A Thesis Entitled Polycyclic Aromatic Hydrocarbon Characterization In

Home , Dibenzofuran, Pentane

A Thesis

entitled

Polycyclic Aromatic Hydrocarbon Characterization

in Otter Creek, Northwest Ohio

Deanna M. Bobak

Submitted as partial fulfillment of the requirements for

the Master of Science Degree in Geology

______Dr. Alison L. Spongberg, Advisor

______College of Graduate Studies

The University of Toledo

May 2010

This document is copyrighted material. Under copyright law, no parts of this document

may be reproduced without the expressed permission of the author.

An Abstract of

Polycyclic Aromatic Hydrocarbon Characterization in Otter Creek, Northwest Ohio

Deanna M. Bobak

Submitted as partial fulfillment of the requirements for the Master of Science in Geology

The University of Toledo May 2010

Polycyclic aromatic hydrocarbons (PAHs) are globally found contaminants, classified into two main groups: petrogenic and pyrogenic. Petrogenic PAHs are introduced naturally into the environment by oil seeps or anthropogenically by spills of crude or refined petroleum product. Pyrogenic PAHs are formed from the incomplete combustion of fossil fuel or biomass and are commonly distributed by atmospheric deposition and urban runoff. The persistence and toxicity of PAHs make them a target for remedial investigations.

Otter Creek, northwest Ohio, is a small tributary to Maumee Bay and has been associated with natural oil and multiple incidents of crude/refined oil spills. As part of the greater Maumee River Area of Concern, PAH contamination has been documented, but never attributed to sources. Knowledge of PAH type may help in remedial actions planned for the creek.

A modified EPA Method 8270 was used for the analysis of 23 Otter Creek sediment samples by gas chromatography/mass spectrometry (GC/MS). A suite of 44 non-alkylated and alkylated PAHs was identified and quantified in selected ion

iii

monitoring mode. PAH source allocation was accomplished through compilation of diagnostic data, including distribution histograms of homologous PAH series, key diagnostic ratios, and principal component analysis.

Total PAH concentrations in Otter Creek ranged from 10.9 to 3015.0 mg kg-1.

Sites located at the headwaters had relatively low concentrations compared with downstream sites associated with PAH point sources and were considered baseline values of PAH contamination. Pyrogenic PAHs associated with urban background were prevalent in the headwaters section of the creek. As proximity to known point sources increased, total PAH concentrations increased and sediments exhibited mixing of petrogenic and pyrogenic sources. Mixed signatures were predominant over the refinery and residential stretches of the creek. Furthest downstream, the industrial portion of

Otter Creek contained the highest PAH concentrations and mainly petrogenic features.

This work is dedicated to Donald D. Swicegood, Jr. Great minds inspire those around them. I miss you Daddy.

Acknowledgements

I would like to thank Dr. Alison Spongberg for her unending guidance, support, and patience during this process. I never could have done this without her confidence in me. I would also like to thank my committee members, Dr. Daryl Dwyer and Dr. David

Krantz for continued support on this project and their interest in serving on my committee.

Funding for this project was provided from United States Department of Agriculture grant

#2003-38894-02032 and 2006-38894-03732.

Helder Costa from BBL Associates, Dr. Scott Stout from Newfields, LLC., and Liz

Porta from Alpha Woods Hole Analytical Laboratory have all been very generous in sharing their expertise and knowledge in the field of environmental forensics.

Cooperation from Mark Klemmer, Arcadis, is greatly appreciated in obtaining access to

Otter Creek on CSX property.

Many, many thanks to GESS lab technician, Jason Witter, who has been an integral part in field sampling, laboratory guidance, and has always had an ear to lend or a comment to give. Thank you also to Alycia Pittenger for her sampling help and to my work peeps, Jordan Rofkar and Kris Barnswell, for not kicking me out of the office as I whined my way through the writing of this document.

Last, but certainly not least, thank you to my friends and family who have held me up when I could not stand. You have been the best personal cheering squad anyone could ask for.

Table of Contents

Abstract iii

Acknowledgements vi

Table of Contents vii

List of Tables ix

List of Figures xi

1 Introduction 1

1.1 Preamble ……………………………………………………………….. 1

1.2 Objectives and Scope ………………………………………………….. 2

1.3 Hypotheses …………………………………………………………...... 3

1.4 Site Description and History ………………………………………...... 3

2 Literature Review 8

2.1 General PAH Characteristics ………………………………………….. 8

2.2 PAH Sources ………………………………………………………...... 9

2.3 Controls on PAH Assemblages ...…………………………………...... 12

3 Materials and Methods 15

3.1 Materials and Analytical Methods 15

3.1.1 Sample Collection …………………………………………….. 15

3.1.2 Moisture Content Analysis …………………………………… 18

vii

3.1.3 Total Organic Carbon Analysis ………………………………. 19

3.1.4 Particle Size Analysis ………………………………………… 20

3.1.5 Forensic Analysis …………………………………………...... 20

3.1.5.1 Soxhlet Extraction …………………………………. 21

3.1.5.2 Solid Phase Extraction …………………………….. 22

3.1.5.3 Gas Chromatography/Mass Spectrometry (GC/MS) Method .………………………………..... 23

3.1.5.3.1 Method Development for Non-Alkylated PAHs ………………… 23

3.1.5.3.2 Method Development for Alkylated PAHs …………………….... 27

3.1.5.3.3 Method Development for Samples …… 28

3.2 Statistical Methods (Principal Component Analysis) …………………. 31

4 Results and Discussion 32

4.1 General Trends ……………………………………………………...... 32

4.2 Petrogenic and Pyrogenic Distributions ……………………………….. 37

4.3 Diagnostic Source Ratios …………………………………………….... 41

4.4 Principal Component Analysis ……………………………………...... 51

5 Conclusions 57

References 62

Appendices

A Individual PAH Concentrations 68

B PAH Distribution Histograms 73

viii

List of Tables

1-1 Possible industrial sources of pollution currently along Otter Creek and

their locations. .…………………………………………………………… 4

1-2 Ohio EPA documented releases to Otter Creek between 1978 and 2005. .. 6

2-1 Targeted PAHs in environmental investigations and their source

allocation. ………………………………………………………………… 10

3-1 Sampling dates and location in Otter Creek. ……………………………... 16

3-2 Target PAHs for Otter Creek study. ……………………………………… 24

3-3 Elution times for non-alkylated target and surrogate compounds. ……….. 26

3-4 Calibration standard peak areas and calculated response factors for

non-alkylated PAHs. ……………………………………………………... 27

3-5 Program for DSIMHOT. …………………………………………………. 29

3-6 Program for DSIMNP. …………………………………………………… 29

3-7 Program for DSIMFFP. …………………………………………………... 29

3-8 Program for DSIMDC. …………………………………………………… 30

4-1 Summary of chemical and physical data for each sampling site in Otter

Creek. …………………………………………………………………….. 36

4-2 Distribution patterns for the five homolog groups: naphthalenes (N0-4),

fluorenes (F0-3), dibenzothiophenes (D0-4), fluoranthenes (FL0-3), and

chrysenes (C0-4). ………………………………………………………… 38

4-3 Source ratio data for the Otter Creek dataset. ……………………………. 44

5-1 Final summary for source allocation in Otter Creek. …………………….. 58

List of Figures

1-1 Oily sheen produced from bottom sediment disturbance in March 2005. .. 6

2-1 Representative patterns for petrogenic and pyrogenic homolog

distributions. ……………………………………………………………… 11

3-1 Study area and sampling locations in Otter Creek, Northwest Ohio. ……. 17

4-1 Profile of total PAH concentrations observed over the length of the creek

from headwaters to mouth. ……………………………………………….. 33

4-2 Total PAH concentrations normalized to percent organic carbon content. . 36

4-3 Typical distribution patterns found in the headwaters section of Otter

Creek. …………………………………………………………………….. 39

4-4 Sites typical of both petrogenic and pyrogenic inputs. …………………... 40

4-5 Petrogenic signatures were abundant in the heavily industrialized section

of the creek. ………………………………………………………………. 41

4-6 Double-plot ratios of Otter Creek samples in relation to calculated

averages of literature values for common PAH sources. ………………… 49

4-7 Principal component scores for Otter Creek sample sites. ……………….. 52

4-8 Component loadings for Otter Creek sites. ………………………………. 53

4-9 Component scores for Otter Creek sites and reference sources. …………. 54

4-10 Component loadings for Otter Creek sites and reference sources. ……..... 56

Chapter 1

Introduction

1.1 Preamble

The largest source of terrestrial, marine, and groundwater contamination is from petroleum or refined petroleum products (Douglas and Uhler, 1993). Strict regulations regarding the reporting of oil spills were enacted in the 1970s. The new requirement that potentially responsible parties (PRPs) pay for the cost of remediation resulted in the need for a defensible strategy to accurately and fairly determine the extent of the responsibility of each PRP. Thus, much of the geochemical work regarding oil exploration became the basis for the environmental forensic process used today (Boehm et al., 1997).

The environmental forensics investigator has a number of tools to consider when dealing with hydrocarbon-contaminated sites, but one of the most useful is a modification of EPA Method 8270, Semivolatile Organic Compounds by Gas Chromatography/Mass

Spectrometry (GC/MS). This technology is sensitive enough to detect polycyclic aromatic hydrocarbons (PAHs), which constitute only ~5% of crude oil but are one of the most important classes of compounds in hydrocarbon source allocation (Boehm et al.,

1997; Sauer and Uhler, 1994-1995; Stout et al., 2002).

Knowledge of PAH source can be instrumental in remediation efforts, as PAHs from different sources partition uniquely and predictably in sediments. PAHs associated with soot particles tend to accumulate in sediments and be less bioavailable to aquatic organisms, while PAHs associated with spilled crude oil or natural oil seeps tend to be more easily dissolved and mobile within the water column (Latimer and Zheng, 2003).

Treatment strategies can be designed to hinder or impede point source contamination from re-contaminating a newly restored site and can consider the persistent accumulation of non-point source PAHs from an urban background contribution.

1.2 Objectives and Scope

The overall objective of this study was to duplicate the GC/MS method used by environmental forensics investigators to quantify and allocate PAH contamination in

Otter Creek to petrogenic or pyrogenic sources. Specific objectives of this study included:

- Develop a method for GC/MS identification and quantification of PAH

analytes

- Quantify PAH concentrations in sediments of Otter Creek at selected sites

- Determine sediment grain-size and total organic carbon content and

examine relationships with PAH concentrations at each site

- Allocate PAH contamination to source by inspection of PAH distribution

histograms, diagnostic source ratios, and principal component analysis

1.3 Hypotheses

Otter Creek sediments should exhibit signatures for both petrogenic and pyrogenic sources. Most samples should be impacted by pyrogenic, urban background.

Petrogenic signatures should be seen in sediments adjacent to or downstream of known point sources of PAH contamination, i.e., the refinery operations and oil pipeline.

1.4 Site Description and History

Otter Creek is a small tributary east of the Maumee River in Northwest Ohio.

Stretching approximately 7.9 miles (12.71 km) long and draining an area of approximately 8 square miles (21 square km), the headwaters begin in the city of

Northwood and continue flowing in a northeasterly direction through the cities of Toledo and Oregon to be the last tributary to empty into the Maumee River before it flows into

Maumee Bay and Lake Erie (Prater and Anderson, 1977).

Otter Creek and oil have always been historically intertwined. Historical accounts cite Native Americans using pitch from natural oil seepages along the creek banks to tip their arrows (TMACOG, 2007). The most famous documentation of oil in the area occurred in the lowlands between Otter Creek and Duck Creek near river mile 2.0. Here, the “Klondike Gusher” erupted in July of 1897, producing 100 barrels an hour. However, the Gusher was reported as being “somewhat of a frost” in the Toledo Blade in

September later that year (Downes, 1954). This preceded a boom in oil exploration in

Lucas County. While maps from the late 1800s and early 1900s display an enormous number of oil wells (that have since been closed), the area quickly became realized for its refining activities more than its crude production abilities.

The first documented industry in the vicinity of Otter Creek was the Crystal Oil

Company, which opened in early 1895 as the Diamond Oil Company and was renamed the Sun Oil Company later that year (Fassett, 1961). The 14-acre facility produced kerosene, gas, oil, and fuel oil and has since grown to over 270 acres. Standard Oil

Company (now BP) built its first facility in 1919, and its 465-acre facility produces gasoline, jet fuel, kerosene, petroleum coke, and low sulfur diesel fuel (OEPA, 1999).

Smaller refineries that are no longer in operation were managed under Craig Oil and Gulf

Oil (formerly Paragon). Other industrial entities along the creek include Libbey-Owens

Ford (now Pilkington North America, Inc.), a glass manufacturing plant with waste ponds, the Toledo Wastewater Treatment Plant whose sludge water impoundments discharge to Otter Creek under National Pollutant Discharge Elimination System

(NPDES) permit, Envirosafe Services of Ohio, Inc. which contains extensive hazardous waste landfills with a history of suspected leachate releases, a large oil pipeline operated by Buckeye Partners, L.P., and the CSX loading docks used for transportation of coal

(Table 1-1). Two commercial and residential landfills are also closely situated near the creek, as are approximately sixteen formerly uncontrolled hazardous waste landfills

(Maumee RAP, 1997).

Table 1-1: Possible industrial sources of pollution currently along Otter Creek and their locations. Location on creek Industry Possible pollution input (river miles/km) Libbey Owens Ford (Pilkington) Heavy metals RM 6.6-6.7 (10.6-10.8 km) Sun Oil Refinery Hydrocarbon/petroleum RM 5.0-5.4 (8.0-8.6 km) Toledo Wastewater Treatment Plant Thermal RM 2.3-3.0 (3.7-4.8 km) Envirosafe Services of Ohio, Inc. Various hazardous waste RM 2.0-2.3 (3.2-3.7 km) Buckeye Pipeline Hydrocarbon/petroleum RM 1.9-2.2 (3.1-3.5 km) BP Oil Refinery Hydrocarbon/petroleum RM 0.5-1.8 (0.8-2.9 km) CSX loading docks Hydrocarbon/coal RM 0-2.1 (0-3.4 km)

As part of the greater Maumee River Area of Concern, designated by the United

States Environmental Protection Agency, attention is focused on Otter Creek due to its reputation as an “industrial sewer”, with oil soaked banks and a suite of detectable organic and inorganic contaminants in its waters and sediments (Maumee RAP, 1999).

Otter Creek is deemed a Limited Resource Waterway (LRW) upstream of river mile 6.4 and a Modified Warm-water Habitat (MWH) downstream of river mile 6.4 (OEPA,

1989). LRWs are waters that do not support a diverse aquatic community and their potential for recovery is essentially null due to natural background or irreversible human- induced conditions. MWHs include waters that are incapable of maintaining a balanced aquatic community due to irreversible habitat modification (OAC 3745-1). Causes of impairment with high to moderate magnitude include oil and grease, siltation, flow and habitat alteration, and known and unknown toxicities. The main sources of impairment were named as major industrial point sources, landfills, urban run-off/storm sewers, channelization, and streambank/riparian habitat modification (TMACOG, 2006).

Formerly part of the Black Swamp, Otter Creek supported a diverse warm-water fish habitat prior to industrialization (OEPA, 1976). Sediment and water column studies have detected alarming concentrations of contaminants detrimental to the aquatic community, as well as those hazardous to humans. Many of these compounds are associated with oil (Maumee RAP, 1997). Several reports cite oil sheens floating on the surface of the creek, sulfur or gasoline odors coming from the creek, and an appearance of oily sheens when sediment is disturbed (OEPA, 1976; Maumee RAP, 1997). Figure 1-

1 illustrates an oily sheen that surfaced when sediments were disturbed during sampling in March 2005. Table 1-2 details documented releases of oil and oily wastewater to the

creek between 1978 and 2005. Over 52,000 gallons of oil products have been released to the creek; however, this number only represents amounts from roughly half of those spills. Of the 97 known releases, 49 spills are of unknown amounts.

Figure 1-1: Oily sheen produced from bottom sediment disturbance in March 2005. (Photo by: Deanna Bobak)

Table 1-2: Ohio EPA documented releases to Otter Creek between 1978 and 2005 (OEPA, 2006). Total known amount Number of spills with Product spilled Total spills spilled (gallons) amount unknown Crude oil 9 1667 2 Decant oil 1 unknown 1 Diesel fuel 1 1800 -- Fuel oil 3 700 1 Fuel oil #2 2 127 -- Gasoline 5 32,302 2 Hydraulic oil 1 unknown 1 Hydrocarbons 1 20 -- Kerosene 1 1000 -- Light oil 1 unknown 1 Lube oil 1 unknown 1 Oil 49 1109 35 Petroleum substances 4 200 3 Waste oil 16 486 5 Wastewater oil 2 12,720 1 TOTAL 97 52,131 49

Sediment characterization in Otter Creek has occurred several times over the last four decades (ChemRisk, 1999; SulTRAC, 2007). The majority of the studies were part of a greater Area of Concern sampling program, and if PAHs were included in the analyses, relatively few were analyzed by the standard EPA Method 8270 (ChemRisk,

1999). PAH data that were generated focused only on the EPA Priority Pollutants and sampling sites usually did not span the entire length of Otter Creek. To compensate for the deficiency in these studies, samples collected from the entire length of Otter Creek in

2007 were analyzed for an extended PAH list including 34 analytes, but the scope of that study was not to interpret PAH sources. Results from this project will be the first to interpret the primary sources of PAH contamination in Otter Creek and may have implications for future remedial work within the watershed.

Chapter 2

Literature Review

2.1 General PAH Characteristics

Polycyclic aromatic hydrocarbons (PAHs), also known as polyarenes or polynuclear aromatic hydrocarbons, are ubiquitous contaminants in the environment.

PAHs constitute a very large group of organic compounds that contain two or more fused benzene rings (Harvey, 1997). Theoretically, PAHs could have infinite benzene rings, and the arrangement of those benzene rings could produce a copious number of isomers.

However, PAHs most studied contain eight or less rings, with the most environmentally significant ones having seven or less (Harvey, 1997; Boehm et al., 2002). PAHs may also be alkylated (substituted), meaning straight-chained hydrocarbons are attached to the rings at one or more points (Boehm et al., 2002). PAHs form homologous series, containing a non-alkylated PAH known as the parent compound and alkylated PAHs that have the parent compound as their base. PAHs attached by one alkyl chain are classified as C1-members, those with two alkyl chains are classified as C2-members, those with three alkyl chains are C3-members, and so on. Commonly associated with PAHs are heterocycles, compounds in which an atom of nitrogen, sulfur or oxygen has replaced one carbon atom in a ring (Boehm et al., 2002).

2.2 PAH Sources

PAHs typically occur in assemblages and are classified into three main sources: petrogenic, pyrogenic, and biogenic (Neff, 1979; Page et al., 1999). Petrogenic PAHs are formed over long periods of time in low temperature (100-300°C), high pressure environments where ancient organic matter is converted into petroleum and coal (Stout et al., 2001a; Boehm et al., 2002). Petrogenic assemblages are enriched in 2- and 3-ring, lower molecular weight compounds and their constituent homolog groups exhibit a predominance of alkylated members to non-alkylated parents (Table 2-1 and Figure 2-1).

Pyrogenic PAHs typically have 4- to 6-rings and are rapidly formed during incomplete combustion of fossil fuel or biomass at temperatures >500°C (Table 2-1). At these higher temperatures, the more reactive, alkylated PAHs tend to be destroyed, so homolog groups in pyrogenic assemblages are dominated by non-alkylated, parent compounds, with subsequently less alkyl-members as the degree of alkylation increases (Figure 2-1)

(Boehm et al., 2002; Stout et al., 2001). Biogenic PAHs are synthesized by plants and animals or formed during early diagenesis in sediments. Unlike petrogenic and pyrogenic PAHs, biogenic PAHs generally are found individually or in very simple mixtures.

PAHs in the environment are an ecological and human-health concern. Of the one-hundred twenty-six Environmental Protection Agency Priority Pollutants listed by the Clean Water Act, sixteen are PAHs, with seven being known carcinogens (Table 2-1)

(U.S. EPA, 2009). An understanding of the different mechanisms in which PAHs are introduced into the environment can help limit exposure to them.

- to 3 to

rings. thatAnalytes are listed as having mixed sources commonare petrogenic in and pyrogenic sources. - ringedin and molecular lower weight (MW), pyrogenic while compounds higher are molecular in weight and have 4 6 to Targeted inPAHs environmental investigations and their source allocation. Petrogenic compounds usually are 2

1: 1: -

Table 2

Figure 2-1: Representative patterns for petrogenic and pyrogenic homolog distributions. C0 is the parent, non-alkylated member, and degree of alkylation increases from 1 to 4 (adapted from Sauer and Uhler, 1994-1995).

Pathways of entry into the environment for both petrogenic and pyrogenic PAHs include both natural and anthropogenic mechanisms and occur on local, regional, and global scales. Natural oil seeps and the erosion of source rocks, such as oil shales, are a global phenomenon. In this manner, PAHs adsorb to the natural sediments and become part of the environmental background (Boehm et al., 2002). It is estimated that approximately 10% of the hydrocarbons in the marine environment are from natural oil seeps (Volkman et al., 1992). Petrogenic PAHs are also released into the environment by accidental spills of crude oil or its refined products while in production or transport.

While oil spills attract much media attention, the contribution of PAHs to the environment in this manner is considered to be relatively minor due to the low percent of

PAH in crude oil (Boehm et al., 2002). Most of the input of petrogenic PAHs to the environment is attributed to urban runoff and wastewater outfalls. Mixing of uncombusted lubricating oil drippings with stormwater runoff and illegal/unintentional discharge of waste oil are eventually discharged to larger water bodies through municipal waste systems (Eganhouse et al., 1982).

Pyrogenic PAHs are naturally established in the environment by forest and grassland fires, but the largest pathway of pyrogenic PAHs comes from anthropogenic activities (Neff, 1979). Pyrogenic PAHs are released into the atmosphere by incomplete or inefficient combustion of fossil fuels. Sediment cores across the world have shown a substantial increase in PAH concentration with the onset of industrialization (Hostettler, et al., 1999; Brenner et al., 2002; Kannan et al., 2005). PAHs attach to soot particles and have been known to settle over distances far from their original source (Aboul-Kassim and Simoneit, 1995; Lima et al., 2005). As soot and atmospheric particulates settle, they adsorb to soil, enter the water column of lakes and streams directly, or accumulate with road dust and mix in stormwater and urban runoff.

Pyrogenic PAHs are linked with industrial processes such as aluminum smelting, coke production, manufactured gas production, and creosote preservation. These processes are pathways of entry into the environment via wastewater streams, by-product residues, and atmospheric emissions (Boehm et al., 2002). Vehicle emissions contain a mixture of PAHs from the original oil, their combustion, those trapped in the lubricating oil, or found in the exhaust system. Their assemblages in vehicle emissions may or may not closely resemble the original oil that was combusted, as many confounding factors exist between vehicles (Lima et al., 2005). PAHs are also derived from domestic heating in coal, oil, or wood burning furnaces and/or ovens (Oanh et al., 1999).

2.3 Controls on PAH Assemblages

PAH assemblages observed in the environment are a result of three main controls:

(1) petroleum genesis, (2) refining effects, and (3) environmental factors. Differences

among crude oils are a result of depositional environments and geologic history. The source rock and type of organic matter that formed the petroleum are the two largest factors contributing to differences in crude oils. Thermal maturity of the source rock and compositional changes during oil migration are also important (Stout et al., 2002). The distillation of crude oil into distinct factions for use as different refined products is based on boiling point. Often, refinement of crude oil will lower or distort ratios of diagnostic compounds and will confound the ability to correlate refined products to the source oil

(Stout et al., 2002). To determine differences between refined products, PAHs are used in conjunction with biomarkers and other diagnostic hydrocarbons, especially where middle-distillate fuels (jet fuel, fuel oil, and diesel #2) are suspected.

PAHs in the environment are subjected to a number of environmental controls that can alter their assemblages. Lower molecular weight compounds (2- and 3-ring), such as naphthalene and phenanthrene, are most affected by weathering controls. Lower molecular weight PAHs generally have higher vapor pressures than higher molecular weight PAHs (4- to 6-ring); therefore, these lighter PAHs are preferentially evaporated from the environment (Stout et al., 2002). Lower molecular weight PAHs also tend to have higher water solubilities than higher molecular weight compounds, and are prone to water-washing, or solubilization. Generally speaking, as the degree of alkylation increases across a homologous series, solubility decreases (Douglas et al., 1992).

Concordant with solubility, lower molecular weight compounds and less-alkyl compounds are preferentially biodegraded (Douglas et al., 1992). Due to these general characteristics, heavily weathered PAH assemblages are characterized by the following pattern across any homologous series: C0 < C1 < C2 < C3 < C4.

Forensic investigators use histograms of PAH homolog distributions, coupled with ratios of key analytes to categorize environmental PAH assemblages into their respective source groupings. Distribution histograms and diagnostic ratios were investigated in this study to discern the sources of PAH assemblages in Otter Creek.

Chapter 3

Materials and Methods

3.1 Materials and Analytical Methods

3.1.1 Sample Collection

Sediment samples were collected on two different days from a total of twenty- three collection sites along Otter Creek (Table 3-1 and Figure 3-1). Sites were chosen based on accessibility to the creek and were generally sampled from the upstream sides of culverts, overpasses, etc. Samples were collected by grab-sample methodology.

Approximately two hundred grams of sediment from the top fifteen centimeters were collected with a sterilized hand trowel and/or modified hand auger and placed in pre- washed, dark brown glass jars with Teflon lids, to reduce photo-reactivity. All sample jars were placed in an iced cooler until return to the laboratory. Samples were then stored at -4°C in the dark until analysis.

Shortly before analysis, sediments were drained of excess water and allowed to air dry for 12-24 hours. Samples were divided into aliquots for: (1) moisture content analysis by the American Society for Testing and Materials (ASTM) D2216 method, (2)

Total Organic Carbon by a modified ASTM D 2974-87 method, (3) grain size analysis by

laser diffraction using the Fraunhofer standard operating procedure on a Malvern

Mastersizer 2000, and (4) quantification and identification of polycyclic aromatic hydrocarbons (PAHs) using a modification of U.S. EPA Method SW-846 8270 utilized in forensic investigations.

Table 3-1: Sampling dates and location in Otter Creek. River Mile (km) Date Sampled from mouth Site Description March 18, 2005 7.9 (12.7) Wales Rd./Tracy Rd. 7.8 (12.6) Wales Rd. 7.1 (11.4) Broadway St. before Andrus St. 6.0 (9.7) Oakdale St./Sunshine St. 5.6 (9.0) Pickle Rd. cemetery, upstream end 5.5 (8.9) Pickle Rd. cemetery, mid-way 5.4 (8.7) Pickle Rd. cemetery, downstream end 5.0 (8.0) Navarre Ave. 4.7 (7.6) East Moreland St. park, upstream end 4.6 (7.4) East Moreland St. park, mid-way 4.5 (7.2) East Moreland St. park, downstream end March 25, 2005 4.2 (6.8) Wheeling St. Lutheran home, upstream 4.1 (6.6) Wheeling St. Lutheran home, downstream 3.7 (6.0) N. Berlin St. park, upstream 3.6 (5.8) N. Berlin St. park, mid-way 3.5 (5.6) N. Berlin St. park, downstream 3.1 (5.0) Corduroy Ave./Torch St. apartments 2.3 (3.7) York St. 2.1 (3.4) Millard Ave. 0.5 (0.8) CSX property; furthest upstream 0.3 (0.5) CSX property; upstream 0.2 (0.3) CSX property; downstream 0.0 (0.0) CSX property; mouth of Otter Creek

Figure 3-1: Study area and sampling locations in Otter Creek, Northwest Ohio.

3.1.2 Moisture Content Analysis

This analysis ran concurrently with PAH forensic analysis. Pre-folded foil packets were weighed prior to filling to determine the mass of the foil packet (Mc). At least 20 grams of sediment were placed in each foil packet and re-weighed to measure the mass of the container and “wet” sediment (Mcw). Each foil packet was placed in a drying oven at 105°C for 24 hours. The packets were removed, allowed to cool to room temperature in a dessicator, and were re-weighed to determine the mass of the packet and

“dry” sediment (Mcd).

The mass of water (Mw) in each sample was determined by the following calculation:

Mw = Mcw-Mcd

The total percent moisture in each sample was determined by the following calculation:

% Moisture = [Mw / (Mw+Ms)] x 100

The calculated value of percent moisture in each sample was used to correctly normalize the concentration of PAH components on a dry weight basis. This was accomplished by taking the mass of the Soxhlet sample and multiplying it by the percent moisture value. The calculated result was subtracted from the Soxhlet sample mass to determine dry sediment weight.

3.1.3 Total Organic Carbon Analysis

This analysis was run independently from any other analysis. Individual foil packets were constructed and weighed to determine the individual mass (Mp). Each packet was filled with approximately 10-20 grams of sediment and placed in a drying oven at 105°C for 24 hours to eliminate atmospheric and inter-granular moisture. After

24 hours, packets were removed from the oven and placed in a dessicator to cool to room temperature. The packets were re-weighed to determine the dry mass of the sample

(Mdp). Samples were placed in a muffle furnace for five hours at a temperature of

500°C to burn any organic matter present in the samples. After five hours, the samples were allowed to cool to room temperature, were re-wetted with distilled water, and placed in a drying oven at 105°C for another 24 hours. Upon removal from the drying oven, the samples were cooled to room temperature in a dessicator and re-weighed

(Mbp).

The mass of sample minus sediment moisture (Ms) was determined by the following calculation:

Ms= Mdp-Mp

The mass of sample (Msb) after ashing was determined by the following calculation:

Msb= Mbp-Mp

The mass of total organic carbon (TOC) per sample was determined by the following calculation:

TOC= Ms-Msb

3.1.4 Particle-size Analysis

This analysis was run independently from any other analysis. The method for particle size preparatory work was designed by Amber Lahners, of the GLASS Lab of Dr.

Timothy Fisher at the University of Toledo.

Samples were unconsolidated using a mortar and pestle, but were not overly crushed as to preserve grain-size integrity. Representative samples of approximately 1-2 grams were placed in individual Nalgene tubes. Each sample was immersed in acetic acid for carbonate removal and allowed to sit for 24 hours under a hood. Upon carbonate removal, each sample was immersed in 15 mL of 2% sodium hypochlorite (bleach).

Treatment in bleach of 24 hours oxidized all organic matter present in the samples. To encourage dispersion of particles, each sample was immersed in 15 mL of a 4% sodium hexametaphosphate solution, shaken, and allowed to sit for approximately 24 hours.

Samples were analyzed on a Malvern Mastersizer 2000 (Malvern Instruments,

Ltd., Worchestershire, UK), using the Fraunhofer standard operating parameters for laser diffraction analysis, with classification of particle size using ASTM parameters of less than 5.00 μm, between 5.00 and 74.00 μm, and between 74.00 and 200.00 μm for clay, silt, and sand, respectively (Sperazza et al., 2004).

3.1.5 Forensic Analysis

Samples were removed from the freezer and allowed to air-dry overnight under cover to remove excessive moisture. An aliquot of approximately 20 grams was removed for moisture content analysis (See Section 3.1.2). Approximately 2-5 grams were removed from the remaining sediment for Soxhlet extraction of organic compounds.

Additional sample clean-up was accomplished by solid phase extraction (SPE). SPE extracts were analyzed using gas chromatography/mass spectrometry (GC/MS) for PAH identification and quantification. All samples were processed in triplicate.

3.1.5.1 Soxhlet Extraction

Empty cellulose thimbles were weighed, filled with 2-5 grams sediment, and re- weighed. Each sample was spiked with 0.5 mL of 10 parts per million (ppm) surrogate standard solution containing deuterated compounds not expected to be seen in environmental samples: naphthalene-d8, acenaphthene-d10, phenanthrene-d10, chrysene- d12, and perylene-d12 (Semivolatile Internal Standard (SIS) Mix (2000 μg/mL), lot number LB21066, Supelco, Inc., St. Louis, MO). This prepared standard served as both a surrogate and recovery standard to account for the amount of compounds lost throughout the extraction and clean-up procedures.

An activated pre-rolled ball of copper turnings (2 grams) was added to each thimble to complex any sulfur present in the sediments. Each ball was activated 1-3 hours before use according to the method outlined in Battelle standard operating procedure #5-192-05 (Battelle Memorial Institute, 2001). The balls were placed in a glass beaker to which equal parts distilled nanopure water and 12 N hydrochloric acid were added. The mixture was stirred for 2-3 minutes or until the balls became a bright, copper color. The turnings were rinsed 8 times in distilled water, 3 times in acetone, and

2 times in methylene chloride.

The thimbles were placed in one of two rapid Soxhlet extraction units (Tecator

Soxtec System HT6 1043 or Foss Soxtec System 2043 Extraction Unit; Foss, Eden

Prairie, MN), operated on an au tomated program in which samples were immersed in methylene chloride at temperatures between 110-125°C for 45 minutes. Samples were refluxed with solvent vapor for an additional 10 minutes. Solvent recovery occurred over approximately 25-45 minutes at a constant temperature under air pumped conditions.

Extracts were collected in pre-weighed glass extraction cups, allowed to cool to room temperature, and re-weighed for final extract mass.

3.1.5.2 Solid Phase Extraction

Solid phase extraction was used to separate non-target species from sample extracts. Silica extraction columns (LC-Si tubes, 3 mL; Supelco, Inc., St. Louis, MO) were attached to a Visiprep Solid Phase Extraction Vacuum Manifold (Supelco, Inc., St.

Louis, MO) and were conditioned with 1 mL of hexane. Extracts were reconstituted with

2 mL of 1:1 methylene chloride:hexane solution and were transferred to the silica columns via disposable glass transfer pipette. The vacuum manifold stopcock was opened and samples drained, gravity driven, through the pre-packed column and emptied into collection tubes. Two mL of 1:1 methylene chloride:pentane solution were added to the silica column and allowed to percolate, gravity driven, to collect in the collection tube. An additional 1 mL of 1:1 methylene chloride:pentane solution was added to the column and allowed to percolate, gravity driven, to collect in the collection tube (Qian et al., 1998).

Elutant was evaporated to approximately 2 mL using a controlled stream of nitrogen gas with the Visidry Drying Attachment (Supelco, Inc., St. Louis, MO). The amount of extracted liquid was measured and transferred to brown 2 mL GC vials via a

1 mL disposable graduated glass pipet with 0.1 divisions marked. Recovery of surrogate standards for this method averaged 52%.

3.1.5.3 Gas Chromatography/Mass Spectrometry (GC/MS) Method

Target analytes included twenty-two non-alkylated PAHs and twenty-two alkylated homolog members (Table 3-2). Isolation, identification, and quantification of the 16 Priority Pollutants via EPA Method SW-846 8270 follows a standard procedure of organic extraction, sample clean-up, and analysis using GC, with MS capabilities. The expansion of the target analyte list to include alkylated PAH compounds, for which analytical standards are not available, required a significant effort in method development.

3.1.5.3.1 Method Development for Non-alkylated PAHs

Calibration standards included 8270 LCS Mix 1 (Supelco, Inc., St. Louis, MO; lot number LB21442). This mix of semivolatile compounds included the following target analytes: acenaphthene, acenaphthylene, anthracene, benzo(a)anthracene, benzo(a)pyrene, benzo(b)fluoranthene, benzo(g,h,i)perylene, benzo(k)fluoranthene, chrysene, dibenz(a,h)anthracene, dibenzofuran, fluoranthene, fluorene, indeno(1,2,3-c,d)pyrene, naphthalene, phenanthrene, pyrene, 1-methylnaphthalene, and

2-methylnaphthalene. This prepared solution contained a concentration of 100 μg/mL of all compounds mixed in a solution of 90:10 methylene chloride:acetone. Neat standards of the remaining non-alkylated target analytes, benzo(e)pyrene (Supelco, Inc., St. Louis,

MO; lot number LB21553), biphenyl (Supelco, Inc., St. Louis, MO; lot number

LB15806), dibenzothiophene (Supelco, Inc., St. Louis, MO; lot number LB20805), and perylene (Supelco, Inc., St. Louis, MO; lot number LB20731) were prepared to a 10 ppm solution by adding 1.00 mg of each to methylene chloride to fill a 100 mL flask.

Table 3-2: Target PAHs for Otter Creek study. Abbreviations are used in subsequent tables. Analyte/Analyte Group Abbr. Analyte/Analyte Group Abbr. Naphthalene N0 Pyrene PY C1-Naphthalenes N1 C1-Fluoranthenes/pyrenes FL1 C2-Naphthalenes N2 C2-Fluoranthenes/pyrenes FL2 C3-Naphthalenes N3 C3-Fluoranthenes/pyrenes FL3 C4-Naphthalenes N4 Benz(a)anthracene BAA Biphenyl BPH Chrysene C0 Acenaphthylene ACL C1-Chrysenes C1 Acenaphthene ACE C2-Chrysenes C2 Dibenzofuran DBF C3-Chrysenes C3 Fluorene F0 C4-Chrysenes C4 C1-Fluorenes F1 Benzo(a)fluoranthene BAF C2-Fluorenes F2 Benzo(b)fluoranthene BBF C3-Fluorenes F3 Benzo(j,k)fluoranthene BKF Anthracene AN Benzo(e)pyrene BEP Phenanthrene P0 Benzo(a)pyrene BAP C1-Phenanthrenes/anthracenes P1 Perylene PER C2-Phenanthrenes/anthracenes P2 Indeno(1,2,3-c,d)pyrene ID C3-Phenanthrenes/anthracenes P3 Dibenzo(a,h)anthracene DA C4-Phenanthrenes/anthracenes P4 Benzo(g,h,i)perylene BGP Dibenzothiophene D0 C1-Dibenzothiophenes D1 Naphthalene-d8 Nd8 C2-Dibenzothiophenes D2 Acenaphthene-d10 Ad10 C3-Dibenzothiophenes D3 Phenanthrene-d10 Pd10 C4-Dibenzothiophenes D4 Chrysene-d12 Cd12 Fluoranthene FL Perylene-d12 Pd12

Each standard (surrogate, LCS 8270 Mix, and prepared neat) was analyzed using a Hewlett Packard 6980 Gas Chromatograph coupled to a Hewlett Packard 5972 Mass

Selective Detector (Hewlett Packard L.P., Palo Alto, CA) in full-scan mode.

The GC instrument specifications followed those published by Stout et al. (2001b):

Injection volume: 1.0 µL Syringe size: 10.0 µL Pre-injection washes, solvent A (MeCl): 0 Pre-injection washes, solvent B (MeCl): 1 Post-injection washes, solvent A (MeCl): 2 Post-injection washes, solvent B (MeCl): 1

Column: RTX-5 (Restek, Bellefonte, PA) 60m x 0.25mm x 0.25μm capillary column (max temp. 330˚C) Flow: 1.0 mL/min, constant Pressure: 16.08 psi Initial temperature: 40˚C (hold for 1.00 minute) Ramp: 6˚C to 300°C (hold for 30 minutes) Carrier gas: helium Total run time: 74.33 min.

Elution times for all non-alkylated target and surrogate compounds were confirmed through replicate analyses in which the elution time for each individual component remained consistent (±0.1 min) (Table 3-3). Once the elution times were identified, the PAHs were confirmed with comparison of mass-to-charge (m/z) ratios to library database values (NBS-75K, 1995).

Quantification of compounds was not performed using this method. The sensitivity of full-scan mode is markedly decreased due to the full range of m/z ratios being scanned. Instead, full-scan was used as a qualitative assessment of the whole sample and as a way to evaluate elution times of non-alkylated compounds.

Table 3-3: Elution times for non-alkylated target and surrogate compounds. Elution Time Elution Time Compound (minutes) Compound (minutes) Naphthalene-d8 20.86 Pyrene 39.51 Naphthalene 20.94 Benz(a)anthracene 44.25 Biphenyl 25.48 Chrysene-d12 44.33 Acenaphthylene 27.25 Chrysene 44.42 Acenaphthene-d10 27.85 Benzo(b)fluoranthene 49.04 Acenaphthene 27.98 Benzo(j,k)fluoranthene 49.16 Dibenzofuran 28.60 Benzo(e)pyrene 50.53 Fluorene 29.98 Benzo(a)pyrene 50.81 Dibenzothiophene 33.33 Perylene-d12 51.15 Phenanthrene-d10 33.73 Perylene 51.29 Phenanthrene 33.82 Indeno(1,2,3-cd)pyrene 58.61 Anthracene 34.00 Dibenz(a,h)anthracene 58.68 Fluoranthene 38.60 Benzo(g,h,i)pyrene 60.82

Non-alkylated and surrogate compounds were quantified under the same GC parameters using the selected ion monitoring (SIM) mode for mass spectrometer detection. SIM replaces the full-scan mode and differs in that the MS is programmed to detect only a limited number of m/z ratios (unique to each target analyte) over specified time periods, thus increasing the sensitivity of the instrument and improving detection limits. Peak area for each designated quantification m/z ratio was integrated using the

RTE Integrator setting in the ChemStation GC/MS software and baselines were manually adjusted for correct placement. Average response factors were calculated for each PAH using the standard solutions at concentrations of 10, 1, 0.1, 0.01, and 0.005 ppm.

Response factors were calculated from the peak areas of either five-point, four-point, or three-point calibration curves (dependent upon the detection limit and working range for that particular compound) and were later used to quantify each PAH within the sample

(Table 3-4).

Table 3-4: Calibration standard peak areas and calculated response factors for non-alkylated PAHs. PPM Nd8* N0 Bph Acl Ad10 Ace Dbf 0.005 95 248 130 182 66 142 161 0.010 339 453 352 289 142 218 327 0.100 4783 7501 3659 5101 2008 3128 4385 1.000 67081 92359 44892 71807 26045 42062 59383 10.000 1112523 1147999 620974 1034705 469117 599994 810386 Response 110809 114574 61925 103152 46702 59819 80820

PPM F0 D0 Pd10 P0 AN FL PY 0.005 120 132 55 240 88 169 170 0.010 208 266 131 328 125 233 273 0.100 3031 2378 1735 3990 2945 3322 3449 1.000 43666 35468 26813 60509 53697 58959 60029 10.000 632109 509687 577015 918798 896715 1019618 1022000 Response 63014 50813 57392 91564 89309 101529 101776

PPM BaA Cd12 C0 Bbf BkF BeP BaP 0.005 22 - 38 - - - - 0.010 41 - 55 34 35 - - 0.100 523 137 693 162 158 171 80 1.000 18809 5038 35571 3528 11497 2639 3546 10.000 731295 310869 742218 304227 458433 154715 237595 Response 72585 30826 73832 30154 45499 15343 23557

PPM Pd12 Per ID DA BgP 0.005 - - - - - 0.010 - - - - - 0.100 39 48 39 24 48 1.000 748 1043 580 378 535 10.000 71659 100379 77869 60954 68879 Response 7102 9948 7715 6038 6824 *abbreviations defined in Table 3-2

3.1.5.3.2 Method Development for Alkylated PAHs

With the exception of two C1-naphthalene compounds (1-methylnaphthalene and

2-methylnaphthalene), standards were not available for alkylated target compounds. A proprietary sample of North Slope, Alaskan crude oil was used to aid in identification of alkylated compounds. The crude oil was diluted in methylene chloride and analyzed by

GC/MS under the same instrument parameters as the non-alkylated target compounds.

Replicates of this sample were analyzed in full-scan mode and m/z ratios were compared

with library databases to identify the groupings of alkylated compounds. Response factors for the alkylated PAHs could not be calculated due to the lack of reference standards; therefore, response factors from their respective parent compounds were used

(Douglas and Uhler, 1993; Stout et al., 2001a).

Once this was accomplished, elution times for all compounds (alkylated, non- alkylated, and surrogate) were compared and overlapping groupings were split into separate GC/MS programs to make identification and quantification of PAHs within samples easier.

3.1.5.3.3 Method Development for Samples

All samples were analyzed by GC/MS under the previously described instrument parameters. Each sample was analyzed using four separate programs utilizing the selected ion monitoring (SIM) mode. Tables 3-5, 3-6, 3-7, and 3-8 list each program and the mass-to-charge ratio(s) used to identify and quantify the compound(s). Analytes targeted within a certain time range were distinguished from each other by elution time within that time period and m/z.

Table 3-5: Program for DSIMHOT. This method targets the non-alkylated compounds and the surrogate standards. Analytes (*surrogate standards) Time (minutes) m/z Naphthalene, Naphthalene-d8* 19.00-23.99 128, 136 Biphenyl 24.00-26.49 154 Acenaphthylene 26.50-27.49 152 Acenaphthene, Acenaphthene-d10* 27.50-28.19 154, 162 Dibenzofuran 28.20-29.49 168 Fluorene 29.50-32.99 166 Dibenzothiophene 33.00-33.54 184 Anthracene, Phenanthrene, Phenanthrene-d10* 33.55-36.99 178, 188 Fluoranthene, Pyrene 37.00-42.99 202 Benz(a)anthracene, Chrysene, Chrysene-d12 43.00-47.99 228, 240 Benzo(a)fluoranthene, Benzo(a)fluoranthene, 48.00-55.99 252, 264 Benzo(a)fluoranthene, Benzo(a)pyrene, Benzo(e)pyrene, Perylene, Perylene-d12 Indeno(c,d-1,2,3)pyrene, Dibenzo(a,h)anthracene, 56.00-end of 276, 278 Benzo(g,h,i)pyrene, method

Table 3-6: Program for DSIMNP. This method targets the alkylated naphthalenes and phenanthrenes/ anthracenes. Analytes Times (minutes) m/z (*used for quantification) C1-naphthalenes 23.00-24.99 142*, 141 C2-naphthalenes 25.00-27.99 156*, 141 C3-naphthalenes 28.00-29.99 170*, 155 C4-naphthalenes 30.00-34.99 184*, 169 C1-phenanthrenes/anthracenes 35.00-36.99 192*, 191 C2-phenanthrenes/anthracenes 37.00-38.99 206*, 191 C3-phenanthrenes/anthracenes 39.00-40.74 220*, 205 C4-phenanthrenes/anthracenes 40.75-end of 234*, 219, 191 method

Table 3-7: Program for DSIMFFP. This method targets the alkylated fluorenes and fluoranthenes/pyrenes. Analytes Time (minutes) m/z (*used for quantification) C1-fluorenes 31.00-32.99 180*, 165 C2-fluorenes 33.00-35.49 194*, 179 C3-fluorenes 35.50-39.99 208*, 193 C1-fluoranthrenes/pyrenes 40.00-41.99 216*, 215 C2-fluoranthrenes/pyrenes 42.00-43.99 230*, 215 C3-fluoranthrenes/pyrenes 44.00-end of method 244*, 229, 215

Table 3-8: Program for DSIMDC. This method targets the alkylated dibenzothiophenes and chrysenes. Analytes Time (minutes) m/z (* used for quantification) C1-dibenzothiphenes 34.00-35.99 198*, 197 C2-dibenzothiphenes, 36.00-44.99 212* C3-dibenzothiphenes, 226* C4-dibenzothiphenes 240* C1-chrysenes 45.00-46.99 242*, 241 C2-chrysenes 47.00-48.99 256*, 241 C3-chrysenes, 49.00- end of method 270* C4-chrysenes 284*, 269

The mass of compound (mg) per kilogram of soil in each sample was calculated by dividing the area counts under each peak by the average response factor for that compound (or its parent), multiplying this number by the extract volume in liters, and then multiplying by the dry mass of sample in kilograms to achieve a final mass of compound per unit weight of dry soil. PAH analytes were corrected for surrogate recovery.

Standard quality control measures were taken to validate data integrity, as follows. Matrix blanks were run between each sample and batch standards were run to confirm elution time and response factors. Detection limits were compound specific, and were set at the lowest concentration of the calibration curve where each individual component was identified, with adequate signal to noise ratio for confidence. Due to the use of selected ion monitoring mode, peaks were not limited to a 3:1 signal to noise ratio for identification, but were identified based on a consistent elution time from sample to sample. All extracts were analyzed by GC/MS in triplicate. All data in subsequent tables and figures are represented as means.

3.2 Statistical Methods (Principal Component Analysis)

Principal component analysis (PCA) is a variable reduction procedure that reduces the dimensionality of a dataset. Essentially a dataset with a large number of correlated variables is compressed into a few “principal components” that are completely uncorrelated. These principal components account for a large amount of variance within the dataset and are used to assess relationships between multivariate samples

(Johnson et al., 2002).

Prior to PCA, all data was transformed by the sample normalization procedure described by Johnson et al. (2002) to minimize concentration effects between sites. Data were entered into a spreadsheet with analytes as the columns and sites and/or reference standards as rows. Each analyte was normalized to total site concentration by dividing the analyte concentration by the total concentration for that site (row) and multiplying the result by 100. PCA was performed twice with two datasets. The first dataset included only the 23 sites from Otter Creek with all 44 analytes as variables. The second data set included the 23 sites from Otter Creek and an additional 13 reference standards from

Burns et al. (1997). Only 36 analytes were published by Burns et al. (1997); therefore, the Otter Creek data was adjusted to 36 analytes as well. Biphenyl, dibenzofuran, C4- dibenzothiophenes, C2- and C-3 fluoranthenes/pyrenes, benzo(a)fluoranthene, benzo(e)pyrene, and perylene were omitted from the second PCA. PCA was conducted using the PRINCOMP procedure in SAS 9.1 (SAS Institute, Inc., Cary, NC).

Chapter 4

Results and Discussion

4.1 General Trends

A profile of total PAH (TPAH) concentrations over the length of Otter Creek is depicted in Figure 4-1. TPAH concentrations ranged from 10.9 mg kg-1 to

3015.0 mg kg-1. Individual PAH concentrations are presented in Appendix A. The lowest concentrations were found in the headwater segment (from RM 7.9 to RM 6.0), a relatively less urbanized area in Otter Creek. The average TPAH across these first two river miles was 27.5 mg kg-1, consistent with a 2007 study that found TPAH in this area to be 29 mg kg-1 (SulTRAC, 2007). While these concentrations are relatively low for

Otter Creek, they are over twice the average TPAH concentration found by Stout et al.

(2004) at sites impacted by only urban background (TPAH= 12.0 mg kg-1).

Sites at RM 5.6, 5.5, and 5.4 were located in a cemetery upstream of the first refinery that Otter Creek passes, and TPAH concentrations in these three sites were considerably higher than in the headwaters section, peaking at 297.9 mg kg-1. The first site sampled immediately downstream of the refinery contained 697.6 mg kg-1 TPAH.

Concentrations in this stretch were expected to be high when compared to the headwaters sites due to the proximity to a point source of petroleum input to the creek. TPAH

concentrations (284.0 mg kg-1) were also elevated in this stretch of Otter Creek during sampling in 2007 (SulTRAC, 2007).

3500

3000

2500

2000

1500

1000

500 PAH concentration (mg/kg) concentration PAH 0 7.9 7.8 7.1 6.0 5.6 5.5 5.4 5.0 4.7 4.6 4.5 4.2 4.1 3.7 3.6 3.5 3.1 2.3 2.1 0.5 0.3 0.2 0.0 River mile location (headwaters to mouth)

Figure 4-1: Profile of total PAH concentrations observed over the length of the creek from headwaters to mouth.

TPAH concentration dropped to 45.0 mg kg-1 at RM 4.7; however, sites in the same vicinity maintained relatively high er concentrations (404.6 mg kg-1 and 574.3 mg kg-1), remaining consistent with high values found immediately after the refinery. This anomaly in concentration may be due to variability in sampling. This sample was possibly taken from a sediment pocket with relatively little PAH contamination. This sample was close to the bank of the creek, and could possibly be bank sediment that had recently settled to the bottom of the creek. TPAH concentrations leveled between 94.4 mg kg-1 and 156.6 mg kg-1 over the residential portion of the creek (RM 4.2-RM 2.3).

These concentrations are approximately eight to thirteen times higher than single-source urban background sites (Stout et al., 2004). The residential portion is likely impacted by a mixing of both urban background and the petrogenic point source inputs (sources of

known spills) that bracket it both upstream in the refinery section and downstream in the heavily industrialized portion of Otter Creek.

Concentrations spiked at 3015.0 mg kg-1 over the heavily industrialized portion of the creek where a pipeline crosses (site of major spill in 1992), the second oil refinery, and the railroad coal loading docks. Near the mouth of the creek, concentrations dipped to 32.9 mg kg-1. Similar trends were documented in the 2007 sampling of Otter Creek:

TPAH levels ranged lower (37.0- 59.0 mg kg-1) over the residential portion of the creek, spiked in the heavily industrialized portion (108.0 to mg kg-1), and fell at the mouth (54.0 kg-1) (SulTRAC, 2007).

Concentrations found in this study were consistently higher than values found in the 2007 study, though most values were within the same magnitude. Some of this difference may be explained by the inconsistency of exact sampling sites and variability in sampling. Data compared between studies were taken in the same stretch of the creek, but not at the same locations. Within this study, one order of magnitude variability can be seen between sites that are 0.1 miles apart (i.e., RM 4.7 and 4.6); increasing the distance between site comparisons may compound this effect. Also, this study reports

TPAH as the sum of 44 analytes, while the 2007 study reports TPAH as the sum of 34 analytes, omitting the entire dibenzothiophene homologous series, the C1- through C-3 alkyl fluoranthenes/pyrenes, biphenyl, dibenzofuran, and benzo(a)fluoranthene.

Instrument sensitivity was greater in this study, as PAHs were detected in selected ion monitoring mode as opposed to full-scan mode.

In other waterways, concentrations of PAHs found in sediments have been correlated with percent fine particles (silt + clay) and percent organic matter (Boehm and

Farrington, 1984; Colombo et al., 1989). Organic carbon and finer-grained particles have an affinity for binding chemical contaminants and are likely to accumulate in fluvial areas of lower water energy, where sedimentation rates and biological activity are higher

(Uhler et al., 2000). Only twelve of the twenty-three samples were predominantly comprised of fine-grained particles (Table 4-1). Correlation coefficients (Spearman rank) were weak when comparing percent fine-grained particles to percent total organic carbon

(%TOC) and TPAH concentrations (rs= 0.10 and rs= -0.55, respectively). Correlation between %TOC and TPAH was also weak (rs= 0.34). This trend was consistent with results presented by Stout et al. (2003) in a similar study involving these variables and indicated that at this site, particle size is not a good indicator of %TOC or TPAH content.

Since abundant organic matter in sediments provides more adsorption sites for

PAHs, a stronger correlation between %TOC and TPAH was expected. As PAHs adsorb to organic matter, adsorption sites become “occupied” and PAHs are left to exist in interstitial areas between sediment particles, forming a “PAH soup” (Boehm et al., 2002).

Sites with greater %TOC will have more adsorption sites available for PAH molecules, and by normalizing PAH concentrations to %TOC, a determination can be made whether sites have overloaded the natural ability to sequester PAHs. Some sites with elevated

PAH loadings may be affected by factors other than %TOC that could be responsible for high contamination levels, such as point sources (Figure 4-2). Sites at RM 0.5, 0.3, and

0.2 are in close proximity to an oil pipeline, and the site at RM 5.0 is immediately downstream of an oil refinery.

Table 4-1: Summary of chemical and physical data for each sampling site in Otter Creek. TPAH Grain size distribution River Mile -1) % TOC (mg kg % >sand % sand % silt % clay 7.9 60.0 - 47.8 32.8 19.3 3.9 7.8 23.4 - 26.6 49.6 23.8 2.8 7.1 15.6 - 72.8 21.6 5.6 1.4 6.0 10.9 - 27.3 47.0 25.7 3.3 5.6 297.9 22.9 70.3 5.8 1.0 2.3 5.5 140.2 - 59.2 34.0 6.8 3.9 5.4 113.3 - 24.9 58.2 16.9 4.5 5.0 697.6 34.4 54.8 9.7 1.1 2.6 4.7 45.0 - 20.9 46.5 32.6 2.3 4.6 404.6 - 72.6 20.2 7.3 2.5 4.5 574.3 - 70.3 24.2 5.4 3.5 4.2 156.6 - 73.7 17.8 8.5 2.8 4.1 134.1 - 49.7 35.6 14.8 7.8 3.7 115.7 - 49.9 31.2 18.9 4.1 3.6 151.0 - 53.2 34.5 12.3 9.2 3.5 100.5 - 39.5 40.0 20.5 8.7 3.1 94.4 - 45.3 40.8 13.9 9.0 2.3 100.7 - 46.0 40.1 13.9 7.3 2.1 649.6 - 49.8 42.5 7.8 4.6 0.5 1970.9 - 71.6 25.8 2.6 6.3 0.3 3015.0 - 77.4 20.7 1.9 4.6 0.2 1636.3 - 75.9 20.7 3.4 7.1 0.0 32.9 42.4 57.4 0.2 - 1.7

Figure 4-2: Total PAH concentrations normalized to percent organic carbon content.

Like many other urban streams, Otter Creek does not have a pristine section to compare levels of contamination within the creek itself. The headwaters section of Otter

Creek is most representative of baseline values in Otter Creek today, as that section is the least urbanized, has the least amount of PAH contamination, and has no known petroleum inputs to the area. Evidence to support that the headwaters sediments are only impacted by urban background is in the following sections.

4.2 Petrogenic and Pyrogenic Distributions

PAH distributions are innovative interpretive tools in hydrocarbon fingerprinting

(Sauer and Uhler, 1994-1995). Relative abundances of non-alkylated PAHs to their alkylated counterparts can visually help identify sediment impacted by petrogenic or pyrogenic sources. Petrogenic distributions are characterized by a “bell-shaped” appearance, in which the C-1, C-2, and C-3 alkylated members of the homologous series are more abundant than the C-4 and non-alkylated parent member, while pyrogenic distributions have an “inverse slope” or “skewed” appearance, in which the parent compound is most abundant and abundance decreases with increasing alkylation in the homologous group (refer back to Figure 2-1) (Sauer and Uhler, 1994-1995; Stout et al.,

2001a; Wang and Brown, 2009). Select distribution patterns are presented here, and a full compilation of distribution patterns at each site is presented in Appendix B.

The dominant distribution pattern from the “cleanest” stretch of the creek, the headwaters section, is that of a pyrogenic character. One to three homolog groups at each site displayed pyrogenic patterns, and no petrogenic patterns were identified (Table 4-2).

Headwaters sites were dominated by fluoranthene, pyrene, and benzo(e)pyrene and

contained more 4- to 6-ring PAHs than 2- or 3-ring PAHs (Figure 4-3). Abundance of these compounds, coupled with a high percentage of high molecular weight compounds

(4- to 6- rings) is characteristic of a pyrogenic, urban background source, most likely runoff or atmospheric fallout of combustion products (Stout et al., 2001a; Menzie et al.,

2002; Stout et al., 2003).

Table 4-2: Distribution patterns for the five homolog groups: naphthalenes (N0-4), fluorenes (F0-3), dibenzothiophenes (D0-4), fluoranthenes (FL0-3), and chrysenes (C0-4). River Mile N0-4 F0-4 P0-4 D0-3 FL0-3 C0-4 T LPAH T HPAH 7.9 - - pyro - pyro - 7.8 52.2 7.8 - - pyro - pyro pyro 8.9 14.5 7.1 - - - - pyro - 7.4 8.2 6.0 - - - - pyro - 5.4 5.5 5.6 - - - - pyro pyro 58.2 239.8 5.5 petro - petro petro pyro pyro 92.9 47.3 5.4 petro - pyro - pyro pyro 48.7 64.7 5.0 petro - - - pyro - 255.9 441.8 4.7 - - - - pyro pyro 8.9 36.1 4.6 - - - - pyro pyro 62.1 342.5 4.5 - - petro petro petro - 346.8 227.5 4.2 - - - - pyro - 36.0 120.6 4.1 - - - - pyro - 19.9 114.2 3.7 ------45.4 70.3 3.6 - - - - pyro - 89.0 62.0 3.5 - pyro - - pyro - 60.7 39.8 3.1 - - - - pyro - 22.1 72.3 2.3 - - - - pyro - 23.8 76.9 2.1 - - petro petro petro petro 308.6 340.9 0.5 petro - petro petro petro - 925.7 1045.3 0.3 petro - petro petro petro - 1251.1 1763.9 0.2 petro - petro petro petro - 946.0 690.3 0.0 - petro petro - petro - 21.8 11.1 petro= petrogenic T LPAH= total 2-3 ring mg kg-1 pyro= pyrogenic T HPAH= total 4-6 ring mg kg-1

- denotes no identifiable pattern

Figure 4-3: Typical distribution patterns found in the headwaters section of Otter Creek. Pyrogenic homolog groups are labeled.

As proximity to the first refinery on Otter Creek increases (RM 5.6- 5.0), petrogenic signatures were identified in lower molecular weight homolog groups, especially the naphthalenes. Pyrogenic distributions were displayed in higher molecular weight compounds, suggesting a mixture of pyrogenic (urban background) and petrogenic (refinery) inputs. At RM 5.5, lower molecular weight compounds dominated, though pyrogenic distributions in the 4-ringed compounds were apparent (Figure 4-4).

The next three sites (RM 4.7, 4.6, and 4.5) are in proximity to the refinery and were expected to display some residual petrogenic signature from the refinery input; however, only RM 4.5 exhibited petrogenic signatures and an abundance of 2- and 3-ring compounds. As discussed earlier, the sample from RM 4.7 may not necessarily be representative of that section of the creek. The lack of a petrogenic signature at RM 4.6 could be due to an increase in water-washing, as lower molecular weight compounds tend to solubilize more readily than higher molecular weight compounds (Stout et al., 2002), or RM 4.5 may be a site of greater deposition than RM 4.6, as sedimentological trends

have been linked to PAH levels (Boehm and Farrington, 1984). However, an evaluation of the hydrodynamics of Otter Creek was beyond the scope of this study.

Figure 4-4: Sites typical of both petrogenic and pyrogenic inputs. Petrogenic and pyrogenic distributions are labeled.

Over the course of the residential section (RM 4.2- 2.3), no petrogenic signatures were expected or found, though the abundances of individual PAHs were more variable than in the headwaters section (Table 4-2). Most sites were dominated by 4- to 6- ring compounds, such as fluoranthene, pyrene, C-1 and C-2 fluoranthenes/pyrenes, benzo(e)pyrene, and benzo(b)fluoranthene (Appendix B). Again, the distributions at the residential sites suggest a pyrogenic source, though concentrations are an order of magnitude higher than reported values for sites impacted exclusively by urban background. Interestingly, sites at RM 3.6 and 3.5 do not follow this trend; they are dominated by 2- and 3-ring compounds, namely C-2 naphthalenes and acenaphthylene.

The dominance of “petrogenic” compounds and the lack of a petrogenic signature at

these sites is confounding and allocation of PAH sources might better be explained with diagnostic ratios and principal component analysis.

As expected, petrogenic signatures were prominent at sites within the heavily industrialized section of the creek, where many documented spills have occurred.

Petrogenic signatures were evident in three to four homolog groups at each site, though high percentages of 4- to 6-ring compounds at RM 2.1-0.3 are found, indicating the presence of some pyrogenic input (Figure 4-5). Lower molecular weight compounds were dominant at the furthest sites downstream, RM 0.2 and at the mouth.

Figure 4-5: Petrogenic signatures were abundant in the heavily-industrialized section of the creek.

4.3 Diagnostic Source Ratios

Visual inspection of PAH distributions can give a general overview of PAH contamination at a site, but ratios of specific PAH parent and alkylated compounds can further characterize and allocate sources. Some ratios are based on the assemblage of parent PAHs and their alkylated counterparts found in petrogenic and pyrogenic sources,

termed „source ratios‟ while others are based on knowledge of preferential weathering and thermodynamic stability between specific PAH isomers (Douglas et al., 1996;

Yunker et al., 2002; Costa and Sauer, 2005; Boehm, 2006). Usage of these ratios can be applied across many different datasets, while others are more site-specific or source- specific. The more commonly cited ratios were applied to the Otter Creek dataset and compared with literature values to further discern petrogenic and pyrogenic sources.

A simple allocation method to quantify petrogenic inputs was developed by

Boehm and Farrington (1984). The Fossil Fuel Pollution Index (FFPI) estimates the amount of fossil fuel (petrogenic) PAH to total PAH by assigning select compounds

(namely 2-and 3-ring compounds) into pyrogenic, petrogenic, or mixed categories based on their common occurrence.

FFPI is calculated as:

FFPI= Σ naphthalenes(C0-C4)+ Σ dibenzothiophenes(C0-C3)+ ½Σ phenanthrenes(C0-C1)+ Σ phenanthrenes(C2-C4)/ Σ PAH where the homologous groups of naphthalenes and dibenzothiophenes are considered purely petrogenic, parent phenanthrene and the C1-phenanthrenes are considered evenly split between petrogenic and pyrogenic origins, the C2-C4 phenanthrenes are considered petrogenic, and ΣPAH equals the total of all PAH analyzed (Boehm and Farrington,

1984). Crude oil has an FFPI value of approximately one, although environmental samples classified as having largely petrogenic input have been shown to have values around 0.5, while pyrogenic site values were closer to zero (Boehm and Farrington,

1984).

Overall, FFPI values agreed with PAH distribution histograms. All sites on Otter

Creek that exhibited petrogenic distributions in at least two of the five homolog groups

had relatively higher FFPI values, ranging from 0.33-0.53, consistent with previously reported petrogenic environmental samples (Table 4-3, column C) (Boehm and

Farrington, 1984). As expected, sites within the industrial portion of the creek (RM 2.1 to the mouth) and two sites near the first refinery fell within this category. Unexpectedly,

RM 3.6 and RM 3.5 exhibit high FFPI values, though no petrogenic signatures were apparent (Appendix B). Sites that exhibited at least one petrogenic distribution or had a predominance of low molecular weight compounds had intermediate FFPI values (0.22-

0.29), while sites that exhibited only pyrogenic signatures and had a dominance of high molecular weight compounds had relatively low FFPI values (0.06-0.18). Most sites in the residential section were classified as pyrogenic according to the FFPI. Interestingly,

FFPI values for two of the four headwaters sites, previously thought to be exclusively impacted by urban background, indicate mixed sources.

A modification of the FFPI assigns the entire suite of PAH compounds to either petrogenic, pyrogenic, mixed, or biogenic classifications (Stout et al., 2004). Briefly, all

2- and 3-ring compounds are considered petrogenic, with the exception of phenanthrene and C1-phenanthrenes, which are considered 50% petrogenic and 50% pyrogenic. Most

4- to 6-ring compounds are considered pyrogenic, except the C2- and C3-fluoranthenes and C2-, C3-, and C4 -chrysenes. Perylene is the only biogenic marker. Refer back to

Table 2-1 for a complete list of classifications. Generally, biogenic contributions were extremely small (<1.4%), except at RM 5.6, where perylene accounted for 7.3% of TPAH

(Table 4-3, column E). Formation of perylene is attributed to early diagenic processes in anoxic conditions (Boehm et al., 2002). The sample at RM 5.6 was taken from sediment with an overlying layer of abundant plant detritus, possibly explaining the elevated level

of perylene. Regardless, the abundance of perylene at this site was negligible for classification purposes.

Table 4-3: Source ratio data for the Otter Creek dataset. A B C D E F G River LPAH/ Mile TPAHd EPAHd FFPI HPAH %Petro/Pyro/Bio FL/PY P0/AN 7.9 60.0 42.5 0.06b 0.15b 12.7/86.2/1.1b 1.29b 9.46b 7.8 23.4 13.9 0.16b 0.61b 35.2/63.9/0.9b 1.36b 16.32a 7.1 16.0 7.1 0.26c 0.90b 45.0/55.0/ - b 1.06b - 6.0 10.9 4.6 0.24c 0.99b 47.9/51.1/1.0b 1.10b 12.74a 5.6 297.9 198.1 0.12b 0.24b 18.4/74.3/7.3b 1.11b 6.77b 5.5 140.2 43.1 0.53a 1.97a 65.1/34.3/0.6a 1.22b 8.23b 5.4 113.4 66.4 0.27c 0.75b 42.1/56.6/1.3b 1.21b 8.24b 5.0 697.6 406.9 0.22c 0.58b 36.0/63.2/0.8b 1.08b 2.68b 4.7 45.0 30.3 0.11b 0.25b 19.0/79.9/1.1b 1.24b 16.69a 4.6 404.6 291.8 0.10b 0.18b 16.2/82.7/1.1b 1.17b 17.52a 4.5 574.3 120.3 0.47a 1.52a 68.7/30.9/0.4a 0.83a 2.79b 4.2 156.6 94.0 0.16b 0.30b 28.3/71.1/0.6b 1.10b 12.63a 4.1 134.1 90.8 0.09b 0.17b 16.6/82.3/1.1b 1.20b 12.51a 3.7 115.7 33.2 0.29c 0.65b 58.1/41.6/0.3a 0.81a - 3.6 151.0 57.5 0.42a 1.43a 61.4/37.8/0.8a 1.09a 4.35b 3.5 100.5 44.6 0.38a 1.53a 60.8/38.4/0.8a 1.23b 8.46b 3.1 94.4 55.8 0.15b 0.31b 26.3/72.8/0.9b 1.17b 8.05b 2.3 100.7 53.3 0.18b 0.31b 34.6/64.9/0.5b 0.91a 11.59a 2.1 649.6 67.1 0.40a 0.91b 75.7/24.1/0.2a 0.48a 2.24b 0.5 1970.9 557.2 0.37a 0.89b 53.4/46.3/0.3a 0.29a 9.01b 0.3 3015.0 690.0 0.33a 0.71b 56.5/43.4/0.1a 0.23a 5.28b 0.2 1636.3 295.0 0.45a 1.37a 67.7/32.1/0.2a 0.25a 6.85b 0.0 32.9 6.5 0.47a 1.97a 67.9/32.1/ - a 0.44a 9.51b aindicates petrogenic classification bindicates pyrogenic classification cindicates petrogenic and pyrogenic mixture dmg kg-1 - analyte(s) below detection limit

Sites were designated as petrogenic or pyrogenic based on the dominant grouping for that site (Table 4-3, column E). Most sites were clearly petrogenic or pyrogenic,

meaning that the dominant grouping was >60%, such as RM0.2 (67.7% petrogenic) or

RM7.9 (86.2% pyrogenic). All four headwaters sites were classified as pyrogenic. Two sites (RM 7.1 and 6.0) were relatively close (50% ±10%) in percentages of petrogenic and pyrogenic compounds, implying that these two sites may have an input other than urban background. Two sites (RM 0.5 and 0.3) in the industrial portion of the creek were also close in petrogenic and pyrogenic grouping, suggesting that these sites are not single-source impacted sites.

An alternative allocation method compares the ratio of low molecular weight

PAH (LPAH, 2- and 3-ring) compounds to the total of high molecular weight (HPAH, 4- to 6-ring) compounds (Stout et al., 2001a; Barriera et al., 2007). Sites with values <1 were categorized as pyrogenic and >1 as petrogenic (Table 4-3, column D).

Classifications from these three methods generally were in agreement, although at RM

3.7, 2.1, 0.5, 0.3, and 0.2, classifications were conflicting, depending on method. While the LPAH/HPAH index can give a general feel for the source at a site, it is not discriminatory enough to truly classify each compound into its appropriate class, i.e., phenanthrene is not considered mixed source, some 4- to 6-ring alkylated compounds are actually petrogenic in nature. FFPI and its modification are compound specific, but the modification takes into account all compounds analyzed.

The modified FFPI approach is considered the best representation of source allocation for Otter Creek. According to this allocation, all headwaters sites are pyrogenic in nature, and all industrial sites are petrogenic in nature. RM 5.5 and 4.5 are considered petrogenic, as supported by the presence of petrogenic homolog distributions and proximity to a petrogenic point source. Most residential sites exhibit pyrogenic

sources, with the exception of RM 3.7- 3.5. These sites are indexed as petrogenic, with no apparent petrogenic input.

The 16 EPA Priority Pollutants (refer to Table 2-1 for designations) are all non- alkylated compounds. In pyrogenic distributions, non-alkylated compounds predominate over alkylated compounds; thus, high percentages of these Priority Pollutants could indicate pyrogenic sources. Stout et al. (2004) previously found that sites dominated by pyrogenic urban background sources exhibit two general trends: 1) 96% of sites contained less than 200 mg kg-1 of EPA Priority Pollutant PAH (EPAH) and b) 96% of those sites contained less than 300 mg kg-1 TPAH. Sediments that contain more than these amounts may be impacted by non-background, or point sources. Using these guidelines, sixteen of the twenty-three sites sampled in Otter Creek suggest an urban background signature (Table 4-3, column A and B). Sites at RM 5.0, 4.6, 0.5, 0.3, and

0.2 exceed both criteria, and sites at RM 4.5 and 2.1 exceed at least one of the criteria.

All the aforementioned sites are either located adjacent to or directly downstream of an oil refinery or pipeline. This suggests that Otter Creek sediments are impacted by both a pyrogenic, “background” non-point source, as well as petrogenic point source(s).

The ratio of some high molecular weight PAH isomers can also be used to help differentiate sources. The ratio of fluoranthene to pyrene (FL:PY) is influenced by the temperature of formation process. A FL:PY ratio less than one is typical for petrogenic sources (which are formed at low temperatures and high pressure), but some lower temperature refinery processes such as carbureted water gas processes will also exhibit this ratio. Ratios greater than one are related to high temperature processes, such as coal carbonization and combustion in automobile engines (McCarthy et al., 2000; Costa et al.,

2004; Neff et al., 2004). Fifteen of the twenty-three sites display FL:PY ratios >1 (Table

4-3, column F). RM 4.5, 3.7, and all sites from RM 2.3 to the mouth imply petrogenic sources. For sites furthest downstream (RM 0.5 to the mouth), these results are consistent with findings from the modified FFPI calculations and EPAH and TPAH comparisons. Though RM 2.1 and 2.3 have been grouped with pyrogenic sources using

EPAH and TPAH comparisons, they could be impacted from movement of petrogenic contamination from downstream sites upstream via seiche events (SulTRAC, 2007).

Another well-studied ratio involves the three-ringed compounds phenanthrene and anthracene. Anthracene is a thermodynamically less stable isomer of phenanthrene.

During the combustion process, anthracene is produced more favorably than phenanthrene, thus the phenanthrene: anthracene (P0:AN) ratio can serve as an indicator of PAH origin (Lima et al., 2005). P0:AN values >10 imply petrogenic sources, while values <10 can suggest pyrogenic sources (Budzinski et al., 1997). In this case, the

P0:AN ratio contradicts many classifications set forth by other ratios already calculated for this dataset (Table 4-3, column G). At RM 7.8, 4.7, 4.6, 4.2, and 4.1, the P0:AN ratio suggests a petrogenic source, while all other ratios and distributions suggest a pyrogenic source.

While the P0:AN ratio is well-documented, it has been ranked low in terms of the relative stability between isomers; thus, it may not be a reliable indicator (Yunker et al.,

2002). Budzinski et al. (1997) recommend that P0:AN is more robust when used in a double-plot ratio with FL:PY. Lima et al. (2005) have found that some diesel emissions display ratios above the threshold of 10, appearing as petrogenic sources; while some petrogenic sources of gasoline and diesel display P0:AN ratios <10, mistakenly

indicating a pyrogenic source. Reasonably, RM 7.8 could have a preponderance of diesel soot associated with its sediments from a surrounding major interstate, though this would be expected to be seen at more than one site. Though not exhibited in other diagnostic features, RM 4.7, 4.6, 4.2, and 4.1 likely have residual petrogenic compounds due to their location close downstream from a refinery. At RM 4.5, 3.6, 2.1, 0.2, and 0.0, the P0:AN ratio suggests a pyrogenic source, while all other diagnostic ratios and distributions suggest a petrogenic source. Urban runoff and atmospheric fallout will occur at all points along the creek; thus, pyrogenic features will be present at all locations along the creek.

Sites that are predominantly affected by petrogenic input, will also possess an underlying pyrogenic faction, which may indicated by this ratio.

Diagnostic ratios are often plotted against each other to aid in visually identifying source groupings. Averages of common literature values (Yunker et al., 2002) for select sources are plotted along with Otter Creek sample location values for comparison (Figure

4-6a and 4-6b). The FL/FL+PY (fluoranthene: fluoranthene + pyrene) ratio can distinguish petrogenic and pyrogenic sources from each other at a value of 0.40.

Pyrogenic sources can be split into two general groupings: FL/FL+PY ratios between

0.40 and 0.50 are indicative of petroleum combustion and values greater than 0.50 are classified as wood, coal, or grass combustion. The isomers indeno(c,d)pyrene and benzo(ghi)pyrene (ID/ID+BgP) can also be used to separate petrogenic compounds from the two different types of combustion classifications.

Figure 4-6a and 4-6b: Double-plot ratios of Otter Creek samples in relation to calculated averages of literature values for common PAH sourcesa aYunker et al., 2002

In Figure 4-6a, refinery, residential, and headwaters sites were classified from both ratios as pyrogenic and clustered around the dividing lines for vehicle combustion and other combustion, near source values of urban air, wood and coal combustion.

Industrial sites plotted in the petroleum section of the graph for FL/FL+PY. The

ID/ID+BgP ratio signaled a pyrogenic (vehicle/crude combustion) ratio for most

industrial sites. Industrial values were similar to average values for diesel, suggesting the type of petrogenic contamination found in this stretch of the creek. Interestingly, the diesel ratio for ID/ID+PY falls into a pyrogenic category, though diesel is a known petrogenic product. This trend is similar to diesel values that have been reported as pyrogenic for the P0:AN ratio (Lima et al., 2005). This may be a unique signal for identifying diesel contamination in Otter Creek.

Ratios involving benz(a)anthracene and chrysene (BaA/BaA+C0) are often used because the ratio will remain relatively constant over large concentration ranges and weathering conditions (Costa and Sauer, 2005). The BaA/BaA+C0 ratio shows similar trends to the ID/ID+PY ratio. Most Otter Creek sites were pyrogenic in nature, but a greater separation can be seen between the refinery sites and the residential and headwaters sites. Refinery sites plot closer to values for wood combustion and wood soot than urban air; however, the clustering of all the pyrogenic source values makes it difficult to allocate pyrogenic contamination to multiple sources. Industrial sites, like diesel and crude oil reference values, exhibit petrogenic features with the FL/FL+PY ratio and are assigned a pyrogenic classification from the BaA/BaA+C0 ratio. Clearly the reference oils are petrogenic; the similarities in ratios to the industrial sites suggest residual diesel or crude oil may be the dominant contamination source.

For this study, the FL/FL+PY ratio effectively separates petrogenic from pyrogenic sites, while ID/ID+BgP and C0/C0+BaA does not. Compounds at molecular weight 276 (benzo(ghi)perylene, indeno(1,2,3-cd)pyrene), 202 (pyrene, fluoranthene), and 252 (benzo(e)pyrene, benzo(a)pyrene), perylene, benzo(b)fluoranthene, benzo(j)fluoranthene, and benzo(k) fluoranthene) were ranked greatest in range of

isomeric stability and potential for source allocation, while compounds at molecular weights of 178 (phenanthrene, anthracene), 278 (dibenz(a,h)anthracene and others not used in this study), and 228 (chrysene and benz(a)anthracene) retained the least potential

(Yunker et al., 2002). Interestingly, the double-plots using the BaA/BaA+C0 (ranked worst) and the ID/ID+BgP (ranked best) ratios yielded similar clusters within the pyrogenic classifications, though they did not discern petrogenic from pyrogenic sources.

The contradictory results using different ratios support using a large suite of ratio data when interpreting PAH source. No specific set of ratios has been found to be consistently reliable, nor will every ratio be applicable in all situations. Ratios are analyzed on a case- by-case basis with interpretations being supported by the majority of the data.

4.4 Principal Component Analysis (PCA)

Principal component analysis (PCA) has been demonstrated as a useful tool in environmental forensic investigations (Burns et al., 1997; Stout et al., 2001a and 2001b;

Johnson et al., 2002; Barreira et al., 2007; Mudge, 2007). This variable reduction procedure reduces the size of a large dataset into a few “principal” components that can explain a large amount of variance within the dataset. The first component explains the largest amount of variance, and each successive component accounts for the next largest portion of variance (Johnson et al., 2002). The individual analytes responsible for differentiation between sources can be determined based on their loadings on each component. Each sample is assigned a component score and graphed. Samples with similar chemical compositions will cluster together or near reference standards (Burns et al., 1997; Stout et al., 2001a; Mudge, 2007).

PCA was run in two subsequent analyses. The first analysis included sample sites only from Otter Creek and incorporated all 44 PAHs analyzed (Figure 4-7). The first

(PC-1) and second components (PC-2) were responsible for 46% and 21% of the variance, respectively. A third component accounted for another 14%. Sites at RM 5.5,

4.5 3.7, 0.5, 0.3, 0.2, and 0.0 (petrogenic) clustered together positively on PC-1 and negatively on PC-2. PAH histogram distributions, FFPI values, and FL/PY ratios have indicated that these sites, with the exception of RM 3.7, are of petrogenic origin. RM 3.7 has shown both petrogenic and pyrogenic features from other diagnostic data (Table 4-3).

Figure 4-7: Principal component scores for Otter Creek sample sites.

A second cluster (pyrogenic) was evident and included sites at RM 7.9, 7.8, 7.1,

6.0, 5.6, 5.4, 4.7, 4.6, 4.2, 4.1, and 2.3. These sites have all been previously classified as pyrogenic sites from the majority of data, with the exception of RM 7.1, 6.0, 5.6, and 5.4.

These sites have shown both petrogenic and pyrogenic features, and suggest a mixing of both petrogenic and pyrogenic inputs. Sites at RM 3.6, 3.5, and 3.1 show a relationship

with each other, but did not appear to be associated with either the petrogenic or pyrogenic clusters. In other diagnostic examinations, RM 3.6 and 3.5 have shown mixed features, while RM 3.1 has exhibited purely pyrogenic features.

A plot of the component loadings for each individual PAH analyte can reveal information about the chemical causes underlying the groupings (Stout et al., 2001a).

Component loadings mark which PAHs are most responsible for “pulling” the data towards one group or the other (Figure 4-8). Two- and three-ringed alkylated PAHs loaded positively on PC-1 and negatively on PC-2. The greater abundance of these compounds in petrogenic sources indicates that sites that have overall scores that load on the principal components in this manner would be from petrogenic sources. Likewise, 4- to 6-ring compounds are responsible for the clustering of most other sites into the pyrogenic category. The positions of RM 3.5 and 3.6 seem to be heavily influenced by acenaphthylene, which could be a unique indicator for a source at these two sites.

Figure 4-8: Component loadings for Otter Creek sites.

Suspected source materials could not be obtained for this study. Similar to the study conducted by Iqbal et al. (2008), reference data from Burns et al. (1997) was included in the second iteration of the PCA (Figure 4-9). The first and second principal components were responsible for 32 and 16 percent of the variance in the data, respectively, compared to 46 and 21 percent from the first iteration. A third component accounted for an additional 9 percent. The decrease in explained variance could be caused by systematic error between the two datasets or a decrease in key analytes from those used in the first iteration. Recognizing that reference samples from Burns et al.

(1997) are not sources for Otter Creek sites, the datasets were combined to see how closely Otter Creek samples plotted with typical source samples.

Figure 4-9: Component scores for Otter Creek sites and reference sources. Sources are reference sources from Burns et al. (1997), while unknown, petrogenic, and pyrogenic points are source designations for Otter Creek sites from the first PCA (Figure 4-7).

Component scores plotted in Figure 4-9 reveal that most samples taken from Otter

Creek are not solely from one source. No sample plotted close to reference samples for fresh or lightly weathered crude or diesel oil. All samples that displayed solely

petrogenic distribution signatures (RM 4.5, 2.1-0.0) exhibited similarity to moderately to heavily weathered crude oil. RM 5.5 was classified as pyrogenic in the first PCA, but plotted among the petrogenic sources. Pyrogenic reference samples plotted in proximity to each other. Few Otter Creek samples (only RM 7.9, 5.4, and 3.1) plotted close enough to the pyrogenic reference samples to discern pyrogenic impacts as their sole source.

Most sites in Otter Creek fall within the area between the heavily weathered petrogenic and pyrogenic groupings, including those that exhibited mixed petrogenic and pyrogenic diagnostic features. Within this “mixed” zone, RM 7.1, 6.0, 4.2, 3.7, and 2.3 plotted closer to the petrogenic cluster and RM 7.8, 5.6, 5.0, 4.7, 4.6, and 4.1 plotted closer to the pyrogenic cluster. Sites that were classified as pyrogenic by PAH distributions, FFPI, and/or diagnostic ratios should plot more closely to the pyrogenic reference sources, but no trend was evident.

Two sites, RM 3.5 and 3.6, fall between fresh petrogenic inputs and pyrogenic clustering. These two sites are situated in the residential portion of the creek, where no suspected point source occurs for petrogenic input. The component loading plot shows 2- and 3-ringed non-alkylated PAHs are the most influential on these two sites

(Figure 4-10). These sites could be impacted by some anthropogenic input previously not documented, or a natural oil seep may be responsible for the underlying petrogenic inferences at these sites, as Otter Creek was known to be associated with natural oil.

Further sampling and/or vertical sediment profiles at these sites would need to be established to further support either claim.

Figure 4-10: Component loadings for Otter Creek sites and reference sources.

More variation within the data was explained in the first iteration and more closely defined clustering of petrogenic and pyrogenic sites was displayed in the iteration using only the Otter Creek sites (Figures 4-7 and 4-8). However, the iteration involving a comparison dataset served as reference for inferring similarity between Otter Creek sites and representative sources. Ideally, it is best to have data of suspected sources within the study site for comparison.

Chapter 5

Conclusions

For the first time, PAH source allocation was successfully accomplished in Otter

Creek. Analysis of 23 sediment samples from Otter Creek by a modified version of EPA

Method 8270 and examination of histogram distributions, diagnostic ratios, and principal component analyses revealed that PAH contamination in the creek comes from both petrogenic and pyrogenic sources (Table 5.1). PAH data from Otter Creek can be divided into four groups: 1) headwaters section; 2) refinery section; 3) residential section; and 4) industrial section.

1) Sites within the headwaters section were predominantly impacted by an urban

background signature, as evidenced by predominance of 4- and 6-ringed

compounds, PAH distribution histograms, and PCA analysis. The headwaters

were considered a baseline value as relatively “clean” in comparison to other

stretches of the creek.

Final summaryfor source allocation Otter in Creek.

1: - able 5 T

2) As expected, some refinery sites had higher TPAH than background sites, and

show mixtures of urban background and petrogenic input. Sampling variability

and/or hydrodynamics may be responsible for the lack of mixing at RM 4.7 and

4.6, as they are bracketed by sites that are affected by petrogenic inputs, but only

display petrogenic features in the P0:AN ratio. This ratio was determined to be

not applicable to this study.

3) The downstream residential sites showed reduced TPAH compared with the

refinery sites, and exhibited urban background signatures, except at RM 3.7, 3.6,

and 3.5. Here, petrogenic features were displayed in an area where no known

point source has ever been documented. The abundance of 2- and 3-ring PAHs

at these sites suggest a petrogenic input, but it is undetermined whether the

source may be a natural oil seep or input from some anthropogenic means.

4) Industrial sites showed the highest levels of TPAH and strong evidence of

petrogenic inputs. TOC values from these sites suggest an overloading of PAHs

in these sediments from a point source. While no reference sources specific to

Otter Creek could be obtained for this study, plots of these sites along with

literature values in double-plot ratios and principal component analysis imply,

but are do not confirm, crude or diesel inputs.

Determinations for probable source were based mainly on the PAH distribution histograms, the strongest diagnostic ratios (% Petro/Pyro/Bio and FL/PY), and principal component analysis with the Otter Creek dataset (PCA-1). The %Petro/Pyro/Bio

allocated each specific compound of the study into a definitive classification, whereas

FFPI and LPAH/HPAH were more ambiguous. When compared to the results of other ratios, P0:AN often gave contradictory results, and may not be an appropriate indicator for use in this study. To avoid the probability of systematic error when combining two datasets, results from PCA-2 [Otter Creek and Burns et al. (1997)] were not taken into consideration for source allocation. PCA with the combined datasets did support a third grouping of sites that appear to have a source different from sites exhibiting pyrogenic background or petrogenic industrial inputs.

Stout et al. (2002) cautions, “…no single PAH source indicator is universally applicable to all situations. In fact, any forensic investigation requires that the forensic investigator use groups of indicator compounds and ratios systematically, and in concert with other pyrogenic features and historical facts, before reaching a defensible source identification conclusion.” No specific set of diagnostic ratios will yield the same results in every forensic investigation, nor will one specific ratio illustrate a complex system. A preponderance of evidence must be used in source allocation. This study supports the analysis of both non-alkylated and alkylated PAHs to characterize PAH sources and the use of multiple diagnostic ratios to reach those source conclusions.

In 2000, stakeholders in northwest Ohio formed the Duck and Otter Creek

Partnership (DOCP), dedicated to promoting human and ecological health through education, protection, and restoration (DOCP, 2010). Initiatives of the DOCP include the identification of sites along Otter Creek with the potential for wetland restoration, and a large-scale human-health and ecological risk assessment. Proposed remediation within the creek has included dredging, capping, and wetland enhancement. Identification of

PAHs and characterization of their sources within Otter Creek gives insight into areas of high contamination and provides a means for understanding where point source contributions are high and which sediments are impacted heavily by urban background.

Petrogenic and pyrogenic PAHs react differently in the environment. Two- and three-ring compounds are more bioavailable because they are more soluble in the water column, while four through six-ring compounds are more persistent and pose greater toxic effects. Future remedial activities in the creek need to consider total PAH concentrations at Otter Creek sites and the respective inputs for those sites. Treatment strategies should balance disturbance that will release soluble, petrogenic PAHs to the water column with effective remediation of persistent, pyrogenic PAHs and should compensate for the continuing input of pyrogenic, urban background contamination.

References

Aboul- Kassim, T.A. and Simoneit, B.R.T. 1995. Aliphatic and aromatic hydrocarbons in particulate fallout of Alexandria, Egypt: sources and implications. Environmental Science and Technology. 29: 2473-2483.

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Appendix A

Individual PAH Concentrations

The following pages detail the individual PAH concentrations found at each Otter

Creek site. Abbreviations are listed in Table 3-2.

Appendix B

PAH Distribution Histograms

The following pages detail the PAH distribution histograms for each Otter Creek site. Abbreviations are listed in Table 3-2.

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

3 FP

2 FP

1 FP

D2 D1

Compounds D0

RM 7.9 RM

DbF

Ace

Acl

Bph

N1 N0

9 8 7 6 5 4 3 2 1 0

10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 7.8 RM

DbF

Ace

Acl

Bph

N1 N0 4 3 2 1 0

4.5 3.5 2.5 1.5 0.5 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 7.1 RM

DbF

Ace

Acl

Bph

N1 N0 2 1 0

2.5 1.5 0.5 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 6.0 RM

DbF

Ace

Acl

Bph

N1 N0 2 1 0

2.5 1.5 0.5 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 5.6 RM

DbF

Ace

Acl

Bph

N1 N0

60 50 40 30 20 10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 5.5 RM

DbF

Ace

Acl

Bph

N1 N0

8 6 4 2 0

14 12 10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 5.4 RM

DbF

Ace

Acl

Bph

N1 N0

8 6 4 2 0

16 14 12 10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 5.0 RM

DbF

Ace

Acl

Bph

N1 N0 0

80 60 40 20

120 100 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds

RM 4.7 RM

DbF

Ace

Acl

Bph

N1 N0

8 7 6 5 4 3 2 1 0 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 4.6 RM

DbF

Ace

Acl

Bph

N1 N0 0

80 60 40 20

120 100 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 4.5 RM

DbF

Ace

Acl

Bph

N1 N0

60 50 40 30 20 10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 4.2 RM

DbF

Ace

Acl

Bph

N1 N0

5 0

40 35 30 25 20 15 10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds

RM 4.1 RM

DbF

Ace

Acl

Bph

N1 N0

5 0

25 20 15 10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 3.7 RM

DbF

Ace

Acl

Bph

N1 N0

8 6 4 2 0

16 14 12 10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 3.6 RM

DbF

Ace

Acl

Bph

N1 N0

5 0

45 40 35 30 25 20 15 10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 3.5 RM

DbF

Ace

Acl

Bph

N1 N0

5 0

30 25 20 15 10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 3.1 RM

DbF

Ace

Acl

Bph

N1 N0

8 6 4 2 0

14 12 10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 2.3 RM

DbF

Ace

Acl

Bph

N1 N0

5 0

25 20 15 10 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 2.1 RM

DbF

Ace

Acl

Bph

N1 N0 0

80 60 40 20

120 100 Concentrations (mg/kg) Concentrations

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 0.5 RM

DbF

Ace

Acl

Bph

N1 N0 0

80 60 40 20

180 160 140 120 100 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 0.3 RM

DbF

Ace

Acl

Bph

N1 N0 0

500 450 400 350 300 250 200 150 100 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 0.2 RM

DbF

Ace

Acl

Bph

N1 N0 0

80 60 40 20

160 140 120 100 Concentration (mg/kg) Concentration

BgP

Per

BaP

BeP

BkF

BbF

BaF

BaA

FP3

FP2

FP1

D2 D1

Compounds D0

RM 0 RM

DbF

Ace

Acl

Bph

N1 N0 3 2 1 0

2.5 1.5 0.5 Concentration (mg/kg) Concentration