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 phosphates Halogenated 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 phosphatesShort -hand

Halogenated alkyl phosphatesShort -hand

Triethyl phosphate  TEP Tris(2-chloroethyl) phosphate TCEP

Tripropyl phosphate TPrP Tris(2-chloroisopropyl) phosphate TCPP

Tributyl phosphate TBP Tris(1,3-dichloro-2-propyl) phosphate TDCPP

Tris(2-ethylhexyl) phosphate TEHP Tris(2,3-dibromopropyl) phosphate TDBPP

Aryl phosphates Other phosphates

Triphenyl phosphate TPP Tris(2-butoxyethyl) phosphate TBEP

Tricresyl phosphate TCrP Tetrakis(2-chlorethyl) dichloroisopentyldiphosphate

V6

2-Ethylhexyl-diphenyl phosphate EHDPP

•Target compounds selected based on:CMP = Chemical management planTSCA = EPA's Toxic Substances Control ActSFEI = San Francisco Estuary InstituteCECBP = California Environmental Contaminant biomonitoring program listLeading 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

2H15-TEP

2H12-TCEP

2H21-TPrP

2H18-TCPP

2H15-TDCPP

13C15-TPP

2H27-TBP

Recovery

2H15-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

min6.0 10.0 14.0 18.0 22.0

327> 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 8.59

9.69

13.14

13.31

14.24

16.08

16.67

Rela

tive

resp

onse

min

17.0 21.0 25.0 29.0

435 >99

TEHP

27.07

min

363 > 251

EHDPP22.01

min

369> 165

TCrP

min

399 > 299

TBEP

18.85

min

267 > 99

TBP

17.98

min

716 >99

TDBPP

17.25

20.89

Rela

tive

resp

onse

Column: - Waters C18MS

Mobile phase: - 0.1% HCOONH4 + 0.1%

HCOOH MeOH

Two MRMs per compound

All Analytes Separated by Mass and Time

TEP TCEP TPrP TCPP V6 TDCPP TPP TDBPP TBP TBEP TCrP EHDPP TEHP0

20

40

60

80

100

120

140

Recovery from reagent water and sediment samples

Aqueous Solids

Perc

ent r

ecov

ery

TEP

TCEP

TP

rP

TCPP

V6

TDCPP

TPP

TDBPP

TBP

TBEP

TC

rP

EHDPP

TEHP

0

20

40

60

80

100

120

140

MS recovery from influent, effluent and biosolids

Effluent Influent Biosolid

Perc

ent r

ecov

ery

Reporting limits: - 0.1 to 5 ng/L for a 0.5 L sample - 0.01 to 0.5 ng/g for a 5 g wet sample

Quantitative measurement of OPFRs from aqueous samples, sediment and biosolids

PRIMARY EFFLUENT Surface

WatersPrimary Clarifier

Aeration Basin

Secondary Clarifier Disinfection

Bio-Gas Digestion Dewatering Land Application

PRIMARY

SLUDGE

WASTE ACTIVATED SLUDGE

TREATED BIOSOLIDS

RAWINFLUE

NT

SECONDARY EFFLUENT

Sampling Throughout the Process

= Sample points at the study site

TPrPTCEP

TBPTCrP

TEHP

TDCPPTBEP

0

200

400

600

800

4 6 52 61 69 75 92172181219

2,236Biosolids

Conc

. ng/

g, d

ry

TPrP

EHDPP TCrP

TPP

TDCPP TCPP

TBP 0

200

400

600

800

1 1 26 27133

227344

440547581

Final Effluent

Conc

. ng/

L

3292

3807

ND ND ND

TPrP

TEHP

EHDPP TPP

TEP TCEP

TBEP 0

200

400

600

800

16 33 40 41 90

227299

380

598

7473,293Raw Influent

Conc

. ng/

L

ND

ND

ND

ND

TPrP V6TEHP

TCrP TEPTBP

TBEP0

200

400

600

800

8 2172 76 98 106139168174196

1,069Primary Sludge

Conc

. ng/

g, d

ry

ND 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.

Influent Effluent Biosolid0

1000

2000

3000

4000

5000

6000

7000

5763

2328

3167

210 12

1,900

46 19 268

Comparison of ∑OPFR conc. vs. other FRs in sec-ondary wastewater treatment

∑ OPFRs ∑ BDE BTBPE DBDPE

Tota

l con

c. ng

/L, o

r ng/

g, d

ry

• 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 MethodSummary 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: mwoudneh@axys.com

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

Aftercleanup

Clean extracts were achieved with NH2 and silica columns

SeaStar Water Effluent Influent Soil Bio-solid0

50

100

150

200

Matrix effects for 0.5 L reagent water, 0.5 L effluent and in-fluent, 10 g soil and biosolid samples

TEP TCEP TPrP TCPP V6 TDCPP TPP TDBPP TCrP EHDPP TEHP

Perc

ent r

ecov

ery * *

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

0 20 40 60 80 100 120 140 1600

20

40

60

80

100

120

Stability of TEP, TCEP, TPrP

TEP TCEP TPrP

Number of Days in storage

Perc

ent r

ecov

ery

0 20 40 60 80 100 120 140 1600

20

40

60

80

100

120

Stability of TPP, TDBPP, TBP

TPP TDBPP TBP

Number of Days in storage

Perc

ent r

ecov

ery

0 20 40 60 80 100 120 140 1600

20

40

60

80

100

120

Stability of TCPP, V6, TDCPP

TCPP V6 TDCPP

Number of Days in storage

Perc

ent r

ecov

ery

0 20 40 60 80 100 120 140 1600

20

40

60

80

100

120

Stability of TBEP, TCrP, EHDPP, TEHP

TBEP TCrP EHDPP TEHP

Number of Days in storage

Perc

ent r

ecov

ery

OPFR Standard Stability in Methanol

OPFR Extract Stability

TEP

TCEP

TP

rP

TCPP

V6

TDCPP

TPP

TDBPP

TBP

TCrP

EHDPP

TEHP

020406080

100120140

Reagent water extracts after 4 months in storage

SeaStar Water

Perc

ent r

ecov

ery

TEP

TCEP

TP

rP

TCPP

V6

TDCPP

TPP

TDBPP

TBP

TCrP

EHDPP

TEHP

0

20

40

60

80

100

120

140

Effluent extracts after 4 months in storage

Effluent

Perc

ent r

ecov

ery

TEP

TCEP

TP

rP

TCPP

V6

TDCPP

TPP

TDBPP

TBP

TCrP

EHDPP

TEHP

020406080

100120

Influent extracts after 4 months in storage

Influent

Perc

ent r

ecov

ery

TEP

TCEP

TP

rP

TCPP

V6

TDCPP

TPP

TDBPP

TBP

TBEP

TC

rP

EHDPP

TEHP

020406080

100120

Sediment and bio-solid extracts after 4 months in storage

Sediment Biosolid

Perc

ent r

ecov

ery

MRM transitionsTarget Analyte Typical Retention

Time (minutes)Parent Ion

Mass 1DaughterIon Mass 1 Quantified Against

TEP 8.68 183 99.0 (127) d15-TEPTCEP 9.79 284.9 222.9 (63.1) d12-TCEPTPrP 13.32 225.1 99.0 (140.9) d21-TPrPTCPP 13.50 329.1 (327.1) 99.0 (99.0) d18-TCPPV6 14.38 582.8 360.8 (99.0) d15-TDCPPTDCPP 16.28 430.9 99.0 (208.9) d15-TDCPP TPP 16.86 327.1 152.0 (77.1) 13C18-TPPTDBPP 17.44 715.5 (698.6) 99.0 (99.0) d15-TDCPPTBP 18.17 267.1 99.0 (155.0) d27-TBP

TBEP 19.09 399.1 299.0 (199.0) d27-TBP

TCrP 21.08 369.1 165.0 (91.0) 13C18-TPP

EHDPP 22.20 363.1 250.9 (77.1) 13C18-TPP

TEHP 27.21 435.3 99.0 (211.0) d27-TBP

Surrogate Standard

d15-TEP 8.55 198.0 102 (134) d15-TPPd12-TCEP 9.70 297.0 231.9 (67.1) d15-TPPd21-TPrP 13.10 246.2 102 (150.1) d15-TPPd18-TCPP 13.41 347.0 (345) 102 d15-TPPd15-TDCPP 16.18 446.0 102 (216) d15-TPP13C18-TPP 16.86 345.0 164.1 (83.1) d15-TPPd27-TBP 17.93 294.2 102 (166.1) d15-TPP

Recovery Standard

d15-TPP 16.67 342.1 160.1 (82.1) d15-TPP

LC Gradient

Time (min) Flow mixture 1 LC Flow Rate Program

GradientCurve

0.015% solvent A

85% solvent B0.2 mL/min 1

3.050% solvent A

50% solvent B0.2 mL/min 6

1070% solvent A

30% solvent B0.2 mL/min 6

1885% solvent A15% solvent B

0.2 mL/min 6

2195% solvent A5% solvent B

0.2 mL/min 4

3095% solvent A

5% solvent B0.25 mL/min 6

3115% solvent A

85% solvent B0.2 mL/min 1

3515% solvent A

85% solvent B0.2 mL/min 1

Calculated Log KD vs Log Kow for OPFRs

TCEP

TB

P TE

P V6

TCPP

TDCPP

TPrP

TDBPP

TPP

TBEP

TE

HP

EHDPP

TCrP

0.01.02.03.04.05.06.07.08.09.0

10.0

Log Kd log Kow log Koc