3. Wastewater characterization (1/6) 3.

1 Wastewater characterization (2/6)

3.1 Generalities 3.1 Generalities

3.2 Biodegradability 3.2 Biodegradability Biodegradable

3.3 Toxicity
3.1. Generalities 3.3 Toxicity
Effluents

3.4 Chemical analysis

3.2 Biodegradability assessment 3.4 Chemical analysis
Wastewater Characterization
3.5 Strategy 3.5 Strategy
• charge
3.3 Toxicity assessment • flow Biorecalcitrant
• composition Effluents
• toxicity
3.4 Chemical Characterization • biodegradability

3.5 General strategy for wastewater characterization

Toxic and/or not
before and during the treatment biodegradable
effluents

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Causes of biorecalcitrance or non biodegradability
3.1 Generalities 3.1 Generalities
Causes of biorecalcitrance or non biodegradability Biological causes:
3.2 Biodegradability 3.2 Biodegradability

3.3 Toxicity 3.3 Toxicity

• Inability of organisms to metabolise substrate, caused by:

Biological causes: - failure to induce enzymes

3.4 Chemical analysis 3.4 Chemical analysis
- inability to enter the cell
3.5 Strategy 3.5 Strategy
- failure to serve as substrate for the enzymes
Chemical causes: - production of toxic compounds that disrupt metabolism
• Absence of appropriate organisms
• Predation, parasitism or poor viability
Environmental causes: • Loos of genetic potential; genetic instability
• Absence of extra-cellular enzymes
• Failure to involved appropriate metabolic attack
• Inadequate acclimation or adaptation
Chemical causes: Environmental causes:

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Causes of recalcitrance
3.1 Generalities 3.1 Generalities
Chemical causes: Causes of biorecalcitrance or not biodegradability
3.2 Biodegradability 3.2 Biodegradability
• Refractory to enzymatic and/or non-enzymatic attack because of:
3.3 Toxicity 3.3 Toxicity
- molecular size, charge, shape Environmental causes:
3.4 Chemical analysis 3.4 Chemical analysis
- presence of stable bonds or residues • Inappropriate temperature, pH, water potential, pO2, redox
3.5 Strategy 3.5 Strategy
- number, type and position of substituent groups
potential, light or pressure
• Concentration too low, too high, or too variable
• Unsuitable ionic conditions, including salinity
• Inadequate or slow solubility in water or lipids
• Presence of toxins or other inhibitory or competing
• Failure to form emulsions compounds
• Unsuitable physical state; solid, liquid or gas • Physical inaccessibility
• Covalent or other bounds to organic or inorganic materials
• Adsorption to surfaces

Biological causes: Environmental causes: Biological causes: Chemical causes:

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3.2 Biodegradability assessment (1/14) 3.2 Biodegradability assessment (2/14)

Biodeg.
3.1 Generalities 1 2 3 3.1 Generalities

Conditions Ready Inherent Simulation 1 positive:

3.2 Biodegradability
Chemicals rapidly
3.2 Biodegradability
Readily biodegradability test
Inoculum
3.3 Toxicity Conc. low high low biodegradable in the 3.3 Toxicity
environment
3.4 Chemical analysis 3.4 Chemical analysis
Acclimatisation no possible yes Inherent biodegradability test
3.5 Strategy 3.5 Strategy
2 positive:
Mineral media no yes natural Chemicals inherently
biodegradable under
Target compound optimal conditions Simulation biodegradability test
low high low
conc.
Other sources of
no possible yes
organic carbon 3 positive:
Chemicals biodegradable
Natural substrates
no no yes in simulated conditions
(sediments, algae)

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Readily biodegradability tests – OECD 301 Biochemical Oxygen Demand

3.1 Generalities 3.1 Generalities
The mass concentration of dissolved oxygen
3.2 Biodegradability
CH3CH2OH + 3 O2 2 CO2 + 3 H20
3.2 Biodegradability
consumed under specific conditions by the
3.3 Toxicity 3.3 Toxicity biological oxidation of organic and / or inorganic
COD O2 Formation matter in water.
3.4 Chemical analysis consumption CO2 3.4 Chemical analysis

3.5 Strategy Respirometric 3.5 Strategy

Methods
AFNOR: Evolution of DOC This parameter constitutes a good indicator of the Organic
STURM: Evolution of CO2 amount of biodegradable organic matter of a water. pollutant
Respirometric
MITI: O2 consumption
methods
FILOLE: O2 consumption
Principle: Micro-organisms
Method Poorly Volatile
soluble • BOD is measured after 5 days (= BOD5), at 20°C
Mineral
(favourable temperature for the activity of the O2-
NO NO medium
AFNOR OCDE No 301 A consuming microorganisms) and in darkness (to avoid
STURM YES NO OCDE No 301 B all parasitic photosynthesis).
MITI YES Yes/NO OCDE No 301 C
FIOLE NO SI OCDE No 301 D

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3.1 Generalities 3.1 Generalities

Inherent Biodegradation
3.2 Biodegradability 3.2 Biodegradability

3.3 Toxicity 3.3 Toxicity

OECD 302B Zahn-Wellens Semicontinuous activated

3.4 Chemical analysis 3.4 Chemical analysis
sludge (SCAS)
3.5 Strategy 3.5 Strategy Biodegradability test applied in
(industrial) wastewater treatment Biodegradability in the
plants. environment)

•Mineral medium (synthetic) • Mineral medium natural

(domestic wastewater)
•Target substances as only
The OxiTop system is a patented method of • Others sources of carbon
source of carbon
determining biochemical oxygen demand, which offers
numerous advantages over traditional methods. In • Target substance at low
concentration
addition, it meets all testing requirements of Standard (COD: 20 mg/l)
Method 5210-D.
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3.1 Generalities 3.1 Generalities Zahn-Wellens-Biodegradability test

3.2 Biodegradability 3.2 Biodegradability

3.3 Toxicity 3.3 Toxicity

3.4 Chemical analysis 3.4 Chemical analysis

3.5 Strategy 3.5 Strategy

OECD, Guidelines for testing of Chemicals

Vol 2, test 302B. 1996

Organization for Economic Cooperation and Development

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3.1 Generalities Zahn – Wellens biodegradability test 3.1 Generalities Zahn – Wellens biodegradability test
3.2 Biodegradability 3.2 BiodegradabilityIn this method, the measure of the concentration of
3.3 Toxicity
25° C + 3° C 3.3 Toxicity dissolved organic carbon or the chemical oxygen
Thermometer demand is used to assess the ultimate biodegradability of
Air
3.4 Chemical analysis 3.4 Chemical analysis
Temperature Aeration the test substance.
3.5 Strategy 3.5 Strategy

The method is applicable only to those organic test

microscopic substances which, at the concentration used in the
Observations
test:
-are soluble in water under the test conditions,
-have negligible vapor pressure under the test conditions,
-are not inhibitory to bacteria,
pO2
pH Agitator -are adsorbed within the test system only to a limited
extent,
-are not lost by foaming from the test solution.
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3.1 Generalities Zahn – Wellens biodegradability test 3.1 Generalities Zahn – Wellens biodegradability test
3.2 Biodegradability
The amount of degradation attained at the end of the test is
3.2 Biodegradability
Favorable Conditions
3.3 Toxicity
reported as the 'Biodegradability in the 3.3 Toxicity

Zahn - Wellens test':

• Air flow
3.4 Chemical analysis 3.4 Chemical analysis

• Agitation
3.5 Strategy
Dt(%) = (1 – (Ct – CB) / CA) *100 3.5 Strategy

where: • Room Temp.

Dt = biodegradation (%) at time T, • Nutritious
CA = DOC (or COD) values in the test mixture measured three hours after
the beginning of the test (mg/l) (DOC = Dissolved Organic Carbon, • Substances
COD = Chemical Oxygen Demand),
• High biomass concentration (1-2 g/l)
CT = DOC or COD values in the test mixture at time of sampling (mg/l),
CB = DOC or COD value of the blank at time of sampling (mg/l), Zanh-wellens test

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3.1 Generalities 3.1 Generalities Comparison BOD5 and Z-W

3.2 Biodegradability 3.2 Biodegradability
Compound BOD5 (%COD) Z-W (%COD)
Adipic acid 83 100
3.3 Toxicity 3.3 Toxicity
Simulation tests 2-Aminobenzoic acid 90 97
3.4 Chemical analysis 3.4 Chemical analysis 3-Aminobenzoic acid 11 100

3.5 Strategy 3.5 Strategy

Aniline 90 100
tert-Butanol 0 96
Citric acid 90 85
Wastewater Treatment Plant Natural Ecosystems
2-Chlorobenzoic acid 0 94
2-Chloroaniline 0 94
n-Dodecylsulfate 85 97
rivers soils oceans Dyethilene glycol 80 95
EDTA 3 37
3-Nitrophenol 2 84
4-Nitrobenzoic acid 48 96
Triethanolamine 9 82

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 3.3 Toxicity assessment (1/7) 3.3 Toxicity assessment (2/7)

3.1 Generalities Toxic action 3.1 Generalities In order to evaluate the impact of the toxic and
3.2 Biodegradability 3.2 Biodegradability
biorefractory compounds on municipal activated
Organisms? Selection of the most appropriate sludge plants, various toxicity screening procedures
3.3 Toxicity
biological organism. 3.3 Toxicity
have been proposed in the specialized literature and
3.4 Chemical analysis
Ecological representativity 3.4 Chemical analysis are based on the detection of the following variables
indicative of the bacterial activity:
3.5 Strategy
Contest ? Biological criteria 3.5 Strategy

LD50 (lethal dose-50)

• growth rate (viability of cells, substrate uptake, etc.),
Acute or chronic? Definition of work conditions: • enzymatic activity (dehydrogenase, adenosine
pH triphosphate, etc.),
Temperature
Appropriate test ?
Light
• bacterial luminescence (Microtox® test),
Oxygen concentration • metabolic heat production (microcalorimetric
Alimentation of used techniques), and
microorganisms • respiration rate.
Chemical Hazards Handbook
Introduction to applied toxicology.
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3.1 Generalities •Small fish 3.1 Generalities Microtox test

•Daphnia The Microtox Acute
3.2 Biodegradability 3.2 Biodegradability
•Protozoa (Colpidium) Toxicity Test is a 15
3.3 Toxicity
Acute •Bacteria
3.3 Toxicity
minute exposure
3.4 Chemical analysis Growth 3.4 Chemical analysis
metabolic inhibition test
Specific enzymatic activities
3.5 Strategy
Cellular Energy
3.5 Strategy which uses
Toxicity Bioluminescence
Testing freeze-dried
luminescent bacteria
•Algae (growth inhibition)
(Vibrio fischeri)
Chronic •Daphnia and fish (reproduction,
growth, survival and accumulation
effects)
to assess the acute toxicity of water, soil or sediment samples.

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Respirometic test Respirometic test

3.1 Generalities 3.1 Generalities

3.2 Biodegradability 3.2 Biodegradability

OECD method 209.
OECD method 209. The respirometer consists of a glass respirometric cell (1 l
3.3 Toxicity 3.3 Toxicity
working volume) equipped with a water jacket connected to a
Respirometer:
3.4 Chemical analysis thermostat to maintain a constant reaction temperature of
3.4 Chemical analysis

3.5 Strategy A closed batch reactor characterized by a high 3.5 Strategy 20±1°C.
liquid/gas volume ratio (VL/VG=5) operating without The content of the cell is thoroughly stirred by a magnetic mixer.
liquid flow and with negligible oxygen mass transfer: A glass diffuser feeds pure oxygen.
according to the definition proposed by the Task
Three openings are located in the reactor cover and are used
Group IAWQ [10] it is an LSS respirometer (liquid
for inserting the oxygen electrode (WTW OXI cell 325), the
phase principle, static gas, static liquid).
aeration frit (located at the bottom of the cell) and for dosing the
sample.
The oxygen electrode is interfaced with an IBM PC for
automatic recording of data processed by specialized software
(FIG P for Windows).
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3.3 Toxicity assessment (7/7) 3.4 Chemical Characterization (1/3)

Simplicity and realistic approach of the

3.1 Generalities
biotest
3.1 Generalities
TOC/DOC
3.2 Biodegradability 3.2 Biodegradability

3.3 Toxicity enzymatic 3.3 Toxicity global ions

Ideal
Simplicity of the biotest

particular
functions
3.4 Chemical analysis
Biotest 3.4 Chemical analysis

3.5 Strategy
Physiologic
mono-specific 3.5 Strategy
COD
Multigenerational
ions
Chemical
Communities
multi-specific Analysis
Acute toxicity test artificial Communities
multi-specific
naturals
GC /GC-MS
(microcosms) Artificial
Ecosystems
(mesocosms)
natural
specific HPLC
Ecosystem
Chronic toxicity test (macrocosms)
UV
Realistic approach GC/MS and flash animations of other analytical techniques
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3.1 Generalities Chemical Oxygen Demand (COD) 3.1 Generalities Chemical Oxygen Demand (COD)
3.2 Biodegradability
The Chemical Oxygen Demand (COD) test is used to estimate the 3.2 Biodegradability

3.3 Toxicity
amount of organic matter in a sample. 3.3 Toxicity
The equivalent amount of
It is a measurement of the oxygen equivalent of the materials present in
3.4 Chemical analysis 3.4 Chemical analysis
oxygen required to oxidize the
the sample subject to oxidation by a strong chemical oxidant, in this
3.5 Strategy
case dichromate. 3.5 Strategy organic matter to CO2 is equal
to the COD of the wastewater
and is determined from the
COD of organic matter in wastewater is measured by allowing it to react with a strong amount of dichromate
chemical oxidizing agent (potassium dichromate) in an acidic solution. consumed in the COD test.

The test is carried out at high temperatures and in the presence of a catalyst (silver sulfate).

The amount of dichromate remaining at the end of the test is measured and subtracted
from the amount of dichromate originally added to the wastewater. CH2 + Cr2O7= + 8H+ Î CO2 + 2Cr(III) + 5H2O
yellow/orange green
440 nm 600 nm
Chemical Oxygen Demand
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3.1 Generalities Total Organic Carbon (TOC) 3.1 Generalities

3.2 Biodegradability 9Total Organic Carbon (TOC) analysis is a well-defined and 3.2 Biodegradability COD/TOC HPLC Microtox
commonly used methodology that measures the carbon content BOD5
3.3 Toxicity 3.3 Toxicity
of dissolved and particulate organic matter present in water.
3.4 Chemical analysis 3.4 Chemical analysis ions UV GC
3.5 Strategy 9TOC analysis consists of inorganic carbon removal, oxidation 3.5 Strategy Zhan Wellens
of the organic carbon into CO2, and quantification of the CO2.

COD/TOC Indication about oxidation state

Total Organic Carbon (TOC)
AOS = 4(TOC – COD)/TOC
Dissolved Organic Carbon (DOC)

Total Carbon (TC) BOD5/COD Indication about biodegradability

Domestic wastewater Î~0.4
Inorganic Carbon (IC).

TC = TOC + IC TOC-5000A
TOC analyzers
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3.1 Generalities 3.1 Generalities Environmental Analytical Water Quality

Criteria Minimal detectable
3.2 Biodegradability 3.2 Biodegradability signal:
S/N = 3
3.3 Toxicity Relation COD/TOC 3.3 Toxicity
Sensibility Quantification limit:
3.4 Chemical analysis 3.4 Chemical analysis S/N = 10
Substance COD/TOC Applicability
Resolution power:
3.5 Strategy
CH4 5.33 AOS 3.5 Strategy
Specificity R = Num. of signals *100
Types of molec.
CH3CH2OH 4.00 Quality of the
measurement Absolute Error : Ea
CH3COCH3 3.56 -4 +4 Ea = xi - xt
C6H6 3.34 CH4 CO2 Accuracy
Relative Error : Er
Er = (xi -xt)/xt *100%
C6H5COOH 2.86
Reliability Repeatability
(COOH)2 0.67 Under the same
Experimental standard
conditions of
Precision measurement Deviation: s

IUPAC compendium of analytical nomenclature Reproducibility s = √(∑(xi -x)2/(N-1))

Different conditions Variance: s2

Statistics for analytical chemistry

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3.5 General strategy for characterization (3/3)

3.1 Generalities Environmental Analytical Water Quality

Criteria Seuil de détection:
3.2 Biodegradability S/N = 3

3.3 Toxicity Seuil de quantification:

Sensibilité S/N = 10
3.4 Chemical analysis
Applicabilité
Pouvoir de résolution:
3.5 Strategy
Specificité R = Nb de signaux *100
Types de moléc.
Qualité de la
mesure Erreur absolue : Ea
Ea = xi - xt
Exactitude
Erreur relative : Er
Er = (xi -xt)/xt *100%
Fidélité Répètabilité
Mêmes conditions
expérimentales Déviation standard: s
Précision
s = √(∑(xi -x)2/(N-1))
IUPAC compendium of analytical nomenclature Reproductibilité
Variance: s2
Différentes conditions
Statistics for analytical chemistry

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