Lecture 2 – Pollutant properties and fate

Pollutant properties and degradation 污染物物理化学特性 - 污染物分布 - 重

要特性（溶解度、Kow、吸附、蒸气
压、KH pKa ......） - 有机化合物特性
评估的实验数据或预测（EpiSuite 或类
• Pollutant phys-chem properties – pollutant distribution 似数据）

- Important properties (solubility, Kow, sorption, vapour pressure, KH pKa…)

- Experimental data or predicted (EpiSuite or similar) for property
estimation of organic compounds
• Pollutant degradation - biodegradation 污染物降解 - 生物降解 - Ready biodegra
dability à OECD 301 - 土壤中的生物降
- Ready biodegradability à OECD 301 解 à OECD 307

- Biodegradation in soils à OECD 307

• Degradation kinetics 降解动力学 - 一阶 - 复合动力学 - Mon
od 动力学 - 经验法则
- First order
- Composite kinetics
- Monod kinetics
- Rules of thumb
• Pollutant fate in aquatic or terrestrial systems à modelling
- Box models, e.g. for a lake (not treated here) 水生或陆地系统中的污染物归宿 à 建
模 - 盒模型，例如对于湖泊（此处未
- Soil-water-plant-atmosphere models 处理）-土壤-水-植物-大气模型
 污染物分布 第 2 讲 – 污染物性质和归
Lecture 2 – Pollutant properties and fate 宿 污染物的物理化学性质是确定污染
物分布、生物利用度/降解和毒性的基
本参数。当我们设计和解释降解实验
Pollutant distribution 时，这些特性很重要。实验性的或由
QSAR 估计的
The phys-chem properties of pollutants are basic parameters for determining
pollutant distribution, bioavailability/-degradation and toxicity. These properties are
important when we design and interpret degradation experiments. Experimental or
estimated by QSAR

Ex. 1: Distribution - Distribution of a solute (A)

between water, air and soil
From SWP1

• Distribution coefficient:
Kd (Koc and foc)
• Henrys Law coefficient:
KH
mAtot • Bio-concentration factors
[A]w = KBCF
(n − θ ) ⋅ KH + θ + ρb ⋅ Kd
Lecture 2 – Pollutant properties and fate

Soil-water-plant-atmosphere models
Soil systems: Inputs, outputs (leaching, volatiles, biomass), transformations.
Calculations start with modelling of the soil water balance, and hence water transport.
Climate, soil and plant cover (evapotranspiration) are main controlling factors
土壤系统：输入、输出（浸出、挥发
物、生物量）、转化。

计算从土壤水分平衡模型开始，然后
是水传输模型。

气候、土壤和植物覆盖（蒸发蒸腾）
是主要控制因素

DAISY: Comprehensive model developed at UCPH over last 30 years. Intensively used
for nitrate, pesticide and carbon modelling. See: https://daisy.ku.dk
Lecture 2 – Pollutant properties and fate

Example: Daisy modelling of nitrate leaching

Nitrate fertilizers added + mineralization of nitrogen

Lecture 2 – Pollutant properties and fate

Biotoxin leaching to surface and groundwater

生物毒素渗入地表水和地下水

生物毒素的产生、转移、归宿和运输
到土壤水系统中的水库。需要吸附、
溶解度、水解、生物降解、光降解…
…的数据

Biotoxin production, transfer, fate and transport to water reservoirs in in soil-

water system. Needs data for sorption, solubility, hydrolysis, biodegradation,
photodegradation….
 Lecture 2 – Pollutant properties and fate

The OECD Guidelines for the Testing of

Chemicals
• Section 1: Physical Chemical Properties
https://www.oecd.org/
• Section 2: Effects on Biotic Systems
• Section 3: Environmental Fate and Behaviour
• Section 4: Health Effects
• Section 5: Other Test Guidelines

Section 1 - examples
Kow, Koc, water solubility, vapour pressure, Henrys Law constant, acidity constants,
alkalinity, hydrolysis
Section 2 – examples
Fish acute toxicity, bumblebee acute contact toxicity, earthworm reproduction test
Section 3 – examples
Ready biodegradability, phototransformation, bioaccumulation, leaching in soil
columns, transformation in soil
Section 4 – examples
Pesticide residues, pesticide metabolism in crops.
Lecture 2 – Pollutant properties and fate

Octanol-water partitioning coefficient (Kow)

- HPLC method, OECD 117 (corrected June 2022) -

The guideline describes the principle of the

methods, the ingredients to be used, the
method in detail (equipment), the calculus,
uncertainty and reporting 该指南描述了方法的原理、要使用的
成分、详细的方法（设备）、微积分
、不确定性和报告

On the HPLC column determine the retention

time and capacity factor k: 在 HPLC 列上确定保留时间和容量因
子 k：

or other
Then: functional
relation
Example of reference curve.
Methanol/water as eluent using af
Lichrosphere RP-18 column 参考曲线示例。
Kaune et al. (1998) 使用 af Lichrosphere RP-18 柱的甲醇/水
作为洗脱液
Lecture 2 – Pollutant properties and fate

Octanol-water partitioning coefficient (Kow)

- HPLC method, OECD 117 (corrected June 2022) -

OECD 117 中推荐的参考化合物子集 -- Comparison of

---------三嗪除草剂 + 代谢物的测量 K
ow 值与文献 Kow 值的比较。 measured and literature
Subset of reference compounds Kow values for triazine
recommended in OECD 117 herbicides + metabolit.
Kaune et al. (1998)
Lecture 2 – Pollutant properties and fate

Bio-degradation
微生物（酶促）过程。例如： 水解
还原 氧化 聚合

Microbial (enzymatic)
processes. E.g.:
• Hydrolysis
• Reduction
• Oxidation
• Polymerization
Lecture 2 – Pollutant properties and fate

Aerobic/Anaerobic transformation in soil (OECD 307)

• Test substance: Often 14C labelled substance used, sometimes in addition to
unlabelled substances (isotopic label in the most stable part (other isotopes than
最稳定部分的同位素标记（可以使用
carbon can be used)) 碳以外的其他同位素）
• Analytical method: Sensitivity (LOD, LOQ), recovery, repeatability. 14CO2,
灵敏度（LOD、LOQ）、回收率、重
degradation products. 复性。 CO2，降解产物
• Test system type: Static closed systems or flow-through system. 静态封闭系统或流通系统
• Application of test substance: as solution, as solid mix, as commercial
formulation (not recommended). Careful with organic solvent added! Relevant
application levels (e.g. pesticides). Check for uniform distribution!
• Soils: Pathway (one soil) and rates (three soils). Key variables such a C, pH, clay,
microbial biomass, storage of soils. Appropriate moisture content
• Soil sampling: Fresh (!) or stored for a max. of three months. Pre-incubation of 2
– 28 days.
• Test: Darkness, 20 oC (depending on relevance), moisture control (pF 2.0 – 2.5),
duration up to 120 days. Include sterile controls and normal controls. Mass
balances! Duplicates.
• Analysis: Duplicate analysis at different time intervals (minimum of 6 times).
Extraction of soil, and if separate water phase that should also be analysed.
• Rate data fitted by 1st order equation if possible. If carried out at different
temperatures, then determine activation energy using Arrhenius equation.
如果可能，用一阶方程拟合。如果在
不同温度下进行，则使用 Arrhenius 方
程确定活化能。
Lecture 2 – Pollutant properties and fate

Setups for degradation studies

Flow-through apparatus for degradation studies

Test flask method

2: Gas washing bottle
3: Filter to keep microorganisms out
4: Soil reactor
CO2 sorber
5: Ethylene glycol trap for organic volatiles
6: Sulphuric acid trap for alkaline volatiles Sorber for
7 and 8: Alkaline traps for acid volatiles (CO2, etc) organic volatiles
9: Flow-meter
Soil
 Lecture 2 – Pollutant properties and fate

Mineralisation 放射性标记的底物混入土壤中。捕获
在 NaOH 溶液中的不断演变的 14C-C
- by use of 14C labelled substrate - O2 测量：在不同的时间牺牲样品瓶。
移除检测孔，加入闪烁液并使用闪烁
计数器测量 14 C-CO2 的量

Radioactive labelled substrate mixed into

soil. The evolving 14C-CO2 trapped in the
NaOH solution
Measurement: At different times a
sample vial is sacrificed. The detection-
well is removed, added scintillation liquid
and the amount of 14C-CO2 measured
using a scintillation counter

Semi-automatic Incubation setup

to study mineralization by use of
14C labelled substrate.

使用 14C 标记底物研究矿化的半自动 Mineralization of

孵化装置 cypermethrin
insecticide in a clayey
soil, 25 oC.
Lecture 2 – Pollutant properties and fate 污染物特性和归宿 挥发性物质在生物
降解试验期间逸出，使生物降解数据
不可靠。需要替代设置。四氢化萘和
癸烷的例子
Biodegradation of volatile substances
Volatile substances escape during the biodegradation test making biodegradation
data unreliable. Alternative setups needed.
Example of tetralin and decane

P: parent compound, M:
metabolites, V: volatile
Test system setup 4 different
First-order degradation.
soils
with Tenax sorbent
in the outlet tube. O2
flow through the
system. Fv: volatilization fraction
Shresta et al. (2019)
Lecture 2 – Pollutant properties and fate

OECD 307

• The guideline cannot be used for substances with high volatility. Why not?
• The sampling and analytical method should have a high recovery. What does that
mean?
• The test substance may be added to the soil using an organic solvent. When
would we use an organic solvent? What are the concerns if we do it that way?
What can we do to remove the organic solvent?
• The test chemical should be “unknown” to the test soils. Why?
• It is recommended to do a sterile control. What can we learn from that?
• What is the quantity of test substance that should be added to the soil?
• If we use 14C labelled substrates, how do we determine the amount of non-residue
(NER) at end of incubation?
• Why are mass balances important?
• Major metabolites should be determined. How and why?
Lecture 2 – Pollutant properties and fate

OECD 301 – Ready biodegradability

Principle: A solution or suspension of the test substance is inoculated and
incubated under aerobic conditions in a ”mineral” (growth supporting) solution at
room temperature (22 oC), pH 7. Blanks (only inoculum, and without inoculum) and
test substance in parallel. The degradation may be followed by measurements of
DOC, CO2 evolution, or O2 uptake. Also specific chemical analysis can be used for
the test substance. Typical test period of 28 days. At least duplicates. The
inoculum typically from activated sludge
Prior info: Phys-chem properties such as solubility, vapour pressure….If the test
substance is volatile or has low solubility in water, then special requirements apply.
Controls like aniline, sodium acetate and sodium benzoate.
Test methods:
• DOC die away: DOC (10 – 40 mg/L) is measured in the flasks
• CO2 evolution: CO2 accumulated in NaOH or Ba(OH)2 and back-titrated
• O2 consumption (BOD): Respirometry (dissolved O2 or manometric in which
case the CO2 produced is sorbed (so it does not affect pressure))
Pass level (for ready biodegradability): 70 % removal of DOC over 10 days window
during the 28 days period.
Lecture 2 – Pollutant properties and fate

Kinetics - Basic rate equations

Differential form Linear form
used in
d[A]
1st order = -k[A] ln[A] = -kt+ln[A]0 95 % of
dt
all cases
2nd order d[A] 1 1
= -k[A]2 = kt +
dt [A] [A]0

Example of first-order degradation

Mycotoxin degradation
Parameter estimation:
100
Degradation of Zearalenone
Zearalenon nedbrydning i jord in soil - use logarithmic form:
) Kg-1)

90
80 OH O CH3 ln[Zea]t = ln[Zea]o – k・t
(µg

70 - or use non-linear fitting

-1

O
Indhold (ug kg

60
- Half-life: t1/2 = ln 2/k
Concentration

HO O Lammefjord
50
Jyndevad
40
Askov
30
20 Soil k (d-1) t1/2 (d)
10 Jyndevad 0.067 10
0
0 20 40 60 80 100 120 140 Askov 0.037 19
Tid (dage)
Time (days) Lammefjord 0.099 7
Lecture 2 – Pollutant properties and fate

Deviation from与一级动力学的偏差：双相动力学或
first-order kinetics: Bi-phasic kinetics or
other types 其他类型 通常会观察到最初的快速反
应，然后是较慢的反应。

为什么？
Often an initial fast reaction followed by a slower reaction is observed.
Why?

• The bioavailable fraction decreases over time due to sorption

• Many compartments in the soil – some with faster degradation than others
• Less activity of microbial degraders during later parts of degradation
• Changes in availability of moisture, temperature, substrates, nutrients etc.
由于吸附，生物可利用部分随着时间
during degradation 的推移而减少 土壤中的许多隔室 - 一
些隔间比其他隔间降解更快 在降解后
期微生物降解剂的活性较低 水分、温
度、底物、养分等可用性的变化。
A lag phase may be seen. Reasons may
降解过程中
be:
• Time needed for microbial population to
可能会看到滞后期。原因可能是： 微
grow 生物种群生长需要时间 底物对降解物
有毒，微生物群需要适应
• The substrate is toxic to degraders and
microbial consortium need to adapt
Lecture 2 – Pollutant properties and fate

First-Order Multi Compartment Model (FOMC)

The soil is divided into

multiple compartments
each with different first-
order degradation rates; if
the distribution of rate
constants follow a
gamma-distribution (∼
normal distribution) à
FOMC equation
土壤被分成多个隔间，每个隔间具有
不同的一级降解率；如果利率常数的
分布服从伽马分布（ 正态分布）à F
OMC 方程

Example of degradation according to the

FOMC model demonstrating the effects of α
and β parameters.
• Increasing β: More ”flat” curves
• Increasing α: Increasing initial rates
Lecture 2 – Pollutant properties and fate

First-Order Multi Compartment Model (FOMC)

Clay loam

Degradation of the pesticide

propoxycarbazone in soils.
Sandy clay loam

Vargas-Perez et al. (2019)

Lecture 2 – Pollutant properties and fate

Hockey-Stick Model

Two sequential first-

order reactions

k1 k2

基板首先被一个过程降解，然后终止
，另一个较慢的过程接管。

k1 增加：初始降解率增加 k2 增加：
缓慢降解率增加

The substrate is first being degraded

by one process, which then
terminates and another slower
process takes over.

• k1 increases: Initial degradation

rate increases
• k2 increases: Rate of slow
degradation increases
Lecture 2 – Pollutant properties and fate

Bi-exponential Model 双指数模型

k1 k2

Two simultaneous first-order reactions

referring to two different reactivity pools
of the substrate (e.g. solution and
sorbed)

Example of degradation according to

the bi-exponential model showing the
significance of k1, k2 and M1 parameters
(M2 = 100-M1)
Lecture 2 – Pollutant properties and fate

Bi-exponential Model – Sum of two first order reactions

- Example for Ptaquiloside biotoxin from Bracken -
Topsoil, sandy
HO CH3 O 100 100

CH3 75 75

Degradation (%)

Degradation (%)
H 3C
Glucose Base
Normal
50 50
Ptaquilosid
HO CH3 O
25 sterilized 25
(a)
Syre CH3
0 0
H3C 0 100 200 300 400 500 600 700 0
Ptaquilosin Time (hours)

CH3 O Syre PTA(t) = a(1 - e -k1F ×t ) + (100 - a)(1 - e -k1S ×t )

PTA-degr.
HO
100 100
CH3

H3C
Parameter sterile untreated
75 75

Degradation (%)
Degradation (%)

Pterosin B Non-toxic k1F (h-1) 1.5 1.0

50
product k2S (h-1) 50 0.2 1.8
a ”fast” (%)
25 46 20 25
(c)
0 0
Lecture 2 – Pollutant properties and fate

Lag-phase models
Modified Hockey-

No reaction at t < tb
Stick model
Logistic Model
 Lecture 2 – Pollutant properties and fate

Logistic Model
Lag phase
Example of Logistic Model with use of
different a0 and r parameters (amax =
0.08)
• Increasing r: less lag phase
• Increasing a0: less lag phase

When a0 is equal to amax the model

1st order reduces to a simple first-order equation

Farhan et al. (2021)

Mineralization of MCPA in a sandy soil Degradation of chlorpyrifos in soil
 酶促降解适用于大多数有机化合物，
Lecture 2 – Pollutant properties and fate 即由于微生物产生的酶（包括胞外酶
）。

必须存在能够降解化学品的生物体（
Degradability 存在编码正确酶的降解基因）
污染物可以充当底物（能源）或者它
Rules of thumb - I 们与作为主要底物的另一种物质共同
降解（共代谢）

• Enzymatic degradation apply to most organic compounds, i.e. due to

microbially produced enzymes (incl. exoenzymes).
• Organisms capable of degrading the chemical has to be present (presence of
degrader genes coding for the right enzymes) 1污染物必须是生物可利用的；因此，
强烈吸附的污染物不容易降解 2 亚基
• Contaminants may act as substrates (energy sources) or they are的毒性或防止微生物降解
co- 3 适当的环
境条件，如水分、氧化剂（通常是 O2
degraded with another substance being the primary substrate ）、pH、温度和营养素是微生物呼吸
的重要要求4 脂肪族化合物比芳香族
(cometabolism) 化合物更快速地5降解 卤素（F、Cl、
Br）和有时的硝基取代基会延缓降解
• The contaminant has to be bioavailable; hence, strongly sorbed contaminants
are not easily degraded
• Toxicity of the substrate retards or prevents microbial degradation
• Proper environmental conditions such as moisture, oxidants (usually O2), pH,
temperature, and nutrients are important requirements for microbial
respiration
• Aliphatic compounds are more rapidly degraded than aromatic compounds
• Halogens (F, Cl, Br) and sometime nitro substituents retard degradation
Lecture 2 – Pollutant properties and fate

Degradability
Rules of thumb - II

• Alkanes: medium size (C10 – C18) most rapidly degraded

Branching decreases biodegradability
Alkenes and alkanes of similar degradability
Cycloalkanes less readily degraded than non-cyclic but faster than
aromatics
• Aromatics: BTX compounds relatively fast to degrade
PAHs degrade slowly, degradation rates decreasing with number
of fused rings
• Heteroaromatics (e.g. containing N, O): Very slow to degrade
• Ethers: May be very slow (e.g. in case of MTBE)
烷烃：中等大小 (C10 – C18) 降解最
快 支化降低生物降解能力 烯烃和具有
类似降解能力的烷烃 环烷烃比非环烷
烃不易降解，但比芳烃快
芳烃：BTX 化合物降解速度相对较快
PAHs 降解缓慢，降解速率随着稠合环
数
杂芳烃（例如含有 N、O）：降解非
常缓慢
醚类：可能非常缓慢（例如 MTBE）
Lecture 2 – Pollutant properties and fate

Degradability
Rules of thumb – III/ a ”grading” system

• Polymers are generally less biodegradable than monomers

+1 point for MW 500 - 1000
+2 points for MW > 1000
• Aromatic compounds are less biodegradable than aliphatics
+1 point for each aromatic ring
• Substitutions decrease biodegradability
+0.5 points for each substitution, except
+1 point for each halogen or nitro-group substitution
• Double or triple bonds decrease biodegradability
+1 point for each internal double or triple bond (NA to aromatic rings)
• Oxygen and Nitrogen bridges decrease biodegradability
+1 point for each O or N bridge (–, =)
• Branches (secondary or tertiary compounds) are generally less biodegradable
than corresponding primary compounds
+0.5 point for each branch
Lecture 2 – Pollutant properties and fate

Degradability
Rules of thumb – III – cont’d

No. Points Rate of Comment

Biodegradation
< 1.5 High > 90% removal by biological treatment

2–3 Moderate 10–90% removal rate by biological

treatment. BOD10 = 0.1–0.9 x TOD
3.5 – 4.5 Slow < 10% removal by biological treatment
BOD10 ∼ 0.1 TOD
5 – 5.5 Very slow < 1% removal by biological treatment. 90%
biodegradation in soil or water takes more
than 6 months
>6 Refractory Half-life in soil or water several years
 环境归宿估计的起点：隔间（土壤、
水、气体、生物群）之间的分布 溶解
Lecture 2 – Pollutant properties and fate 度、吸附、降解数据的最终目标……
整个系统的建模 --> 风险评估、安全
、监管、决策制定！ 经合组织准则。
Summary 确保标准化的测试条件，使数据能够
用于监管、化学品批准、安全计算 O
ECD 307：土壤中化学品的生物降解（
示例）。请转到幻灯片 10 了解摘要
• Starting point for environmental fate estimation: Distribution between
compartments (soil, water, gas, biota)
• Ultimate goal of data for solubility, sorption, degradation…à modelling for
whole systems --> risk assessment, safety, regulation, decision making!
• OECD guidelines. To ensure standardized testing conditions that enables data
for use in regulation, chemicals approval, safety calculations
• OECD 307: Biodegradation of chemicals in soil (example). Go to slide 10 for a
summary.
> which soils, analytical methods, application of test chemical, which dose
(calc.), pathway and kinetics, degradation and mineralization, temperature
effects, mass balances, flow-through and test flask methods. 哪些土壤、分析方法、测试化学品的
应用、剂量（计算）、途径和动力学
• Kinetics of degradation/mineralization 、降解和矿化、温度影响、质量平衡
、流通和测试烧瓶方法。
> First order (ln[A] versus time linear, k and t1/2) 降解/矿化动力学 > 一阶（ln[A] 与时
> First order multi compartment (FOMC) 间线性关系，k 和 t1/2） > 一阶多室 (F
OMC) > 曲棍球棒：两个连续的一阶 >
> Hockey-stick: two sequential first-order 双指数：两个平行的第一-order > 滞
> Bi-exponential: two parallel first-order 后期模型：改进的 Hockey-stick 和 Logi
stic 可降解性。受芳香族/烯烃、双键
> Lag-phase models: Modified Hockey-stick and Logistic 、支化、硝基/卤素取代、N 和 O 桥、
环影响的相对降解率。
• Degradability. Relative degradation rates as affected by aromatic/olefinic,
double bonds, branching, nitro/halogen substitution, N and O bridges, rings.