Analysis and Relative Distribution of OPFR Compounds in Wastewater Treatment Million B. Woudneh, Coreen Hamilton, Guanghui Wang, Richard Grace, John R. Cosgrove Sidney B.C., Canada, V8L 5X2

 OPFRs are commercial additives widely used to inhibit, suppress or delay the production of flames. Trialkyl phosphatesHalogenated phosphatesTriaryl phosphates Restrictions on polybrominated diphenyl ethers (PBDEs) has increased demand for OPFRs. Since they are not chemically bonded agents, OPFRs use as additives results in easy release to the environment. Importance of Organophosphate Flame Retardants (OPFRs)

Alkyl phosphates Short - hand Halogenated alkyl phosphates Short - hand Triethyl phosphate TEPTris(2-chloroethyl) phosphateTCEP Tripropyl phosphateTPrPTris(2-chloroisopropyl) phosphateTCPP Tributyl phosphateTBPTris(1,3-dichloro-2-propyl) phosphateTDCPP Tris(2-ethylhexyl) phosphateTEHPTris(2,3-dibromopropyl) phosphateTDBPP Aryl phosphatesOther phosphates Triphenyl phosphateTPPTris(2-butoxyethyl) phosphateTBEP Tricresyl phosphateTCrP Tetrakis(2-chlorethyl) dichloroisopentyldiphosphate V6 2-Ethylhexyl-diphenyl phosphateEHDPP Target compounds selected based on: CMP = Chemical management plan TSCA = EPA's Toxic Substances Control Act SFEI = San Francisco Estuary Institute CECBP = California Environmental Contaminant biomonitoring program list Leading researchers in the field Target Analytes of Interest to Regulators & Scientists

Matrices: Influent, Effluent, Soil, Sediment, Biosolid Solids + surrogates Extraction: Soxhlet, DCM:EtAOc Cleanup: -NH2 SPE, Silica Solvent exchange to MeOH Analysis by LC-MS/MS Aqueous + surrogates Extraction: L-L with DCM Cleanup: -NH2 SPE Surrogates 2 H 15 -TEP 2 H 12 -TCEP 2 H 21 -TPrP 2 H 18 -TCPP 2 H 15 -TDCPP 13 C 15 -TPP 2 H 27 -TBP Recovery 2 H 15 -TPP Initial and ongoing QCs: 5 replicates MS/MSD QC samples: 1 Lab blank 1 Spiked sample 1 Duplicate Isotope labeled surrogates: 7 Surrogates 1 Recovery Method Makes Use of Eight Isotope- Labeled Standards

min > 152 TPP min 431> 99 TDCPP min 583 >361 V6 min 329> 99 TCPP min 225> 99 TPrP min 285 > 223 TCEP min 183 > 99 TEP Relative response min >99 TEHP min 363 > 251 EHDPP min 369> 165 TCrP min 399 > 299 TBEP min 267 > 99 TBP min 716 >99 TDBPP Relative response Column: - Waters C18MS Mobile phase: - 0.1% HCOONH % HCOOH MeOH Two MRMs per compound All Analytes Separated by Mass and Time

Reporting limits: to 5 ng/L for a 0.5 L sample to 0.5 ng/g for a 5 g wet sample Quantitative measurement of OPFRs from aqueous samples, sediment and biosolids

P RIMARY E FFLUENT Surface Waters Primary Clarifier Aeration Basin Secondary Clarifier Disinfection Bio-GasDigestion DewateringLand Application P RIMARY S LUDGE W ASTE A CTIVATED S LUDGE T REATED B IOSOLIDS R AW I NFLUENT S ECONDARY E FFLUENT Sampling Throughout the Process = Sample points at the study site

ND 1069 OPFRs survive secondary wastewater treatment Distribution of OPFRs

BDE influent and effluent data from Kim, et al., Water Research 47, no. 7: 2213–21. BDE influent and effluent data from Kim, et al., Water Research 47, no. 17: 6496–6505. BTBPE and DBDPE data from Grace et al. BFR 2011, Boston. Final effluent and biosolid from secondary wastewater treatment are significant sources of OPFRs. Relative to brominated diphenyl ethers, OPFRs have a much higher affinity for the aqueous phase. OPFR Concentrations Significantly Higher than Other FRs

AXYS OPFR Method Summary and Conclusions  A reliable method for quantitative measurement of OPFRs from aqueous, sediment and biosolid samples.  The method makes use of eight isotope-labeled surrogates.  OPFRs survive secondary wastewater treatment.  Final effluent and biosolids from secondary wastewater treatment are significant sources of OPFRs.  OPFRs have much more affinity for the aqueous phase relative to BDEs  OPFRs are present at significantly higher concentrations than BDEs and other FRs. For Follow-up:

Acknowledgements  Dr. Shirley Anne Smyth and her research group at Environment Canada are thanked for their assistance in sample collection and water quality data analysis.  Dr. Heather M. Stapleton is thanked for providing ‘V6’ technical standard for the initial method development.

Supplemental Information

NH2 1 g SPE - Condition: - with 2 x col. vol of HXN and 2 x col. Vol of DCM - Load: 1 mL extract in Hexane - Wash: 5 mL 20:80 DCM:Hexane (DISCARD) - Elute: 12 mL of 1:9 EtOAc:DCM (COLLECT) Silica, 8 g, 5% deactivated - Pack column in hexane - Load: 1 mL DCM extract - Wash: 20 mL 15: 85 DCM:Hexane (DISCARD) - Elute : 100 mL of 1:2 DCM:ETOAc (COLLECT) Before cleanup After cleanup Clean extracts were achieved with NH2 and silica columns

* * Samples sizes: 0.25 L for influent 0.50 L for effluent and 2.5 g wet for biosolid 10 g for biosolid These samples sizes were demonstrated to have no matrix effects. No significant matrix effects Reagent

OPFR Standard Stability in Methanol

OPFR Extract Stability

MRM transitions Target Analyte Typical Retention Time (minutes) Parent Ion Mass 1 Daughter Ion Mass 1 Quantified Against TEP (127)d15-TEP TCEP (63.1)d12-TCEP TPrP (140.9)d21-TPrP TCPP (327.1)99.0 (99.0)d18-TCPP V (99.0)d15-TDCPP TDCPP (208.9)d15-TDCPP TPP (77.1) 13 C 18 -TPP TDBPP (698.6)99.0 (99.0)d15-TDCPP TBP (155.0)d27-TBP TBEP (199.0)d27-TBP TCrP (91.0) 13 C 18 -TPP EHDPP (77.1) 13 C 18 -TPP TEHP (211.0)d27-TBP Surrogate Standard d15-TEP (134)d15-TPP d12-TCEP (67.1)d15-TPP d21-TPrP (150.1)d15-TPP d18-TCPP (345)102d15-TPP d15-TDCPP (216)d15-TPP 13 C 18 -TPP (83.1)d15-TPP d27-TBP (166.1)d15-TPP Recovery Standard d15-TPP (82.1)d15-TPP

LC Gradient Time (min)Flow mixture 1 LC Flow Rate Program Gradient Curve % solvent A 85% solvent B 0.2 mL/min % solvent A 50% solvent B 0.2 mL/min % solvent A 30% solvent B 0.2 mL/min % solvent A 15% solvent B 0.2 mL/min % solvent A 5% solvent B 0.2 mL/min % solvent A 5% solvent B 0.25 mL/min % solvent A 85% solvent B 0.2 mL/min % solvent A 85% solvent B 0.2 mL/min1

Calculated Log KD vs Log Kow for OPFRs