In Plant
Contamination - Modelling and Simulation of Organic Chemical
Processes; Trapp, S., McFarlane, J. C., Eds.; Lewis Publishers:
Boca Raton, FL, 1995.
(30) Briggs, G. G.; Bromilow, R. H.; Evans, A. A. Relationships Between
Lipophilicity and Root Uptake and Translocation of Non-Ionized
Chemicals By Barley. Pestic. Sci. 1982, 13, 495-504.
(31) Trapp, S.; Matties, M. Generic one compartment model for the
uptake of organic chemicals by foliar vegetation. Environ. Sci.
Technol. 1995, 29, 2333-2338.
(32) Hsu, F. C.; Marxmiller, R. L.; Yang, A. Y. S. Study of Root Uptake
and Xylem Translocation of Cinmethylin and Related-Compounds in Detopped Soybean Roots Using a Pressure Chamber
Technique. Plant Physiol. 1990, 93, 1573-1578.
(33) Burken, J. G.; Schnoor, J. L. Predictive relationships for uptake
of organic contaminants by hybrid poplar trees. Environ. Sci.
Technol. 1998, 32, 3379-3385.
(34) Doucette, W. J.; Chard, J. K.; Moore, B. J.; Staudt, W. J.; Headley,
J. V. Uptake of sulfolane and diisopropanolamine (DIPA) by
cattails (Typha latifolia). Microchem. J. 2005, 81, 41-49.
(35) Barak, E.; Jacoby, B.; Dinoor, A. Adsorption of Systemic Pesticides
on Ground Stems and in the Apoplastic Pathway of Stems, as
Related to Lignification and Lipophilicity of the Pesticides. Pestic.
Biochem. Physiol. 1983, 20, 194-202.
(36) Uchida, M. Affinity and Mobility of Fungicidal Dialkyl Dithiolanylidenemalonates in Rice Plants. Pestic. Biochem. Physiol.
1980, 14, 249-255.

(37) McCrady, J. K.; McFarlane, C.; Lindstrom, F. T. The Transport

and Affinity of Substituted Benzenes in Soybean Stems. J. Exp.
Bot. 1987, 38, 1875-1890.
(38) Topp, E.; Scheunert, I.; Attar, A.; Korte, F. Factors Affecting the
Uptake of C-14-Labeled Organic Chemicals by Plants from Soil.
Ecotoxicol. Environ. Saf. 1986, 11, 219-228.
(39) OConnor, G. A.; Kiehl, D.; Eiceman, G. A.; Ryan, J. A. Plant
Uptake of Sludge-Borne PCBs. J. Environ. Qual. 1990, 19, 113118.
(40) Smelt, J. H.; Leistra, M. A. E. Hexachlorobenzene in soils and
crops after soil treatment with pentachloronitrobenzene. Agric.
Environ. 1974, 1, 65-71.
(41) Schroll, R.; Scheunert, I. Uptake of the lipophilic model
compound hexachlorobenzene by different plant species during
the vegetation period. Fresenius Environ. Bull. 1992, 1, 334338.
(42) White, J. C.; Wang, X. P.; Gent, M. P. N.; Iannucci-Berger, W.;
Eitzer, B. D.; Schultes, N. P.; Arienzo, M.; Mattina, M. I.
Subspecies-level variation in the phytoextraction of weathered
p,p-DDE by Cucurbita pepo. Environ. Sci. Technol. 2003, 37,
(43) Karickhoff, S. W. Semiempirical Estimation of Sorption of
Hydrophobic Pollutants on Natural Sediments and Soils.
Chemosphere 1981, 10, 833-846.
(44) Chiou, C. T.; McGroddy, S. E.; Kile, D. E. Partition characteristics
of polycyclic aromatic hydrocarbons on soils and sediments.
Environ. Sci. Technol. 1998, 32, 264-269.
(45) Cooke, C. M.; Bailey, N. J.; Shaw, G.; Lester, J. N.; Collins, C. D.
Interaction of formaldehyde with soil humic substances: Separation by GFC and characterization by H-1-NMR Spectroscopy
Bull. Environ. Contam. Toxicol. 2003, 70, 761-768.
(46) Reiderer, M. Estimating partitioning and transport of organic
chemicals in the foliage/atmosphere system: Discussion of a
fugacity based model. Environ. Sci. Technol. 1990, 24, 829-837.
(47) Tolls, J.; McLachlan, M. S. Partitioning of Semivolatile Organic
Compounds between Air and Lolium multiflorum (Welsh Ray
Grass). Environ. Sci. Technol. 1994, 28, 159-166.
(48) Hauk, H.; Umlauf, G.; McLachlan, M. S. Uptake of Gaseous
DDE in Spruce Needles. Environ. Sci. Technol. 1994, 28, 23722379.
(49) Meneses, M.; Schuhmacher, M.; Domingo, J. L. A design of two
simple models to predict PCDD/F concentrations in vegetation
and soils. Chemosphere 2002, 46, 1393-1402.
(50) Welsch-Pausch, K.; McLachlan, M. S.; Umlauf, G. Determination
of the Principal Pathways of Polychlorinated Dibenzo-p-Dioxins
and Dibenzofurans to Lolium multiflorum (Welsh Ray Grass).
Environ. Sci. Technol. 1995, 29, 1090-1098.
(51) Bacci, E.; Gaggi, C. Chlorinated Hydrocarbon Vapors and Plant
Foliage - Kinetics and Applications. Chemosphere 1987, 16,
(52) Bacci, E.; Cerejeira, M. J.; Gaggi, C.; Chemello, G.; Calamari, D.;
Vighi, M. Bioconcentration of Organic-Chemical Vapors in PlantLeaves - the Azalea Model. Chemosphere 1990, 21, 525-535.
(53) Bacci, E.; Calamari, D.; Gaggi, C.; Vighi, M. Bioconcentration of
Organic Chemical Vapors in Plant Leaves - Experimental
Measurements and Correlation. Environ. Sci. Technol. 1990,
24, 885-889.
(54) Komp, P.; McLachlan, M. S. Octanol/air partitioning of polychlorinated biphenyls. Environ. Toxicol. Chem. 1997, 16, 24332437.
(55) Wang, M. J.; Jones, K. C. Uptake of Chlorobenzenes by Carrots
from Spiked and Sewage Sludge Amended Soil. Environ. Sci.
Technol. 1994, 28, 1260-1267.
(56) Buckley, E. H. Accumulation of Airborne Polychlorinated
Biphenyls in Foliage. Science 1982, 216, 520-522.
(57) Schreiber, L.; Riederer, M. Determination of diffusion coefficients
of octadecanoic acid in isolated cuticular waxes and their
relationship to cuticular water permeabilities. Plant Cell Environ.
1996, 19, 1075-1082.
(58) Muller, J. F.; Hawker, D. W.; Connell, D. W. Calculation of
Bioconcentration Factors of Persistent Hydrophobic Compounds in the Air/Vegetation System. Chemosphere 1994, 29,
(59) Simonich, S. L.; Hites, R. A. Organic Pollutant Accumulation in
Vegetation. Environ. Sci. Technol. 1995, 29, 2905-2914.
(60) Komp, P.; McLachlan, M. S. Interspecies variability of the plant/
air partitioning of polychlorinated biphenyls. Environ. Sci.
Technol. 1997, 31, 2944-2948.
(61) Schreiber, L.; Schonherr, J. Uptake of Organic Chemicals in
Conifer Needles - Surface Adsorption and Permeability of
Cuticles. Environ. Sci. Technol. 1992, 26, 153-159.

(62) McCrady, J. K.; Maggard, S. P. Uptake and Photodegradation of

2,3,7,8-Tetrachlorodibenzo-p-Dioxin Sorbed to Grass Foliage.
Environ. Sci. Technol. 1993, 27, 343-350.
(63) Kao, A. S.; Venkataraman, C. Estimating the Contribution of
Reentrainment to the Atmospheric Deposition of Dioxin.
Chemosphere 1995, 31, 4317-4331.
(64) Chamberlain, A. C. Radioactive Aerosols; Cambridge University
Press: Cambridge, 1991; Vol. 3.
(65) Riederer, M.; Daiss, A.; Gilbert, N.; Kohle, H. Semi-volatile organic
compounds at the leaf/atmosphere interface: numerical simulation of dispersal and foliar uptake. J. Exp. Bot. 2002, 53, 18151823.
(66) Nakajima, D.; Yoshida, Y.; Suzuki, J.; Suzuki, S. Seasonal Changes
in the Concentration of Polycyclic Aromatic Hydrocarbons in
Azalea Leaves and Relationship to Atmospheric Concentration.
Chemosphere 1995, 30, 409-418.
(67) Welschpausch, K.; McLachlan, M. S.; Umlauf, G. Determination
of the Principal Pathways of Polychlorinated Dibenzo-p-Dioxins
and Dibenzofurans to Lolium multiflorum (Welsh Ray Grass).
Environ. Sci. Technol. 1995, 29, 1090-1098.
(68) Hetrick, D. M.; McDowell-Boyer, L. M. A users manual for TOXSCREEN: A multimedia screening-level program for assessing
the potential fate of chemicals released to the environment; Oak
Ridge National Laboratory: Oak Ridge, TN, and U. S. EPA, Office
of Toxic Substances: Washington, DC, 1984.
(69) Dreicer, M.; Hakonson, T. E.; White, G. C.; Whicker, F. W.
Rainsplash as a Mechanism for Soil Contamination of Plant
Surfaces. Health Phys. 1984, 46, 177-187.
(70) Guggenberger, G.; Pichler, M.; Hartmann, R.; Zech, W. Polycyclic
aromatic hydrocarbons in different forest soils: mineral horizons. Z. Pflanz. Bodenk 1996, 159, 565-573.
(71) Kaupp, H.; Blumenstock, M.; McLachlan, M. S. Retention and
mobility of atmospheric particle-associated organic pollutant
PCDD/Fs and PAHs in maize leaves. New Phytol. 2000, 148,
(72) Little, P.; Wiffen, R. D. Emission and deposition of petrol engine
exhaust Pb-I. Deposition of exhaust Pb to plant and soil
surfaces. Atmos. Environ. 1977, 11, 437-447.
(73) Pinder, J. E., III; McLeod, K. W.; Lide, R. F.; Sherrod, K. C. Mass
loading of soil particles on a pasture grass. J. Environ. Radioact.
1991, 13, 341-354.
(74) Hulster, A.; Marschner, H. Transfer of PCDD/PCDF from
Contaminated Soils to Food and Fodder Crop Plants. Chemosphere 1993, 27, 439-446.
(75) Hung, H.; Mackay, D. A novel and simple model of the uptake
of organic chemicals by vegetation from air and soil. Chemosphere 1997, 35, 959-977.
(76) Chiou, C. T.; Sheng, G. Y.; Manes, M. A partition-limited model
for the plant uptake of organic contaminants from soil and
water. Environ. Sci. Technol. 2001, 35, 1437-1444.
(77) USDA. USDA National Nutrient Database for Standard Reference,
Release 16; U.S. Department of Agriculture: Washington, DC,
2005. http://www.nal.usda.gov/fnic/foodcomp/Data/SR16/
(78) Flindt, R. Biologie in Zahlen. Eine datensammlung in tabellen
mit uber 10 000 Einzelwerten; Gustav Fischer Verlag: Stuttgart/
New York, 1988.
(79) Tam, D. D.; Shiu, W. Y.; Qiang, K.; Mackay, D. Uptake of
chlorobenzenes by tissues of the soybean plant: Equilibria and
kinetics. Environ. Toxicol. Chem. 1996, 15, 489-494.
(80) Burken, J. G. Uptake and metabolism of organic compounds the green liver model. In Phytoremediation - Transformation
and Control of Contaminants; McCutcheon, S. C., Schnoor, J.
L., Eds.; John Wiley and Sons: Hoboken, NJ, 2003; p 987.
(81) Huckelhoven, R.; Schuphan, I.; Thiede, B.; Schmidt, B. Biotransformation of pyrene by cell cultures of soybean (Glycine max
L.), wheat (Triticum aestivum L.), jimsonweed (Datura stramonium L.), and purple foxglove (Digitalis purpurea L.). J. Agric.
Food Chem. 1997, 45, 263-269.
(82) Shang, T. Q.; Gordon, M. P. Transformation of C-14 trichloroethylene by poplar suspension cells. Chemosphere 2002, 47,
(83) Ugrekhelidze, D.; Korte, F.; Kvesitadze, G. Uptake and transformation of benzene and toluene by plant leaves. Ecotoxicol.
Environ. Saf. 1997, 37, 24-29.
(84) Nepovim, A.; Hubalek, M.; Podlipna, R.; Zeman, S.; Vanek, T.
In-vitro degradation of 2,4,6-trinitrotoluene using plant tissue
cultures of Solanum aviculare and Rheum palmatum. Eng. Life
Sci. 2004, 4, 46-49.


(85) Hannink, N. K.; Rosser, S. J.; Bruce, N. C. Phytoremediation of

explosives. Crit. Rev. Plant Sci. 2002, 21, 511-538.
(86) Chekol, T.; Vough, L. R.; Chaney, R. L. Plant-soil-contaminant
specificity affects phytoremediation of organic contaminants.
Int. J. Phytoremediat. 2002, 4, 17-26.
(87) Schnabel, W. E.; Dietz, A. C.; Burken, J. G.; Schnoor, J. L.; Alvarez,
P. J. Uptake and transformation of trichloroethylene by edible
garden plants. Water Res. 1997, 31, 816-824.
(88) Niu, J. F.; Chen, J. W.; Martens, D.; Henkelmann, B.; Quan, X.;
Yang, F. L.; Seidlitz, H. K.; Schramm, K. W. The role of UV-B
on the degradation of PCDD/Fs and PAHs sorbed on surfaces
of spruce (Picea abies (L.) Karst.) needles. Sci. Total Environ.
2004, 322, 231-241.
(89) Trapp, S.; Matthies, M. Generic One-Compartment Model For
Uptake of Organic Chemicals By Foliar Vegetation. Environ.
Sci. Technol. 1995, 29, 2333-2338.
(90) McFarlane, C.; Pfleeger, T.; Fletcher, J. Effect, Uptake and
Disposition of Nitrobenzene in Several Terrestrial Plants.
Environ. Toxicol. Chem. 1990, 9, 513-520.



(91) Collins, C. D.; Bell, J. N. B.; Crews, C. Benzene accumulation in

horticultural crops. Chemosphere 2000, 40, 109-114.
(92) Ma, X. M.; Burken, J. G. TCE diffusion to the atmosphere in
phytoremediation applications. Environ. Sci. Technol. 2003, 37,
(93) Hong, M. S.; Farmayan, W. F.; Dortch, I. J.; Chiang, C. Y.;
McMillan, S. K.; Schnoor, J. L. Phytoremediation of MTBE from
a groundwater plume. Environ. Sci. Technol. 2001, 35, 12311239.
(94) Trapp, S., MaFarlane, J. C., Eds. Plant Contamination Modelling and Simulation of Organic Chemical Processes; Lewis
Publishers: London, 1995.

Received for review April 28, 2005. Revised manuscript received October 11, 2005. Accepted October 19, 2005.