Specific Identification of Organic Pollutants in the Catawba Watershed Wastewater Treatment Plant Effluents John Turner Faculty Mentor: Dr. Pat Owens Faculty Committee Members: Dr. Pat Owens, Dr. Cliff Calloway, Dr. Peter Phillips

Threats posed by organics Many are toxic (harmful), carcinogenic (cancer- causing), or mutagenic (capable of altering genetic material) at very low levels Many organics have been shown to be endocrine disruptors capable of altering the sex of fishes May have the capability for bioaccumulation in the fat cells and membranes

From The Charlotte Observer October 4 th, 2004

The Status Quo Presently there is no testing of organic levels in WWTP effluents within the Catawba watershed Furthermore, testing of drinking water and river water for organics is not being done. Some organics such as 1,4-dioxane have been known to travel as far as 200 kilometers from its point of entry in a watershed. (Draper et al, 2000)

Research Design and Methods Collection of WWTP samples –Bottle washing procedure Glass bottles (approximately 600 mL) were used with glass stoppers 2 mL HPLC grade Methanol added to each bottle Stoppers returned to bottle and rolled and flipped so that Methanol completely covered the inside of the bottle This was then repeated using reagent grade Ethyl Acetate Unstopped bottles were placed in a dry oven at 108 o C for 24 hours

Research Design and Methods Collection of WWTP samples –Samples were collected from 2 WWTPs Samples were collected from Rock Hill (Manchester Creek) and Fort Mill WWTPs. The samples that were collected were composite samples

WWTP Locations Rock Hill WWTP (Manchester) Fort Mill WWTP

Research Design and Methods Organic Extraction –The Basic Apparatus Glass Funnel Membrane Filter Vacuum Hose Erlenmeyer Vacuum Flask

Research Design and Methods Organic Extraction –Empore C 18 Membrane Filter Preparation An Empore C 18 Membrane Filter was placed at the bottom of the Glass Funnel on the screen. 25 grams of Empore Filter Aid glass beads were added to the funnel The disk was washed with 20 mL of elution solvent (methanol or ethyl acetate) and then allowed to dry 20 mL of methanol was used to condition the disk

Research Design and Methods Organic Extraction –Concentration of Organics onto the filter The sample was decanted into the glass funnel. The vacuum was then applied and the sample was allowed to run through the disk. The vacuum was allowed to continue to run for 5-20 minutes so that residual water was removed.

Research Design and Methods Organic Extraction –Organic Elution The organics were eluted from the disk into an organic phase, and they were collected by placing a Kuderna-Danish flask under the membrane filter. 10 mL of elution solvent was then added (either methanol or ethyl acetate) so that it covered the disk completely The vacuum was used to pull the solvent that now contains the organics into the Kuderna-Danish flask.

Research Design and Methods Organic Extraction –Concentration of Elution Once the organics were concentrated into 10 mL of solvent they were further concentrated for analysis. Two mL of elution solvent was removed. Then the remaining 8 mL of elution solvent was evaporated down to approximately 1 mL. This was done by blowing a gentle stream of nitrogen that passed through an organic carbon filter over the solvent.

Research Design and Methods Organic Extraction –Analytical Plan 500 mL of Water 10 mL of extract 1 mL of extract 1/50th 1/500th 1 μL analyzed

Research Design and Methods 1 μl of extract was injected into the GC column The extract contains a mixture of the organics that were originally found in the wastewater but have now been concentrated into an organic phase. The GC column separated the analytes based upon their polarity. The MS measured the molecular weight of the various ions and a chromatogram was produced. Run Gas Chromatography-Mass Spectrometry

Results Siloxanes Phenol Phthalates

Siloxanes – D5 Rock Hill WWTP min with Ethyl Acetate min with Methanol

D5 (cyclopentasiloxane)

Library Match for D5

Siloxanes – D6 (cyclohexasiloxane) Rock Hill WWTP min with Ethyl Acetate

Library Match for D6 Loss of 88

Siloxanes – D7(cycloheptasiloxante) Rock Hill WWTP min with Ethyl Acetate

Library Match for D7

Siloxanes – D8 (cyclooctasiloxane) Rock Hill WWTP min with Ethyl Acetate

Siloxanes – D9 (cyclononasiloxane) Rock Hill WWTP & min with Ethyl Acetate

Siloxanes – D10 (cyclododecasiloxane) Rock Hill , , , , , , , and min with Ethyl Acetate min with Methanol Fort Mill and min with Ethyl Acetate min with Methanol

Library Match for D10

Risks Associated with Siloxanes Bioaccumulation in tissues and organs Capable of inhibiting estrogen, leading to reproductive problems

Phenol Rock Hill WWTP min with Ethyl Acetate min with Methanol

Library Match for Phenol

Risks Associated with Phenol Most affects are related to the exposure to large quantities over time Probably not a major problem unless found in substantial quantities

Bis (2-ethylhexyl) phthalate Rock Hill WWTP min with Methanol Fort Mill min with Methanol

Library Match for a Phthalate

Risks Associated with Bis (2-ethylhexyl) phthalate Has been shown to bioaccumulate in tissue Affects the abilities of males to father babies

Conclusions GC-MS was shown to be an effective means of identifying organics in wastewater Within the organics identified, the siloxanes and phthalates have serious health effects at very low levels This research established a basic understanding of the use of GC-MS in identifying organics in wastewater and has accumulated information to fuel further research in this area in the future at Winthrop