M e t h o d s t o m e a s u r e m a s s t r a n s f e r k i n e t i c s , p a r t i t i o n

r a t i o s a n d a t m o s p h e r i c f l u x e s o f o r g a n i c c h e m i c a l s

i n f o r e s t s y s t e m s

Damien Johann Bolinius

Methods to measure mass transfer ki-netics, partition ratios and atmospher-ic fluxes of organic chemicals in for-est systems

Damien Johann Bolinius

©Damien Johann Bolinius, Stockholm University 2016

ISBN print 978-91-7649-593-3

ISBN PDF 978-91-7649-594-0

Cover illustration by Chris Madden, used with the author’s permission.

Printed in Sweden by US-AB, Stockholm 2016

Distributor: Department of Environmental Science and Analytical Chemistry (ACES)

La science, mon garçon, est faite d’erreurs, mais d’erreurs qu’il est bon de commettre, car elles mènent peu à peu à la vérité.

Science, my lad, has been built upon many errors; but they are errors which it was good to fall into, for they led to the truth.

From: A journey into the centre of the Earth, by Jules Verne

i

Abstract

Vegetation plays an important role in the partitioning, transport and fate of

hydrophobic organic contaminants (HOCs) in the environment. This thesis

aimed at addressing two key knowledge gaps in our understanding of how

plants exchange HOCs with the atmosphere: (1) To improve our understand-

ing of the uptake of HOCs into, and transfer through, leaves of different

plant species; and (2) To evaluate an experimental approach to measure

fluxes of HOCs in the field. The methods presented in papers I, II and III

contribute to increasing our understanding of the fate and transport of HOCs

in leaves by offering straightforward ways of measuring mass transfer coef-

ficients through leaves and partition ratios of HOCs between leaves, leaf

lipids and lipid standards and reference materials like water, air and olive oil.

The passive dosing study in paper III in particular investigated the role of the

composition of the organic matter extracted from leaves in determining the

capacity of the leaves to hold chemicals and found no large differences be-

tween 7 different plant species, even though literature data on leaf/air parti-

tion ratios (Kleaf/air) varies over 1-3 orders of magnitude. In paper IV we

demonstrated that the modified Bowen ratio method can be extended to

measure fluxes of persistent organic pollutants (POPs) if the fluxes do not

change direction over the course of the sampling period and are large enough

to be measured. This approach thus makes it possible to measure fluxes of

POPs that usually require sampling times of days to weeks to exceed method

detection limits. The experimental methods described in this thesis have the

potential to support improved parameterization of multimedia models, which

can then be evaluated against fluxes measured in the field using the modified

Bowen ratio approach.

ii

iii

Sammanfattning

Vegetationen spelar en viktig roll för hydrofoba organiska föroreningars

(HOCs) öde i miljön, hur de fördelar sig och transporteras. I denna

avhandling adresseras två viktiga kunskapsluckor i vår förståelse av växters

utbyte av HOCs med atmosfären: (1) att studera upptaget av HOCs till, och

transporten igenom blad från olika växter; och (2) att utvärdera en

experimentell fältmetod för att mäta flöden av HOCs. De metoder som

används i papper I, II och III bidrar till att öka vår förståelse för transporten

av HOCs i blad genom att erbjuda enkla sätt att mäta

masstransferkoefficienter genom blad och fördelningskoefficienter av HOCs

mellan löv, lövlipider, lipid standarder och referensmaterial såsom vatten,

luft och olivolja. En passiv doseringsstudie användes i papper III för att

undersöka hur kompositionen av det organiska materialet som extraherats

från blad påverkar bladens kapacitet att hushålla kemikalier. Vi fann inga

stora skillnader mellan 7 olika växtarter, trots att fördelningskoefficienter

mellan blad / luft (Kblad/luft) i litteraturen varierar mellan 1-3

storleksordningar. I papper IV visade vi att den modifierade Bowen ratio

metoden kan utvidgas till att mäta flöden av långlivade organiska

föroreningar (POPs) om flödena inte ändrar riktning under

provtagningsperioden och är tillräckligt stora för att uppmätas. Detta

tillvägagångssätt gör det möjligt att mäta flöden av POPs som vanligtvis

kräver provtagningar i dagar /veckor för att överskrida metod

detektionsgränsen i labbet. De experimentella metoder som beskrivs i denna

avhandling har potential att bidra till bättre parameterisering av

multimediamodeller, som sedan kan utvärderas mot flöden uppmätta i fält

med hjälp av den modifierade Bowen ratio metoden.

iv

v

Samenvatting

Vegetatie speelt een belangrijke rol in het lot en transport van hydrophobe

organishe contaminanten (HOCs) in het mileu. Deze thesis was gericht op

twee lacunes in onze kennis van oppervlakte-atmosfeer fluxen van HOCs:

(1) Het verbeteren van onze kennis van de opname van HOCs in en door

gebladerte van verschillende soorten planten; en (2) het evalueren van een

nieuwe methode om verticale fluxen van semi-vluchtige stoffen te meten in

het veld. The methodes gepresenteerd in artikels I, II en III dragen bij tot een

verbeterde kennis van het lot en transport van HOCs in gebladerte doormidel

van simpele methoden die gebruikt kunnen worden voor het meten van mass

transfer coefficients door en in gebladerte en partitie ratios van gebladerte,

vetten geextraheerd van gebladerte en vet standaarden. De passive dosing

studie in artikel III, in het bijzonder, onderzocht de invloed van de

samenstelling van de vetten in gebladerte op de opname capaciteit voor

HOCs en vond geen grote verschillen tussen gebladerte van 7 verschillende

planten soorten, hoewel waarden van Kblad/lucht van verschillende

plantensoorten in de literatuur tot 3 log eenheden van elkaar verschillen. In

artikel IV, hebben we gedemonstreerd dat de gemodificeerde Bowen ratio

methode gebruikt kan worden voor het meten van verticale fluxen van HOCs

indien de richting van de flux constant blijft gedurende de staalname en dat

de fluxen groot genoeg zijn om te meten. Deze methode maakt het dus

mogelijk om fluxen van HOCs te meten, voor welke normaal lange

staalnames nodig zijn om detectielimieten te bereiken. De experimentele

methoden beschreven in deze thesis hebben potentieel om de parameterisatie

van bestaande multimedia fate models te verbeteren, welke dan geevalueerd

kunnen worden met de gemodificeerde Bowen ratio methode.

vi

vii

List of papers

Paper I

Hamid Ahmadi*, Damien Johann Bolinius*, Annika Jahnke, Matthew Mac-

Leod (2016), Mass transfer of hydrophobic organic chemicals between sili-

cone sheets and through plant leaves and low-density polyethylene, Chemo-

sphere, 164: 683-690.

* Shared first authorship

Paper II

Damien Johann Bolinius, Matthew MacLeod, Michael McLachlan, Philipp

Mayer, Annika Jahnke (2016), A passive dosing method to determine the

fugacity capacity of leaves, Environmental Science: Processes & Impacts,

18: 1325-1332.

Paper III

Damien Johann Bolinius, Matthew MacLeod, Francesco Iadaresta, Jan

Holmbäck, Annika Jahnke, Sorptive capacities of leaf lipids for hydrophobic

organic chemicals: Lipid characterization and passive dosing experiments.

Manuscript in preparation.

Paper IV

Damien Johann Bolinius, Annika Jahnke, Matthew MacLeod (2016), Com-

parison of eddy covariance and modified Bowen ratio methods for measur-

ing gas fluxes and implications for measuring fluxes of persistent organic

pollutants, Atmospheric Chemistry and Physics, 16: 5315-5322.

viii

Author contributions

Paper I

H.A. performed the lab work and made the first analysis of the data, which is

published in his Bachelor thesis at Stockholm University entitled: Measuring

diffusive mass transfer of organic chemicals through plastic and vegetation.

D.J.B. came up with the idea for the study, planned the experiment, super-

vised bachelor student H.A., redid the data analysis and took the lead in writ-

ing the manuscript based on H.A.’s thesis. M.M. was the main supervisor of

the project and together with A.J. helped write the paper.

Paper II

A.J and P.M. came up with the initial idea for this study, D.J.B. developed

the idea further and planned the experiment together with A.J. and M.M.

D.J.B. did the lab work and data analysis and took the lead in writing the

manuscript with contributions from the co-authors.

Paper III

A.J. and M.M. came up with the initial idea for this study. D.J.B. developed

the idea further and modified the existing passive dosing setup, designed the

experiment, did the leaf extractions and passive dosing experiments and

analyzed the data. F.I. and J.H. developed the nuclear magnetic resonance

method and did the principal component analysis of the results; D.J.B took

the lead in the writing of the manuscript, supported by contributions from the

co-authors.

Paper IV

M.M. had the initial idea for this study, D.J.B and M.M developed the idea

further, designed the study and D.J.B. gathered the data, performed the anal-

ysis and prepared the manuscript with contributions from A.J and M.M.

ix

Abbreviations

ACES – Department of Environmental Science and Analytical Chemistry

c.v. – Coefficient of variance

DGDG – Digalactosyldiacylglycerol

DW – Dry weight

EC – Eddy covariance

EI – Electron impact

EOM – Extractable organic matter

F – Fugacity

FW – Fresh weight

GC – Gas chromatography

H – Henry’s law constant

HCH – Hexachlorocyclohexane

HOC – Hydrophobic organic contaminant

k – Mass transfer coefficient

Kaw – Air / water partition ratio

KEOM/olive oil – EOM/olive oil partition ratio

Koa – Octanol / air partition ratio

Kow – Octanol / water partition ratio

LDPE – Low-density polyethylene

LRT – Long-range transport

MBR – Modified Bowen Ratio

MDL – Method detection limit

MQL – Method quantification limit

MS – Mass spectrometry 1H-NMR – Proton nuclear magnetic resonance

PAH – Polycyclic aromatic hydrocarbon

PC – Phosphatidylcholine

PCA – Principal component analysis

PCB – Polychlorinated biphenyl

PDMS – Polydimethylsiloxane

POP – Persistent Organic Pollutant

PTV – Programmed temperature vaporizing injector

QA/QC – Quality assessment / Quality control

QSPR – Quantitative structure property relationship

QuEChERS – Quick-Easy-Cheap-Effective-Rugged-Safe

x

REA – Relaxed eddy accumulation

SIM – Selective ion monitoring

Std. Dev. – Standard deviation

VR – Vetenskapsrådet

Z – Fugacity capacity

xi

Contents

Abstract ............................................................................................................ i

Sammanfattning ............................................................................................. iii

Samenvatting .................................................................................................. v

List of papers ................................................................................................. vii

Author contributions ......................................................................................viii

Abbreviations ................................................................................................. ix

Contents ......................................................................................................... xi

1 Introduction 1.1 Fate and transport of hydrophobic organic chemicals in the environment .......... 1 1.2 Importance of understanding air-surface exchange of HOCs ............................. 2 1.3 Mass transfer coefficients and equilibrium partitioning into leaves ...................... 4 1.4 Measuring surface-atmosphere fluxes in the environment .................................. 6 1.5 Objectives of this thesis ...................................................................................... 8

2 Methods 2.1 Chemicals .......................................................................................................... 9 2.2 The “silicone sandwich” (Papers I and II) .......................................................... 10 2.3 Passive dosing via the headspace using spiked olive oil (Paper III) ................. 12 2.4 Comparison of the modified Bowen ratio method and the eddy covariance

technique (Paper IV) ........................................................................................ 15

3 Results and discussion ....................................................................... 17

4 Conclusion .......................................................................................... 24

5 Outlook ................................................................................................ 25

6 Acknowledgements ............................................................................. 27

7 References .......................................................................................... 29

1

1 Introduction

1.1 Fate and transport of hydrophobic organic chemicals in the environment

Chemicals have played a vital role in society since the industrial revolution

and are being developed at an ever increasing rate. Now, roughly 15,000

new substances are registered every day1. Not all chemicals are without con-

cern however and it is important to assess which chemicals pose a potential

threat to the health and reproduction of organisms, including ourselves. An

important part of this risk assessment is to predict what happens to a chemi-

cal once it is released into the environment, i.e.: will it stay where it has been

emitted, which environmental matrix will it mainly partition to and which

transformation processes will it undergo? Can it be transferred through the

food chain? How long can it survive in the environment before it is broken

down? The answers to these and related questions are what we define as the

fate and transport of a chemical2,3

.

The fate and transport of a chemical in the environment are determined by

the chemical’s partitioning properties and its persistence, which are both

dependent on the physicochemical properties of the chemical and that of the

matrix in which the chemical is located3. Environmental factors such as tem-

perature, precipitation, presence/absence of certain biota and advective flux-

es also play an important role3,2

. Persistent chemicals are of particular con-

cern as they can build up in concentration in the environment if emissions

continue. Frank Wania divided persistent organic chemicals into 4 groups,

according to their transport characteristics4. There are “fliers”, “single hop-

pers”, “multiple hoppers” and “swimmers”. The multiple hoppers are a par-

ticularly interesting group of chemicals. This group consists of semi-volatile

chemicals with air/water partition ratios (log Kaw) between -4 and 0 and oc-

tanol/air partition ratios (Log Koa) values between 6.5 and 10. These chemi-

cals can bind strongly to organic matter but can also be transported through

the atmosphere, potentially “grasshopping” to polar regions over multiple

cycles between the atmosphere and the terrestrial surface (Fig. 1).

2

A well-known group of chemicals that is both persistent and has a large po-

tential for long-range transport (LRT) is the persistent organic pollutants

(POPs) for which the production and usage has been banned or severely

restricted under the Stockholm convention3. POPs can accumulate in remote

arctic areas, far away from major sources due to a combination of so-called

“cold condensation” (decreasing volatility of chemicals with decreasing

temperature) and reduced degradation in cold climates6. Almost all POPs are

semi-volatile persistent hydrophobic organic chemicals (HOCs), which have

been the focus of this thesis.

1.2 Importance of understanding air-surface exchange of HOCs

Having a good understanding of the air-surface exchange of HOCs is essen-

tial to understand their fate for several reasons. First of all, the atmosphere

covers the entire globe and is in contact with soils, freshwaters, oceans and

vegetation. The atmosphere serves as a transport medium for HOCs to areas

far beyond their point of release7. Secondly, the exchange of a chemical

between air, water, soil or sediment can have a large impact on its lifetime in

the environment and therefore its potential for LRT. Degradation in air is

much faster than in soil and sediment for the majority of organic chemicals

due to the presence of highly reactive hydroxyl radicals in the atmosphere3.

Finally, it is important to understand the air-surface exchange of persistent

Figure 1: Figure displaying the pathways through which persistent organic chemi-cals can be fractionated across the earth. Figure redrawn by Matthew MacLeod, based on Wania and Mackay (1996)

5.

3

HOCs in order to make accurate mass balance calculations of chemicals in

the environment8,9

.

Vegetation covers 80 % of the terrestrial surface area10

and plays an im-

portant role in the uptake and transfer of HOCs from the atmosphere to the

terrestrial environment10

. The transfer of HOCs from the atmosphere to

plants that are subsequently consumed by herbivores is thought to be a major

uptake pathway in humans for many HOCs11–13

. Vegetation can also influ-

ence the fate and transport of HOCs in the environment through a process

referred to as the forest filter effect14,15

, which leads to increased deposition

of HOCs to forested soils in comparison with non-forested soils. This elevat-

ed deposition on the soil, caused by litter fall and wax shedding from the

foliage, is most pronounced for chemicals with a log Koa between 7 and 11

and a log Kaw larger than -616–18

. Forest soils are generally assumed to func-

tion as sinks for HOCs from the atmosphere due to their high abundance and

large capacity to sorb HOCs16

. It has been hypothesized that climate change

and reduced primary inputs of many HOCs, however, could turn forest soils

and vegetation into secondary sources of HOCs to the atmosphere, thereby

acting as a buffer to maintain atmospheric levels and hence prevent decreas-

ing atmospheric concentrations of HOCs19,20

.

Modelling studies have shown that the forest filter effect has the potential to

cause a significant reduction of the LRT potential of HOCs but at the same

time result in an increase of the overall persistence of HOCs in the environ-

ment as degradation of these chemicals is often slower in soil than in air or

water21

. Boreal forested areas21

and forested mountain regions22,23

in particu-

lar play an important role due to their nearness to chemical sources, high tree

coverage and low temperatures which lead to cold condensation and reduced

degradation rates.

In the case of HOCs, direct uptake into plants from the soil is limited due to

the high capacity of the soil to store the chemicals (high fugacity capacity, Z)

and the low solubility of these chemicals in water (high octanol/water parti-

tion ratios, Kow). In most plant species, the majority of HOCs are instead

deposited from the atmosphere onto the leaves24–28

. This deposition is

thought to occur mainly by sorption into the cuticle, the outermost waxy

layer of the leaf, which is in direct contact with the atmosphere29

. The cuticle

is a complex heterogeneous structure that consists of (1) extractable lipids

such as waxes and (2) polymeric lipids such as cutin and cutan. The poly-

meric lipids are intertwined with proteins, waxes and sugars (Fig. 2), and

cannot be extracted from the leaves with conventional solvent extraction

methods30–32

.

4

The structure of the cuticle can differ widely between leaves from different

species31

, leaves from the same species31

and even different parts of the cuti-

cle on the same leaf33,34

. While it has been suggested that stomata could play

an important role in the uptake of HOCs from the gas phase35

, studies on the

uptake of phenanthrene into leaves showed no significant uptake via this

pathway36,37

. A better understanding of the uptake of HOCs into leaves

would allow for more accurate predictions of the fate of these chemicals in

the environment and the related risks19

. This understanding can be achieved

by generating more data on: (1) partition ratios of HOCs between leaves, air

and water; (2) kinetics of transfer of HOCs into and through leaves; and (3)

the variability of these partition ratios and transfer rates across plant species.

1.3 Mass transfer coefficients and equilibrium partitioning into leaves

Studies on the kinetics of mass transfer of organic chemicals into leaves

were popular in the 1980ies and led to some important developments in the

field of agrochemicals. An example is the development of surfactants for use

in pesticide formulations38

. There is only limited information available from

the literature on mass transfer coefficients of HOCs through intact leaves

however, as the majority of the literature data on mass transfer kinetics

through leaves focuses on the transfer through isolated cuticles39–41

. The

approach of isolating cuticles is work-intensive but has been demonstrated to

yield similar results as with intact leaves, with the added benefit that the

cuticles are rather stable and can be stored for weeks to months before use39

.

It has been reported however that thinner cuticles can easily be damaged

during the isolation process, which can influence their permeability for

chemicals42

.

Figure 2: Cross section of a leaf (A) and detailed structure of the cuticle (B). The cross section was taken from Wikimedia commons and was designed by user Zeph-yris. The cuticle was redrawn from a figure by Peter J. Holloway

31.

5

The main barrier that slows down the uptake of HOCs in leaves is thought to

be the crystalline waxes in the outer layer of the cuticle. Experiments meas-

uring the mass transfer rates of chemicals through isolated cuticles with and

without waxes found that the transfer rates were substantially higher if the

waxes had been removed43

. For the chemical 2,4-dichlorophenoxy acid, for

example, the differences in mass transfer rates across the cuticle spanned 4

orders of magnitude, before and after removal of the epi-cuticular waxes by

solvent dipping43

. The epi-cuticular waxes lining the surface of the cuticle

(Fig. 2) can form various complex structures depending on their chemical

composition44

and have been shown to become more amorphous over time

and when located close to sources of air pollution45

.

Using two-photon excitation microscopy, Wild and colleagues were able to

visualize the uptake of phenanthrene from the atmosphere into the leaves of

maize and spinach, and observed that phenanthrene diffused through the

cuticles of the leaves within 24-48 hours34

. The authors observed that while

the most important uptake pathway for phenanthrene was through gaseous

uptake, this chemical was also taken up from particles that were found em-

bedded into the epi-cuticular waxes of the leaves34,46

. It has been demon-

strated that particulate matter smaller than 10.4 µm can become encapsulated

in the wax, after being deposited on the leaves, so that it is kept from being

washed off by precipitation46

. The leaves from maize and spinach showed

different transport pathways, as phenanthrene was found mainly in the cell

walls of maize while in spinach most of the phenanthrene was found in the

cytoplasm (Fig. 2)34

. In a follow-up paper, the authors suggested that this

fast uptake through the cuticle could mean that the cuticle was not the main

reservoir for HOCs36

, in contrast with many other studies29

.

Multimedia fate models that include a vegetation compartment use a range

of different strategies to estimate the uptake of HOCs in leaves. One ap-

proach is to assume that the uptake capacity of leaves is dominated by the

lipid fraction of the leaves and can be approximated by the use of an equiva-

lent volume of octanol (e.g. in models based on the BETR framework47

).

Another modeling approach is to use reported values for Kleaf/air, derived

from deposition measurements onto leaves, and assume that leaves from all

other plant species have similar uptake capacities (e.g. CoZMo-POP48

uses a

model parameterization based on measured deposition to 2 types of forests,

beech/oak and spruce16

).

Model parameterization is challenging, however, since Kleaf/air data reported

in the literature differ by up to three orders of magnitude between plant spe-

cies (Fig. 3).

6

While part of this variability is likely due to differences in the methods that

were used across studies, as reviewed below, the study by Kömp and

McLachlan49

shows a difference of up to a factor 20 in Kleaf/air measured for 5

grass species for a range of PCB congeners, using the same method. The

authors suggested that the observed differences were likely caused by the

composition of the leaf lipids rather than the lipid fraction as normalizing

Kleaf/air to the lipid fraction of the leaves did not decrease the variability be-

tween plant species49,50

. Normalizing to the thickness of the cuticle has not

been found to decrease the variability between Kleaf/air from different plant

species either50

. So far, to the best of our knowledge, the hypothesis that the

composition of leaf lipids drives variability in sorption capacity for chemi-

cals has not been systematically tested. However, Chen et al.51

recently sug-

gested that differences observed in the literature for Kleaf/air might be caused

by the fraction and accessibility of the cutin.

1.4 Measuring surface-atmosphere fluxes in the environment

High quality data on surface-atmosphere fluxes are needed to check the va-

lidity of multimedia fate and transport model outputs in the field9. It is not

yet feasible to measure the fluxes of HOCs in the environment using the

eddy covariance (EC) technique, which is currently the preferred approach

to measure fluxes of CO2, H2O, heat and recently also mercury. The EC

technique is based on high frequency measurements (5-10 Hz) of the chemi-

cal of interest together with measurements of the wind speed and direction.

Analysis of co-variance in these measurements provides an estimate of the

Figure 3: Plant/air partition ratios (log Kpa) from different reports in the literature plotted versus the chemicals’ log Koa. Data marked with * originate from fugacity meter measurements by Kömp and McLachlan,

49 and those marked with # were

derived from deposition fluxes by McLachlan and Horstmann.16,49

Bacci et al.52

reported Kleaf/air for azalea leaves and Su et al.

53 derived their Kpa from deposition

fluxes in a deciduous Canadian forest.

7

chemical fluxes. Unfortunately, there are no existing analytical methods that

are fast enough to apply the EC approach for HOCs. Conventional sampling

techniques for airborne HOCs instead rely on the use of (1) large-volume air

sampling or (2) passive sampling that can take hours to months before suffi-

cient amounts of the chemicals have been collected to overcome existing

detection limits associated with the analysis of these chemicals7.

An alternative method that could be used to measure fluxes of samples that

require longer sampling times is the relaxed eddy accumulation (REA) tech-

nique. This approach uses high frequency measurements of the wind speed

and direction in combination with fast switching valves to split the incoming

airflow according to the dominating wind direction55

. The sampled air can

then be bulked in reservoirs56

or passed through denuders or sorbents57

until

sufficient amounts of analytes have been captured to overcome method

quantification limits. Unlike the MBR method, however, the REA technique

has not seen any recent use to measure fluxes of HOCs, possibly because it

is technically challenging to apply.

Instead of applying the REA or EC techniques to measure the fluxes of

HOCs, recent field experiments have relied on more indirect methods to

estimate atmospheric fluxes. One of the methods that has been used is the

modified Bowen ratio (MBR) method, which is based on the assumption that

under conditions of neutral atmospheric stratification, turbulent atmospheric

transport occurs to the same extent for all scalar quantities, such as fluxes of

heat and chemicals58

. This assumption allows us to estimate the flux of a

chemical of interest (Fx) by measuring its concentrations (Cx) at two differ-

ent heights (Z) and using the mass transfer coefficient (ky) derived from

measuring heat flux over the same height interval (Eq. 159

).

Eq. 1: 𝐹x = −𝐾y ∗ ∆𝐶x

∆𝑍

The MBR method has recently been applied to measure the dip in atmos-

pheric concentrations of HOCs in the field associated with the onset of leaf

development in spring60

. In that study, concentrations of polycyclic aromatic

hydrocarbons (PAHs) were measured in the atmosphere, with sampling

times of 24 hours. A similar approach, but with shorter sampling times was

also used to estimate fluxes of polychlorinated biphenyls (PCBs) from Lake

Superior61

. While both studies showed promising results, it is not clear from

either of them if the MBR is really suitable for such long sampling times.

8

1.5 Objectives of this thesis

The primary objective of this thesis was to improve our understanding of

surface-atmosphere fluxes of HOCs, in particular related to exchange of

HOCs between air and vegetation. The development of relatively simple and

fast methods to study the mass transfer kinetics of HOCs through leaves and

partition ratios between leaves and different matrices was the main focus of

papers I, II and III included in this thesis. In paper IV, we evaluated the va-

lidity of the modified Bowen ratio method that has recently been used to

measure surface-atmosphere fluxes of HOCs in the field60,61

.

Our goal in paper I was to test an existing passive dosing method62,63

to

measure mass transfer kinetics of HOCs from a silicone donor phase on one

side through intact leaves to a silicone acceptor phase. This method clearly

offers an interesting tool when measuring, for instance, the fluxes of chemi-

cals transferred through the husks of protected produce into the fruit (e.g.

corn). Our main aim, however, was to determine how fast chemicals would

pass through leaves and if there were any clear differences in the mass trans-

fer kinetics between the species tested as opposed to a simple polymer, i.e.,

low-density polyethylene (LDPE).

In paper II our goal was to modify the method used in paper I to use a sili-

cone donor phase on each side of the leaf, and to determine concentrations of

chemicals in leaves at equilibrium with the silicone. Such a method would

provide a straightforward way to measure the fugacity capacities of intact

leaves which could then easily be converted into Kleaf/air and partition ratios

between leaves and water, Kleaf/water. A similar experimental approach using

isolated cuticles was recently published and gave us a good reference for

discussion64

.

In paper III, we aimed to improve upon an existing passive dosing setup, and

use it to measure the partitioning properties of leaf extracts from various

plant species with foliage of widely differing characteristics. Our goal was to

test the hypothesis that the large differences reported for Kleaf/air values in the

literature were reflected in the partitioning properties of the extractable or-

ganic matter of leaves from different species.

In paper IV, we evaluated how well the vertical fluxes in the atmosphere

obtained using the MBR method correspond with measurements made by the

more commonly used EC approach to study if we can extend the MBR

method to accommodate for the long sampling times of days to months re-

quired for most HOCs in air. Our goal in paper IV was therefore to evaluate

the MBR method using CO2 and H2O as proxies for HOCs and to see how it

compares with measurements of fluxes made with the EC approach.

9

2 Methods

2.1 Chemicals

The chemicals used in this study are listed in Table 1, together with relevant

physicochemical data. These chemicals were chosen for study in this thesis

because their properties have been well studied and their partition ratios

cover a wide range of values typical for semi-volatile chemicals. Table 1: List of chemicals that were the focus of this thesis. The partition ratios for the PCBs were obtained from Schenker et al. using QSPRs

65, all the others were

obtained from Epiweb 4.1. , using the reported experimental values.

Chemical CAS number Log Kaw Log Kow Log Koa Used in

PCB 3 2051-62-9 -1.99 4.77 6.88 Papers II, III

PCB 4 13029-08-8 -1.42 5.24 6.86 Papers II, III

PCB 28 7012-37-5 -2.00 5.65 7.91 Papers I, II, III

PCB 52 35693-99-3 -1.81 6.10 8.25 Papers I, II, III

PCB 101 37680-73-2 -2.01 6.53 8.95 Papers I, II, III

PCB 118 31508-00-6 -2.39 6.51 9.30 Papers I, II,III

PCB 138 35065-28-2 -2.20 6.96 9.64 Papers I, II, III

PCB 153 35065-27-1 -2.20 6.96 9.64 Papers I, III

PCB 180 35065-29-3 -2.40 7.39 10.34 Papers I, II, III

Naphthalene 91-20-3 -1.75 3.30 5.20 Paper I

Fluorene 86-73-7 -2.41 4.18 6.79 Paper I

Phenanthrene 85-01-8 -2.76 4.46 7.22 Paper I

Anthracene 120-12-7 -2.64 4.45 7.55 Paper I

Pyrene 129-00-0 -3.31 4.88 8.80 Paper I

Fluoranthene 206-44-0 -3.44 5.16 8.88 Paper I

Monochlorobenzene 108-90-7 -0.90 2.84 3.31 Paper III

1.2-Dichlorobenzene 95-50-1 -1.11 3.43 4.36 Paper III

1,2,4- Trichloroben-

zene

120-82-1 -1.24 4.02 4.95 Paper III

1,2,3,4-

Tetrachlorobenzene

634-66-2 -1.51 4.60 5.64 Paper III

Pentachlorobenzene 608-93-5 -1.54 5.17 6.71 Paper III

Hexachlorobenzene 118-74-1 -1.16 5.73 7.38 Paper III

10

2.2 The “silicone sandwich” (Papers I and II)

To measure the transfer kinetics of HOCs through leaves and partition ratios

between leaves and different media, we used a passive dosing approach us-

ing sheets of the silicone polydimethylsiloxane (PDMS). This method was

originally developed in a study to quantify the effects of medium composi-

tion on the mass transfer rates of HOCs through unstirred boundary layers62

and was later used to measure the mass transfer rates of HOCs through slices

of potato and carrot63

. The original method used one sheet of PDMS that was

spiked with the HOCs of interest (the donor) and one sheet of PDMS that

was not spiked, and which was used as an acceptor phase. In both studies,

replicate samples showed a very high degree of precision. A modification of

the method was introduced by Kim and colleagues64

, who exchanged the

acceptor phase with a second donor phase and used the modified system to

measure cuticle/PDMS partition ratios (Kcuticle/PDMS) instead of transfer kinet-

ics64

. The precision in the study by Kim et al. was lower than that reported

previously and indicated a larger variability associated with analysis of

chemicals within the leaf samples than in the silicone matrices used in the

studies of Mayer et al.60

and Trapp et al.62,63

.

Previously used methods to measure the mass transfer coefficients of organic

chemicals through isolated cuticles, were based on several types of transport

chambers in which isolated cuticles were mounted between two cham-

bers41,42,66

, one containing radioactively labelled chemicals of interest in an

aqueous based buffer solution and the second chamber containing a phos-

pholipid suspension (to keep concentrations in the second chamber close to

zero). A technique that was used to measure the mass transfer coefficients

into intact leaves used glass cylinders that were fixed to the leaf surface us-

ing silicone rubber. These cylinders were filled with a buffer solution con-

taining radioactively labelled chemicals of interest. Samples of the leaves

were then analyzed at certain time points to determine the mass transfer rate.

However, the very low solubility of most HOCs in water means that using

this approach for HOCs is often done at low analyte concentrations, which

can be an issue when the use of radioactively labelled chemicals is not pos-

sible39

. An exception to this is a recent study by Li et al.40

in which they

managed to measure mass transfer rates of unlabeled phenanthrene through

isolated cuticles. The advantage of the “silicone sandwich” approach is that

it is easy to work with (sample preparation under 30 min), has a high

throughput and due to the high capacity of silicone for hydrophobic chemi-

cals, it is possible to work at high concentrations of HOCs.

In our studies, we have used both versions of the “silicone sandwich” to

reach our goals as described above and illustrated in Fig. 4. To measure the

mass transfer kinetics of HOCs through leaves and LDPE (paper I), we used

11

the “silicone sandwich” system with one donor and one acceptor PDMS

phase and analyzed only the concentration of HOCs in the PDMS donor and

acceptor phases. To measure the fugacity capacities of leaves (Zleaf) (paper

II) we used the system with 2 donor phases and analyzed the concentrations

of HOCs both in the leaves and in the combined PDMS donor phases.

Differences between our approach and those from the literature62–64

include:

A glass sheet was inserted between the “sandwich” setup and the

magnets or clamps to distribute the pressure more equally

PDMS had the same thickness but was obtained from a different

vendor

A different procedure was used to load the PDMS with chemicals

To calculate the mass transfer coefficients (kc) in paper I we used GraphPad

Prism to fit a one-phase association curve (Eq. 2) to the data in order to cal-

culate the rate constant k (h-1

) and then used these values in combination

with the thickness of the matrices to derive kc (m h-1

). In Paper II, the same

curve was fitted to the leaf/PDMS concentrations ratios and was then used to

determine the concentration ratio at equilibrium (Kleaf/PDMS) which was used

to calculate Zleaf (see paper II for detailed calculations67

).

Eq. 2: Y = Ymax (1-e(-kt)

)

An alternative method to measure Zleaf of intact leaves would have been to

dose the leaves first by placing them in a contamination chamber with the

chemicals of interest and then measure the fugacity (f) through the use of a

fugacity meter in which air is passed over the contaminated leaves at a slow

enough flow rate so that equilibrium between the air and leaves is reached68

.

At equilibrium, the fugacity in the air is equal to that in the leaves which can

easily be determined from Eq. 3 by measuring the concentration in the out-

going air as the fugacity capacity of air for most organic chemicals is equal

to 1/ RT with R being the gas constant (8.314 m3 Pa K

-1 mol

-1) and T the

temperature of air (K)2.

Figure 4: Example of the setup used in paper I (1 donor + 1 acceptor sheet) and

paper II (2 donor sheets).

12

Eq. 3 f = concentration / Z

When the fugacity in air, and thus that in the leaves is known, Eq. 3 can then

be used to back calculate Zleaf by using the concentrations measured in the

leaves at equilibrium. The fugacity meter has successfully been used to

measure Kleaf/air for PCBs in different plant species49

. Its use is technically

much more challenging however than the PDMS “sandwich” used in our

studies which makes it difficult to scale up to large number of samples.

2.3 Passive dosing via the headspace using spiked olive oil (Paper III)

To investigate potential differences in the sorptive capacities of leaves, a full

lipid extraction69

of foliage from 7 different plant species was performed,

yielding the extractable organic matter (EOM). These EOM samples were

then introduced into a passive dosing system that is illustrated in Fig. 5. In

this system, spiked olive oil was used as the donor phase to measure the

partition ratios between the EOM and olive oil, KEOM/olive oil, for a set of

HOCs (Fig. 5). The passive dosing system used in this study was introduced

in 201070

to determine the sorptive capacities of matrix-immersed PDMS-

coated fibers and then extended in 201571

to analyze KEOM/olive oil for EOM

from 5 different animal species. The passive dosing system consists of a 500

mL glass jar which contains olive oil that is spiked at high levels (~1 mg mL-

1) with the compounds of interest and to which 5 mL vials containing the

matrix to be dosed (such as EOM or model lipids) are introduced. The sys-

tem is left to equilibrate via the headspace over time, after which the concen-

trations in both matrices are measured and Kmatrix/olive oil can be determined.

Using passive dosing through the headspace offers the advantage that we

avoid the necessity of separating the donor and acceptor phases upon analy-

sis. Furthermore, as both the matrix and the olive oil are similar in their

composition, we can use the same extraction method for both matrices,

avoiding a process-related bias.

An alternative method that could have been used for the more volatile chem-

icals, if we had a headspace sampler would be through the analysis of the

headspace in two different vials; one containing the EOM and the other con-

taining water. By then taking the ratios of the areas measured for the chemi-

cal of interest in either headspace (assuming a linear detector response and

equilibrium between the air and EOM or water72

), Klipid/water can be deter-

mined by Eq. 473

. Aw and AEOM are the peak areas for the chemicals detected

13

in the headspace of the vials with water and EOM respectively, M is the

amount of chemical in the water and EOM.

Eq. 4 𝐾EOMwater⁄

= 𝐴w

𝐴EOM 𝑀tot.EOM

𝑀tot.w

In this study, the previously reported method using the spiked olive oil71

was

modified in two ways: (1) A fan was introduced into the lid of each jar to

increase the turbulence in the system and thereby the mass transfer kinetics

via the headspace towards the samples to be dosed; and (2) The vials inside

each jar were placed on a metal grill to increase the contact area between the

air and the donor oil and to avoid losses of the donor oil when the vials were

taken out of the jar for analysis (Fig. 5).

The plant species were selected to represent a range of plant lipid character-

istics based on differences in their visual appearance, and their availability in

a nearby botanical garden (Bergianska Trädgården). Leaves were collected

from three deciduous species, two conifers, one shrub and one grass species

(Fig. 6).

Following the sample collection, leaves were transported directly to the lab

where they were rinsed with de-ionized water (to remove particulate matter),

cut into small pieces, mixed together to randomize the source of the material

and then extracted using an exhaustive solvent extraction, the so-called Jen-

sen extraction (modification 2)69

. In total three extractions were performed

with roughly 10 g of material for each of the plant species.

Figure 5: Experimental setup for paper III. Image composed from vector drawings from openclipart.org.

Figure 6: Plant species used in paper III. Pictures were taken from Wikimedia.org.

14

The motivation for choosing the modified version of the Jensen extraction

was based on its superior yield of lipids from low-fat samples compared to

the original Jensen method69

. The Jensen extraction method has also been

shown to give EOM yields comparable to those of the more “classical” Bligh

and Dyer74

and Folch75

methods which have been used for total lipid extrac-

tions, but without the use of halogenated solvents thereby making it safer to

work with69

, and preferable from the perspective of green chemistry. In addi-

tion to EOM extracts from leaves of 7 different plant species, we also in-

cluded a set of lipid standards as reference materials (olive oil (as a surrogate

for storage lipids, consisting of 99 % triglycerides)71

, digalactosyldiacyl-

glycerol (DGDG, most abundant membrane lipid in higher plants76

) , l-a-

phosphatidylcholine (PC, membrane lipid in plants and animals), nonaco-

sane (long chain paraffin present in plant waxes) and octanol (well used

proxy for lipids3)).

Subsamples of the EOM and lipid standards were taken before exposure to

the donor oil and at 4 different time points over the course of two weeks. At

each time point, three replicates were taken for the EOM and lipid standards

and one for the donor oil. Each sample was extracted directly upon sampling

using a purge-and-trap method and then analyzed using gas chromatography

coupled to mass spectrometry (GC-MS)71,77

.

Equilibrium between the chemical concentrations in the donor oil and those

in the EOM and lipid standards was evaluated by comparing the 95 % confi-

dence intervals of the concentration ratios in the EOM/lipid and olive oil

over time. The partition ratio between EOM and olive oil, KEOM/olive oil, and

between a lipid standard and olive oil, Klipid/olive oil, were then calculated by

taking the average of all the replicates that reached equilibrium. In case equi-

librium was not reached during the 2-week exposure time, KEOM/olive oil and

Klipid/olive oil were calculated by extrapolation using a one-phase association

curve in GraphPad Prism (Eq. 2). The EOM of each plant species and the

lipid standards were characterized using a screening method based on proton

nuclear magnetic resonance (1H-NMR)

78 to evaluate potential correlations

between lipid composition and KEOM/olive oil and Klipid/olive oil.

15

2.4 Comparison of the modified Bowen ratio method

and the eddy covariance technique (Paper IV)

To evaluate how well the results of the MBR method compare with those

from EC measurements, we used a publicly available dataset from

FLUXNET79

which collects data from flux towers around the world. The

FLUXNET datasets contain measured concentrations and EC-derived flux

measurements of CO2 and H2O at defined heights on a tower averaged over

30 minute intervals (Fig 7).

In this case we chose to use the 2009 dataset from Borden, a mixed decidu-

ous forest in Ontario, Canada. An earlier version of this dataset was used

also by Choi et al.60

to derive mass transfer coefficients that they used to

estimate fluxes of PAHs, thereby providing a reference point to validate our

Figure 7: Illustration of the FLUXNET tower and the measurements that were used in paper IV (from Bolinius et al.

80).

16

calculations. Based on the assumption that CO2 and H2O are good proxies

for micro pollutants such as HOCs, the concentration measurements in the

Borden FLUXNET data were pooled over selected time periods to simulate

the long sampling times needed to sample HOCs. The mass transfer coeffi-

cient of heat (kheat), available from the same dataset, was used to specify the

eddy diffusivity in the MBR method.

17

3 Results and discussion

In paper I, we demonstrated the use of a passive dosing method to measure

mass transfer coefficients through intact leaves. Out of 13 analytes tested in

this study, only two (fluorene and phenanthrene) were measured in samples

above the method quantification limit (MQL) for 3 out of 4 matrices tested

(Fig. 8).

Transfer through rhododendron leaves was too slow to result in any of the

analytes reaching the receptor silicone at levels above MQL. These observa-

tions indicate that analyte concentrations in this passive dosing experiment

should be increased in future experiments.

The measured mass transfer coefficients were within the range of those re-

ported in the literature for (2,4-dichlorophenoxy) acetic acid41

but 3 orders of

magnitude lower than those reported for phenanthrene in isolated cuticles of

Figure 8: Percentage of compounds transferred through a matrix to the acceptor PDMS over time for two target compounds with analyte levels in the acceptors > MQL for diffusion through hydrangea leaves, romaine lettuce leaves and LDPE. All acceptors were MDL were used to fit the curve (from Ahmadi et al.)

81.

18

green pepper40

.The study provided a proof-of-concept for the approach and a

set of recommendations for further improvements. The measurements ob-

tained in this study gave a first indication that mass transfer through rhodo-

dendron leaves is very slow. Another interesting result from this study was

that we found differences of a factor 12 between our measured mass transfer

kinetics of phenanthrene between two sheets of PDMS in direct contact and

those found in the literature for a very similar experiment62

. This difference

is too large to be caused solely by different types of PDMS used82

and we

speculate that the difference could reflect lower pressure on our system ex-

erted by the magnets compared to those used in previous work, as our study

used glass sheets to spread the pressure more equally and a higher leaf sur-

face area relative to the surface area of the magnets (Fig. 4).

In paper II we modified the method used in paper I as described above and

showed that it could also be used to measure fugacity capacities of leaves,

which can then easily be converted into the partition ratio of interest or be

used as a direct input into multimedia fate models. The analytes in the sys-

tem reached 76 to 99 % of equilibrium within the 4 days of exposure of the

leaves to the loaded PDMS sheets which is comparable to the kinetics re-

ported for PAHs in a similar setup64

. The results obtained for the partition

ratios in this study were well within the range of uncertainty that is found in

the literature, with Kleaf/water and Kleaf/air increasing with increasing log Kow of

the analytes (Table 2, Fig 3).

Table 2: Rate constants (k in h

-1), log Kleaf/PDMS and values for Zleaf, log Kleaf/water and

log Kleaf/air. Values of k and Zleaf are given with standard deviations which show the uncertainty of the extrapolation using the nonlinear regression. For log Kleaf/PDMS, log Kleaf/water and log Kleaf/air, the uncertainty was propagated from the input data and is given as a range of ± 1 standard deviation of the measurements (Bolinius et al.

67).

Compound k (h-1) Log Kleaf/PDMS Zleaf (mol m-3 Pa-1) Log Kleaf/water Log Kleaf/air

PCB 3 0.045 ± 0.022 -0.4 (-0.5 to -0.4) 146 ± 64 3.5 (3.3-3.7) 5.6 (5.3-5.7)

PCB 4 0.015 ± 0.012 -0.8 (-1.1 to -0.6) 40 ± 24 3.6 (3.2-3.8) 5.0 (4.6-5.2)

PCB 28 0.029 ± 0.017 -0.7 (-0.8 to -0.6) (1.6 ± 0.7) x 103 4.7 (4.4-4.8) 6.6 (6.3-6.8)

PCB 52 0.021 ± 0.012 -1.0 (-1.0 to -0.9) (1.4 ± 0.7) x 103 4.6 (4.3-4.8) 6.6 (6.3-6.7)

PCB 101 0.023 ± 0.014 -1.1 (-1.2 to -1.0) (5.7 ± 2.8) x 103 5.1 (4.8-5.2) 7.2 (6.9-7.3)

PCB 118 0.028 ± 0.017 -0.9 (-1.0 to -0.8) (2.2 ± 1.1) x 104 5.4 (5.1-5.6) 7.8 (7.5-7.9)

PCB 138 0.022 ± 0.012 -1.0 (-1.2 to -0.9) (1.7 ± 0.8) x 104 5.7 (5.4-5.8) 7.6 (7.3-7.8)

PCB 180 0.024 ± 0.016 -1.1 (-1.3 to -1.0) (8.1 ± 4.1) x 104 5.8 (5.5-6.0) 8.3 (8.0-8.5)

19

Furthermore, by normalizing our results to estimated fractions of cuticle in

rhododendron leaves, a close fit was found to cuticle/water partition ratios

(Kcuticle/water) from the literature29,64

, allowing us to propose a 1-parameter

regression model to estimate log Kcuticle/water from log Kow (Fig. 9).

The method presented in paper II provides a valuable tool to measure fugaci-

ty capacities of leaves from a wide range of plant species and sheds more

light on how these capacities differ between species.

In paper III, we used another passive dosing system based on spiked olive oil

to measure the partition ratios of the EOM of leaves from 7 different plant

species and olive oil (KEOM/olive oil) and Klipid/olive oil. While we expected large

variations between the species due their appearance and literature data (Fig.

3), we only observed significant differences in the sorptive capacities of leaf

extracts for HOCs in a few cases and those were all below a factor 2 (Fig.

10).

Figure 9: Comparison of the data presented in paper II, normalized to the estimat-ed fraction of cuticle in the leaf and cuticle/water partition ratios from Kim et al.

64 ,

who reported their own measurements, a collection from the literature and meas-urements from Riederer et al.

29 The regression equation through all data points (n

= 75), forced to a slope of 1, is logKcuticle/water = logKow + 0.0963 (R2 = 0.95), or

more conveniently: Kcuticle/water = 1.25 Kow (from Bolinius et al.67

).

20

Fig

ure

10

: M

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83).

21

The fact that Kolive oil/olive oil measured in our system was consistently close to

1.0 is an indication that the system either reached equilibrium or that the

concentration ratio at equilibrium could be derived by extrapolation of the

data. Klipid/EOM could not be obtained for octanol as most of the octanol was

lost from the sample vials within 24 hours. It was also not possible to meas-

ure Klipid/EOM for nonacosane due to very slow uptake rates. This is likely

caused by the crystalline structure of the nonacosane.

This similarity in KEOM/olive oil could be an indication that either the EOM was

not representative of the leaf lipids, possibly because the structure of the

lipids had been significantly altered during the extraction procedure64

, or that

the extractable leaf lipids did not represent the major sorptive capacity of the

leaves. It is interesting, however, that our measurements for Zrhododendron,

which were measured in paper II, fit well with the measurements of KEOM/olive

oil for rhododendron leaves from paper III and so did estimations of Koc-

tanol/olive oil, based on partition ratios calculated with poly-parameter linear free

energy relationships from the literature73

. The values of KEOM/olive oil measured

in paper III were also similar to those measured for animal extracts71

. It is

therefore possible that the sorptive capacity of EOM, being a “bulk lipid”

that contains a variety of lipids, waxes and co-eluting compounds, does not

vary substantially across species and kingdoms and might be usefully ap-

proximated by the use of octanol in models.

Our results also showed a clear difference among the Klipid/olive oil measured

for the pure lipids with the data for DGDG being on average a factor 2.1

smaller than that of olive oil (triglycerides) and larger than that of PC by a

factor 1.6. The averaged data for KEOM/olive oil fell between those of DGDG

and olive oil. That the sorptive capacities of PC and triglycerides, often used

as representatives of membrane lipids and storage lipids, respectively, differ

is not a surprise. Physiologically based pharmacokinetic models already take

into account the differences in those types of lipids84

. However, the differ-

ence in sorptive capacities of PC and DGDG has not yet been reported in the

literature. Membrane lipids in higher plants can contain up to 75 % DGDG76

,

so distinguishing between the two types of membrane lipids has the potential

to improve our estimations of leaf uptake capacities.

In paper IV we demonstrated that the MBR method could be used to esti-

mate fluxes of HOCs between the atmosphere and the earth’s surface under

two conditions: (1) The measured gradient between the concentrations at two

heights must be strong enough to be measured accurately; and (2) The flux

does not change direction during the sampling. If fluxes do change direction

during the sampling time, it is possible to introduce a bias in the flux size

and direction when using long sampling times and hence this needs to be

avoided. The potential for bias when the flux direction changes during the

22

sampling period is demonstrated in Fig. 11, where estimated fluxes for three

different sampling durations are plotted against time. In this simulation, the

flux data have not been separated according to their flux direction. The im-

pact of this procedure on the fluxes measured with the MBR over 1 hour

intervals is limited compared to the EC measurements (half hour averages of

fluxes determined on timescales less than a second). Over longer time inter-

vals, however, the fluxes estimated with the MBR method are (1) in the op-

posite direction and (2) almost a factor 2 higher than those measured with

the EC technique.

If the two conditions stated above are met then our case study shows an

agreement between the results of the MBR method and those measured us-

ing the EC technique within a factor 3 for CO2 and within a factor 10 for

H2O (Table 3).

Figure 11: Illustrating the potential impact of changing flux directions during the sampling period of fluxes measured with the MBR method. The green curve (EC) represents half hourly data from eddy covariance data based on samples collected on timescales of less than a second, the blue and red curves represent measurements made with the MBR for hourly and daily pooled concentrations, respectively (from Bolinius et al.

80).

23

An interesting observation made in this study was that under the conditions

mentioned above, the duration of the sampling (1 h to 1 week) did not have a

large influence on the fluxes calculated with the MBR method. It also did

not seem to have an effect if we used an averaged value for the mass transfer

coefficient (in this case kheat) or hourly values, making the MBR method a

very useful approach if no high frequency data to characterize the mass

transfer coefficient can be obtained.

Table 3: Cumulative fluxes for 8 h periods representing day and night across the 2

month periods representing spring, summer, fall and winter. Fluxes measured by the

MBR method that are in the opposite direction than those EC results is based on the

geometric mean of the MBR results divided by the EC result. The MBR fluxes for the

1-week sampling period were left out in the calculation of the geometric mean dur-

ing the day in winter for CO2 and during the night in fall for H2O. This table shows

fluxes calculated with a fixed value for Kheat (from Bolinius et al.80

).

24

4 Conclusion

In this thesis, we have aimed at providing both methods and data that can be

used to improve our understanding of the role on vegetation of the fate of

HOCs in the environment. With paper I we have shown proof of concept for

a straightforward method that can be used to measure mass transfer of HOCs

through leaves. In paper II we have demonstrated a method to measure fu-

gacity capacities of leaves which can be used to calculate partitioning ratios

between leaves and a variety of different media. Values for Kleaf/air, derived

with this method using rhododendron leaves, were well within the range

reported for Kleaf/air for different plant species in the literature. As these val-

ues range by up to three orders of magnitude however, it is clear that more

research needs to be done to understand what drives this variability. The

method from paper II is easy to upscale and could be used to measure Kleaf/air

for a variety of plant species. With paper III we have delved deeper into the

issue of the variability in the sorptive capacities of leaves for HOCs and

found no link between the lipid composition of leaf EOM and its sorptive

capacity. While it was not clear yet if and to what extent the sorptive capaci-

ty of the leaf lipids is affected by the homogenization process, the results did

give an indication that the variability in Kleaf/air could be due to differences in

the fraction and accessibility of the cutin, which was not included in the

EOM. Finally in paper IV, we have evaluated the use of the MBR method to

measure fluxes of HOCs in the field and found that results measured with

the MBR method were within a factor 3 of those measured with the EC

technique for CO2 and within an order of magnitude for H2O. Two condi-

tions that need to be met however are that the fluxes should not change di-

rection during the sampling period and that fluxes are large enough to be

measured. The methods and data presented in this thesis can potentially lead

to improved parameterizations of multimedia models. The results of these

multimedia models can then be evaluated against fluxes measured in the

field using the modified Bowen ratio approach.

25

5 Outlook

As a direct extension of this research, the passive dosing approach developed

in paper II should now be applied to measure the fugacity capacity of leaves

from a wide variety of plant species to quantify inter-species differences in

fugacity capacities of intact leaves. If possible, this should be done by focus-

ing on the same set of species that were studied in paper III so that differ-

ences from the EOM can be evaluated against differences of intact leaves.

This experiment could be extended to measuring the fugacity capacities of

leaves with and without epicuticular waxes (which can be removed by strip-

ping85–87

). Another option would be to compare the sorptive capacities of

intact leaves and those of isolated cuticles. It has been shown that the mass

transfer coefficients of organic chemicals through isolated cuticles is similar

to those measured for intact leaves39

, yet the same has not yet been investi-

gated for the sorptive capacities for HOCs.

To increase the range of application of the passive dosing setup from paper

III, it is necessary to study the stability of the EOM and the lipid standards

over time, in order to see if it is possible to extend the passive dosing exper-

iment from paper III in time so that equilibrium can also be reached for less

volatile compounds. A solvent extraction instead of the purge-and-trap

method could increase the recoveries for less volatile compounds from the

EOM and lipid standards.

Another key gap in our understanding is what happens when leaves that have

accumulated substantial amounts of semi-volatile pollutants fall to the forest

floor and start decomposing. An unknown fraction of the chemicals will

likely be incorporated into the organic matter of the soil while the remainder

could be released into the atmosphere or metabolized. It has been suggested

that the fugacity of the chemicals sorbed to the leaves may increase during

decomposition due to the loss of sorption sites and decreasing fugacity ca-

pacities of the organic matter, in a process analogous to the uptake of HOCs

from food in the gastrointestinal tract of animals88

. It seems instead that the

litter has a capacity to sorb HOCs that is equal to that for fresh leaves, possi-

bly caused by complex aging effects that compensate for the loss of more

rapidly degradable components in the litter20,89.

Using the methods presented

in this thesis, it would be possible to study the decomposition of leaves in

more detail by measuring Zleaf at different stages of decomposition with the

26

silicone sandwich described in paper II. This approach enables focusing on

changes in Zleaf over time.

The MBR method should be validated by making field measurements with

large-volume samplers, low-volume samplers and passive samplers to get a

range of measurements with different sampling times. Atmospheric levels of

HOCs in background environments are likely too low to make fast meas-

urements; instead they could be measured in areas with high constant fluxes

such as urban areas (e.g. with siloxanes) or sites known to contain high lev-

els of HOCs. An idea would be to test the use of the MBR method to meas-

ure horizontal atmospheric fluxes of HOCs. A proof-of-concept for this ap-

proach could be tested in a similar way as was done in paper IV, but with a

selection of sampling sites across a horizontal transect, rather than measure-

ments taken at different heights on a tower. This method could provide valu-

able data on the impact of point sources of HOCs, such as the occurrence of

so-called “urban pulses” in which concentrations of certain HOCs are found

in high concentrations within urban areas and decrease with increasing dis-

tance of the city center90

.

Finally, it would be interesting to see if the MBR method can be extended to

measure fluxes of HOCs between sediments and the overlying water col-

umn. This setup would allow us to verify estimations made through mass

balance studies91

or equilibrium sampling using polymer-coated glass jars92

.

The successful use of the EC approach has recently been demonstrated for

flux measurements of O2 under water93

which could be a first step in that

direction.

27

6 Acknowledgements

Writing this thesis would not have been possible without the support, inspi-

ration and company from many people. As there are way too many of you to

list here, I will pick out just a few: My parents, who have always made sure

that I had good food, warm clothes and a sturdy roof over my head. Who

have paid for my entire education, including all the pub crawls, without ever

asking for anything in return and have supported me in every decision that I

have made, even when these meant going abroad repeatedly. They taught me

the value of hard work and honesty. My sister, the person that probably

knows me better than anyone and who I miss the most when being abroad, I

look forward to those few days of silliness each year. Sonni, the only guy in

the world that can make me cry from laughter 1500 km away and who is

travelling all the way to Stockholm just to see my defence. My friends in

Belgium, who still think that I work with mussels but are nice enough to take

me out for beers and road trips on short notice whenever I make it to Bel-

gium ;-). You always make it feel like I’ve never left, for which I am very

grateful. My colleagues at ACES, both old and new, for creating such an

amazing work environment. Whether students or staff, it always feels like

we’re all in it together and it has been a good inspiration to be surrounded by

so many people that are much smarter than me. Michael once asked me dur-

ing my job interview at ACES which scientist I looked up to the most. While

baffled at the time, I think I now have quite a long list of names to pick

from! My supervisors: Annika, Matt and Michael who have always left their

door open for me and who gave me the chance to make (a lot of) mistakes as

well. I have learned a lot during these years. In particular Annika and Matt,

thank you for guiding me towards this thesis. It has been great working with

you through all these tight deadlines and to learn from your feedback. I hope

that you can get some rest now as well :). Last but not least, thank you Maria

for not getting tired of my company yet and for all your support. It has been

an amazing three years!

"Stay hungry, stay healthy, be a gentleman, believe strongly in your-

self and go beyond limitations."

Arnold Schwarzenegger, Childhood Hero

28

29

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