Biotic and abiotic degradation studies of organic contaminants

Transcription

1 Biotic and abiotic degradation studies of organic contaminants Manuela Peschka Ankara, 8th ctober 2007 utline rganic pollutants Selected compounds Simulation of abiotic/biotic degradation ixed bed bioreactor Bottle test Suntest Analysis of micopollutants and identification of transformation products Sample pretreatment Analytical Instrumentation Results: Case study I: barbiturates Case study I: photodegradation of bentazone Case study II: biodegradation of a new fluorosurfactant Case study IV: biotransformation of clotrimazole 1

2 R 3 R 1 C H H S 2 R 2 bentazone barbiturates pharmaceuticals monitoring abiotic pesticides degradation rganic rganic Micropollutants Micropollutants Cl clotrimazole biodegradation surfactants S 10-(trifluoromethoxy) decane-1-sulfonate Degradation Tests: 2

3 ixed Bed Bioreaktor sampling aerator fixed bed 10 L storage tank pump Bottle Test 5 L storage tank aerator 3

4 Suntest simulation of abiotic photodegradation E = h ν xenon arc lamp quartz tubes Sample Pretreatment: 4

5 sorbent 200mbar AIS HLB Reversed Phase material with Hydrophilic Lipophilic Balance A = 810 m 2 /g pore diameter = 80 Å V = 1,3 cm 3 /g particle diameter = 60, 30, 25, 15 und 5 µm hydrophilic monomer lipophilic monomer 1 atm 1 atm 5

6 AIS MCX Mixed-Mode Cation exchange and Reversed Phase 1,0 meq/g S 3 H Ion exchange capacity S 3 - S 3 - H2 H AIS MAX Phasenchemie Mixed-Mode Anion exchange and Reversed Phase CH 2 R + H 3C + CH 3 C 4H 9 S C - CH 3 6

7 Elution (for non polar SPE) typical solvents: methanol methanol with acid methanol with base acetonitrile acetic acid TH acetone dichlormethan, chloroform hexane increasing elution potential A B Analysis Analytical Instrumentation: 7

8 General setup of a mass spectrometer chromatographic separation Torr injector ion source mass analyzer detector vacuum system signal convertion result GC-MS 8

9 Advantage: utilization of mass spectra libraries strong fragmention gives structural information Disadvantage: single MS EI only in combination with GC LC-MS 9

10 Advantage: - analysis of polar, thermolabile compounds - very robust for routine analysis Disadvantage: - no spectral libraries - matrix effects Quadrupole Time of light Mass Spectrometer 1 E kin = * m * v² = z * e * U * m * ² = z * e * U 2 2 m t² = ( 2 * e * U) * z L² L = ca. 2m 10

11 Advantage: - high resolution, e.i. determination of up to the fourth dezimal (e.g amu) - determination of the elemental composition -MS² Disadvantage: - not sensitive QqLIT - user can select various modi, e.g. Q1MI, MRM, product ion scan - enhanced modi: Q3 acts as trap resulting in higher sensitivity Advantage: - structure elucidation of unknown compounds - high sensitivity Disadvantage:? 11

12 Ion chromatography -determination of inorganic paramter, e.g. S 4 2-, Cl -, - - mineralization of a molecule flow direction A + E + - E + - A + S S H 3 C CH 3 stationary phase monitoring structure elucidation QqT LC-MS QqLIT GC-MS (IC) 12

13 Case studies: Barbiturates conc. [µg/l] 5 5,3 5,4 R 3 pentobarbital butalbital secobarbital phenobarbital 4,55 4 R 1 C H 3 2,08 R 2 2, ,21 0,49 0,28 0,10 0,09 0,12 0,10 0,31 0,30 0,29 0,39 0,28 0,30 0,19 0,40 0,22 0,51 0,27 0,98 0,88 1,5 1,41 0,86 0,3 0, concentration [ng/l] mix aprobarbital pentobarbital butalbital secobarbital hexobarbital phenobarbital 1500 ixed Bed Bioreactor Exposion to sunlight Hydrolysis (ph 2 12) time [h] groundwater in Berlin 13

14 Bentazone one of the most applied herbicides in the Ebro river delta 31 µg/l in application time from May to August (mean value) (Ebro 0.27 µg/l) comparable high concentrations river waters of the Tiber region in Italy not biodegradeable in a fixed bed bioreactor passes through membrane bioractor present in ground- and surface water E = h ν xenon arc lamp quartz tubes H S bentazone S3-bentazone bentazone Area Area (bentazone) (bentazone) Area (S3-bentazone) A distilled water Time Time [h] [h] bentazone S3-bentazone bentazone Area Area (bentazone) Area (S3-bentazone) B - moderately hard water Time Time [h] [h] bentazone 250 bentazone S3-bentazone bentazone 200 Area Area (bentazone) Area (S3-bentazone) C moderately hard water plus 5 µg/ml humic acids Time Time [h] [h] 14

15 observed mass (m/z) ± elemental composition actual mass C10H1124S m/z = mda -0.8 C7H523S m/z = H H S C10H1122 m/z = H S C7H42 m/z = observed mass (m/z) ± ± elemental composition actual mass C10H1324S m/z = C10H132 m/z = C6H6 m/z = mda H S H H present in drainage channels in the Ebro delta luorosurfactants blood Anzahl der Publikationen (Primärliteratur) properties: - persistent to hydrolysis, oxidation, reduction and biodegradation - accumulate in liver, kidneys, mussel tissue and bind to blood proteins S 15

16 Biodegradation in a ixed Bed Bioreaktor 10-(trifluormethoxy)decane-1-sulfonate β = 100 mg/l V = 5 L surface water ph 7 T = room temperature (20 C) Analytical methodologies Q-Trap Trap MS³-experiments Ion Chromatography -, S 2-4, TC Q-To high resolution mass spectrometry 16

17 mass spectrum of 10-(trifluormethoxy) (trifluormethoxy)- decane-1-sulfonate m/z = 85 3 C [M-H] - m/z = 305 (CH 2 ) 10 S 3 S m/z = 80 S S C oxygenase S C oxidation H 3 C (CH 2 ) x (CH 2 ) x - S 3 H S 3 3 C C H beta-oxidation -C 2 3 C CH -C 2 HC S 3 beta-oxidation -C 2 HC S 3 3 C CH S C 2 3 C H -H C 2-17

18 Monitoring of TC, - and S 2-4 c ( -, S 4 2-, TC) [mg/l] TC - S 4 2- primary degradation metabolites (m/z = 85) Time [d] 1,20 1,00 0,80 0,60 0,40 0,20 0,00 c/co (Trifluormethoxydecansulfonat) S C S 3 oxygenase 3 C H 3 C (CH 2 ) x (CH 2 ) x - S 3 oxidation 3 C C H beta-oxidation -C 2 3 C CH? -C 2 3 C CH after 28 days: 3 C H -H - primary degradation: 100% -> 90 % S concentration theoret. -58 % - - concentration theoret. C 2, - 18

19 6,E+04 5,E+04 4,E+04 area 3,E+04 3 C (CH 2 ) x (CH 2 ) x S 3-2,E+04 1,E+04 0,E Time [d] S 4 2-1,20 1,00 c ( -, S 4 2-, TC) [mg/l] ,80 0,60 0,40 0, Time [d] 0,00 19

20 * * * C 11 H 18 3 S 5-3 C (CH 2 ) x (CH 2 ) x S 3 - * * * * * [M-H] - MS² MS³ S C -C S 3 S [M-H] oxidation in omega-position! 20

21 3 C S 3 -- oxygenase 3 C (CH 2 ) x (CH 2 ) x - S 3 H S 3 HC S 3 beta-oxidation -C 2 HC S 3 -C 2 S 4 2- C S 3 HC S C 6 H 11 S 5-21

22 3 C S 3 S oxygenase 3 C oxidation H 3 C (CH 2 ) x (CH 2 ) x - S 3 H S 3 3 C C H beta-oxidation -C 2 3 C CH HC S 3 beta-xidation -C2 3 C -C 2 CH <10% slow HC S 3 S C 2 3 C H -H C 2,- > 90% fast Cl Mode of action: Inhibitor of ergosterol synthesis resulting in the damage of cell membrane MW [g/mol] logk W 4.1 S water [mg/l] 0.49 Vp [Pa] 3.31*10-7 mp [ C] bp [ C]

23 alse positive signal in next four measured samples (solvent - acetone)! approx. 2 ng/l 6 Log K W versus ph 5 Cl 4 log KW 3 H Cl ,00 2,00 4,00 6,00 8,00 10,0 12,0 14,0 ph 23

24 adapted SPE-Method Acidification to ph 2 (stirring 30 min) of samples before filtration SPE-Material asis HLB 3cc (60 mg), Waters conditioning: 1 x 2 ml n-hexan, 3 x 2 ml methanol 5 x 2 ml groundwater (ph 2) 2 SPE: 100 ng Atrazin D5 (internal standard) flow approx. 20 ml/min drying: 45 min ( 2 ) elution: 4 x 1 ml acetone evaporation: to 150 µl external standard: 100 ng luazifop-buthyl (ESTD) and fill up to 200µL final volume with acetone Adsorption hinders standardized biodegradation tests 1,2 BBR 1 c/c0 0,8 0,6 two hours after BBR passage 0,4 0, t [d] 24

25 5 L storage tank aerator reaction with electrophiles phase I H phase II CR z.b. glucoronic acid SH reaction with nucleophiles 3,50E+06 3,00E+06 2,50E+06 CLT-metabolite clotrimazole Area 2,00E+06 1,50E+06 1,00E+06 5,00E+05 0,00E Time [d]

26 478.1 EPI H H3 Cl H H3 Cl Cl H Cl

27 Summary: no biotic nor abiotic degradation persist in the aquatic enironment for several years biotransformed further investigations barbiturates clotrimazole ate & Degradation new fluorosurfactant bentazone no biotic degradation but undergoes photolysis degradable in sunlight exessable compartments undergoes biodegradation potential substitute for the harmful fluorosurfactants presently used Acknowledgement: AQUATERRA (Project number GCE) Spanish Ministry of Education and Science (Project number CTM E) Merck KGaA Thank you for your attention! 27