Effect of micron- and nanosized titanium dioxide on algal growth inherent inhibitory effects and as modifying factor on cadmium

m toxicity
nanoECO, Monte Verit, 6th March 2008

Nanna B. Hartmann Anders Baun

nah@env.dtu.dk

Why look at interaction?

Hypothesis: Nanoparticles may act as carriers of coexisting contaminants (nanovectors) No exposure = No risk! Low exposure = low risk? Trojan horse / piggy bag effect / nanovector Supported by existing studies: C60 present: Increased toxicity of phenanthrene in algae and daphnia. Sorbed phenanthrene was bioavailable1. TiO2 present: Enhanced uptake of Cd / As in carp2, 3
1Baun

Andersen et al., 2008

et al. (2008), Aquatic Toxicol, 86 (2008) 379387 et al. (2007), Chemosphere, 67, 160-166.

2Zhang 3Sun

et al. (2007) Water Air and Soil Pollution, 178, 245-254

Preliminary study
Cd Cd + TiO2 P25
Distribution of cadmium in algae

Cd + TiO2 P25 Cd + TiO2 UV100 Cd + TiO2 LW-S

Mixture Toxicity to algae Experimental work

Cd
Single compound toxicity to algae

TiO2 P25 TiO2 UV100 TiO2 LW-S

Influence of light intensity

- TiO2 (control) TiO2 P25

Sorption of cadmium onto

TiO2 LW-S

TiO2 UV100

TiO2 particles
Particle diameter [nm] P25 (Degussa) 25 Crystalline BET SAA Aggregate size in algal test phase [m2/g] media [nm] 80% anatase, 20% rutile 100% anatase 100% anatase 47 180** Zeta potential -25.8 mV**
-

Hombikat UV100 (Sachtleben) Hombitan LWS (Sachtleben)

* As stated by producer

100

288

300

11.5

** Measured by Frank von der Kammer, University of Vienna, Austria

Methods
TiO2 suspended in ISO algal medium Cd concentrations measured by AAS Toxicity and accumulation single compounds and mixtures Algal growth (Freshwater green algae Pseudokirchneriella subcapitata)

TiO2 in water + metals & algae

Intercellular/extracellular concentrations
Control studies light intensity UV pre-illumination Shading

Preliminary study
Cd Cd + TiO2 P25
Distribution of cadmium in algae

Cd + TiO2 P25 Cd + TiO2 UV100 Cd + TiO2 LW-S

Mixture Toxicity to algae Experimental work

Cd
Single compound toxicity to algae

TiO2 P25 TiO2 UV100 TiO2 LW-S

Influence of light intensity

- TiO2 (control) TiO2 P25

Sorption of cadmium onto

TiO2 LW-S

TiO2 UV100

Effects of TiO2 on algal growth

Compound Degussa P25 Humbikat UV100 Hombitan LW-S EC10 (mg/L) 0.9 [0.5;1.9]95% 2.7 [0.5;13]95% 11 [1.2;100]95% EC20 (mg/L) 3.2 [1.9;5.2]95% 15 [4.9; 43]95% 42 [7.1;244]95% EC50 (mg/L) 33 [25;44]95% 364 [101;1310]95% 546 [64;4640]95%

Reproducibility issue

Preliminary study
Cd Cd + TiO2 P25
Distribution of cadmium in algae

Cd + TiO2 P25 Cd + TiO2 UV100 Cd + TiO2 LW-S

Mixture Toxicity to algae Experimental work

Cd
Single compound toxicity to algae

TiO2 P25 TiO2 UV100 TiO2 LW-S

Influence of light intensity

- TiO2 (control) TiO2 P25

Sorption of cadmium onto

TiO2 LW-S

TiO2 UV100

Toxicity of Cd(II) to algae

Dose-response curve for Cd(II) in the absence/presence of 2 mg/L TiO2 P25

Dose-response curve for Cd(II) in the absence/presence of 2 mg/L TiO2 LW-S

Toxicity of Cd(II) to algae

With TiO2 P25 EC50,72h = 71 g/l [67;74]95% EC50,72h = 67 g/l [62;73]95%

Without TiO2 P25 Sorbed = bioavailabe?

EC10<2mg/L<EC20

Toxicity of Cd(II) to algae

Dose-response curve for Cd(II) in the absence/presence of 2 mg/L TiO2 P25

Dose-response curve for Cd(II) in the absence/presence of 2 mg/L TiO2 LW-S

Toxicity of Cd(II) to algae

TiO2 LW-S

Without TiO2 LW-S EC50,72h = 61 g/l [59;64]95% EC50,72h = 78 g/l [72;83]95%

With TiO2 LW-S Reduced bioavailablity through sorption

Reduced bioavailablity through sorption

Preliminary study
Cd Cd + TiO2 P25
Distribution of cadmium in algae

Cd + TiO2 P25 Cd + TiO2 UV100 Cd + TiO2 LW-S

Mixture Toxicity to algae Experimental work

Cd
Single compound toxicity to algae

TiO2 P25 TiO2 UV100 TiO2 LW-S

Influence of light intensity

- TiO2 (control) TiO2 P25

Sorption of cadmium onto

TiO2 LW-S

TiO2 UV100

Sorption of Cd(II) to TiO2

~ 90% sorption

50 g/L Cd(II) 10 mg TiO2/L (Humbikat UV100)

Sorption of Cd (II) to TiO2

Sorption of Cd(II) to TiO2 Freundlich isotherm

Preliminary study
Cd Cd + TiO2 P25
Distribution of cadmium in algae

Cd + TiO2 P25 Cd + TiO2 UV100 Cd + TiO2 LW-S

Mixture Toxicity to algae Experimental work

Cd
Single compound toxicity to algae

TiO2 P25 TiO2 UV100 TiO2 LW-S

Influence of light intensity

- TiO2 (control) TiO2 P25

Sorption of cadmium onto

TiO2 LW-S

TiO2 UV100

Uptake of Cd(II) in P. subcapitata

1. 2. 3. 4. Algae (4.5105 cells/ml; log-phase) Algae + 40 g/L Cd(II) Algae + 40 g/L Cd(II)+ 2 mg/L TiO2 Cd(II) + 2 mg/L TiO2

Samples fractionated after 2h and 24h Fractionation: Centrifugation EDTA washing

Intracellular Sorbed/ extracellular Water

Uptake of Cd(II) in P. subcapitata

2 hours
100% 80% 60% 40% 20% 0% Cd and algae Algae, Cd and TiO2 Cd and TiO2

24 hours
100% 80% 60% 40% 20% 0% Cd and algae Algae, Cd and TiO2 Cd and TiO2

Water

Sorbed/Extracellular

Intracellular

Loss

Reduced intracellular cadmium concentration in the presence of 2 mg/L TiO2

Preliminary study
Cd Cd + TiO2 P25
Distribution of cadmium in algae

Cd + TiO2 P25 Cd + TiO2 UV100 Cd + TiO2 LW-S

Mixture Toxicity to algae Experimental work

Cd
Single compound toxicity to algae

TiO2 P25 TiO2 UV100 TiO2 LW-S

Influence of light intensity

- TiO2 (control) TiO2 P25

Sorption of cadmium onto

TiO2 LW-S

TiO2 UV100

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Speciation of Cd in algal medium

(PHREEQC)

Water phase concentrations

Dirtribution [%]

Modelling of Cd speciation in algal media using PHREEQC

100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0%

Cd distribution

10
Cd2+ CdEDTA2-

20

40

80 160 320

Cd [ug /L]
Other species (mainly CdCl+)

Cd concentrations expressed as Cd2+

Algal toxicity of Cd(II) expressed as Cd2+

Cd2+

TiO2: 2 mg/l (LW-S)

2+ [26;31] EC50,72h = 29 61 g/l Cd Cd(II)[59;64] 95% 95% EC50,72h = 78 26 g/l Cd(II) Cd2+ [22;31] [72;83] 95% 95%

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Algal toxicity of Cd(II) expressed as Cd2+

Cd2+

TiO2: 2 mg/l (P25)

EC EC50,72h = 71 37 g/l g/l Cd(II) Cd2+ [33;42] [67;74] 50,48h = 95% 95% 2+ [7;14] EC EC50,72h = = 67 10 g/l g/l Cd(II) Cd [62;73] 50,48h 95% 95%

% Inhibition

Cd(II) toxicity +/- TiO2 P25

Cd2+ Cd(II)

50

Concentration

[Cd2+]<[Cd(II)] Does not take into account the effect of TiO2

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Summary
Inhibition
Cd2+ effect Measured effect
Changes in effects were caused by: Reduced Cd(II) bioavailability The inherent toxic effect of TiO2 nanoparticles

Inhibition

Cd + TiO2 LW-S

Concentration (g/L) Cd2+ effect TiO2 effect Measured effect

Cd + TiO2 P25

Concentration (g/L)

Conclusions
Effects of TiO2 nanoparticles are larger than the effect of micron sized TiO2 particles The addition of TiO2 nanoparticles to algal tests influenced the toxicity of Cd(II) according to sorption and speciation behaviour Uptake of Cd(II) in algae decreased by addition of TiO2 suspensions No indications of nanoTiO2-facilitated transport of Cd in Pseudokirchneriella subcapitata Know your test system and the influence of TiO2 nanoparticles on toxicity can be predicted! - but how about very low concentrations of TiO2? Presence of HA?

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Acknowledgements
- Laboratory technicians Susanne Kruse & Signe Qualmann - Dr. Frank von der Kammer, Vienna University, Austria

Contact info:
nah@env.dtu.dk

Thank you for your attention!

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