11th Network Conference on

Persistent Organic Pollutants

Sabrina B. Mammana1,2, Daniela A. Locatelli3,4, Alejandra B. Camargo2,3,4, Jorgelina C. Altamirano1,2,*

(1) (2) (3) (4)

INTRODUCTION
Polybrominated diphenyl ethers Blood sample is considered
are Brominated flame retardants Tool for the assessment as an ideal
Can be transported far from matrix
their sites of production Magnifying of a population’s
Nervous
and use through exposure to But is
System
Through the the environmental Invasive!
Added to Environment (air , soils, contaminants Use of non-invasive
polymers of sediments, freshwater…) samples

Human monitoring
PBDEs can Reproductive
Because of their persist and System
physicochemical accumulate in
properties the GREEN nail sample Hair sample Urine sample
Textiles Environment CHEMISTRY
Plastics
They can Complex matrices need to a sample preparation step:
easily leach to GOAL: A novel sample preparation is proposed
to determine PBDEs in human hair samples by
GC-ECD.
Food web Digestion Microextraction
A multivariate approach was used for
Endocrine An alternative to LLME using low understanding and optimizing the effect of
Building materials
System conventional digestions volumes of solvent each pyhiscochemical variable, as well as their
Electronic equippment Among others… Effects on (acidic and basic digestion) interactions.
Wildlife Humans

EXPERIMENTAL RESULTS
Fig. 1. Results of the experimental design Fig. 2. Fractional factorial design (2k-p): Pareto Chart
Variable Lower value Upper value
Waste
BDE-47 BDE-100
Waste Waste (A) Proteolytic digestor 0.03 0.06 Pareto Chart Pareto Chart
BC CE
concentration (mg/mL) 11.58 6.90
(B) Activator concentration (% 0.5 1
CE
v/v) BC
BCE
(C) Digestion time (min) 120 180 8.68 5.17
A
Bonferroni Limit 8.29126
(D) pH of solution 6.5 9.5

t-Value of |Effect|

t-Value of |Effect|
(E) Digestion temperature (°C) 37 60 Bonferroni Limit 4.25951

5.79
(F) Final volume of solution 5 10 3.45

(mL)
(G) Volume of extractant 50 200
t -Value Limit 2.09302
solvent (µL) 2.89 1.72
t-Value Limit 2.57058

(H) Salting-out effect(mg) 0 4

Hair + Hair + proteolytic Salt + solvent +
digestor + activator (J) Extraction time (min) 0 15
peracetic acid manual shaking 0.00 0.00
digestor + pH (K) Extraction temperature 4 40
adjustment (°C) 1 2 3 4 5 6 7 8 9 10 1 2 3 4 5 6 7 8 9 10

Fig. 3. Box Behnken Design: desirability Rank Rank

Variable Lower value Upper value BDE-99 BDE-153

(A) Proteolytic digestor 0.04 0.08 Pareto Chart Pareto Chart
CE
concentration (mg/mL) CE
11.55
9.26

N2 (B) Activator concentration (% 0.5 1

v/v)
(C) Digestion time (min) 60 180 6.94 8.66

(D) Digestion temperature 37 60

t-Value of |Effect|
t-Value of |Effect|

BCE
(°C) BCE
BC
BC
(E) Volume of extractant 50 200 4.63 5.77

solvent (µL) Bonferroni Limit 4.07253

Bonferroni Limit 4.13787

BCG BEG
2.31 2.89
Cartridge for t-Value Limit 2.0639
GC-ECD
clean-up t-Value Limit 2.07961

Sample preparation: experimental conditions 0.00 0.00

1 2 3 4 5 6 7 8 9 10
PROTEOLYTIC DIGESTION AND LIQUID-LIQUID MICROEXTRACTION 1 2 3 4 5 6 7 8 9 10
BDE 47

Human hair sample: 0.2 g Volume of extractant solvent: 110 µL Rank

BDE 100

Rank
BDE 99

BDE 153

Proteolytic digestor concentration: 24 U/mL Solvent type: tetrachloroethilene Fig. 4. GC-ECD spectra obtained from the
Activator volume: 375 µL Salting-out effect: 1 mg analysis of human hair sample spiked
Digestion time: 120 min Extraction time: 1 min, manual shaking with 25 ng g-1 PBDEs.
Solution pH: 8 Extraction temperature: 25°C
Digestion temperature: 60 °C Centrifugation: 10 min, 2000 rpm (350 x g)
Final volume of solution: 10 mL Clean-up: cartridge (up to down) 0.2 g
anhydrous sodium sulfate, 0.8 acidified silica
CONCLUSIONS
44 % (w/w H2SO4), 0.2 activated silica. Elution: 1. A low cost, simple and effective sample preparation has been proposed as an alternative for conventional
9 mL n-hexane:DCM 4:1 v/v
digestions to determine PBDEs at trace levels in hair samples by GC-ECD.
PBDEs determination by GC-ECD: instrumental conditions 2. The resulting sample preparation is suitable for batch preparation using equipment available in most
Equipment Cromatographer HP 5890 Series II coupled to ECD laboratories.
Chromatographic column DB-5ms (30 m x 0,25 mm x 0,25 µm)
3. The novel application of proteolytic digestion for PBDEs determination gave comparable results to those
obtained using conventional conditions.
Injection Volume: 1 µL; splitless mode, temperature: 250 °C
4. The experimental design statistical tool allowed to identify and characterize those variables that had more
Gas carrier Nitrogen, 1.0 mL/min influence on the system, and also their interactions.
90 °C, 1 min 5. The proposed sample preparation methodology has an environmentally friendly character that makes it
Temperature program 90-250 °C a 35 °C/min, 0 min
250-280 °C a 20 °C/min, 20 min
suitable for current green chemistry trends.

ACKNOWLEDGMENTS