WI0400246_INJECTION EVENT RECORD_20121211

RECEIVEDIDENRIDWQ
INJECTION EVENT RECORD DEC 1 1 2012
North Carolina Department of Environment and Natural Resources—Division AV§rggerftion Section
Permit Number WI0400246
Were any wells abandoned during this injection
1. Permit Information event?
YRC Freight ❑ Yes ❑■ No
Permittee If yes,please provide the following information:
Kernersville Terminal Number of Monitoring Wells
Facility Name
1255 NC Highway 66 South, Kernersville, NC Number of Injection Wells
Facility Address Please include two copies of the GW-30 for each
well abandoned.
2. Injection Contractor Information
4. Injectant Information
ARCADIS G&M of North Carolina, Inc.
Injection Contractor/Company Name Sodium Persulfate/Ferrous Sulfate/Citric Acid
Injectant Type
Street Address 801 Corporate Center Drive
Concentration 50 g/L/2.5 g/L/0.5 g/L
Raleigh NC 27607
City State Zip Code If the injectant is diluted please indicate the source
( 919 ) 854-1282 dilution fluid.
Area code—Phone number Total Volume Injected 12,000 gallons
3. Well Information Volume Injected per well Average of 4,000 gallons
Number of wells used for injection 3 5. Injection History
wo11 namnc IW-1 , IW-2, IW-3 Injection tlate(c) 11/11/2012-11/15/2012
Were any new wells installed during this injection Injection number(e.g. 3 of 5) 1 of 2
event?
❑ Yes ❑■ No Is this the last injection at this site?
❑ Yes UI No
If yes,please provide the following information:
Number of Monitoring Wells I DO HEREBY CERTIFY THAT ALL THE
INFORMATION ON THIS FORM IS CORRECT TO
Number of Injection Wells THE BEST OF MY KNOWLEDGE AND THAT THE
INJECTION WAS PERFORMED WITHIN THE
Type of Well Installed (Check applicable type): STA S LAID OUT I THE PERMIT.
❑ Bored ❑ Drilled ❑Direct-Push / 5 !7
❑ Hand-Augured ❑ Other(specify) S 12 OF INJECTION CONTRACTOR D TE/
Please include two copies of form GW-lb for each an Gerber
well installed. PRINT NAME OF PERSON PERFORMING THE INJECTION
Submit the original of this form to the Division of Water Quality within 30 days of injection. Form UIC-IER
Attn:UIC Program, 1636 Mail Service Center,Raleigh,NC 27699-1636,Phone No.919-733-3221 Rev.07/09

North Carolina Secretary of; s Page 1 of 1
North Carolina:
DEP�" ' Elaine Marshall
- - R E -TO Tti
Secretary SECRETARY OF STATE
PO Svcs 24672 Raleigh NC 27626-062? '919 €7-200O
Date:4/18/2012
Click here to:
View Document Filings I File an Annual Report I
Print a pre-populated Annual Report Form I Amended A Previous Annual Report I
Corporation Names
Name Name Type
� C YRC ENTERPRISE LEGAL
SERVICES, INC.
Business Corporation Information
SOSID: 1132080
Status: Current-Active
Effective Date: 1/19/2010
Annual Report Due Date:
Citizenship: FOREIGN
State of Inc.: DE
Duration: PERPETUAL
Annual Report Status: CURRENT
Registered Agent
Agent Name: CT CORPORATION SYSTEM
Office Address: 150 FAYETTEVILLE ST., BOX 1011
RALEIGH NC 27601
Mailing Address: 150 FAYETTEVILLE ST., BOX 1011
RALEIGH NC 27601
Principal Office
Office Address: 10990 ROE AVENUE
OVERLAND PARK KS 66211
Mailing Address: 10990 ROE AVENUE
OVERLAND PARK KS 66211
Officers
This website is provided to the public as a part of the Secretary of State Knowledge Base(SOSKB)system.Version:
2594
http://www.secretary.state.nc.us/corporations/Corp.aspx?PitemId=9422172 4/18/2012

. t
01 ARC D IS RE WED/DENR/DWQ ARCADIS G&M of North Carolina,
Infrastructure Water•Environment•Buildings Inc.
APR0 9 2012 801 Corporate Center Drive
Suite 300
Aquifer protection Section Raleigh
North Carolina 27607
UIC PROGRAM Tel [phone]
Aquifer Protection Section Fax [phone]
North Carolina DENR-DWQ www.arcadis-us.com
1636 Mail Service Center
Raleigh, NC 27699-1636
ENVIRONMENT
Subject:
Application for Permit to Construct Wells for Injection—Roadway Site—1255
Highway 66 South, Kernersville, North Carolina
Date:
April 4, 2012
Dear Mr. Slusser:
Contact
Please see the enclosed Application for Permit to Construct Wells for'Injection for Ryan Gerber
the Roadway Site in Kernersville, North Carolina. The proposed injection wells are
for treatment of soil and groundwater impacted with tetrachloroethene (PCE). Also Phone:
enclosed with this application is a risk assessment evaluation for approval of citric 919.415.2265
acid for injection use in North Carolina. Please let me know if you have any Email:
questions or if you need any additional information. [e-mail]
Sincerely, Our ref:
CL000294.0019.00001
ARCADIS G.&I of North Carolina, Inc.
,--"" .,...(------1-----:' zy//,
Al°
erber, PE
-roject Engineer
Copies:
Mr. Qu Qi—NCDENR- Hazardous Waste Section
Imagine the result
g:lenvlroadwaylkemersville1permits1ulc permit appllcationluic cover letter.doc

State of North Carolina RECENEDIDENRIDWQ
Department of Environment and Natural Resources APR 0 9 2012
Division of Water Quality
APPLICATION FOR PERMIT TO CONSTRUCT AND/OR USE A WELL(SS))�FOR r Protection INJECTION
Type 5I Wells—In Situ Groundwater Remediation/Type 5T Wells—Tracer Injection
• Do not use this form for remediation systems that extract contaminated groundwater,treat it,and reinject the treated groundwater.
• Submit TWO copies of the completed application and all attachments to the address on the last page of this form.
• Any changes made to this form will result in the application package being returned.
Application Number(to be completed by DWQ): 0 DOA£
GENERAL INFORMATION:
1. Applicant's Name(generally the responsible party): YRC Enterprise Services.Inc.
2. Signing Official's Name*: Steve Shinners Title: Senior Manarer
* Signing Official must be in accordance with instructions in part VI on page 7.
3. Mailing address of applicant: 10990 Roe Ave..Mail Stop A605 KCG
City: Overland Park State: KS Zip: 66211
Telephone number: 913-344-3615 Fax number: 913-234-8940
4. Property Owner's Name(if different from Applicant):
5. Property Owner's mailing address:
City: State: Zip:
6. Name and address of contact person who can answer questions about the proposed injection project:
Name: R.an Gerber _ Title: Professional Engineer
Company: ARCADIS G&M of North Carolina.Inc.
Address:. 801 Corporate Center Dr.,Suite 300
City:_ Raleigh State: NC Zip: 27607
Telephone number: 919-415-2265 Fax number: 919-854-5448
Email Address: rvan.eerberarcadis-us.com
II. PERMIT INFORMATION:
1. Project is: (X)New (Modification of existing permit (Renewal of existing permit without modification
( Renewal of existing permit with modification
2. If this application is being submitted for renewal or modification to an existing permit,provide:
existing permit number and the issuance date _
For renewal without modifications, fill out sections I & II only, sign the certification on the last page of this
form,and obtain the property owner's signature to indicate consent(if the applicant is not the owner).
For all renewals,submit a status report including monitoring results of all injection activities to date.
Revised 6/09 UIC-5I/5T Page 1 of 7

APPLICATION FOR PERMIT TO CONSTRUCT AND/OR USE A WELL(S)FOR INJECTION
Type 5I Wells—In Situ Groundwater Remediation/Type 5T Wells—Tracer Injection
III. INCIDENT& FACILITY DATA
A. FACILITY INFORMATION
1. Facility name: Roadwa\ -Kernersville
2. Complete physical address of the facility: 1522 Highway 66 S
City: Kernersville County: Forsyth State: NC Zip:
B. INCIDENT DESCRIPTION
1. Describe the source of the contamination: A leaking used oil UST was removed from the site in 1992.
Leaked oil from the UST was contaminated with chlorinated ethenes from an undocumented source. Chlorinated
ethenes remain in soil and groundwater in the vicinity of the former UST.
2. List all contaminants present in soils or groundwater at the site(contaminants may be listed in groups,e.g.,
gasoline, diesel,jet fuel,fuel oil,chlorinated ethenes,chlorinated ethenes,metals,pesticides/herbicides,etc):
Chlorinated ethenes and low levels of naphthalene and 2-methylnaphthalene
3. Has LNAPL or DNAPL ever been observed at the site(even if outside the injection zone)?
(Yes If yes,list maximum measured separate phase thickness_ feet
(X No If no, list maximum concentration of total VOCs observed at site:Currently 3.400 ug/L in groundwater
4. Agency managing the contamination incident:
(UST Section (Superfund Section(including REC Program and DSCA sites)
(DWQ Aquifer Protection Section (Solid Waste Section
(X)Hazardous Waste Section (Other: _
5. Incident managers name: Ou Qi and phone number:919-707-8213
6. Incident number or other site number assigned by the agency managing the contamination incident:
NCD 046 362 117
C. PERMITS
List all permits or construction approvals that have been issued for the facility or incident, including those not
directly related to the proposed injection operation:
1. Hazardous Waste Management program permits under RCRA: NCD 046 362 117 i Interim Status
2. DWQ Non-Discharge or NPDES permits: None
3. County or DEH subsurface wastewater disposal permits:_ None
4. Other environmental permits required by state or federal law: None _
Revised 6/09 UIC-5I/5T Page 2 of 7

APPLICATION FOR PERMIT TO CONSTRUCT AND/OR USE A WELL(S)FOR INJECTION
Type 5I Wells—In Situ Groundwater Remediation/Type 5T Wells—Tracer Injection
IV. INJECTION DATA
A. INJECTION FLUID DATA
1. List all proposed injectants.
NOTE: Any substance to be injected as a tracer or to promote in situ remediation must be reviewed by the
Occupational and Environmental Epidemiology Section (OEES) of the Division of Public Health, Department of
Health and Human Services. Review the list of approved injectants or contact the UIC Program to determine if
the injectants you are proposing have been reviewed by OEES.
Injectant: Sodium Persulfate
Concentration at point of injection: 50 g/L
Percent if in a mixture with other injectants:
Injectant: Ferrous Sulfate
Concentration at point of injection: 0.5 VI,
Percent if in a mixture with other injectants:
Injectant:_ Citric Acid
Concentration at point of injection: 0.5 g'L.
Percent if in a mixture with other injectants:
Injectant:
Concentration at point of injection:
Percent if in a mixture with other injectants:
2. Source of fluids used to dilute or chase the injectants listed above:
(X)Municipal water supply
Revised 6/09 UIC-5I/5T Page 3 of 7

APPLICATION FOR PERMIT TO CONSTRUCT AND/OR USE A WELL(S)FOR INJECTION
Type 5I Wells—In Situ Groundwater Remediation/Type 5T Wells—Tracer Injection
B. PROPOSED OPERATING PARAMETERS
1. Duration of Injection: Up to 10 days
a. Maximum number of separate injection events: 6
b. Expected duration of each injection event: Up to 10 days
c. Expected duration between events(if more than one event): 4 to 6 months
2. Injection rate per well: 0.5 to 2 gallons per minute(gpm)
3. Total Injection volume: — 1800 gallons per day(gpd); 9000 gallons per event(if separate events)
4. Injection pressure:<5 psi pounds/square inch(psi)
5. Temperature at point of injection; Ambient r 65-75 °F
6. Briefly describe how the above parameters will be measured and controlled: In-line flow totalizers will
record total infection volumes,pressure gauges at each well head will measure injection pressure.thermometers
will be inserted into batch tank prior to injection to confirm temperatures.
7. Estimated hydraulic capacity of the well: 2 gpm each well. Total 6 gpm for three wells
C. INJECTION WELL CONSTRUCTION DATA
1. Injection will be via:
( )Existing well(s)proposed for use as an injection well.Provide the data in(2)through(6)below to the best of
your knowledge.
(X)Proposed well(s)to be constructed for use as an injection well. Provide the data in(2)through(6)below as
proposed construction specifications.
2. Well Drilling Contractor's Name: Parratt Wolff
NC Well Contractor Certification number: 3367-A
3. Date to be constructed: TBD Number of borings: _3 _
Approximate depth of each boring(feet): 29
4. Screened interval/Injection interval of injection wells:
Depth: 14 to 29 feet below ground surface(if multiple intervals,indicate shallowest and deepest depth).
5. Well casing(N/A if injection is through direct push rods):
Type: (X) Stainless steel ( Other: _
Casing depth: 0 to 14 ft.
6. Grout(N/A if injection is through direct push rods):
Type: Cement ( Other:_
Grout depth: 0 to 12 ft.
Revised 6/09 UIC-5I/5T Page 4 of 7

APPLICATION FOR PERMIT TO CONSTRUCT AND/OR USE A WELL(S)FOR INJECTION
Type 5I Wells—In Situ Groundwater Remediation/Type 5T Wells—Tracer Injection
V. ATTACHMENTS
Provide the following items as separate attachments with the given headings:
A. SITE HISTORY
Provide a brief description of the site history including:
(1) site usage historically and present,
(2) origin of the contamination,
(3) previous remedial action(s).
NOTE: G.S. 89E-18 requires that any geologic plans, reports, or documents in which the performance is related to the
public welfare or safeguarding of the environment be prepared by a licensed geologist or subordinate under their
direction. G.S. 89E-13 requires that all drawings, reports, or documents involving geologic work prepared or approved
by a licensed geologist, or a subordinate under their direction, be signed and sealed by the licensed geologist.
B. HYDROGEOLOGIC DESCRIPTION
Provide a hydrogeologic description, soils description,and cross section of the subsurface to a depth that includes the
known or projected depth of contamination. The hydrogeologic description shall include:
(1) the regional geologic setting;
(2) significant changes in lithology;
(3) the hydraulic conductivity,transmissivity,and specific yield of the aquifer to be used for injection,including a
description of the test(s)used to determine these parameters;and
(4) the depth to the mean seasonal high water table.
C. INJECTION FLUID COMPOSITION
Describe the chemical,physical,biological and radiological characteristics of each injectant. Attach the Material Safety
Data Sheet(MSDS)for each injectant. If a private well or a well within'A mile of the injection site is used as the source
well,include chemical analysis of source fluid here.
D. INJECTION RATIONALE
Attach a brief description of the rationale for selecting the injectants and concentrations proposed for injection,including:
(1) goals of the injection project;
(2) explanation and/or calculations of how the proposed injectant volume and concentration were determined;
(3) a description of the reactions between the injectants and the contaminants present including specific breakdown
products or intermediate compounds that may be formed by the injection;and
(4) summary results of modeling or testing performed to investigate the injectant's potential or susceptibility to change
(biological,chemical or physical)in the subsurface.
E. INJECTION PROCEDURE AND EQUIPMENT
Provide a detailed description of all planned activities related to the proposed injection including but not limited to:
(1) construction plans and materials;
(2) operation procedures;
(3) a detailed diagram of the surface and subsurface portions of the system;and
(4) a planned injection schedule.
Revised 6/09 UIC-5I/5T Page 5 of 7

APPLICATION FOR PERMIT TO CONSTRUCT AND/OR USE A WELL(S)FOR INJECTION
Type 5I Wells—In Situ Groundwater Remediation/Type 5T Wells—Tracer Injection
F. MONITORING PLAN
Provide a plan for monitoring the results of the injection,including:
(1) a list of existing and proposed monitoring wells to be used;
(2) a list of monitoring parameters and analytical methods to be used; and
(3) a schedule for sampling to monitor the proposed injection.
NOTE: The selected monitoring wells must be located so as to detect any movement of injection fluids, process by-
products, or formation fluids outside the injection area or zone. The monitoring parameters should include the target
contaminants as well as secondary or intermediate contaminants which may result from the injection and other
parameters which may serve to indicate the progress of the intended reactions, such as pH, ORP, dissolved oxygen, and
other electron acceptors and donors. The monitoring schedule should be consistent with the pace of the anticipated
reactions and rate of transport of the injectants and contaminants.
G. WELL DATA
Provide a tabulation of data on all existing or abandoned wells within 'A mile of the injection well(s) which penetrate the
proposed injection zone, including, but not limited to, monitoring wells and wells proposed for use as injection wells.
Such data shall include a description of each well's use (water supply, monitoring, etc), total depth, screened or open
borehole depth interval,and well construction or abandonment record, if available.
H. MAPS
Attach the following scaled, site-specific maps:
(1) Area map based on the most recent USGS 7.5' topographic map of the area,at a scale of 1:24,000 and
showing the location of the proposed injection site.
(2) Site map including:
a. all property boundaries;
b. all buildings within the property boundary;
c. existing and proposed injection wells or well field(s)
d. any existing sources of potential or known groundwater contamination, including waste storage,
treatment or disposal systems within 'A mile of the injection well or well system;
e. all surface water bodies within'A mile of the injection well or well system; and
f. all existing or abandoned wells within 'A mile of the injection well(s)which penetrate the proposed
injection zone,including,but not limited to,monitoring wells and wells proposed for use as injection
wells.
(3) Potentiometric surface map(s)including:
a. direction of groundwater movement
b. existing and proposed monitoring wells
c. existing and proposed injection wells
(4) Contaminant plume map(s)including:
a. the horizontal extent of the contaminant plume,including isoconcentration lines
b. existing and proposed monitoring wells
c. existing and proposed injection wells
(5) Cross-section(s)to the known or projected depth of contamination,including:
a. horizontal and vertical extent of the contaminant plume, including isoconcentration lines
b. major changes in lithology
c. existing and proposed monitoring wells
d. existing and proposed injection wells
Revised 6/09 UIC-5I/5T Page 6 of 7

i�
1_ APPLICATION FOR PERMIT TO CONSTRUCT AND/OR USE A WELL(S)FOR INJECTION
Type 51 Wells-In Situ Groundwater Remediation/Type ST Wells-Tracer Injection
Vt. CERTIFICATION(to be signed as required below or by that person's authorized agent)
NCAC 15A 2C .0211(b)requires that all permit applications shall be signed as follows:
1. for a corporation: by a responsible corporate officer
2. for a partnership or sole proprietorship: by a general partner or the proprietor,respectively
3. for a municipality or a state, federal, or other public agency: by either a principal executive officer or
ranking publicly elected official
4. for all others: by the well owner.
If an authorized agent is signing on behalf of the applicant,then supply a letter signed by the applicant that
names and authorizes their agent.
I hereby certify under penalty of law that I have personally examined and am familiar with the information
submitted in this document and all attachments therein, and that, based on my inquiry of those individuals
immediately responsible for obtaining said information, I believe that the information is true, accurate, and
complete. I am aware that there are penalties, including the possibility of fines and imprisonment, for submitting
false information. I agree to construct, operate, maintain, repair,and if applicable, abandon the injection well(s)
and all related appurtenances in accordance with the approved specifications and conditions of the Permit.
---_� Printed Name and Title: yD c `----+•►
---e.�.-..;Ailttse
Signature:
Date: Jjr7/Z.---
VI1. CONSENT OF PROPERTY OWNER (if the property is not owned by the applicant)
("Owner"means any person who holds the fee or other property rights in the well being constructed. A well is
real property and its construction on land shall be deemed to vest ownership in the land owner, in the absence of
contrary agreement in writing.)
As owner of the property on which the injection well(s)are to be constructed and operated,I hereby consent to
allow the applicant to construct each injection well as outlined in this application and agree that it shall be the
responsibility of the applicant to ensure that the injection well(s)conform to the Well Construction Standards
(Title 15A NCAC 2C.0200).
Printed Nameand Ti
Signature: �
—— - Date: it 1.."
Submit TWO copies of the completed application package,including all attachments,to:
MC Program
Aquifer Protection Section RECEIVED/DENR,DWQ
North Carolina DENR-DWQ
1636 Mail Service Center APR 0 9 2012
Raleigh,NC 27699-1636
Telephone[phone] Ui fg f pro �R �n
Revised 6/09 UIC-51/5T
Page 7 of 7

Attachment A— Site History
Site Usage Historically and Present
The facility currently functions as a freight terminal and includes three buildings.One of the buildings
serves as the garage, and the other two buildings serve as freight handling and distribution loading docks.
The facility layout is depicted in Figure A-1. The former 1,000 gallon UST was located near the
southwest corner of the garage. The area surrounding the garage and the two freight-handling areas is
paved with a combination of concrete and asphalt. The site is contained by a chain-link fence.
Origin of the Contamination
A former used oil tank which was removed from the Site in 1992. Oil within the tank was apparently
contaminated with chlorinated solvents from an unknown source. The chlorinated solvents release likely
occurred before 1992. Residual concentrations of tetrachloroethene(PCE)are present in saturated
source area soil and groundwater below the former used oil underground storage tank (UST).As a result
of the continuing source material, PCE concentrations in groundwater have remained generally steady at
levels above the North Carolina Administrative Code(NCAC) Subchapter 2L Groundwater Standards
since removal of the leaking UST was conducted in 1992.
Previous Remedial Actions
Monitored Natural Attenuation (MNA) has been employed as a remediation strategy since November
1995. Historical analytical data are displayed in Figure A-2 to allow an evaluation of constituent
concentration fluctuations over time. In a letter dated May 24, 2011 addressed to YRC,the North
Carolina Department of Environment and Natural Resources (NCDENR) noted that PCE concentrations
in groundwater have not decreased over time and that an active remedy should be considered to
expedite clean-up of groundwater impacts.
References
ARCADIS G&M of North Carolina, Inc., 2002,Amended Closure Plan, Roadway Express, Inc. Terminal,
Highway 66 South, Kernersville, North Carolina.August 2002.
ARCADIS G&M of North Carolina, Inc., July 2011 Monitoring Well Sampling Report, Roadway Express,
Inc. Terminal, Highway 66 South, Kernersville, North Carolina, October 2011
North Carolina Department of Environmental and Natural Resources, 2011, Letter Addressed to YRC,
May 24,2011.

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m a- PRJT MANAGER. CHECKED BY. DRAFTER: PROJECT NUMBER: CL000294,2006 FIGURE:
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c NOTES:
•i E ARCADIS G&M of North Carolina, Inc. SITE-LAYOUT ROADWAY EXPRESS TERMINAL SITE LAYOUT A
>z 801 Corporate Center Drive, Suite 300 KERNERSVILLE, NORTH CAROLINA
-p Raleigh, NC 27607 DATE.
0y Tel: [phone] Fax: [phone] 19 SEPT 2006
• �� www.arcadis-us.com J

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�{1�,j 801 Corporate cents Drive, B.Zimmermann CL000294.2011
Infrastructure, environment, facilities Suite30o Concentrations vs.Time /�
Raleigh,NC 27607 DRAFTER: Roadway Express Terminal A-2
R.GERBER Kernersville, North Carolina

Attachment B - Hydrogeologic Description
Regional Geologic Setting
The Site lies in the Piedmont physiographic province. The Piedmont is characterized by igneous and
metamorphic rocks that have been folded,faulted and interstratified with granite or diorite intrusions.
There is little to no lateral or vertical hydraulic continuity in these igneous and metamorphic rock units.
Weathering of the igneous and metamorphic rock has created a layer of clayey saprolite soil at the
surface. The thickness of the saprolite soil is highly variable but tends to be thicker on ridge tops and
thinner in valleys.
Lithologv
Two primary lithologic units were encountered at the Site. The upper unit consists of reddish brown
saprolite which is underlain by granitic rock. The upper saprolitic unit is comprised of silty clay/clayey silt
(50 to 100 percent)with minor amounts of fine to coarse grained quartz rich sand (0 to 50 percent). Minor
granitic rock fragments(pebble to granule size)were also encountered, primarily adjacent to bedrock.
Bedrock at the Site consists of granite and was encountered in the borehole for monitor well GW-7 at a
depth of approximately 51 feet below land surface(ft bls). The bedrock appeared to be highly weathered
from approximately 51 to 64 ft bls based on the drill cuttings which were blown out of the borehole using
the air rotary drilling technique. The bedrock became more competent below approximately 64 ft bls
(Geraghty& Miller 1996).
Hydraulic Properties of the Aquifer System
In-situ hydraulic conductivity of the saturated aquifer material has been calculated from slug-test data
obtained from selected monitor wells (Geraghty& Miller 1996). The slug tests were performed using a
5-foot long cylindrical stainless steel tube to displace the water in the wells while recording the water-level
response with a pressure transducer and data logger. Hydraulic conductivities were calculated using
AQTESOLV software, which utilizes the Bouwer and Rice Method. The results of slug tests performed on
shallow monitoring wells GW-4, GW-5, and GW-8 indicated hydraulic conductivity values of 1.13 x 10"5
centimeters per second (cm/s), 2.79 x 10"5 cm/s, and 1.65 x 10"5 cm/s, respectively. An average value for
the surficial aquifer of 1.86 x 10-5 cm/s(0.053 feet per day[ft/day])was then calculated from these values.
The average interstitial groundwater flow velocity for the surficial aquifer at the site was calculated at
0.0032 ft/day or 1.2 ft/year based on the average hydraulic gradient(0.012 ft/ft),the average hydraulic
conductivity, and an assumed effective porosity of 20 percent(Geraghty&Miller 1996).
Depth to the Water Table
The elevation of the water table roughly mirrors the elevation of the land surface. The elevation of the
groundwater at the site drops from approximately 922 feet above mean sea level(ft msl) at GW-4 on the
western edge of the site to approximately 918 ft msl at GW-9 as it flows east. This corresponds to a
depth to water of approximately 14 feet below ground surface area across the site.
References
Geraghty& Miller, 1996, Groundwater Assessment Report, Roadway Express Terminal, Histwayte44,
�`g
South, Kernersville, North Carolina, January 18, 1996. a®° ((IA CA fin °ei
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Attachment C— Injection Fluid Composition
The injection solution will consist of two separate solutions, one containing a mixture of sodium persulfate
and water(Solution A),and the second containing ferrous sulfate, citric acid, and water(Solution B).
Reaction of the ferrous sulfate and citric acid will result in chelated ferrous-citrate, which will ensure that
iron present in solution remains in the second valence state(Fe(Il))and can react with persulfate anion in
the subsurface. A target sodium persulfate concentration up to 60 g/L will be used for injection. The
second solution will contain sufficient weights of ferrous sulfate and citric acid to generate concentrations
of 0.5 g/L and 0.5 g/L, respectively. These concentrations were selected based on the native geology of
the Site, the observed contaminant concentrations within treatment area groundwater, and are based on
previous injection experience at other sites. The selected injection strengths will be confirmed as part of
a pre-injection reaction study using soil and groundwater collected from the Site. When sodium
persulfate mixes with water it dissociates into sodium cations and persulfate anions. The persulfate anion
(oxidant) is a weak acid with a pKa value of 6.0. The solution will be mixed at the surface and then
pumped into the injection wells. An MSDS for sodium persulfate is included in Appendix C. Relevant
physical and chemical properties of sodium persulfate are included in the table below.
Property Value
Molecular Weight 238.1 g/mole
Physical Form White Crystalline Solid
Solubility Limit 0°C 37 wt.% 1
25°C 42 wt.%
50°C 46 wt.%
Solution Density 10%by wt. 1.067 g/ml
20% bywt. _ 1.146g1m1
30% by wt. 1.237 g/ml
40% by wt. 1.340 g/ml
Source: (Block 2006)
Ferrous sulfate will be added to Solution B prior to injection to activate the chemical oxidation reactions.
The ferrous iron catalyzes the decomposition of the oxidant into free radicals which then react with the
contaminants in the subsurface(Block 2004). Following reaction with persulfate, the Fe(II)present in the
subsurface will be oxidized to ferric iron (Fe(I11))and will precipitate within the native site geochemistry at
a pH above 4 standard units(SU). An MSDS for ferrous sulfate is included in Appendix C. Relevant
physical and chemical properties of ferrous sulfate are included in the table below.
Property Value
Molecular Weight 151.92 g/mole
Physical Form White Crystalline Solid
Solubility Limit 25°C 23 wt.%
Density 3.65 g/cm3
Source: CRC Handbook of Chemistry and Physics
Citric acid will be added to Solution B immediately prior to injection to maintain a low pH and prevent
ferrous iron from precipitating out of solution. An MSDS for citric acid is included in Appendix C. Relevant
properties of citric acid are included in the table below.
Property I Value
Molecular Weight 192.12 glmole
Physical Form White Crystalline Solid
Solubility Limit Very soluble in water
Density 1.67 g/cm' _
Source: CRC Handbook of Chemistry and Physics

References
Block, P.A. (2006) "Peroxygen Talk." FMC Corporation
Block, P.A. (2004) "Novel Activation Technologies for Sodium Persulfate In Situ Chemical Oxidation.
Proceeds of the Fourth International conference on the Remediation of Chlorinated and Recalcitrant
Compounds.
CRC Handbook of Chemistry and Physics, 88th edition.2007-2008.

Attachment D - Injection Rationale
Goals of the Injection Project
Application of an oxidant to the subsurface at the Roadway Kernersville site is intended to decrease the
concentration of contaminants that are currently in the soil and groundwater adjacent to the source area.
The goal of the oxidant injection is to decrease the concentration of contaminants sufficiently so that they
no longer act as a source for downgradient contamination historically detected in the monitor well network
at the Site. The use of oxidant injection is intended to eliminate the existing source mass to expedite the
current monitored natural attenuation strategy for dilute low-level VOCs at the Site.
Volume Calculations
The proposed length of injection screen is 15 feet for each well. The planned injection radius is 7.5 ft
from each injection well, resulting in a total fluid diameter of 15 ft. The estimated mobile porosity in the
aquifer is approximately 0.15(15%). Assuming the injectant solution migrates outward from the injection
well in a cylindrical shape the formula to calculate the required injectant volume in each well is the
following:
Volume= Screen Length (15 ft)x Tr(3.14)x Planned Radius(7.5 ft)2 x mobile porosity(0.15)x 7.48
gallons/cubic ft
A calculated volume of approximately 3,000 gallons per injection well is the solution to the above
equation. The total anticipated injectant volume for the three wells is 9,000 gallons.
Reactions
Sodium persulfate is capable of breaking down a variety of contaminants including chlorinated carbons
and hydrocarbons. When sodium persulfate solid is mixed with water it dissolves and dissociates into
sodium cations and persulfate anions as presented in Reaction 1:
Na2S2O8 -+2Na+ +S2O8_2 (1)
The persulfate anion can then go on to directly react with some compounds via direct electron transfer;
however,these reactions are kinetically slow relative to other oxidation chemistries (i.e., Fenton's
oxidation) (House 1962). It is more common for the persulfate to be activated using a catalyst, in this
case ferrous iron, which transforms the persulfate into sulfate radicals as shown in Reaction 2:
S208-2 +Fe+2 Fe+3 +SO4-2 +SO4* (2)
Following reaction with persulfate,the Fe(li) present in the subsurface will be oxidized to ferric iron
(Fe(III))and will precipitate within the native Site geochemistry at a pH above 4 SU.
The degradation of many types of contaminants present at the Site by either direct electron transfer or
reaction with free radicals has been well established. The degradation pathway of tetrachloroethene, the
main contaminant at the site, is discussed below.
Degradation of chloroethenes typically follows a similar path as reductive dechlorination, namely
tetrachloroethene(PCE)—>trichloroethene (TCE)--,cis-and trans-1,2-dichloroethene(DCE)—,vinyl chloride
(VC)—*chloride. Waldemer et al. (2007)found that near complete degradation of chloroethenes to
chloride is possible, therefore, chloride should be the primary breakdown product of chloroethenes at the
site. The study also noted that trace production of other chlorinated hydrocarbon compounds is possible
when treating TCE, due to the short-term reactions of other radical species (e.g.,chloride radical,
hydroxyl radical)generated during the oxidation approach which can interact with small chained organic
molecules. While the formation of organic byproducts has been previously reported,the generation of
these constituents is transient in nature. Their presence is a direct result of ongoing oxidation reactions

and is linked to available oxidant within the subsurface. As complete reaction with available oxidant
continues, further destruction of any radical intermediates occurs.
Modeling or Testing to Investigate Injectant's Potential to Change in the Subsurface
Past experience with subsurface application of persulfate with ferrous sulfate activation has provided data
on"typical"concentrations of oxidant and base that are used for successful treatment of contaminants at
similar sites. These values were used as starting points to begin calculating the optimum volumes and
concentrations of each injected solution and the predicted changes that would occur in the subsurface.
This past experience provides the baseline for initial injections. Two performance monitoring wells
(GW-13 and GW-14)will be installed in the vicinity of the injection wells will be used to gauge treatment
resulting from injection events. These monitoring wells will be constructed of 2-inch poly-vinyl chloride
(PVC) pipe and installed in the locations depicted in Figure D-1. These monitor wells will be installed to
monitor the most shallow groundwater zone. The monitor wells will have a 15 foot screen length that will
be set from 14 to 29 feet below land surface. Existing monitoring well GW-3 will serve as a third
performance monitoring well during injection events. During injection, grab groundwater samples will be
collected for persulfate, sulfate, and iron (total and dissolved)to confirm sufficient distribution of the
injected reagent and ensure that breakthrough is achieved at the performance monitoring wells. Dose
response observations collected during the preliminary injection event will also be used to optimize the
target injection volumes should subsequent injection events be required. Subsequent groundwater
sampling will be conducted in these wells to confirm constituent concentrations have decreased in the
source area.
References
Block, P.A. (2010)"Activated Persulfate Chemistry: Combined Oxidation and Reduction Mechanisms."
Peroxygen Talk. FMC Corporation
Crimi, M.L., and Taylor, J. (2007). "Experimental Evaluation of Catalyzed Hydrogen Peroxide and Sodium
Persulfate for Destruction of BTEX Contaminants."Soil and Sediment Contamination, 16,29-45.
House, D.A. (1962). "Kinetics and Mechanism of Oxidants by Peroxydisulfate." Chemical Review,62,
185-203.
Huang, K.C., Zhao,Z., Hoag, G.E., Dahmani,A., and Block, P.A. (2005). "Degradation of Volatile Organic
Compounds with Thermally Activated Persulfide Oxidation."Chemosphere, 61, 551-560.
Waldemer, R.H., Tratnyek, P.G., Johnson, R.L., and Nurmi, J.T. (2007). "Oxidation of Chlorinated
Ethenes by Heat-Activated Persulfate: Kinetics and Products."Environmental Science and Technology,
41,1010-1015.

•
18-20
Z 14-16 <4.7
11 ' Gw-1
m APPROXIMATE
m EXCAVATION LOCATION -
OF THE FORMER
el 1,000 GALLON GW-7
USED OIL TANK
1 1
a GW-13 i `=`�
W {+4
5 SB-16 A r 1 A. 18-20
20-22 18-20 1 18-20 18-20 22-24 C 20
18 , <390
17 <420 47,000
SB-10 SITE
> I BUILDING
"' PROPOSED INJECTION I 22-24 (GARAGE)
RADIUS OF INFLUENCE -- 18 20 <330 ��._
= 7.5 FT 88,00o I SB-12 i
N "A 1 18-20 µ
g 1,500
a
I
W GW-2•
18-20 GW-14
' \ 190,000 I
3masil
LEGEND I I1 18-20
W 91 I ® • CONCRETE •
PROPOSED MONITORING WELL I _ _ _ _ _ — v — — f — — — — -_ _I CONTAINMENT
BARRIER
S B-1
g ® PROPOSED INJECTION WELL 22-24 26-28 •
'6 54,000 52,000 •
sa A GEOPROBE SOIL BORING SB-7 SB-2 •
o 14-16 18-20
g • SHALLOW MONITOR WELL 56,000 SB-3 ■ . 180 •
— ---
o▪d 26-28 GW-3
_ DEEP MONITOR WELL
7,800
cg MF-
k$ * EXISTING WELL •
°$
a$ CONCRETE AREA
Or
og APPROXIMATE EXTENT OF SOIL CONTAINING A
ti of I TETRACHLOROETHENE AT CONCENTRATIONS ABOVE
ROADWAY EXPRESS TERMINAL
s i THE SITE-SPECIFIC REMEDIATION GOAL (SSRG). - KERNERSVILLE,NORTH CAROLINA
o i 1 18-20 3 OCTOBER 2010
U' SB-7 BORING NUMBER 190 j
i & 14-16 SAMPLE DEPTH IN FEET BELOW GROUND SURFACE SOIL PCE CONCENTRATIONS
>2 56,000 TETRACHLOROETHENE CONCENTRATION IN Ng/kg AND PROPOSED INJECTION
o g Q SCALEIN FEET AND MONITORING WELLS
FIGURE
--o w 0 5 10
re> I ___. .__--_. ar
D-1
5W

Attachment E— Injection Procedure and Equipment
Construction Plans and Materials
ARCADIS will supervise installation of three injection wells in the areas identified in the investigation that
contain the highest levels of PCE (See Figure D-1). The purpose of the injection wells will be to deliver a
chemical oxidant(sodium persulfate)to the subsurface for the treatment of PCE and other VOCs in soil
and groundwater in a controlled manner. The injection wells will be constructed of 2 inch stainless steel
casing and will be screened from depths of 14 to 29 feet below land surface(ft bls). A sand pack will be
deposited within the bore hole around the well casing from termination depth to two feet above the
screened interval. Neat Portland cement will be added from the top of the sand to approximately 6 inches
below ground surface. The well will be protected within a steel manhole secured in a 2-foot by 2-foot
concrete pad.
ARCADIS will also supervise installation of two additional performance monitoring wells in the vicinity of
the injection wells to use as monitoring points to gauge treatment resulting from the injection events. The
monitoring wells will be constructed similar to the injection wells, but will consist of PVC casing rather than
stainless steel.All wells will be developed after installation to remove sediments and improve
performance of the wells. Existing well GW-3 will serve as a third performance monitoring well during
injection events.
Soil cuttings, purge water, and decontamination water generated during drilling will be placed into 55-
gallon drums and staged onsite pending analysis and disposal. One composite sample of the soil cuttings
will be submitted for laboratory analysis of Toxicity Characteristic Leaching Procedure(TCLP)VOCs,
TCLP Semi-volatile organic compounds (SVOCs), and TCLP Metals. One sample of purge water and one
sample of decontamination water will be used to properly characterize the water for disposal.
Operation Procedures
Prior to an injection event, Solution A will be mixed with water in temporary mixing tanks. Ferrous sulfate
and citric acid will then be mixed in a separate tank (Solution B)and added to the sodium persulfate
solution immediately prior to injection. The persulfate and ferrous sulfate mixture will be pumped into
each of the injection wells. During each injection event, approximately 9,000 gallons of 5.85 (wt.)%
(60 g/L)persulfate solution mixed will be delivered to the impacted soil zones. Each injection well will
receive 3,000 gallons of injectant. During injection, grab groundwater samples will be collected from wells
GW-3, GW-13, and GW-14 for persulfate, sulfate, and iron (total and dissolved)to confirm sufficient
distribution of the injected reagent.
Injection Schedule
The injection wells will be installed within two months of obtaining the UIC permit. The injection event will
be conducted within one month of the well installations and subsequent groundwater monitoring will
conclude within four months of the injection. The remedial performance report will be submitted to
NCDENR within five months of the injection event. The remedial phase will therefore be completed within
one year of approval to proceed. The existing semi-annual groundwater monitoring program will continue
during this time. A second injection event will be scheduled within two years of the initial injection event if
contaminant concentrations do not decrease significantly in subsequent monitoring events.

We S
Attachment F— Monitoring Plan
A list of the existing wells is provided in Table G-1 (Attachment G). The network covers both the shallow
soil aquifer as well as the deeper bedrock aquifer. This monitoring network will be used to evaluate for the
migration of injection fluids outside the injection zone, the presence of secondary contaminants or
reaction end-products (e.g., chloride)from the degradation of target contaminants and other parameters
that can indicate the progress of the intended reactions.
Two performance monitoring wells, GW-13 and GW-14, will be installed in the vicinity of the injection
wells as depicted in Figure D-1; they will be used to gauge treatment resulting from injection events.
These monitor wells will be installed to monitor the most shallow groundwater zone. Existing monitoring
well GW-3 will serve as a third performance monitoring well during injection events. During injection, grab
groundwater samples will be collected from performance monitoring wells for persulfate, sulfate, and iron
(total and dissolved)to confirm that sufficient breakthrough is achieved. Performance monitoring wells
(GW-3, GW-13, and GW-14)will also be sampled during semi-annual sampling events.
Groundwater samples will be collected from GW-13, GW-14, and GW-3 as well as injection wells for VOC
analysis(USEPA Method 8260)before injection, six weeks after injection, and twelve weeks after
injection during a semi-annual sampling event. In addition to VOC analysis, samples will also be
collected for TOC, sulfate, total and dissolved iron and manganese, and chloride. Following injection,
samples will also be collected for persulfate analysis via field test kits. This monitoring program will allow
evaluation of the baseline groundwater conditions relative to concentrations observed following the
injection event, ultimately allowing for evaluation of treatment performance. During collection of the
groundwater samples, a YSI 556 with a flow through cell will be used to monitor pH, ORP, dissolved
oxygen and conductivity. These measurements can provide indirect data regarding the migration of the
applied oxidant and the degradation of the contaminants.
The existing semi-annual groundwater monitoring program will continue to evaluate groundwater
conditions within the Site-wide monitoring network to confirm treatment performance beyond the injection
area. Should additional persulfate injections be required,they will be performed within two years of the
initial injection.

Attachment G —Well Data
During a receptor survey in November,2007 no known water supply wells were discovered within 1,500
feet of the site. The entire area,including the Roadway Facility is used predominantly for freight
trucking. Information regarding the water supply for the area surrounding the site was obtained from Mr.
Nick Seeba of the City of Winston Salem Public Utilities.During a telephone interview on November 12,
2007,Mr. Seeba indicated the entire area has accessible public water. Mr. Seeba further indicated that an
8-inch or 12-inch diameter water main is present running along NC Highway 66. Other water mains that
are connected to this line include Indeneer Drive(6-inch main)and Park Center Drive(8-inch main)west
of NC Highway 66 and also Plaza South Drive(8-inch main)and Brookford Road(6-inch main),both
located south of the site.
Michael Merrell of the Forsyth County Department of Public Health Division of Environmental Health
was contacted to determine if there was any record of wells within 1,500 feet of the site. There are no
wells within 1,500 feet of the site in the Forsyth County record. Forsyth County has kept records of
construction and abandonment of wells since 1988. Mr.Merrell reported from personal knowledge that
there are two bored wells without record in the vacant property, 1200 Highway 66 South,directly across
from the site. He also mentioned that at the Ford Dealership, 1330 Highway 66 South, showed evidence
of a well on a septic tank permit. The dealership was contacted February 14,2012;they had no
knowledge of any wells on site.
By checking with Winston Salem Public Utilities and Forsyth County Department of Health,one can
safely conclude that the entire area surrounding the site has access to public water and that there is no
record of wells used for drinking water in the area.
The only known wells within the vicinity of the former used oil UST are therefore the existing monitoring
wells(GW-1 through GW-12)surrounding the impacted plume. Table G-1 included well construction
details and Table G-2 includes the most recent groundwater data from each of the monitor wells.

Table G.1 Monitor Well Construction Details,Roadway Express Terminal,Kernersville,North Carolina.
Monitoring Wells
Total Well Screened Top of Well
Installation Depth Diameter Interval Casing Elevation
Well No. Date (ft bls) (inches) (ft bls) (ft msl)
GW-1 2/3/1994 20 2 10-20 937.99
GW-2 2/3/1994 20 2 10-20 937.55
GW-3 2/3/1994 20 2 10-20 937.91
GW-4 10/9/1995 25 2 10-25 938.44
GW-5 10/9/1995 25 2 10-25 939.42
GW-6 10/10/1995 24 2 9-24 937.98
GW-7 10/30/1995 81 2 71 -81 937.74
GW-8 10/10/1995 25 2 10-25 938.49
GW-9 10/10/1995 25 2 10-25 938.16
GW-10 6/24/1997 57 2 47-57 938.82
GW-11 9/19/2006 82 2 72-82 938.27
GW-12 9/19/2006 79 2 69-79 939.03
GW-13* TBD 29 2 14-29 TBD
GW-14* TBD 29 2 14-29 TBD
Injection Wells
Total Well Screened Top of Well
Installation Depth Diameter Interval Casing Elevation
Well No. Date (ft bls) (inches) (ft bls) (ft msl)
IW-1* TBD 29 2 14-29 TBD
IW-2* TBD 29 2 14-29 TBD
IW-3* TBD 29 2 14-29 TBD
* Proposed wells
ft bls Feet below land surface.
ft msl Feet above mean sea level.

Table G-2. Groundwater Analytical Results, Roadway Express Terminal,Kernersville,North Carolina.
NCAC 2L
Well ID: Groundwater GW-1 GW-2 GW-3 GW-4 GW-5 GW-6 GW-7 GW-8 GW-9 GW-10 GW-11 GW-12
Date Sampled: Standard 01/04/12 11/08/95 01/04/12 01/04/12 01/04/12 _ 01/04/12 01/04/12 01/04/12 01/04/12 01/04/12 01/04/12 01/04/12
Volatile Organic Compounds
(USEPA Method 8260B) µg/L
Acetone 6,000 <25 <10 <500 <25 <25 <25 <25 <25 <25 <25 <25 <25
Benzene I <1.0 <5 <20 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
2-Butanone 4,000 <10 <10 <200 <10 <10 <10 <10 <10 <10 <10 <10 <10
Carbon disulfide 700 <2.0 <10 <40 <2.0 <2.0 <2.0 <2.0 <2.0 <2.0 <2.0 <2.0 <2.0
Chloroethane 3,000 <1.0 <5 <20 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0
Chloroform 70 <1.0 <5 <20 <1.0 <1.0 <1.0<1.0
<1.0
<5 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <20
Chloromethane 3 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <5 <20 <1.0 <1.0
1,1-Dichloroethane 6 1,1-Dichloroethene 7 <1.0 <5 <20 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
cis-1,2-Dichloroethene 70 14 <5 440 1 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
trans-1,2-Dichloroethene 100 <1.0 <5 <20 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
Ethylbenzene 600 <1.0 <5 <20 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
Methylene chloride 5 <5.0 6.3 i <100 <5.0 <5.0 _ <5.0 <5.0 <5.0 <5.0 <5.0 <5.0 <5.0
Tetrachloroethene(PCE) 0.7 130 - 75 3,700 <1.0 1.2 38 <1.0 [ 20 <1.0 <1.0 <1.0 <1.0
Toluene 600 <1.0 <5 <20 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
1,1,1-Trichloroethane 200 <1.0 <5 <20 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0
Trichloroethene(TCE) 3 13 <5 ( 81 <1.0 <1.0 i 3.6 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
Vinyl chloride 0.03 <1.0 <5 <20 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
Xylenes(Total) 500 <2.0 <5 70 <2.0 <2.0 34 <2.0 <2.0 <2.0 <2.0 <2.0 <2.0
Semivolatile Organic Compounds
(USEPA Method 8270C) µg/L
2-Methylnaphthalene 30 NS <10 <10 NS NS 68 NS NS NS NS NS NS
Naphthalene 6 NS <10 34 • NS NS 91 _ NS NS NS NS NS NS
NCAC 2L North Carolina Administrative Code Title 15A, Subchapter 2L. NA Constituent was not analyzed.
µg/L Micrograms per liter. UJ Constituent concentration is qualified as estimated nondetected.
< Constituent was not detected above the quantitation limit. Bold Constituent concentration that is above the method detection limit.
NE Numerical standard has not been established Indicates the North Carolina Groundwater Standard has been exceeded.

O Health 2
2 Fire 0
• Reactivity 2
Personal E,
Protection
Material Safety Data Sheet
Sodium persulfate MSDS
Section 1: Chemical Product and Company Identification
Product Name: Sodium persulfate Contact Information:
Catalog Codes: SLS1333 Sciencelab.com, Inc.
14025 Smith Rd.
CAS#: 7775-27-1 Houston, Texas 77396
RTECS: SE0525000 US Sales: 1-800-901-7247
International Sales: 1-281-441-4400
TSCA: TSCA 8(b)inventory: Sodium persulfate
Order Online: ScienceLab.com
CI#: Not applicable.
CHEMTREC(24HR Emergency Telephone), call:
Synonym: Peroxydisulfuric acid, disodium salt 1-800-424-9300
Chemical Name: Sodium peroxydisulfate International CHEMTREC, call: 1-703-527-3887
Chemical Formula: Na2S2O8 For non-emergency assistance,call: 1-281-441-4400
Section 2: Composition and Information on Ingredients
Composition:
Name CAS# % by Weight
Sodium persulfate 7775-27-1 100
Toxicological Data on Ingredients: Sodium persulfate LD50: Not available. LC50: Not available.
Section 3: Hazards Identification
Potential Acute Health Effects:
Very hazardous in case of skin contact(irritant, sensitizer), of eye contact(irritant), of inhalation. Hazardous in case of
ingestion. Slightly hazardous in case of skin contact(permeator). Prolonged exposure may result in skin burns and ulcerations.
Over-exposure by inhalation may cause respiratory irritation. Inflammation of the eye is characterized by redness,watering,
and itching. Skin inflammation is characterized by itching, scaling, reddening, or, occasionally, blistering.
Potential Chronic Health Effects:
Very hazardous in case of skin contact(corrosive, irritant, sensitizer), of eye contact(irritant), of inhalation. Hazardous in case
of ingestion. Slightly hazardous in case of skin contact(permeator). CARCINOGENIC EFFECTS: Not available. MUTAGENIC
EFFECTS: Not available. TERATOGENIC EFFECTS: Not available. DEVELOPMENTAL TOXICITY: Not available. The
substance is toxic to blood, lungs. Repeated or prolonged exposure to the substance can produce target organs damage.
Repeated or prolonged inhalation of dust may lead to chronic respiratory irritation.
Section 4: First Aid Measures
p. 1

Eye Contact:
Check for and remove any contact lenses. Immediately flush eyes with running water for at least 15 minutes, keeping eyelids
open. Cold water may be used. Do not use an eye ointment. Seek medical attention.
Skin Contact:
After contact with skin, wash immediately with plenty of water. Gently and thoroughly wash the contaminated skin with running
water and non-abrasive soap. Be particularly careful to clean folds, crevices, creases and groin. Cold water may be used.
Cover the irritated skin with an emollient. If irritation persists, seek medical attention.
Serious Skin Contact:
Wash with a disinfectant soap and cover the contaminated skin with an anti-bacterial cream. Seek medical attention.
Inhalation:Allow the victim to rest in a well ventilated area. Seek immediate medical attention.
Serious Inhalation:
Evacuate the victim to a safe area as soon as possible. Loosen tight clothing such as a collar, tie, belt or waistband. If
breathing is difficult, administer oxygen. If the victim is not breathing, perform mouth-to-mouth resuscitation. Seek medical
attention.
Ingestion:
Do not induce vomiting. Loosen tight clothing such as a collar, tie, belt or waistband. If the victim is not breathing, perform
mouth-to-mouth resuscitation. Seek immediate medical attention.
Serious Ingestion: Not available.
Section 5: Fire and Explosion Data
Flammability of the Product: Non-flammable.
Auto-Ignition Temperature: Not applicable.
Flash Points: Not applicable.
Flammable Limits: Not applicable.
Products of Combustion: Not available.
Fire Hazards in Presence of Various Substances: Not applicable.
Explosion Hazards in Presence of Various Substances:
Risks of explosion of the product in presence of mechanical impact: Not available. Risks of explosion of the product in
presence of static discharge: Not available.
Fire Fighting Media and Instructions: Not applicable.
Special Remarks on Fire Hazards: Dangerous in contact with organic materials.
Special Remarks on Explosion Hazards: Not available.
Section 6: Accidental Release Measures
Small Spill: Use appropriate tools to put the spilled solid in a convenient waste disposal container.
Large Spill:
Oxidizing material. Stop leak if without risk. Avoid contact with a combustible material(wood, paper, oil, clothing...). Keep
substance damp using water spray. Do not touch spilled material. Prevent entry into sewers, basements or confined areas;
dike if needed. Call for assistance on disposal.
Section 7: Handling and Storage
Precautions:
p. 2

Keep away from heat. Keep away from sources of ignition. Keep away from combustible material Do not breathe dust. In case
of insufficient ventilation, wear suitable respiratory equipment If you feel unwell, seek medical attention and show the label
when possible.Avoid contact with skin and eyes Keep away from incompatibles such as reducing agents, organic materials,
metals, acids, moisture.
Storage: Oxidizing materials should be stored in a separate safety storage cabinet or room.
Section 8: Exposure Controls/Personal Protection
Engineering Controls:
Use process enclosures, local exhaust ventilation, or other engineering controls to keep airborne levels below recommended
exposure limits. If user operations generate dust, fume or mist, use ventilation to keep exposure to airborne contaminants
below the exposure limit.
Personal Protection:
Splash goggles. Lab coat. Dust respirator. Be sure to use an approved/certified respirator or equivalent. Gloves.
Personal Protection in Case of a Large Spill:
Splash goggles. Full suit. Dust respirator. Boots. Gloves.A self contained breathing apparatus should be used to avoid
inhalation of the product. Suggested protective clothing might not be sufficient; consult a specialist BEFORE handling this
product.
Exposure Limits: Not available.
Section 9: Physical and Chemical Properties
Physical state and appearance: Solid. (Solid crystalline powder.)
Odor: Odorless.
Taste: Bitter. (Strong.)
Molecular Weight: 238.1 g/mole
Color: White.
pH(1% soln/water): 7[Neutral.]
Boiling Point: Not available.
Melting Point: Decomposes.
Critical Temperature: Not available.
Specific Gravity: 2.6(Water= 1)
Vapor Pressure: Not applicable.
Vapor Density: Not available.
Volatility: Not available.
Odor Threshold: Not available.
Water/Oil Dist. Coeff.: Not available.
lonicity (in Water): Not available.
Dispersion Properties: See solubility in water.
Solubility:
Soluble in cold water, hot water. Insoluble in methanol, diethyl ether, n-octanol.
Section 10: Stability and Reactivity Data
11 3

Special Provisions for Transport: Not available.
Section.15: Other Regulatory Information
Federal and State Regulations: TSCA 8(b) inventory: Sodium persulfate
Other Regulations: OSHA: Hazardous by definition of Hazard Communication Standard (29 CFR 1910.1200).
Other Classifications:
WHMIS (Canada):
CLASS C: Oxidizing material. CLASS D-2A: Material causing other toxic effects (VERY TOXIC).
DSCL(EEC):
R38- Irritating to skin. R41-Risk of serious damage to eyes. R43-May cause sensitization by skin contact.
HMIS(U.S.A.):
Health Hazard: 2
Fire Hazard: 0
Reactivity:2
Personal Protection: E
National Fire Protection Association (U.S.A.):
Health:2
Flammability: 0
Reactivity:2
Specific hazard:
Protective Equipment:
Gloves. Lab coat. Dust respirator. Be sure to use an approved/certified respirator or equivalent. Wear appropriate respirator
when ventilation is inadequate. Splash goggles.
Section 16: Other Information
References:
-Hawley, G.G.. The Condensed Chemical Dictionary, 11e ed., New York N.Y.,Van Nostrand Reinold, 1987. -The Sigma-
Aldrich Library of Chemical Safety Data, Edition II.
Other Special Considerations: Not available.
Created: 10/11/2005 12:36 PM
Last Updated: 11/01/2010 12:00 PM
The information above is believed to be accurate and represents the best information currently available to us. However, we
make no warranty of merchantability or any other warranty, express or implied, with respect to such information, and we assume
no liability resulting from its use. Users should make their own investigations to determine the suitability of the information for
their particular purposes. In no event shall ScienceLab.corn be liable for any claims, losses, or damages of any third party or for
lost profits or any special, indirect, incidental, consequential or exemplary damages, howsoever arising, even if ScienceLab.corn
has been advised of the possibility of such damages.
•
P. 5

0
He a Ith 2
2 O Fire 0
Reactis•ity 0
Personal E
Protection
Material Safety Data Sheet
Ferrous sulfate MSDS
Section 1: Chemical Product and Company Identification
Product Name: Ferrous sulfate Contact Information:
Catalog Codes: SLF1516 Sciencelab.com, Inc.
14025 Smith Rd.
CAS#: 13463-43-9 Houston, Texas 77396
_ RTECS: Not available. US Sales: 1-800-901-7247
TSCA:TSCA 8(b) inventory: No products were found. International Sales: 1-281-441-4400
Order Online: ScienceLab.com
Cl#: Not available.
CHEMTREC(24HR Emergency Telephone), call:
Synonym: Ferrous Sulfate Hydrate; Ferrous Sulfate Dried 1-800-424-9300
Powder
International CHEMTREC,call: 1-703-527-3887
Chemical Name: Ferrous Sulfate
For non-emergency assistance,call: 1-281-441-4400
Chemical Formula: FeSO4.xH2O
Section 2: Composition and Information on Ingredients
Composition:
Name CAS# % by Weight
Ferrous sulfate 13463-43-9 100
Toxicological Data on Ingredients: Ferrous sulfate LD50: Not available. LC50: Not available.
Section 3: Hazards Identification
Potential Acute Health Effects: Hazardous in case of skin contact(irritant), of eye contact(irritant), of ingestion, of inhalation.
Potential Chronic Health Effects:
CARCINOGENIC EFFECTS: Not available. MUTAGENIC EFFECTS: Not available. TERATOGENIC EFFECTS: Not available.
DEVELOPMENTAL TOXICITY: Not available. The substance may be toxic to kidneys, liver, cardiovascular system, central
nervous system (CNS). Repeated or prolonged exposure to the substance can produce target organs damage.
Section 4: First Aid Measures
Eye Contact:
Check for and remove any contact lenses. In case of contact, immediately flush eyes with plenty of water for at least 15
4 minutes. Cold water may be used. Get medical attention.
p. 1

Skin Contact:
In case of contact, immediately flush skin with plenty of water. Cover the irritated skin with an emollient. Remove contaminated
clothing and shoes. Cold water may be used.Wash clothing before reuse. Thoroughly clean shoes before reuse. Get medical
attention.
Serious Skin Contact:
Wash with a disinfectant soap and cover the contaminated skin with an anti-bacterial cream. Seek medical attention.
Inhalation:
If inhaled, remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult, give oxygen. Get medical
attention.
Serious Inhalation: Not available.
ingestion:
Do NOT induce vomiting unless directed to do so by medical personnel. Never give anything by mouth to an unconscious
person. If large quantities of this material are swallowed, call a physician immediately. Loosen tight clothing such as a collar,
tie, belt or waistband.
Serious Ingestion: Not available.
Section 5: Fire and Explosion Data
Flammability of the Product: Non-flammable.
Auto-Ignition Temperature: Not applicable.
Flash Points: Not applicable.
Flammable Limits: Not applicable.
Products of Combustion: Not available.
Fire Hazards in Presence of Various Substances: Not applicable.
Explosion Hazards in Presence of Various Substances:
Risks of explosion of the product in presence of mechanical impact: Not available. Risks of explosion of the product in
presence of static discharge: Not available.
Fire Fighting Media and Instructions: Not applicable.
Special Remarks on Fire Hazards: Not available.
Special Remarks on Explosion Hazards: Not available.
Section 6: Accidental Release Measures
Small Spill:
Use appropriate tools to put the spilled solid in a convenient waste disposal container. Finish cleaning by spreading water on
the contaminated surface and dispose of according to local and regional authority requirements.
LI Large Spill:
Use a shovel to put the material into a convenient waste disposal container. Finish cleaning by spreading water on the
contaminated surface and allow to evacuate through the sanitary system. Be careful that the product is not present at a
concentration level above TLV. Check TLV on the MSDS and with local authorities.
Section 7: Handling and Storage
Precautions:
Do not breathe dust. Wear suitable protective clothing. In case of insufficient ventilation,wear suitable respiratory equipment. If
you feel unwell, seek medical attention and show the label when possible.Avoid contact with skin and eyes.
p. 2

Storage:
Hygroscopic. Keep container tightly closed. Keep container in a cool, well-ventilated area. Do not store above 24°C (75.2°F).
Section 8: Exposure Controls/Personal Protection
Engineering Controls:
Use process enclosures, local exhaust ventilation, or other engineering controls to keep airborne levels below recommended
exposure limits. If user operations generate dust,fume or mist, use ventilation to keep exposure to airborne contaminants
below the exposure limit.
Personal Protection:
Splash goggles. Lab coat. Dust respirator. Be sure to use an approved/certified respirator or equivalent. Gloves.
Personal Protection in Case of a Large Spill:
Splash goggles. Full suit. Dust respirator. Boots. Gloves.A self contained breathing apparatus should be used to avoid
inhalation of the product. Suggested protective clothing might not be sufficient; consult a specialist BEFORE handling this
product.
Exposure Limits: Not available.
Section 9: Physical and Chemical Properties
Physical state and appearance: Solid.
Odor: Not available.
Taste: Not available.
Molecular Weight: 151.9 g/mole+ H2O
Color: Grayish -white to yellowish. (Light.)
pH (1% soln/water): Not available.
Boiling Point: Not available.
Melting Point: Not available.
Critical Temperature: Not available.
Specific Gravity: Not available.
Vapor Pressure: Not applicable.
Vapor Density: Not available.
Volatility: Not available.
Odor Threshold: Not available.
Water/Oil Dist. Coeff.: Not available.
tonicity(in Water): Not available.
Dispersion Properties: See solubility in water.
Solubility: Soluble in cold water.
Section 10: Stability and Reactivity Data
Stability: The product is stable.
1 Instability Temperature: Not available.
P. 3

Conditions of Instability: Incompatibles
Incompatibility with various substances: Not available.
Corrosivity: Not available.
Special Remarks on Reactivity:
Hygroscopic. Loses water at about 300 C. No other information found.
Special Remarks on Corrosivity: Not available.
Polymerization:Will not occur.
Section 11: Toxicological Information
Routes of Entry: Inhalation. Ingestion.
Toxicity to Animals:
LD50: Not available. LC50: Not available.
Chronic Effects on Humans: May cause damage to the following organs: kidneys, liver, cardiovascular system, central
nervous system(CNS).
Other Toxic Effects on Humans: Hazardous in case of skin contact(irritant), of ingestion, of inhalation.
Special Remarks on Toxicity to Animals: Not available.
Special Remarks on Chronic Effects on Humans: May affect genetic material (mutagenic)
Special Remarks on other Toxic Effects on Humans:
Acute Potential Health Effects: Skin: May cause skin irritation. Eyes: May cause eye irritation. Inhalation: May cause
respiratory tract irritation. Ingestion: Harmful if swallowed. May cause gastrointestinal tract distrubances and irritation with
nausea, vomiting, colic, constipation, diarrhea, black stool. May also affect behavior/Central Nervous System (somnolence
-general depressed activity), respiration, cardiovascular system, liver, kidneys (pink urine discoloration). Chronic Potential
Health Effects: Repeated exposure via ingestion may increase iron levels in the liver, and spleen. Damage may occur to
spleen and liver.
Section 12: Ecological Information
Ecotoxicity: Not available.
BOD5 and COD: Not available.
Products of Biodegradation:
Possibly hazardous short term degradation products are not likely. However, long term degradation products may arise.
Toxicity of the Products of Biodegradation: The product itself and its products of degradation are not toxic.
Special Remarks on the Products of Biodegradation: Not available.
Section 13: Disposal Considerations
Waste Disposal:
Waste must be disposed of in accordance with federal, state and local environmental control regulations.
Section 14: Transport Information
DOT Classification: Not a DOT controlled material(United States).
Identification: Not applicable.
p.4

Special Provisions for Transport: Not applicable.
Section 15: Other Regulatory Information
Federal and State Regulations: No products were found.
Other Regulations: OSHA: Hazardous by definition of Hazard Communication Standard (29 CFR 1910.1200).
Other Classifications:
WHMIS(Canada): Not controlled under WHMIS (Canada).
DSCL(EEC):
R36/38-Irritating to eyes and skin. S2-Keep out of the reach of children. S46-If swallowed, seek medical advice immediately
and show this container or label.
HMIS (U.S.A.):
Health Hazard: 2
Fire Hazard: 0
Reactivity: 0
Personal Protection: E
National Fire Protection Association (U.S.A.):
Health: 2
Flammability: 0
Reactivity: 0
Specific hazard:
Protective Equipment:
Gloves. Lab coat. Dust respirator. Be sure to use an approved/certified respirator or equivalent. Splash goggles.
Section 16: Other Information
References: Not available.
Other Special Considerations: Not available.
Created: 10/09/2005 05:33 PM
Last Updated: 11/01/2010 12:00 PM
The information above is believed to be accurate and represents the best information currently available to us. However, we
make no warranty of merchantability or any other warranty, express or implied, with respect to such information, and we assume
no liability resulting from its use. Users should make their own investigations to determine the suitability of the information for
their particular purposes. In no event shall ScienceLab.corn be liable for any claims, losses, or damages of any third party or for
lost profits or any special, indirect, incidental, consequential or exemplary damages,howsoever arising, even if ScienceLab.corn
has been advised of the possibility of such damages.
P. 5

Attachment H
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• • •
PORTS SHALLOW MONITOR WELL
APPROXIMATE ': q LOCATION
EXCAVATION LOCATION OIL/WATER Q 4 ..O
OF THE FORMER IR G NS11 SEPARATOR
TANK FELLE G DEEP MONITOR WELL
I 1,000 GALLON V� • PORTS LOCATION
t WASTE OIL TANK
0 0 PROPOSED SHALLOW
\ o DIESEL GW-12 MONITOR WELL
100 FILL PORTS NS PROPOSED SHALLOW
\ \ MONITOR WELL
\ • GW-8 / / CONCRETE AREA
N.. 18
TETRACHLOROETHENE
GW-10 Q / • 350 CONCENTRATION (}ig/L)
\ 10 <1 / GW-9 TETRACHLOROETHENE
<1 100 ISOCONCENTRATION CONTOUR
IN THE SHALLOW AQUIFER (dug/L)
// \
NS NOT SAMPLED
N. GW-5 SCALE IN FEET
•1 1.4 u 30 60
1 FIGURE:
PRJT MANAGER. CHECKED BY' DRAFTER. ' PROJECT NUMBER. CL000294,2010
ARCAD1S B. ZIMMERMANN J. TILLOTSON R. GERHER
NOTES: DRAWING• PROPOSED INJECTION WELLS AND MONITOR
ROADWAY EXPRESS TERMINAL WELLS WITH TETRACHLOROETHENE H4
o1ACorpora a of
North Carolina,
ISuiten3� TET-07-10 KERNERSVILLE, NORTH CAROLINA CONCENTRATIONS AND SHALLOW AQUIFER
Raleigh, NC 27607 DATE.
Tel: [phone] Fax: [phone] OCT 2010 ISOCONCENTRATION CONTOURS, JULY 2011
www.arcadie—ue.com -

LEGEND 940 A A'GV-4 -940
GW-7 GW-1 GW-3 GW-11 GV-8 GW-10
GV-12 GW-9
PCE CONCENTRATION -
80 •
JULY 20 11 (u
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PCE ISOCONCENTRATION ) Clayi+y.¢nasal:- _ .
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SAM
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. __ _ _ _ - _ . . _ . _ - - _ _-
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-690
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o_ •>' FINE SAND (60%) w� r, •
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i -i SMALL GRAVEL 20°!0) .�..�r�m_c -.yr�ra�� -:ate �. ._
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-860
{_z I 860 �� _ -. - 'NM./ f+.w1 �dw..r _____
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o HORIZONTAL SCALE IN FEET r r
v
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- -
�� -850
t
N 850-
h•ce PRJT MANAGER: CHECKED BY: DRAFTER: PROJECT NUMBER: CL000294.2010 FIGURE:
ii`m R. ZIMMERMAN J. TILLOTSON RGERBER PROPOSED INJECTION AND MONITOR WELLS
.0 N0TES: GEOLOGIC CROSS-SECTION A-A' u
p 1. McDOWELL do ASSOCIATES, PA SURVEY JUNE, 2001, SEPT. 2003, JUNE ROADWAY EXPRESS TERMINAL fl
'�m 2.D4 A JULY Zoos. SHOWING PCE ISOCONCENTRATION CONTOURS
y 3. DATA FOR 12M 2P ARED FROM MAY 2001.R JULY 2011
>o � 3. DATA FOR 12Mw2I AND 12MW31 ARE FROM 2003. KERNERSVILLE, NORTH CAROLINA
v= 4. DATA FOR 12MW10 THROUGH 12MW16 ARE FROM 2004.
°'` 5. DATA FOR ALL OTHERS ARE FROM 2005.

— S c'G4. . Vv..I.o Sao 2 S.Lf
Shrestha, Shristi R
From: McGuire, Theresa <[e-mail]>
Sent: Wednesday, October 24, 2018 9:16 AM
To: Shrestha, Shristi R
Subject: RE: [External] RE:WI0400254 Former Exxon Mobil Facility#46602
CAUTION: • . • 's verified.Send all suspicious email as an attachment to
Resort Siam.
Good afternoon Shristi,
I wanted to let you know that we have had a project manager change. Paul Goodell has been assigned other projects
and no longer manages this site for Exxon. I will be handling the site going forward. My contact information is listed
below if you have any questions.
The new Exxon project manager is:
Ms. Calista Campbell
ExxonMobil Environmental Services
1900 E. Linden Avenue, Building 28a
Linden, NJ 07036
Please let me know if you have any other questions. I look forward to working with you.
Have a great day,
Theresa
Theresa McGuire, P.G. I Certified Project Manager I Theresa.mcquireCa�arcadis.com
Arcadis I Arcadis U.S., Inc., 111-D Sanders Lane, Bluefield,VA 124605 I USA
T. [phone] I M. [phone]
Professional Registration/PG-VA,#001954/PG-PA,#PG005209/PG-NC,#2415/PG-DE,#S4-0001318/
PG-TN,#5785/PG-KY,#PG2604
Connect with us! tivww.arcadis.com I Linkedln
R I tried
From:Shrestha,Shristi R<[e-mail]>
Sent:Tuesday, October 23, 2018 11:58 AM
To:Goodell, Paul <[e-mail]>
Subject: RE: [External] RE:W10400254 Former Exxon Mobil Facility#46602
I sent a letter on September 25th for completion of permit which came back due to wrong address. Please send me
updated name and address for the above permit.
Thank you,
Shristi
Shristi R. Shrestha
Hydrogeologist
Water Quality Regional Operations Section

siAri
1.
e.
•
NORTH CAROLINA
ROY COOPER Environmental Quality
Gyirrmor
MICHAEL S.REGAN
Secretary
LINDA CULPEPPER
irgrrtm Direct
September 25, 2018
Jewel Cox
Former ExxonMobil Facility#46602
1016 W Poplar Ave.
Ste 106 Number 232
Collierville,NC 38017
Subject: Completion of 5I Injection Well Permit WI0400254
Guilford County
Dear Ms. Cox:
On February 15,2013 the Water Quality Regional Operations Section(Former Aquifer Protection
Section) issued a 5I groundwater remediation permit to the above for the remediation of
contaminated groundwater located at 3701 High Point Rd., Greensboro, Guilford County, NC
27407. Expiration date for the permit was January 1, 2018.
A review of the file indicates the Final Project Evaluation (FPE) and injection well construction
have been received, per permit conditions. Therefore, this completes the monitoring and
compliance reporting requirements of your permit.
Please be advised this permit is now inactive.and no further injection activities are permitted. Any
future injection activities must be conducted per the requirements of 15A NCAC 2C .0225
(Revised May 1,2012). You must submit either(1) a Notification of Intent to Install groundwater
remediation wells, or(2) an Application for Permit to Construct Wells for Injection(whichever is
applicable).
E(
North Carolina Department of Environmental Quality I Division of Water Resources I AFOGW Section
512 North Salisbury Street 11636 Mail Service Center I Raleigh,North Carolina 27699-1636
919.707.9129

Please reference permit number WI0400254 in any future correspondence for this site. Operating
an injection well without a valid permit could result in a civil penalty of up to $25,000 per day.
Please contact Shristi Shrestha at 919-707-3662 or Debra Watts at 919-707-3670 if you have any
questions.
Best Regards,
2,: ia.
Shristi Shrestha
Hydrogeologist
Water Quality Regional Operations Section
Division of Water Resources
cc: Sherri Knight, WQROS—Winston-Salem Regional Office
Central Office File,WI0400254
Scott Bostian, scott.bostianarcadis-us.com,
Paul Goodell, Paul.Goodell a,arcadis.com

a
i, 0 ,..„j, , 1 i
NORTH CAROLINA
ROY COOPER Eavisantatatalflity
Govoluor
MICHAEL S.REGAN
Secretary
LINDA CULPEPPER
Inter m ai'rector
October 24,2018
Calista Campbell
ExxonMobil Environmental Services
1900 E. Linden Avenue, Building 28a
Linden,NJ 07036
Subject: Completion of 5I Injection Well Permit W10400254
Guilford County
Dear Ms. Cox:
On February 15,2013 the Water Quality Regional Operations Section(Former Aquifer Protection
Section) issued a 5I groundwater remediation permit to the above for the remediation of
contaminated groundwater located at 3701 High Point Rd., Greensboro, Guilford County, NC
27407. Expiration date for the permit was January 1, 2018.
A review of the file indicates the Final Project Evaluation (FPE) and injection well construction
have been received, per permit conditions. Therefore, this completes the monitoring and
compliance reporting requirements of your permit.
Please be advised this permit is now inactive and no further injection activities are permitted. Any
future injection activities must be conducted per the requirements of 15A NCAC 2C .0225
(Revised May 1, 2012). You must submit either(1) a Notification of Intent to Install groundwater
remediation wells, or(2) an Application for Permit to Construct Wells for Injection(whichever is
applicable).
1DE CO
North Carolina Department of Environmental Quality I Division of Water Resources I AFOGW Section
512 North Salisbury Street 11636 Mail Service Center I Raleigh,North Carolina 27699-1636
919.707.9129

Please reference permit number WI0400254 in any future correspondence for this site. Operating
an injection well without a valid permit could result in a civil penalty of up to $25,000 per day.
Please contact Shristi Shrestha at 919-707-3662 or Debra Watts at 919-707-3670 if you have any
questions.
Best Regards,
Shristi Shrestha
Hydrogeologist
Water Quality Regional Operations Section
Division of Water Resources
cc: Sherri Knight, WQROS—Winston-Salem Regional Office
Central Office File, WI0400254
Theresa McGuire, [e-mail]

1// 3-0L, 0D2c9-
�•V• r L
tei
NORTH CAROLINA_
ROY COOPER Environmental Quality
Coroner
MICHAEL S.REGAN
s-rr ary
LINDA CULPEPPER
interim Director
September 25, 2018
Jewel Cox
Former ExxonMobil Facility#46602
1016 W Poplar Ave.
Ste 106 Number 232
Collierville,NC 38017
Subject: Completion of 5I Injection Well Permit WI0400254
Guilford County
Dear Ms. Cox:
On February 15, 2013 the Water Quality Regional Operations Section(Former Aquifer Protection
Section) issued a 5I groundwater remediation permit to the above for the remediation of
contaminated groundwater located at 3701 High Point Rd., Greensboro, Guilford County, NC
27407. Expiration date for the permit was January 1, 2018.
A review of the file indicates the Final Project Evaluation (FPE) and injection well construction
have been received, per permit conditions. Therefore, this completes the monitoring and
compliance reporting requirements of your permit.
Please be advised this permit is now inactive.and no further injection activities are permitted. Any
future injection activities must be conducted per the requirements of 15A NCAC 2C .0225
(Revised May 1,2012). You must submit either(1) a Notification of Intent to Install groundwater
remediation wells, or (2) an Application for Permit to Construct Wells for Injection(whichever is
applicable).
.,11111k95
North Carolina Department of Environmental Quality I Division of Water Resources I AFOGW Section
512 North Salisbury Street 11636 Mail Service Center I Raleigh,North Carolina 27699-1636
919.707.9129

Please reference permit number WI0400254 in any future correspondence for this site. Operating
an injection well without a valid permit could result in a civil penalty of up to $25,000 per day.
Please contact Shristi Shrestha at 919-707-3662 or Debra Watts at 919-707-3670 if you have any
questions.
Best Regards,
Shristi Shrestha
Hydrogeologist
Water Quality Regional Operations Section
Division of Water Resources
cc: Sherri Knight, WQROS —Winston-Salem Regional Office
Central Office File, WI0400254
Scott Bostian, scott.bostian{-}a,arcadis-us.com,
Paul Goodell, Paul.Goodell{a,arcadis.com

ROY COOPER
Governor
MICHAEL S.REGAN
Secretary
Water Resources LINDA CULPEPPER
Environmental Quality Interim Director
March 21. 2018
CERTIFIED MAIL#7017 0190 0000 1635 4730
RETURN RECEIPT REQUESTED
Jewel Cox
Former ExxonMobil Facility#46602
1016 W Poplar Ave.
Ste 106 Number 232
Collierville,NC 38017
Subject: Notice of Expiration(NOE)
UIC In-situ Groundwater Remediation Injection Permit
Permit No. WI0400254
Guilford County
Dear Ms. Cox,
The Underground Injection Control (UIC) Program of the North Carolina Division of Water
Resources (DWR) is entrusted to protect the groundwater quality and resources of the State of
North Carolina, and is responsible for the regulation of injection well construction and operation
activities within the state. Our records indicate that the above-referenced operating permit for the
groundwater remediation injection well system located at 3701 High Point Rd., Greensboro, NC
27407 was issued on 2/15/2015, and expired on 1/31/2018.
Please note the following:
• If you intend to conduct additional injections or pursue any injection-related activities that
are beyond the scope of the permit referenced above, you should submit an application to
renew the permit (attached) and any well construction records (GW-1) or abandonment
records (GW-30) if not previously,submitted.
• If you do not intend to conduct further injection activities, per 15A NCAC 2C
.0225(k)(2)(C)and the MONITORING AND REPORTING REQUIREMENTS section of
your permit, a Final Project Evaluation (FPE) is due within 9 months after completion
of the injection operation. If it has been 9 months or longer since injection activities were
completed, please submit the required FPE. When all permit conditions are satisfied, a
Permit Completion Letter will be sent and the permit will be closed.
fiNothing Compares..
State of North Carolina I Environmental Quality I Division of Water Resources
Water Quality Regional Operations Section
1636 Mail Service Center I Raleigh,North Carolina 27699-1636
919-707-9129

If any injection well(s) are no longer being used for any purpose, they should be permanently
abandoned per to the regulatory requirements specified in rule 15A NCAC 02C .0240. If any
injection well(s) are permanently abandoned, copies of the GW-30s must be submitted to our
office. NOTE: injection wells using Direct Push/Geoprobe®technology are considered wells and
GW-1 and GW-30s are required to be submitted. However, if the well construction is essentially
the same, only one well construction/abandonment record is needed. Just indicate the number of
wells in the Remarks/Comments line of the form. If injection wells are converted to monitoring
or other use, indicate on Injection Well Status Form (GW-68). These can be downloaded from
DWR's website.
Within 30 days of receipt of this letter, please provide any of the above information and/or your
intentions regarding the permit. Send referenced forms to the address below:
Division of Water Resources
UIC Program
Attn: Shristi Shrestha
1636 Mail Service Center
Raleigh,NC 27699-1636
Please contact Shristi Shrestha, 919-807-6406, [e-mail] if you have any
questions. Please include the permit number on any correspondence, or in the subject line of any
emails regarding this permit.
Best Regards,
Oieksti
Shristi Shrestha
Hydrogeologist
Division of Water Resources
Water Quality Regional Operations Section
Enclosures
cc: Winston-Salem.Regional Office—WQROS w/out enclosures
Central Files - Permit No. WI0400254 w/out enclosures
Scott Bostian, [e-mail]

5I Final Project Evaluation (FPE) Checklist
pa--- k Fat z' 6 6 6 2
Permittee/Applicant: 6t Rw..-1.n -1�J- (-w_ _1 fit t - e d
Permit No.: WI 0 4-0 r; I./-
Reviewer Skyc:sM S}' e& 4
Date: _1 6 / ! 2
1. Permit issued date: 2_J / S / S Expiration date: t/ 3 / /24 / g
2. BIMS: Check to see if there is a current renewal Application for this permit number under
review or newer permit issued(search by facility address). If this is case, no FPE is needed at
this time. p(
3. Intent of Permittee for this site(i.e.,permit complete and no further injection activities planed,
future injection activities are planned,permit to be renewed,
etc.):— G .�
4. FPE or equivalent received? y Date rec'd: ±;t 2-f l g_. If FPE or AI is needed, date
sent: . Date received:
5. Proposed monitoring reports received for most part? 'i
6. Proposed sampling frequency complied with(e.g, monitored for 4 quarters)? y
7. Proposed parameters analyzed? _ '
8. Any clear evidence of daughter products? 11/A- If evident, RO may want to review.
9. GW-l s or equivalent received? '' Date: to/i-s d (Z 'As Built' well construction info
entered into BIMS? Y ; Date:
10. Status of injection wells (i.e., active- to be use as monitoring wells, abandoned,
etc.)_ - fwe E
11. If abandoned, GW-30s Records received?IN/ Date: Status entered/changed into
BIMS? Date:
12. (Note- if we have not received GW-1 s/30s for DPT wells, and obtaining these records is
problematic, injection well status forms (GW-68)may suffice. Bottom line,we need some
kind of a record, even e-mail from the Permittee/consultant,with info on when the well points
were plugged.)
13. Any outstanding NOVs in BIMS?
14. Completion Letter Sent date: 5 iL9- L BIMS updated- `Permit Completed' y
Notes:
w e u -r'a 4 bta e 662,1 c- 4 ,tfeefiG o ft t w .
Revised 9-20-2016

Shrestha, Shristi R
From: Goodell, Paul <[e-mail]>
Sent: Wednesday, September 12, 2018 12:55 PM
To: Shrestha, Shristi R
Subject: [External] RE:WI0400254 Former Exxon Mobil Facility#46602
CAUTION: External email. Do not click links or open attachments unless verified.Send all suspicious email as an attachment to
Hi Shristi,
We'll need to keep the well active and open for monitoring. It's being used for the incident under the UST
Section. If you need anything please let me know.
Thanks
Paul
From:Shrestha,Shristi R<[e-mail]>
Sent:Tuesday,September 11, 2018 10:25 AM
To:Goodell, Paul<[e-mail]>
Subject:WI0400254 Former Exxon Mobil Facility#46602
Good morning,
We have received a final project evaluation report for the above facility. Before we closeout the permit I need to
know the status of the wells. Do you want to keep the well active for monitoring?Or do you want to keep it open for
monitoring?
Shristi
Shristi R. Shrestha
Hydrogeologist
Water Quality Regional Operations Section
Animal Feeding Operations &Groundwater Protection Branch
North Carolina Department of Environmental Quality
919-707-3662 office
shristi.shrestha(wncdenr.rtov
512N. Salisbury Street
1636 Mail Service Center
Raleigh, NC 27699 1636
Email correspondence to and from this address is subject to the
North Carolina Public Records Law and may be disclosed to third parties.
This email and any files transmitted with it are the property of Arcadis and its affiliates.All rights,including without limitation copyright,are reserved.
This email contains information that may be confidential and may also be privileged.It is for the exclusive use of the intended recipient(s).If you are
not an intended recipient,please note that any form of distribution,copying or use of this communication or the information in it is strictly prohibited
and may be unlawful.If you have received this communication in error,please return it to the sender and then delete the email and destroy any
copies of it.While reasonable precautions have been taken to ensure that no software or viruses are present in our emails,we cannot guarantee
♦I...a N.:..........:t........._.a....I.......&:...........[........k........s k......:..a..........a..A.....k........A n.......................�L....:..[........a:....:..sk:..........:I N...a.1......a

ArsiSit
NCDENR
North Carolina-Department of Environment and Natural Resources
Division of Water Quality
Beverly Eaves Perdue Charles Wakild, P.E. Dee Freeman
Governor - Director Secretary
May 15,2012
Steve Shinners—Senior Manager
10990 Roe Avenue,Mail Stop A605 KCG
Overland Park,Kansas 66211
Ref: Issuance of Injection Permit WI0400246
Roadway facility
Kernersville, Forsyth County,North Carolina.
Dear Mr. Shinners:
In accordance with the application received on April 9,2012, and the supporting data received therein, we are forwarding
permit number WI0400246. This permit is to inject sodium persulfate, ferrous sulfate, and citric acid for facilitating the
oxidation of tetrachloroethene at the facility referenced above. This permit shall be effective from the date of issuance
until April 30, 2014, and shall be subject to the conditions and limitations stated therein, including the requirement to
submit a final project evaluation as stated in PART VII—MONITORING AND REPORTING REQUIREMENTS. Please
read the entire permit to ensure that you are aware of all compliance requirements of the permit.
You will need to notify this office by telephone 48 hours prior to initiation of operation of the facility. In order to
continue uninterrupted legal use of the injection facility for the stated purpose, you must submit an application to renew
the permit 120 days prior to its expiration date. Please contact me at 919-807-6352 or at [e-mail] if
you have any questions about your permit.
Best Regards,
1)(2,144;,......7AretAcrj
David Goodrich, L.G.
Hydrogeologist
cc: Sherri Knight, Winston-Salem Regional Office
Qu Qi,Division of Waste Management-Hazardous Waste Section
Ryan Gerber,ARCADIS G&M of North Carolina,Inc. , 801 Corporate Center Drive, Suite 300,Raleigh,NC 27607
WI0400246 Permit File
AQUIFER PROTECTION SECTION
1636 Mail Service Center,Raleigh,North Carolina 27699-1636
Location:512 N.Salisbury St.,Raleigh,North Carolina 27604 One
Phone:[phone] FAX:[phone] NorthCarolina
Internet www.ncwaterqualitvoro
Natul'dl f
An Equal Opportunity\Affirmative Action Employer

•
virmA
Ai;
NCDENR
North Carolina Department of Environment and Natural Resources
Division of Water Quality
Beverly Eaves Perdue Charles Wakild, P.E. Dee Freeman
Governor Director Secretary
April 17,2012
Steve Shinners—Senior Manager
YRC Enterprises Services,Inc.
10990 Roe Avenue
Mail Stop A605 KCG
Overland Par k,KS 66211
Subject: Acknowledgement of
Application No.WI0400246
Roadway-Kernersville
Injection In situ Groundwater
Remediation Well(5I)System
Forsyth County
Dear Mr. Shinners:
The Aquifer Protection Section acknowledges receipt of your permit application and supporting
documentation received on April 9, 2012. Your application package has been assigned the number listed
above, and the primary reviewer is David Goodrich.
Central and Winston-Salem Regional Office staff will perform a detailed review of the provided
application, and may contact you with a request for additional information. To ensure maximum
efficiency in processing permit applications, the Aquifer Protection Section requests your assistance in
providing a timely and complete response to any additional information requests.
Please note that processing standard review permit applications may take as long as 60 to 90 days
after receipt of a complete application. If you have any questions, please contact
David Goodrich at(919) 807-6352 or [e-mail].
Sincerely,
ow4 .
for Debra J.Watts
Groundwater Protection Unit Supervisor
cc: Winston-Salem Regional Office,Aquifer Protection Section
• Ryan Gerber—Arcadis G&M of North Carolina,Inc.
Permit File WI0400246
AQUIFER PROTECTION SECTION
1636 Mail Service Center,Raleigh,North Carolina 27699-1636
Location:512 N.Salisbury St.,Raleigh,North Carolina 27604 One
• Phone:[phone]\FAX:[phone]
Nar•...raiI
lina
Internet www,ncwatefauality o• Na ir�;�
An Equal Opportunity 1 Affirmative Action Employer

AQUIFER PROTECTION SECTION— GROUNDWATER PROTECTION UNIT
REGIONAL STAFF REPORT
Date: 04/30/2012 Permittee (s): YRC Enterprise Services, Inc.
Permit No.: WI0400246
To: APS Central Office County: Forsyth
Central Office Reviewer: David Goodrich Project Name: Roadway -Kernersville
Regional Login No:
I. GENERAL INFORMATION
1. This application is(check all that apply): ❑ SFR Waste Irrigation System ❑UIC Well(s)
®New ❑ Renewal
❑ Minor Modification ❑ Major Modification
❑ Surface Irrigation❑ Reuse❑ Recycle❑ High Rate Infiltration❑Evaporation/Infiltration Lagoon
❑Land Application of Residuals ❑Attachment B included ❑ 503 regulated ❑ 503 exempt
❑Distribution of Residuals ❑ Surface Disposal
❑Closed-loop Groundwater Remediation ® Other Injection Wells(including in situ remediation)
Was a site visit conducted in order to prepare this report? ®Yes or❑No.
a. Date of site visit: 4/27/2012
b. Person contacted and contact information: Ryan Gerber.ARCADIS G&M of North Carolina. Inc. 919-
415-2265 and Cliff Evans, facility manager,Roadway-Kernersville,336-669-9447 ,
c. Site visit conducted by: Shuying Wang
d. Inspection Report Attached: ❑ Yes or®No.
2. Is the following information entered into the BIMS record for this application correct?
® Yes or❑No. If no,please complete the following or indicate that it is correct on the current application.
For SFR Treatment Facilities:
a. Location:
b. Driving Directions:
c. USGS Quadrangle Map name and number:
d. Latitude: Longitude: Method Used(GPS, GoogleTM,etc.);
e. Regulated Activities/Type of Wastes(e.g., subdivision,food processing,municipal wastewater):
For UIC Injection Sites:
(If multiple sites either indicate which sites the information applies to, copy and paste a new section into the
document for each site, or attach additional pages for each site)
a. Location(s):
b. Driving Directions: RECEIVED/MN!
c. USGS Quadrangle Map name and number:
d. Latitude: Longitude: Method Used(GPS,GoogleTM,etc.); �R.�Y f' `
n►,lfor Protection
..ectiI3C?
II. NEW AND MAJOR MODIFICATION APPLICATIONS (this section not needed for renewals or minor modifications,
skip to next section)
APS-GPU Regional Staff Report(Sept 09) Page 1 of 6 Pages

AQUIFER PROTECTION SECTION- GROUNDWATER PROTECTION UNIT
REGIONAL STAFF REPORT
IV. INJECTION WELL PERMIT APPLICATIONS(Complete these two sections for all systems that use injection
wells,including closed-loop groundwater remediation effluent injection wells,in situ remediation injection wells,and heat
pump injection wells.)
Description of Well(s)and Facilities—New,Renewal,and Modification
1. Type of injection system:
❑ Heating/cooling water return flow(5A7)
❑ Closed-loop heat pump system(5QM/5QW)
®In situ remediation(5I)
❑ Closed-loop groundwater remediation effluent injection(5L/"Non-Discharge")
❑ Other(Specify: )
2. Does system use same well for water source and injection? ❑ Yes ®No
3. Are there any potential pollution sources that may affect injection? [ Yes ❑No
0
What is/are the pollution source(s)? CLOY
d/ 4 ' 2 -5441'. :tiv14 1/"."
What is the distance of the injection well(s)from the pollution source(s)? ..-----/ jt4. What is the minimum distance of proposed injection wells from the property boundary?
5. Quality of drainage at site: ❑ Good /1 Adequate ❑ Poor
6. Flooding potential of site: Low ❑ Moderate ❑ High
7. For groundwater remediation systems, is the proposed and/or existing groundwater monitoring program
(number of wells, frequency of monitoring, monitoring parameters, etc.)adequate? ® Yes ❑No. Attach
map of existing monitoring well network if applicable. If No, explain and recommend any changes to the
groundwater monitoring program:
8. Does the map presented represent the actual site(property lines,wells, surface drainage)?® Yes or❑No. If
no or no map,please attach a sketch of the site. Show property boundaries,buildings,wells,potential pollution
sources,roads, approximate scale, and north arrow.
Injection Well Permit Renewal and Modification Only:
1. For heat pump systems, are there any abnormalities in heat pump or injection well operation(e.g.turbid water,
failure to assimilate injected fluid,poor heating/cooling)?
❑ Yes ❑No. If yes, explain:
2. For closed-loop heat pump systems,has system lost pressure or required make-up fluid since permit issuance
or last inspection? ❑Yes ❑No. If}es,explain: _.
3. For renewal or modification of groundwater remediation permits (of any type), will
continued/additional/modified injections have an adverse impact on migration of the plume or management of
the contamination incident?❑Yes ❑No. If yes, explain: unlikely,but need to continue monitorinu the
process.
APS-GPU Regional Staff Report(Sept 09) Page 4 of 6 Pages

AQUIFER PROTECTION SECTION- GROUNDWATER PROTECTION UNIT
REGIONAL STAFF REPORT
4. Drilling Contractor: Name:
Address:
NC Certification number:
5. Complete and attach NEW Injection Facility Inspection Report,if applicable
V. EVALUATION AND RECOMMENDATIONS
1. Provide any additional narrative regarding your review of the Application:
2. Attach new Injection Facility Inspection Form, if applicable
3. Do you foresee any problems with issuance/renewal of this permit? ❑ Yes ®No. If yes,please explain
briefly.
4. List any items that you would like APS Central Office to obtain through an additional information request.
Make sure that you provide a reason for each item:
Item _ Reason _
5. List specific Permit conditions that you recommend to be removed from the permit when issued. Make sure
that you provide a reason for each condition:
Condition Reason
6. List specific special conditions or compliance schedules that you recommend to be included in the permit when
issued. Make sure that you provide a reason for each special condition:
Condition Reason
7. Recommendation: ❑Hold,pending receipt and review of additional information by regional office; ❑ Hold,
I� pending review of draft permit by regional office; ❑ Issue upon receipt of needed additional information;
Issue; ❑ Deny. If deny,please state reasons:
APS-GPU Regional Staff Report(Sept 09) Page 5 of 6 Pages

AQUIFER PROTECTION SECTION- GROUNDWATER PROTECTION UNIT
REGION AL STAFF REPORT
8. Signature of report Preparer(s): ,_541-7/ 4/f4,77
Signature of APS regional supervisor: <<Li.�
Date: 1 l 2_
VI. ADDITIONAL INFORMATION AND SITE MAP(Sketch of site showing house and waste irrkation
system,spray or drip field, location of well(s), and/or other relevant information-SHOWNORTHARRO di 1
APS-GPU Regional Staff Report(Sept 09) Page 6 of 6 Pages

Central Files: APS SWP
04/17/12
Permit Number WI0400246 Permit Tracking Slip
Program Category Status Project Type
Ground Water In review New Project
Permit Type Version Permit Classification
Injection In situ Groundwater Remediation Well (51) Individual
Primary Reviewer Permit Contact Affiliation
david.goodrich Ryan Gerber
Coastal SW Rule 801 Corporate Center Dr
Raleigh NC 27607
Permitted Flow
Facility
Facility Name Major/Minor Region
Roadway-Kernersville Minor Winston-Salem
Location Address County
1522 Hwy 66 S Forsyth
Kernersville NC 27284 Facility Contact Affiliation
Owner
Owner Name Owner Type
Yrc Enterprises Services Inc Unknown
Owner Affiliation
Steve Shinners
10990 Roe Ave
Dates/Events Overland Park KS 66211
Scheduled
Orig Issue App Received Draft Initiated Issuance Public Notice Issue Effective Expiration
04/09/12
Regulated Activities Requested/Received Events
RO staff report requested
RO staff report received
Out#all NULL
Waterbody Name Stream Index Number Current Class Subbasin

AQUIFER PROTECTION SECTION
APPLICATION REVIEW REQUEST FORM
Date: April 18,2012
To: ❑ Landon Davidson,ARO-APS El David May,WaRO-APS
❑ Art Barnhardt,FRO-APS El Charlie Stehman, WiRO-APS
❑ Andrew Pitner,MRO-APS X Sherri Knight, WSRO-APS
El Jay Zimmerman,RRO-APS
From: David Goodrich,Land Application Unit
Telephone: [phone] Fax: [phone]
E-Mail: [e-mail]_
A. Permit Number: WI0400246
B. Owner: YRC Enterprise Services.Inc.
C. Facility/Operation: Roadway-Kernersville
X Proposed ❑ Existing X Facility X Operation
D. Application:
1. Permit Type: ❑ Animal El Surface Irrigation ❑ Reuse ❑ H-R Infiltration
El Recycle El IIE Lagoon X GW Remediation(ND)5I Inj.wells
❑ UIC -(5A7)open loop geothermal
For Residuals: ❑ Land App. ❑ D&M ❑ Surface Disposal
❑ 503 ❑ 503 Exempt ❑ Animal
2. Project Type: X New ❑Major Mod. ❑ Minor Mod. ❑ Renewal ❑ Renewal w/Mod.
E. Comments/Other Information: ❑ I would like to accompany you on a site visit.
Attached, you will find all information submitted in support of the above-referenced application for your
review, comment,and/or action. Within 30 calendar days,please take the following actions:
X Return a Completed APSARR Form. - Please comment
El Attach Well Construction Data Sheet.
❑ Attach Attachment B for Certification by the LAPCU.
El Issue an Attachment B Certification from the RO.*
* Remember that you will be responsible for coordinating site visits and reviews, as well as additional
information requests with other RO-APS representatives in order to prepare a complete Attachment B for
certification. Refer to the RPP SOP for additional detail.
When you receive this request form, please write your name and dates in the spaces below, make a copy
of this sheet, and return it to the appropriate Central Office-Aquifer Protection Section contact person
listed above.
RO-APS Reviewer: Date:
FORM: APSARR 07/06 Page 1 of 1

August 7, 2008
MEMO To Injectant Health Risk Evaluation File
From Qu Qi
Y
gym,. k
Subject: Use Sodium Persulfate and Hydrogen Peroxide to remediate Soil and
Groundwater Contaminated with Chlorinated Solvents
Sodium persulfate was first used at Duracell site(WI0400045)in 2003. Based on a
letter sent to Mr. Thomas Wilson by Evan Kane,dated September. 19,2003, the
information on the proposed injectants—sodium persulfate and H2O2 to be used at
Duracell/Gillette Site was reviewed by Dr. Luanne Williams and she provided a health
risk evaluation on the products in a memo dated September 2, 2003. In the Memo she
did not mention that the products she evaluated were sodium persulfate and H2O2. But
site name and address(Duracell site).were in the subject line;based upon the information
in the file, sodium persulfate and H2O2 were the only injectants to be used. Therefore,
this September 2, 2003 Memo from Dr.Williams must be for the injectants sodium
persulfate and H2O2.
I have sent a request to the Epidemiology Section of the Division of Public Health for a
clarification on the matter. But I am confident that this evaluation is for sodium
persulfate.

Evan Kane
From: Luanne Williams[Luanne.Wiiliams@ncmail,netj
Sent: Monday,October 06,2003 10:10 AM
To: even kane
Subject: use of hydrogen peroxide
1111
Luanne.WiIliams.vcf
I am responding to your request regarding clarification on the hazards
associated with the injection of hydrogen peroxide for soil and
groundwater remediation. As mentioned in previous product reviews,
there is a potential explosion hazard associated with the injection of
hydrogen peroxide into contaminated soil and groundwater. In order to
reduce the explosion hazard risk, it Is recommended to keep the hydrogen
peroxide concentrations at a minimum. The literature suggests that the
explosion hazard increases greatly as the hydrogen peroxide
concentrations approach 30%. Hydrogen peroxide can be safely used by
taking necessary precautions mentioned in the product reviews. In
addition, site-specific conditions should be evaluated to guard against
explosion hazards,
1

d..swev
North Carolina Department of Health and Human Services
Division of Public Health •Epidemiology Section
1912 Mail Service Center•Raleigh,North Carolina 27699-1912
Tel [phone]•Fax [phone]
Michael F-Easley,Governor Carmen Hooker Odom,Secretary
September 2,2003
w �
MEMORANDUM rri"'
rn
TO: Evan Kane cI >o
Groundwater Section Mc
FROM: Luanne K.Williams,Pharm.D.,Toxicologist Oti ry rT
Medical Evaluation and Risk Assessment Unit
� o
Occupational and Environmental Epidemiology Branch
SUBJECT: Use of Products to Remediate Soil Contaminated with Chlorinated
Solvents at the Duracell Battery Manufacturing Facility in Lexington,
North Carolina
I am writing in response to a request for a health risk evaluation regarding the use
of non-biological products to remediate soil contaminated with chlorinated solvents at the
Duracell battery manufacturing facility in Lexington,North Carolina. Based upon my
review of the information submitted,I offer the following health risk evaluation:
1. Some effects reported to be associated with exposure to the proposed chemicals
are as follows:
• .Exposure can cause severe irritation and burning of skin, eyes, nose and throat
• (Meditext—Medical Management by Micromedex TOMEs Plus System CD-
ROM Database,Volume 57,2003;New Jersey Department of Health and
Senior Services Hazardous Substance Fact Sheet TOMEs Plus System CD-
ROM Database,Volume 57,2003).
• Inhalation exposure can cause coughing,wheezing, and/or shortness of breath
including asthma-like allergy.Future exposure can cause asthma attacks with
shortness of breath,wheezing, cough,and/or chest tightness (New Jersey
Department of Health and Senior Services Hazardous Substance Fact Sheet
TOMEs Plus System CD-ROM Database,Volume 57,2003).
• Inhalation exposure can cause severe respiratory irritation and inflammation
leading to shock,coma and seizures(New Jersey Department of Health and
Senior Services Hazardous Substance Fact Sheet TOMEs Plus System CD-
ROM Database,Volume 57,2003).
ter Location:2728 Capital Boulevard•Parker Lincoln Building•Raleigh,INC.27604 An Equal Opportunity Employer

2. Chemicals proposed for use may ignite combustibles such as wood,paper,oil,
etc).The chemicals proposed for use must be stored separately from hydrazine
and organic monomers since violent reactions can occur. The chemicals proposed
for use are not compatible with reducing agents;powdered metals;strong bases
such as sodium hydroxide and potassium hydroxide; alcohols; and hydrocarbon
fuels. Specifically,acetone, a soil contaminant at the injection site,readily forms
explosive peroxides with one of the chemicals being proposed for use. Injection
of the proposed chemicals into the soil contaminated area is likely to result in
an explosion. Based on the information submitted, the nearest property to
the injection area that is not owned and controlled by Duracell is
approximately 350 feet west of the injection area. Measures should be taken
to prevent fires and explosions from occurring to protect workers and
nearby community. A safety plan should be developed in case there is a fire
or an explosion. The addition of a metal as being proposed can act as a
catalyst which can increase the risk of an explosion during decomposition
(Hazardous Substances Data Bank TOMEs Plus System CD-ROM Database,
Volume 57,2003).
3. Store in tightly closed containers in a coot,well-ventilated area away from moist
air and combustibles. Containers may explode when heated.Runoff may create
fire or explosion hazard. Chemicals proposed for use heat up spontaneously when
decomposing and could start fires. (New Jersey Depaitrtient of Health and Senior
Services Hazardous Substance Fact Sheet TOMEs Plus System CD-ROM
Database,Volume 57,2003;Hazardous Substances Data Bank TOMEs Plus
System CD-ROM Database,Volume 57,2003).
4. If the products are released into the environment in a way that could result in a
suspension of fine solid or liquid particles(e.g.,grinding,blending,vigorous
shaking or mixing),then it is imperative that proper personal protective
equipment be used. The application process should be reviewed by an industrial
hygienist to(1)ensure that the injection process is done in a safe manner, and(2)
the most appropriate personal protective equipment is used.
5. Persons working with this product should at least wear goggles or a face shield,
gloves,and protective clothing. Face and body protection should be used for
anticipated splashes or sprays. Again,consult with an industrial hygienist to
ensure proper protection.
6. Eating,drinking, smoking,handling contact lenses, and applying cosmetics
should never be permitted in the application area during or immediately following
application.
7. Safety controls should be in place to ensure that the check valve and the pressure
delivery systems are working properly.
2

$. The Material Safety Data Sheets should be followed to prevent adverse reactions
and injuries.
9. Access to the area of application should be limited to the workers applying the
product. In order to minimize exposure to unprotected individuals,measures
should be taken to prevent access to the area of application.
10. According to the information submitted,there are no wells located on the
Duracell site used for drinking, industrial processes, cooking, or agriculture. The
nearest off-site well is approximately 1,500 feet north of the injection area. This
well is reportedly owned by J&S Motors and is the source of plumbed water. The
direction of groundwater flow from the injection area is also predicted to the
north. The wells are located close to the injection area and are north of the
injection area which is where groundwater flows. Therefore, efforts should be
made to prevent contamination of this existing well or future wells that may be
located near the application area.
11. According to the information submitted, the nearest surface water drainage is an
unnamed intermittent tributary of Fritz Branch that begins approximately 400 feet
north of the injection area. The nearest blue line stream is Fritz Branch,which
begins approximately 750 feet north of the injection area. The nearest
downstream waterbody is Abbotts Creek which runs into High Rock Lake. The
surface water bodies are located close to the injection area and are north of the
injection area which is where groundwater flows. Therefore, efforts should be
made to prevent contamination of the nearby surface waterbodies.
In summary,there are serious human health concerns regarding the use of the
chemicals proposed. Safeguards should be taken to prevent fires and explosions from
occurring. Please do not hesitate to call me if you have any questions at(919) 715-6429
LW:pw
cc: Mr. Stephen H.Barron,Manager
Environmental Health and Safety
305 E US Highway 64
Lexington,North Carolina 27292
Mr.Jack Riggenback
ERM EnviroClean
300 Chastain Center Blvd
Suite 375
Kennesaw,Ga 30144
3

Goodrich, David
From: Rudo, Ken
Sent: Tuesday, May 08, 2012 11:58 AM
To: Goodrich, David
Subject: Citric Acid/ PCE Degradation Request/Haviland Products
Mr. Goodrich I have evaluated the above application for the Citric Acid degradation site use at the Roadway—
Kernersville site. There does not appear to be any drinking water wells in use within 1500 feet of this site. As a result, after
reviewing the application, there should not be any adverse effects from the use of this product at this site. However, since
there was no data that indicated how effective this product would be in reducing tetrachloroethylene(PCE) levels at any
contaminated groundwater site, before this product is utilized at any other sites in North Carolina in the future, I would
recommend that groundwater testing be done at the Roadway-Kernersville site. This testing would be done to determine
the effectiveness of lowering PCE levels as well as any other contamination that may result due to the use of this citric
acid product for purposes of chemical degradation in the future. Sincerely Kenneth Rudo, Ph.D,Toxicologist, OEEB
Email correspondence to and from this address is subject to the North Carolina Public Records Law and may be disclosed to third parties by an authorized State
official.Unauthorized disclosure of juvenile,health,legally privileged,or otherwise confidential information,including confidential information relating to an ongoing
State procurement effort,is prohibited by law. If you have received this e-mail in error,please notify the sender immediately and delete all records of this e-mail.
1

ARCADIS ^ IVE m RID
RECENE�IDEHRIDWQ ARCADIS G&M of North Carolina,
Infrastructure Water-Environment Btail_'^r.�_ pp Inc..
APR0 9 2012 640 Freedom Business Center
Suite 310
Aquifer Protection Section King of Prussia
Pennsylvania 19406
UIC PROGRAM
Tel [phone]
DWQ-Aquifer Protection Section Fax [phone]
1636 Mail Service Center www.arcadis-us.com
Raleigh, NC 27699-1636
ENVIRONMENT
Subject:
RISK ASSESSMENT EVALUATION OF GROUNDWATER OR SOIL REMEDIATION
ADDITIVES CONTAINING Q MICROORGANISMS
Date:
Dear Mr. Slusser: April 5, 2012
I am writing to request for citric acid to be added to the North Carolina Department Contact:
of Environment and Natural Resources (NCDENR) approved injectants list. This Matthew Schnobrich
risk assessment evaluation is submitted concurrently with a UIC Permit Application
for the Roadway—Kernersville site(the Site). Monitored Natural Attenuation (MNA) Phone:
has been employed as a remediation strategy for elevated tetrachloroethene (PCE)
610.768.5813x378
concentrations since November 1995. However, PCE concentrations have not Email:
decreased significantly over time. In a letter dated May 24, 2011, NCDENR noted Matthew.schnobrich@arcadis-
that an active remedy should be considered to_expedite clean-up of groundwater us.com
impacts on the Site. ARCADIS is proposing to inject a solution of sodium
persulfate, ferrous sulfate, and citric acid as a part of an aggressive remedial Our ref:
strategy to expedite treatment within the source area. Citric acid is a critical CL000294.1001.00002
component of this injection solution as it is used to maintain a low pH and ensures
that ferrous iron is present in dissolved form to enable activation of the persulfate
anion to generate the sulfate radical. By comparison, citric acid is a weaker acid
than already approved acids(e.g., phosphoric acid, sulfuric acid)for injection use.
The following information is provided to support the risk assessment evaluation
required to request approval of citric acid for injection use.
Reouired General Information
1. Department of Environment and Natural Resources Aquifer Protection
Section contact person and phone number.
Thomas Slusser
UIC Program Manager
Aquifer Protection Section
512 N Salisbury St.
Raleigh, NC 27604
919-807-6412
Imagine the result
g:lenvlroadwaylkemersvillelpermitslcilric acid risk assessmentlrisk assessment evaluation final.dac

ARCADIS UIC Program
DWQ—Aquifer
Protection Section
April 5,2012
2. Current or future use of site with site contact person, address, and
phone number.
The facility currently functions as a freight terminal and includes three buildings.
One of the buildings serves as the garage, and the other two buildings serve as
freight handling and distribution loading docks. The former 1,000-gallon UST was
located near the southwest corner of the garage. The area surrounding the garage
and the two freight-handling areas is paved with a combination of concrete and
asphalt. The site is contained by a chain-link fence.
Site contact:
Cliff Evans
Roadway Express
1255 NC Highway 66 South
Kernersville, NC 27284
336-669-9447
3. Contractor applying product, contact person, address, and phone
number.
Ryan Gerber, PE
ARCADIS G&M of North Carolina
801 Corporate Center Drive, Suite 300
Raleigh, NC 27607
919-415-2265
4. Distance and likelihood of impact to public or private wells used for
drinking, industrial processes, cooling, agriculture, etc. Is area
served by public water supply? Verification must be provided by the
appropriate Regional Office of the Public Water Supply Section.
The Winston Salem/Forsyth County water system obtains water from two intakes
located on the Yadkin River and also from an intake located on Salem Lake. The
nearest intake,which is located at Salem Lake is approximately 5.5 miles west of the
site.
During a telephone interview on November 12,2007, Nick Seeba of the City of
Winston Salem Public Utilities indicated the entire area surrounding the Roadway
facility has access to public water supply.
During a receptor survey in November,2007 no known water supply wells were
discovered within 1,500 feet of the site. Michael Merrell of the Forsyth County
Department of Public Health Division of Environmental Health was contacted
February 10, 2012 to determine if there was any record of wells within 1,500 feet of
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ARCADIS UIC Program
DWQ—Aquifer
Protection Section
April 5, 2012
the site. There are no wells within 1,500 feet of the site in the Forsyth County record.
Forsyth County has kept records of construction and abandonment of wells since
1988. Mr. Merrell reported from personal knowledge that there are two bored wells
without record in the vacant property, 1200 Highway 66 South, directly across from
the site. He also mentioned that at the Ford dealership, 1330 Highway 66 South,
showed evidence of a well on a septic tank permit. The dealership was contacted
February 14, 2012; they had no knowledge of any wells on site.
Based upon these results, it is not likely that the injections will have an impact to wells
within the area.
5. General description of the contaminants if present in the soil and/or
groundwater at the site.
A former used oil tank which was contaminated with chlorinated solvents was
removed in 1992. The PCE release likely occurred before 1992. Residual
concentrations of tetrachloroethene(PCE) are present in saturated source area soil
and groundwater below a former used oil underground storage tank(UST).As a
result of the continuing source material, PCE concentrations in groundwater have
remained generally steady at levels above the North Carolina Administrative Code
(NCAC) Subchapter 2L Groundwater Standards since removal of the leaking UST
was conducted in 1992.
6. Name, approximate distance, and likelihood of impact to the nearest
body of surface water to the site.
GIS maps for areas adjacent to the Roadway—Kernersville facility revealed the
presence of surface water bodies approximately 2,250 feet east injection location.
This stream is identified as the West Fork Deep River.
An ephemeral storm water drainage ditch is located approximately 1,000 feet west
of the site. The likelihood of impact is very low as the drainage ditch is upgradient of
groundwater flow. In addition, based on the limited migration of PCE over the
historical monitoring period (17 years) and the current position of PCE impacts
entirely on-site, the likelihood of injected reagent migration outside of the immediate
injection area is very low.
As the iron-citrate complex will react with the co-delivered persulfate anion,
complete destruction is anticipated immediately or within weeks of delivery. In
addition, citric acid is biodegradable and will be consumed via the native
microorganisms as the injection solution mixes with and interacts with the native
hydrogeology. Due to both the limited migration via advective groundwater flow and
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G:lENVARoadwaylKernersvillelPermitslCitric Acid Risk Assessment\Risk Assessment Evaluation Final,doc 3/6

ARCADIS UIC Program
DWQ-Aquifer
Protection Section
April 5, 2012
to its short longevity, citric acid injection poses little risk of impact to nearby surface
water bodies.
7. Approximate distance to nearest residence(s)and workplace.
The nearest workplace is located approximately 300 feet southwest(sidegradient)
of the injection points.
Melissa Thompson Agency Inc-Nationwide Insurance
1261 Highway 66 S
Kernersville, NC 27284
The nearest residence is a mobile home park located approximately 2000 ft
southwest of injection points.
The approximate address of the mobile home park is
1308 Shields Rd
Kernersville, NC 27284
Required Product/Process-Specific Information
1. Product manufacturer name, address, phone number, and contact
person.
Terry Thompson
[phone]
Haviland Products Company
421 Ann St. NW
Grand Rapids, MI
2. Identity of specific ingredients (including CAS#) and concentrations
of ingredients contained in the product and purpose of each.*
Citric Acid (77-92-9)
3. Approximate concentration of each ingredient following release into
groundwater or soil.
0.5 g/L
4. Approximate distance and direction of travel for product in
groundwater, the groundwater concentration of each ingredient at
this distance, and distance from this point to the nearest drinking
water source (that is currently used for drinking purposes). These
should be reasonably accurate estimates based on the best available
information and calculations (modeling, if necessary) regarding
aquifer characteristics and flowpaths at the site; where uncertainty
exists in critical aquifer parameters (e.g. effective porosity),
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G:IENVIRoadwaylKernersvillelPermitslCilric Acid Risk AssessmentiRisk Assessment Evaluation Final.doc

•
ARCADIS UIC Program
DWQ—Aquifer
Protection Section
April 5, 2012
conservative assumptions should be made in estimating these values
so that worst-case predictions of travel distances are made.
Approximate distance and direction of travel for citric acid in water:
Groundwater flow at the injection wells is toward the southeast. Based on the
estimated hydraulic conductivity (K)for the targeted geology (1.86 x 10-5 cm/s) and
the observed groundwater gradient across the site(0.012 foot/foot), and an
effective porosity of 20%, the estimated groundwater velocity is 1.17 feet per year.
Based on the estimated groundwater velocity and a conservative citric acid
residence time of one month (30 days), the anticipated travel time of the citric acid
solution is 0.096 feet beyond the target radius of injection (7.5 feet).
There are no known wells used for drinking purposes within 1,500 feet of the Site.
The nearest intake for the Winston Salem/Forsyth County public water supply is
located at Salem Lake which is approximately 5.5 miles west of the site. This
location is upgradient of the flow of water on the Site.
5. Approximate groundwater concentration of each ingredient after
pumping or recovery(if applicable).
Not applicable.
6. If the product is expected to discharge to a nearby surface water,
approximate concentrations of product in the water.
As detailed above, citric acid is not anticipated to discharge to surface water bodies
within the site vicinity.
7. Documentation from authoritative technical references of specific
degradation products expected. (Provide a brief summary of the
referenced material as well as a copy of the referenced material.)
The proposed injection application includes the mixing of citric acid with ferrous
sulfate(FeSO4)prior to co-delivery with sodium persulfate. Reaction of ferrous iron
with citric acid will result in the formation of a tridentate iron complex(Francis,
1992). Once injected, reaction of the ferrous-citrate complex will result in
conversion to a tridentate ferric-citrate complex via oxidation and hydrolysis. The
ferric-citrate further hydrolyzes to a bidentate complex[Fe(OH)2-citrate]Z-, which is
both nontoxic and readily biodegradable (Francis, 1992). Under the oxic conditions
native to the site, rapid conversion of ferrous iron to the ferric form and subsequent
precipitation is anticipated above a pH of 4. Therefore, oxidation of the ferrous-
citrate compound will result in conversion to inert natural byproducts following
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G:1EN ARoadwaylKemersvillelPermits\Citric Acid Risk AssessmentlRisk Assessment Evaluation Final.doc

Jirt'iD'' UIC Program
DWQ—Aquifer
Protection Section
April 5, 2012
reaction with the oxidant and during interaction with the native hydrogeology at the
site. Further, citric acid may be metabolized in both aerobic and anaerobic
conditions, with both reactions terminating in carbon dioxide (Joshi-Tope, 1989).
8. Documentation from authoritative technical references of expected
migratory potential of specific ingredients and degradation products
in soil and groundwater. (Provide a brief summary of the referenced
material as well as a copy of the referenced material.)
The following references are included in Attachment 1:
Francis, A. J., Dodge, C.J., and J. B. Gillow. 1992. Biodegradation of Metal Citrate
Complexes and Implications for Toxic-Metal Mobility. Letters to Nature,
v. 356, p. 140-142.
Joshit-Tope and A.J. Francis. 1989. Mechanisms of Biodegradation of Metal-Citrate
Complexes by Psuedomonas Fluorescens. Journal of Bacteriology. v. 177,
p. 1989-1993.
9. Complete description of the use of the product at the site.
Please see the enclosed application for Permit to Construct Wells for Injection for a
complete description of citric acid use at the Site.
Sincerely,
ARCADIS G&M of North Carolina, Inc.
Matthew Schnobrich, PE
Senior Engineer
Attachments:. RECEIVED
,VE +
Attachment 1 —References APR 0 9 Crf
1OR
Aquifer Protc
Copies: wry
Mr. Qu Qi—NCDENR
Page:
6/6
G.IENVIRoadwaylKernersvillelPermitsTitric Acid Risk Assessment\Risk Assessment Evaluation Final doe

•
Attachment 1
Technical Journals

•
LETTERS TO NATURE
26.Dept of the Environment Waste Management Pap.No 6(HMSO,London,1976). 1.2
27.Abramowitz,D.A.Crit Rev.Biotech.10,241-251(1990).
2a Sedlelc,D.L&Andren,A.W.Ermir.Sat Technot 25,1419-1427(1991).
29.Mackay,D.&Clark,K.E.in Organic Contaminants in the Environment Ch.6(ed.tones,K.C.)
(Elsevier,1991). • • •
ACKNOWLEDGEMENTS.We Mark the AFRO,the NERC and the Warren Spring Laboratory/Department Citric acid
of the Environment for financial support
Biodegradation of metal citrate 0.8
complexes and implications for
toxic-metal mobility 1:2 Cu:Citrate(10 h)
A.J.Francis,C.J.Dodge& J.B.Mow U 0.4 1:2 Cu:Citrate
Department of Applied Science,Brookhaven National Laboratory,Upton,
New York 11973,USA .,
THE presence of synthetic and naturally occurring chelating agents
in nuclear and toxic-metal wastes is a major concern because of S
their potential to enhance mobilization of metal ions away from '' 1:1 Cu:Citrate
the disposal sites'.Of particular interest is citric acid,which is
present in low-level and transuranic radioactive wastes"and in 0'00 10 20 30 40
domestic and industrial wastes(as washing fluids, for instance), Time(h)
as well as being found naturally.Citrate ions form multidentate,
stable complexes with a variety of toxic metals and radionuclides; FIG. 2 Degradation of citrate in the presence of tridentate copper citrate
but biodegradation of these complexes, precipitating the metal complex.A 1:1 copper citrate complex is not biodegraded,The excess free
ions as insoluble hydroxides, oxides or other salts, may retard citrate(0.52 mM)in a 1:2 copper citrate complex Is completely degraded,
migration.Here we report a study of the biodegradation of citrate indicating that the complex is not toxic.
complexes of Ca,Fe(n), Fe(H):), Cd,Cu,Ni,Pb and U.Several
of these complexes were not readily degraded by bacteria,and the
biodegradability depended on the chemical nature of the complex,
not on the toxicity of the metal to the bacteria.This resistance to found by potentiometric titrations using a 1:1 metal citrate
biodegradation implies that citrate complexation may play an complex(containing 0.52 mM of metal and of citric acid).Addi-
important part in migration of these hazardous wastes. tional information on the nature of the complexes was obtained
Mixed cultures of sewage bacteria and axenic cultures of from published data. Calcium, ferric iron and nickel formed
Pseudomonas sp.and P.pseudoakaligenes have been found6 to bidentate, mononuclear complexes with two carboxylic acid
metabolize metal citrate complexes of Ca, Fe, Al and Mg at groups of the citric acid molecule8-13. Copper, ferrous iron,
different rates and extents. Klebsiella sp., also isolated from cadmium and lead formed tridentate, mononuclear complexes
sewage, degrades magnesium citrate but fails to degrade Cd, with citric acid involving two carboxylic acid groups and the
Cu, or Zn citrate complexes'. These studies showed that the hydroxyl group8•14-16. Uranium formed a binuclear complex
lack of biodegradation was not due to toxicity of the metal.The with two uranyl ions and two citric acid molecules involving
nature and biodegradability of the complexes formed between four carboxylic acid groups and two hydroxyl groups17-19
the metal and citric acid were not,however, investigated. We isolated a citrate-metabolizing bacterium, P.fluorescens
Citric acid forms mononuclear,binuclear orpolynuclear com- (ATCC no.55241),from the leachate sample collected from the
plexes depending on the type of metal (Fig. 1). The types of low-level radioactive waste disposal site,West Valley,New York.
complexes formed between equimolar amounts of citric acid The bacterium degraded citrate at a maximum rate of
and Ca', Cd", Cu", Fe2+, Fe3+, Ni2+, Pb24 or UOZ+ were 57 µmol h-1. To investigate its ability to degrade several metal
a CH2cooH b
H,O., .00C OH
HO HO—C—COON _M:
CH2COOH H2O",.• OOCCH2 H2CCOO-
FIG. 1 Types of metal citrate com-
plexes.a,Citric acid;b,bidentate com- Citric acid Bidentate complex
plex involving two carboxylic acids
groups;c tridendate complex involving
two carboxylic acids and the hydroxyl C d O 0
group;cl binuclear complex with UO2+. —O. :'0~_C-
H2C UO2 H I
,OOC 0' `•0�C—CH2C00-
'OOCH2C rCH/ •.•
H2O----MT- -OOC\ C H2 I UO� 1 H2
•,0/C—CH2 COO- O�iC'`�O' 0_
Tridentate complex Binuclear complex
140 NATURE • VOL 356 • 12 MARCH 1992
0 1992 Nature Publishing Group

LETTERS TO NATURE
TABLE 1 Biodegradation of 1:1 metal citrate complexes.
Formation constant Biodegradation rate
Type of complex Metal Formula (log K) (µmol h-1) (%) Reference
Bidentate Calcium (Ca city- 3.5 77 t 10* 100 6,8
Nickel (Ni cit)- 5.4 9 f 1 70 12,13
Fe3+ (Fe(OH)2 cit)2- 1.9-2.6 16 t 1 100 6,11
Tridentate Fe3+ (Fe(OH)city- 9.4 1 t 0 16 6,9
Fe2+ (Fe city' 4.4 ND 0 9,14
Copper (Cu cit)2- 5.9 ND 0 7,8
Cadmium (Cd cit)- 3.7 ND 0 7,15
Lead (Pb cit)- 4.4 ND 0 16
Binuclear Uranium ((UO2)2 cit2)2- 7.4 ND 0 18
ND,none detected.
*Values are mean f1 standard error of the mean(SEM).
citrate complexes we grew the bacterium in a defined medium The tridentate complexes of Cd, Cu and Pb,but not that of
developed in accordance with equilibrium calculations so that Fe(tt), were completely resistant to degradation. Ferrous iron
the metal citrate complex was the predominant species6. The initially formed a tridentate complex (Fe city. Oxidation and
phosphate buffer and inorganic phosphate in the medium were hydrolysis of the ferrous iron resulted in the formation of a
replaced with PIPES buffer and glycerolphosphate to prevent tridentate ferric complex(FeOH cit)-,which was further hydro-
the precipitation of metals.The pH was adjusted to 6.1 and the lysed to a bidentate complex(Fe(OH)2 cit)2-.The rate of degra-
culture was incubated in triplicate at 26°C, Citric acid was dation of the ferrous complex depended on the rate of conver-
analysed by high-pressure liquid chromatography, metals by sion to the more hydrolysed form of the ferric complex. The
atomic absorption spectroscopy and uranium by spectro- presence of bacteria accelerated this conversion.
photometry or by inductively coupled plasma mass spectro- Although the copper citrate complex was not biodegraded
metry. (Fig. 2), adding 1:1 copper citrate complex to the culture
The results in Table 1 show that the bidentate complexes of medium, which contained 0.52 mM of uncomplexed citrate,at
Ca,Fe(m) and Ni were readily degraded,with calcium citrate the start of the experiment and after 10 h affected neither the
showing the fastest rate. Ferric citrate was completely degraded growth nor the rate and extent of degradation of excess free
but at a slower rate. The iron that was released formed a citrate. The lack of degradation of the copper citrate complex
hydroxide precipitate, In contrast, nickel citrate complex was not due to its toxicity.Cadmium-and lead-citrate complexes
showed only 70%degradation. During the degradation of this were not biodegraded,but they have been shown to form poly-
complex the pH of the culture medium increased to 7.6,causing nuclear complexes at higher pH with lead citrate precipitating
changes in the complex structure. At acid to neutral pH the out of solution'5.16,21 22 In the presence of excess citric acid,
complex is present as bidentate (Ni citr. At alkaline pH, these metals also formed 1:2 mononuclear biligand complexes
however,the hydroxyl group was ionized,forming a tridentate such as (Cd cit2)°-.Glucose added to the growth medium was
complex8'20,and was resistant to further biodegradation. readily metabolized by the bacteria, suggesting that these
10`r— — ,
1.2 -a ,— h .
.
•
Citric acid
E S .
E b
0.8 a 10
o •.
a'co
i U •
-0El N
U .
12 U:Citrate(13 h) E 10•
04. o
z 1
1:2 U:Citrate — o— Citric acid
• —lb— 1:2 U:Citrate
—A— 1:2 U:Citrate(13 h)
1:1 U:Citrate
0 00 10 20 30 40 1030 10 20 30 40
Time (h) Time(h)
FIG.3 a,Degradation of citrate in the presence of uranium citrate.The 1:1 6,Increase in bacterial cell number is observed because the available free
complex was not biodegraded. Incomplete degradation in the presence of citrate is metabolized.The uranium citrate complex is not toxic to bacteria.
excess free citrate is due to the formation of a 2:3 uranium citrate complex. Error bars are f1 standard error on the mean.
NATURE • VOL 356 • 12 MARCH 1992 141
0 1992 Nature Publishing Group

LETTERS TO NATURE
complexes too are non-toxic. lack of metabolism of the complex by the bacteria14,24-2e In
Uranium formed a binuclear complex((UO2)2 cit2)2-and was addition to its presence in hazardous and mixed wastes, citric
not degraded by the bacteria(Fig.3a).In the presence of excess acid can be produced from the degradation of organic con-
citrate, —35% of the citrate was metabolized regardless of stituents of the waste. Citric acid can thus facilitate transport
whether the 1:1 uranium citrate complex was added after 0 h of certain toxic metals and radionuclides from the waste disposal
or 10 h of growth.The number of bacteria increased,indicating sites by forming stable complexes which are resistant to bio-
the absence of toxicity,but growth was limited by the presence degradation. 0
of uncomplexed citrate (Fig. 3b). It seems that U formed a
polynuclear 2:3 complex in the presence of excess citrate, Received 12 November 1991;accepted 17 January 1992.
thereby limiting the amount of free citrate available for biodegra- 1.Means,J.L.,Crerar,D.A.&Duguid J.O.Science 200,1477-1481(1978).
dation 3. Further experiments with varying uranium:citrate 2.Francis,A.J.in Solt Reclamation Processe&McroblologicalAnallyses end Applications(eds Tate,
ratios also confirmed this observation. R.L.&Klein,D.L.)279-331(Marcel Dekker,New York,1985).
3.Francis,A.J.Experlenda 45,840-851(1990).
Several other bacterial cultures isolated from wastes and 4.DOE Office of Energy Research DOE/ER-0492T(IS Department of Energy.Washington DC,1991).
leachates containing toxic metals and radionuclides degraded 5.DOE Office of Energy Research,OOE/ER-0419(US Department of Energy,Washington DC,1989).
III) and Ni, but not those of Cd,of Ca, Fe citrate loom taxes 6.Madsen,E.&Alexander,M.Appt emir.Microbial.50,342-349(1985).
P ( 7.Brynhildsen,L.&Rosswall,T.Appl,envN.Microbial 55,1375-1379(19E19).
Cu,Pb and U. On the basis of the formation constant(log K) 8.Cempi,E.,°staeoll,G.Meirone,M.&Salni,G.1 bang nucl.Chem.26,553-554(1964).
of the metal citrate (Tablebiodegradation complexes 1),biode radation should a Ham,R.E,Shull,C.M&Grant,D.M.1 Am.chem.Soc.78,2111-2114(1954).
10.Strouse,A.Layton,S.W.&Strouse,C.E..1 Are ahem Sec.99,562-571(1977).
follow the order Fe(nz)(OH)2>Ca>Cd>Pb=Fe(ir)>Ni> ii.Dhar.S.K.&Slot,S.I inorg nucL Men,41,126-127(1979).
Cu>U(vz)>Fe(m)OH. Instead we observed the following 12.Li.N.C..Linder&aum,A.&White,J.IL.1 inorg nice Chem.12,122-128(1959).
13.Hedwlg,G.R,Liddle,J.R.&Reeves.R.D.Austr.A Chem.33,1685-1693(1980).
order:Ca>Fe(ttr)(OH)2>Ni>Fe(nt)OH,and the complexes 14.Villafrenca,1 J.&Mtldvan,A.S.I Not Chem.247,3454-3463(1972).
Fe(xi), Cu,Cd, Pb,and U(vt)were not degraded. 15.Capone.S.,DeRaberUs,A,DeStefeno,C.&Sammartano,S.Talents 33,763-767(1986).
These results show that the t YP e of cam lex formed between 16.Ekstrom,L.G.&Olin,A.Chem Script 18,10-15(1978-79).
P 17.Rajan,K.S.&Martell.A.E.lnorg.Chem 4,462-469(1965).
the metal and citric acid is important in determining its bio- 18.Nunes,M.T.&Gill,V.M.S.Morg claim Acta 129,283-287(1987).
degradability.The presence of a free hydroxyl group of the citric 19.Sillen,L.G.&Martell,A.E.Stability Constants of Metal-ion Complexes Spec Publ.No.25,SuppL
No.1.(Chemical Society.London,1971).
acid seems to be the key to biodegradation of the metal complex. 20.Still,E R.&wilcberg,P.hag enlm Acts 46,153(1980).
For example,Ca,Fe(III)and Ni formed mononuclear bidentate 21.Grenthe,I.,Wiltberg,P.&SUI1 E.R.berg.shim Acta 91,25-31(1984).
22.Battari,E Ann.Chin.66,593-607(1975).
complexes and were readily biodegraded, whereas Ca, Cu, 23.Heltner,C.&Bobtelaky,M.Bull Soo Chlm.France 356,23(1954).
Fe(rr)and Pb,which formed mononuclear tridentate complexes, 24.Bergsema,J.&Konings,W.N.fur. elocbem 184,151-156(1983).
and U, which formed a binuclear complex involvingthe 25.WilieckGlusite J.P. he them. Oehr, 345-352 ern.2.
P 26,willedce,K.Ghee,E M&Oehr,P. blot Chem.248,807-834(1973).
hydroxyl group of the citrate, were not biodegraded. The lack
of degradation of tridentate and binuclear complexes was not ACKNOWLEDGEMENTS.This research was performed under the auspices of the Environmental
Sciences Division's Subsurface Science Program,Office of Health and Environmental Research,Office
due to toxicity, but probably limited by the transport and/or of Energy Research,US Department of Energy.
A shear-strain anomaly following resolution and long-term stability better than 100 ne per years.
Raw data from the instrument consist of diameter changes in
the Loma Prieta earthquake three directions at 120° to each other in the horizontal plane.
These are reduced to areal strain ea, and engineering shear
R. L.Gwyther, M.T.Gladwin* & R.H.G.Hart strains y, and y2 (roughly parallel to and at 45°to the fault,
respectively). The strain meter is grouted into the surrounding
rock, and this instrument inclusion is softer to shear than to
Department of Physics,University of Queensland,4072,Australia compression. Observed strain components are therefore scaled
BOREHOLE tensor strain instruments deployed along the San by hole coupling parameters4 determined by tidal calibration.
Andreas fault for the past ten years have provided sufficient Data from borehole inclusions are initially dominated by
resolution and stability to sample regional tectonic processes, grout compression of the instrument, by thermally controlled
potentially enhancing earthquake prediction capability. Data decay as the instrument site re-establishes equilibrium with its
obtained from the instrument at San Juan Bautista,in the near- surroundings and by an exponential recovery of the virgin stress
field region of the 17 October 1989 Loma Prieta earthquake field relieved at the borehole during the drilling process .The
(Ma=7.1) provide the first opportunity to observe shear strain exponential signals have no relevance to the monitoring of
processes associated with a large earthquake.We previously repor- regional strain changes and were removed by an exponential
lath a prominent shear-strain anomaly in those data for more than least-squares fit over the interval January 1984 to February 1988
a year before the earthquake. This anomaly ceased immediately (ref. 1).These residuals were then reduced to the strains ea. YS
after the earthquake,but,as we report here,a new and higher rate and y2 shown in Fig. 1 a.
of fault-parallel shear accumulation(2 microstrain per year)was The dominant signals present are the coseismic strain steps
established about four months later and has continued to the of the Loma Prieta earthquake which in Fig. lb are removed
present.Associated changes in creep rate are apparent at a number from the data to make long-term trends more apparent. As
of sites on the surface trace of the fault within 30 km of the strain previously reported,an anomalous change in y1 is apparent by
meter.We propose that the observed strain accumulation results late 1988, showing a remarkably linear strain accumulation of
from increased slip around a nearby locked section of the fault, 1 µt per year'.A smaller change of^0.3 Ike per year was also
this slip arising from loading by the failed Loma Prieta source evident from mid-1988.The azimuth of maximum shear for the
region to the north.This model is consistent with suggestions of accumulating shear strain was roughly parallel to the local San
an increased probability of a moderate earthquake near San Juan Andreas strike.
Bautista9"O, and with evidence12 that interactions between fault Immediately after the earthquake,the strain rate returned for
regions are important in earthquake processes. —10 days to its value before the anomaly, then decreased for
A Gladwin borehole tensor strain meter2 installed near the two months.By May 1990,following the Chittenden aftershock
San Andreas fault at San Juan Bautista in late 1983 has provided sequence,the present linear shear accumulation rate of—2 Ile
continuous areal and shear strain data with sub-nanostrain(ne) per year with the original sense had been established. During
the whole of the period since mid-1988,medium-term changes
in Ea and y2 have been less than 250 ne per year. The strain
•To whom correspondence should be addressed. data indicate that by May 1990, the accumulating strain was
142 NATURE • VOL 356 • 12 MARCH 1992
0 1992 Nature Publishing Group

•
JOURNAL OF BACTERIOLOGY,Apr.1995,p. 1989-1993 Vol.177,No.8
0021-9193/95/$04.00+0
Copyright©1995,American Society for Microbiology
Mechanisms of Biodegradation of Metal-Citrate
Complexes by Pseudomonas fluorescens
GEETA JOSHI-TOPE AND AROKIASAMY J. FRANCIS*
Department of Applied Science, Brookhaven National Laboratory, Upton, New York 11973
Received 3 October 1994/Accepted 4 February 1995
Biodegradation of metal-citrate complexes by Pseudomonas fluorescens depends on the nature of the complex
formed between the metal and citric acid. Bidentate Fe(HI)-, Ni-, and Zn-citrate complexes were readily
biodegraded,but the tridentate Cd-and Cu-citrate,and U-citrate complexes were not.The biodegradation of
Ni-and Zn-citrate commenced after an initial lag period;the former showed only partial (70%)degradation,
whereas the latter was completely degraded. Uptake studies with 14C-labeled citric acid and metal-citrate
complexes showed that cells grown in medium containing citric acid transported free citric acid at the rate of
28 nmol min-1 and Fe(HI)-citrate at the rate of 12.6 nmol min-1 but not Cd-, Cu-, Ni-, U-, and Zn-citrate
complexes.However,cells grown in medium containing Ni-or Zn-citrate transported both Ni-and Zn-citrate,
suggesting the involvement of a common,inducible transport factor.Cell extracts degraded Fe(III)-,Ni-,U-,
and Zn-citrate complexes in the following order: citric acid = Fe(HI)-citrate > Ni-citrate = Zn-citrate >
U-citrate.The cell extract did not degrade Cd-or Cu-citrate complexes.These results show that the biodeg-
radation of the U-citrate complex was limited by the lack of transport inside the cell and that the tridentate o
Cd-and Cu-citrate complexes were neither transported inside the cell nor metabolized by the bacterium.
0
Citric acid is a multidentate ligand and forms stable corn- U-, and Zn-citrate complexes by P.fluorescens to provide an co
plexes with various metal ions (19). The rate and extent of explanation for the observed differences in the biodegradation c
biodegradation of several metal-citrate complexes by microor- of metal-citrate complexes. 3
ganisms vary.For example,Pseudomonas pseudoalcaligenes de-
graded Mg-citrate at a much lower rate than Ca-,Fe(III)-,and
Al(III) citrate (29). Studies with a Klebsiella sp. showed that MATERIALS AND METHODS c
citric acid and Mg-citrate were readily degraded,whereas Cd-,
Culture conditions.P.fluorescens biovar II(ATCC 55241)capable of metab- cr
Cu-,and Zn-citrate were resistant to biodegradation(9).Both olizing citric acid as a sole source of carbon was grown in modified Simmon's
these studies also showed that metal toxicity was not respon- citrate medium or in a defined mineral salts medium.The modified Simmon's
sible for the lack of or the lower rate of degradation of certain citrate medium (pH 6.2)contained(per liter)citric acid,2 g;MgSO4,0.2 g; 0
metal-citrate complexes but gave no other explanation. Bio- NIi4CI.1 g;K2HPO4,1 g;and NaCI.5 g.The defined mineral salts medium
degradation studies with Pseudomonas fluorescens showed that contained(per liter)NHaCI,35.8 mg;CaC12 2H2O,2.75 mg;MgCl,•6H2O,
6.25 mg;PIPES [piperazine-N,N'-bis(2-ethanesulfonic acid)]buffer(disodium
bidentate complexes of Fe(III)-, Ni-, and Zn-citrate were salt;Sigma Chemical Co..St.Louis,Mo.).1.47 mg;glycerol phosphate,1.74 mg;
readily biodegraded, whereas complexes that involve the hy- FeSO4 7H20, 1.49 mg; MnSO4•H2O, 1.155 mg; CuC12.2H20, 0.101 mg;
droxyl group of citric acid, the tridentate Cd- NaMoO4.2H20,0.0945 mg;ZnSO4.H2O,0.103 mg;and CoC12.6H20,0.151
and Cu-citrate mg.The final concentration of citric acid or metal-citrate complex in the medium ccoo
complexes,and U-citrate complex were not biodegraded(17). was 0.52 mM.The ionic strength of the medium was adjusted to 0.1 M by the
No relationship between biodegradability and stability of the addition of KCI,and the pH was adjusted to 6.1 with KOH.The composition of
complexes was observed. The tridentate Fe(ll)-citrate corn- the defined mineral salts medium was developed in accordance with equilibrium
plex, although recalcitrant,was readily biodegraded after oxi- calculations to ensure the integrity of the complex so that the cation complex was
the predominant species(29).
dation and hydrolysis to the bidentate Fe(III)-citrate form, Metal-citrate complexes.Stock solutions.13.0 mM of citric acid and each of
denoting a structure-function relationship in the metabolism of the following metals,were prepared in deionized water:Cd(NO2)2.41420(Alfa
the complex(16). Products); Ni(N0)2.6H20 (Alfa Products); CuCl2.21-120 (Mallinckrodt);
Microorganisms have selective transport systems for the up Fe(No,),•9H2O(Mallinckrodt);Pb(NO,)2(Mallinckrodt);UO2(NOa)2•6H2O
take of metals of known biological function(11,23,28).Heavy (sDH Chemicals,Analar,Poole,England);and ZnSO4 H2O(Matheson).Met-
g ) al-citrate complexes were prepared by adding equimolar amounts of citric acid
metals are transported into cells by preexisting transport sys- and metal solutions and diluting the mixture to 5.2 mM.The ionic strength was
tems and are subsequently either chemically neutralized, se- adjusted to 0.1 M by adding KCI,and the pH was adjusted to 6.1.
questered, or removed from the cells by rapid efflux (6, 35). Speciation of the metal in the defined mineral salts medium (pH 6.1)was
verified by using the MINTEQA2 program (1). Glycerol phosphate was not
Metals affect the transport of citric acid in bacteria.For exam- found in the database and was not included.However,it is present at a very low
ple, the presence of divalent cations influences citrate trans- concentration(5.7 p,M)relative to citric acid(0.52 mM)and is not expected to
port in Bacillus subtilis(5,35).The inability of microorganisms have any effects.PIPES buffer was not included because it does not complex with
to metabolize certain metal-citrate complexes, such as triden- am
metals function(20).of ThepH d(istributH2.0 to 9ion of.0).theThermodynamic major metal speciesconstants alsoin wasthe detMINTerminedEQA2 as
p
tate Cd- and Cu-citrate complexes or the U-citrate complex, database were used in the calculations.The following formation constants were
may be due to lack of transport of the metal-citrate complexes used:for Fe(III),log K=9.4 for[Fe(OH)Citl-,where Cit is citrate,and log K
inside the cell or the inability of the citric acid-degrading en- = 2.6 for[Fe(OH)2Cit]2-; for U-citrate, log K = 7.4 for the mononuclear
zymes to utilize the complexes as substrate or both.We inves- (UO2Cit)-and log Kf=18.9 for the binuclear[(UO2)2(Cit)212-(16,22,32,33).
tigated the transport and metabolism of Cd-,Cu-,Fe(III)-,Ni-, Biodegradation of metal-citrate complexes.Biodegradation of metal-citrate
complexes was determined by aseptically adding 20 ml of 5.2 mM filter-sterilized
metal-citrate complex to 180 ml of the defined mineral salts medium in 500-m1
Erlenmeyer flasks.The final concentration of the metal-citrate complex in the
medium was 0.52 mM.Two milliliters of log-phase cells(18-h culture)grown in
*Corresponding author.Phone:[phone].Fax:[phone]. the defined mineral salts medium containing 0.52 mM citric acid was added to(i)
1989

1990 JOSHI-TOPE AND FRANCIS J.BACFERIOL.
defined mineral salts medium containing 0.52 mM citric acid and(ii)defined TABLE 1. Speciation of 1:1 metal-citrate complexes in defined
mineral salts medium containing 0.52 mM metal-citrate complex.Uninoculated mineral salts medium at pH 6.1 by MIN TEQA2(1)
medium containing the metal-citrate complexes was used as a control. The
samples were incubated in triplicate,at 26±1°C in the dark,on a rotary shaker. Complex Metal species %of total
Aliquots were withdrawn periodically,filtered through a 0.22-p.m-pore-size Mil- metal
lipore HA filter,and analyzed for citric acid.At the end of the experiment,1-ml
aliquots were centrifuged at 13,000 X g for 5 min at room temperature.The Cd-citrate (CdCit)- 56.5
supernatant and pellet were separated,acidified with HCI,and analyzed for the Cd2+ 42.2
metal. CdCI+ 1.1
Biodegradation of the metal-citrate complexes also was studied by using rest-
ing cells. Cells were grown in the defined mineral salts medium overnight, Cu-citrate (CuCit)- 93.6
harvested by centrifugation,washed,and resuspended in the defined mineral Cu" 6.0
salts medium without citric acid and NH4CI.The density of the cell suspension
was adjusted to an optical density of 0.2 at 600 nm.Ten-milliliter aliquots of the Fe(III)citrate (FeOHCit)- 68.2
cell suspension were then incubated with 0.52 mM metal-citrate complex in 2_
triplicate,at 26±1°C in the dark.Aliquots were removed periodically,filtered, [Fe(OH)2Cit] 31.3
and analyzed for citric acid.
Adaptation of bacteria to Ni-and Zn-citrate.To determine whether cells Ni-citrate (NiCit)- 88.0
grown in media containing Ni-or Zn-citrate could be adapted to biodegrade Ni- Ni" 12.0
and Zn-citrate without a lag,the following experiment was performed:(i)2 ml of
citric acid-grown log-phase cells(16-h culture)was inoculated into 200 nil of U-citrate [(UO2)2(Cit)2]2- 58.0
mineral salts medium containing 0.52 mM citric acid,0.52 mM Ni-citrate,or 0.52 [UO2Cit]- 17.6
mM Zn-citrate;(ii)2 ml of Ni-citrate-grown log-phase cells(48-h culture)was [(UO2)3oH]5- 22.8
inoculated into 200 ml of mineral salts medium containing 0.52 mM citric acid,
0.52 mM Ni-citrate,or 0.52 mM Zn-citrate;and(iii)2 ml of Zn-citrate-grown
log-phase cells(48-h culture)was inoculated into 200 ml of mineral salts medium Zn-citrate (ZnCit)- 79.4
containing 0.52 mM citric acid, 0.52 mM Ni-citrate, or 0.52 mM Zn-citrate. Zn2+ 20.5
Samples were incubated in triplicate at 26±1°C in the dark on a rotary shaker.
Aliquots were withdrawn at periodic intervals,filtered through a 0.22-p.m-pore- a
size Millipore HA filter,and analyzed for citric acid. o
In addition,we investigated whether the adaptation was reciprocal by inocu-
lating cells grown in the presence of Ni citrate into medium containing Zn- degradation was determined by high-pressure liquid chromatography(HPLC).
For this purpose,500-µl aliquots of the extract were incubated with 14 µM m
citrate,and vice versa. a
MnSO4,40 p.M NADP,and 2.08 mM metal citrate complex in 30 mM Tris-HCI CD
Transport of metal-citrate complexes.Transport of metal citrate complexes by (pH 7.4)at a final volume of 3.0 ml.At periodic intervals,250-µl aliquots were n
the bacterium was determined by monitoring the uptake of 14C-labeled metal- removed, acidified with 750 µl of 0.003 N H2SO4,and centrifuged, and the p
citrate complexes(21,37).Labeled complexes with a specific activity of 0.192 supernatants were analyzed for citric acid by HPLC.The boiled extract as well as 3
mCi/mmol were prepared by mixing a stock solution of the metal with a stock reagents without the cell extract was used as a control. The experiment was `v
solution of[1,5- C]citric acid(Sigma Chemical Company).Cells grown over- repeated three times. coas
night in modified Simmon's citrate medium were harvested by centrifugation, To determine the fate of the metal,250-µl aliquots of the reaction mixture 3
washed,and resuspended in the mineral salts medium without citric acid.The were removed at the end of incubation and centrifuged at 13,000 X g for 5 min b
density of the cell suspension was adjusted to an optical density of 0.15 at 600 nm, at room temperature.The supematant and the pellet were separated,acidified, tn3
and aliquots of the cell suspension were used in transport assays for the metal- and analyzed for the metal. cr
citrate complexes.Nine milliliters of the cell suspension was dispensed into a Analyses.Metals were determined by atomic absorption spectroscopy,and o
15-m1 scintillation vial,which was placed in the side arm of a biometer flask m
iron was determined by the o-phenanthroline method(2).Uranium was deter-
(Bellco,Vineland,N.J.).The main reaction vessel of the biometer flask was filled mined by a colorimetric method(24)or inductively coupled plasma mass spec- nO
with 25 ml of 0.2 N KOH to absorb CO2,including CO2,evolved from the trometry._Citric acid was analyzed by HPLC with an Amines HPX-87H column o
metabolism of the complex. 14C-metal-milliliter of a 5.2 mM solution of the equipped with a UV-visible detector set at 210 nm.The amount of protein in the m
citrate complex was added to the cell suspension to obtain a final concentration cell extract was determined as described by Bradford(8),with reagents obtained ry
of 0.52 mM.At periodic intervals. 1-ml aliquots of the cell suspension were
withdrawn,filtered under vacuum through a Millipore HA filter(0.45-pm-pore from Bio-Rad.
size),and washed three times with the mineral salts medium without citric acid. No
The filters were air dried,and the radioactivity was counted in a liquid scintil- RESULTS co
Iation counter(Wallac 1410)with Hi-safe scintillation cocktail.The data ob-
tained as counts per minute(cpm)were converted to nanomoles of citric acid Speciation of 1:1 metal-citrate complexes.Table 1 shows the
from the specific activity of[ C]citric acid used in the experiment.
We also investigated the transport of 14C-labeled Ni-or Zn-citrate complexes distribution of the major metal species for equimolar concen-
to confirm that the lag observed before biodegradation of Ni-and Zn-citrate trations of metal and citric acid in the defined medium at pH
complexes was due to the induction of transport factors. Cells grown in (i) 6.1. All the metals are present predominantly in the (metal
defined medium containing citric acid and(ii)defined medium containing Ni-or citrate)- form, except for U-citrate which exists as two major
Zn-citrate were used in the transport assay as described above. species,(UO Cit)-and[(UO ) (Cit)2]2-.At P H 6.1,75.6%of
Preparation of cell extract.Bacteria grown in the modified Simmon's citrate S P 2 2 2
medium for 18 h were harvested by centrifugation,and the cell pellet was stored the uranium was complexed with citric acid, 17.6% as the
frozen at-20°C after its wet weight was recorded.Later,the cells were thawed monomeric species and 58% as the dimeric species. For cad-
and resuspended in lysis buffer(50 mM Tns-HCI, 50 mM EDTA[pH 7.6]). mium,56.5%was present as(CdCit) ,and 42.5%was present
Lysozyme(1 mg/g)was added to the cell suspension,and the cells were allowed z+ion.
lyse at room temperature for 30 min.The lysate was clarified by centrifugation, as the Cd
and the supernatant was fractionated by ammonium sulfate precipitation(35 to Speciation of the metals in 1:1 metal-citric acid in the de-
50%).The ammonium sulfate fraction was dialyzed against two changes of 30 fined medium as a function of pH showed maximum complex
mM Tris-HCI at 4°C overnight, and the dialysate (cell extract)was used to formation of Cd, Cu,Ni,and Zn with citric acid at near neu-
determine the degradation of the metal-citrate complexes.The absence of whole
and viable cells in the extract was checked by direct microscopy and by plating on trality.More than 90%of ferric iron was complexed with citric
solid Simmon's citrate agar,respectively. acid between pH-3.0 and 9.0. Uranium exhibited maximum
Aconitase activity. The cell extract was assayed for aconitase as described complex formation at pH 5.0 (92%), and complex formation
elsewhere(3).All reagents for the aconitase assay were obtained from Sigma decreased with further increase in pH.
Chemical Company.Aconitase converts citric acid to isocitric acid via a cis-
aconitate intermediate. The cis-aconitate absorbs at 240 ran,While citric and Biodegradation of metal-citrate complexes. Figure 1 shows
isocitric acids show no absorbance at this wavelength.In this assay,one unit of the degradation of Cd-, Cu-,Fe(III)-,Ni-,U-, and Zn-citrate
aconitase is defined as that amount which causes an initial rate of increase in complexes by P. fluorescent. Citric acid was degraded com-
optical density of 0.001 per minute. t
Degradation of metal-citrate complexes by cell extract.The spectrophotomet- pletely in 15 h at the rate of 56.8 µmol h- ,whereas Fe(III)-
ric assay for aconitase activity could not be used to study the degradation of citrate was degraded completely in 60 h at the rate of 16 µmol
metal-citrate complexes because the metals interfered with the assay;hence,such h-r.All the iron released after citrate degradation precipitated

Vol,. 177, 1995 BIODEGRADATION OF METAL-CITRATE COMPLEXES 1991
a • 0.5
0.5
OA
0.3 `
0.4 02 r`
e 0.3 0 a �����-
EA 0 20 ao 60 80
m
pi
E 0.5 b —48
8 0.20d0.4
Urate am 0.30.1 v —A— Niulcacid
• _v 0.2 -
S —:7— Ni Cftrdte
and u-aerate• c0.i —A— Zn-citrate -
0.0 ■ a ■ U 0
0 20 40 60 80 0 20 40 60 80
Hours 0.5 C
FIG. 1. Degradation of metal-citrate complexes by P.fluorescens in defined 0.4
mineral salts medium containing 0.52 mM metal-citrate complex.Values are the 0 3
means of triplicate determinations. Variability between experiments was less
than 10%. 0.2 p
0
o-i 3
0
0 0.
0 20 40 60 80 m
from the solution as a dark orange-red precipitate, and the a
Hours a
final pH of the medium was 7.2.Ni-and Zn-citrate complexes
were degraded after a lag of 20 h. Zn-citrate was completely FIG. 2. Degradation of Ni-and Zn-citrate by P.,fluoreccens grown in citric 3
acid(a),Ni-citrate(b),and Zn-citrate(c).Values are the means of triplicate
degraded by the bacterium,forming a white precipitate which determinations.Three independent experiments showed less than 8%variability. y
contained ---70% of the zinc, and the pH of the medium in- 3
creased from 6.1 to 7.5.The bacterium degraded only 70%of b
the Ni-citrate,with a concurrent rise in pH from 6.1 to 7.8,and cr
all the nickel was detected in solution.Cd-,Cu-,and U-citrate times, and the variability between experiments was less than 0
were not degraded by the bacterium, and the metals were 10%. o
present in solution. The pH of the medium remained un- Table 2 shows the transport of Ni- and Zn-citrate by cells 0
changed for Cd-and Cu-citrate,whereas for U-citrate,the pH grown in media containing Ni-and Zn-citrate. Cells grown in m
increased to 6.5. medium containing citric acid transported free citric acid,but
Resting cells degraded only citric acid and Fe-citrate com- not Ni-or Zn-citrate,at the rate of 30±2 nmol min-1.Normal
pletely in 2 and 4 h,respectively. Cd-, Cu-, Ni-, U-, and Zn- transport of citric acid was observed by cells grown in medium o
citrate showed no degradation. containing Ni-citrate, while Ni- and Zn-citrate were trans- co
Adaptation of bacterium to Ni-and Zn-citrate.Cells grown ported at the rate of 23 ± 3 and 22 ± 2 nmol min-1,respec-
in medium containing citric acid degraded citric acid corn- tively. Similar results were obtained with cells grown in me-
pletely in 16 h without a lag,whereas Ni-and Zn-citrate were dium containing Zn-citrate. Analysis of 14CO2 produced
degraded after a lag of 20 h (Fig. 2a). No lag was observed
during the degradation of citric acid,Ni-citrate,or Zn-citrate
by cells grown in medium containing Ni-citrate(Fig.2b).Sim-
ilarly, cells grown in medium containing Zn-citrate degraded t Citric acid
citric acid, Ni-citrate, and Zn-citrate without a lag (Fig. 2c). 700 --0— Cd-,Cu-,Ni-,U-,and zncdrate I. -
Only 70% of the Ni-citrate was degraded by cells grown in B —o— Fanuw[rale 1
medium containing citric acid,as well as cells grown in medium 0
containing Ni- or Zn-citrate,while Zn-citrate was completely o 600- .
degraded. rq
Transport of metal-citrate complexes. Cells grown in me- cS 400 - 1
dium containing citric acid showed uptake of[14C]citric acid at -4.
the rate of 28 nmol min-1,whereas the[14C]citrate complex of E 300-
Fe(III)was transported at the rate of 12.6 nmol min-1(Fig.3). c 200 -
The lower rate of uptake of the latter complex was consistent
with the rate of degradation of Fe(III)-citrate by the bacte- too f-
rium. During transport assays with citric acid and Fe(III)- - 0
citrate, 123,275 and 100,600 cpm of 14CO2i respectively,were o 0 10 20 30
detected. This corresponded to 58 µmol of citric acid and 47 Minutes
p.mol of Fe(III)-citrate being metabolized.The bacterium did FIG. 3. Uptake of ^C-labeled metal-citrate complexes by P.Jluorescens.Re-
not transport Cd-, Cu-,Ni-, U-, or Zn-citrate complexes, and sults are the means and error bars indicate+standard errors of the means for
no 'CO, was detected. This experiment was repeated three three independent experiments.

1992 JOSHI-TOPE AND FRANCIS J.BACrERIOL.
TABLE 2. Transport of Ni-and Zn-citrate by adapted cells 3:2 complex at pH values above 6.5, and its stability is not
Uptake(nmol min')° known (32); the binuclear species has a log K of 18.9 (33).
Source of cells The lag observed before the biodegradation of Ni-and Zn-
Citric acid Ni citrate Zn-citrate citrate by cells grown in medium containing citric acid reflects
Citric acid grown 30 2 ND ND the time needed for induction of the transport factor for these
Ni-citrate grown 29±2 23±3 22±2 complexes,while the reciprocity of induction suggests that a
Zn-citrate grown 27±1 21±1 22±2 common transport factor is involved. Although inducible sys-
tems for Ni and Zn uptake are known (25, 30), there is no
°Results are means±standard errors of the means.ND,not detected. direct evidence for reciprocity of their uptake. Indirect evi-
dence for their reciprocity may be inferred from the inhibition
of energy-dependent Ni transport systems by Zn in Clostridium
during transport assays with Ni-citrate(121,925 cpm)and Zn- pasteurianum and Bradyrhizobium japonicum JH (10, 18).We
citrate(118,900 cpm)indicated that 57 limo]of Ni-citrate and did not determine whether Ni- and Zn-citrate were trans-
56 µmol of Zn-citrate were metabolized.Cells grown in Ni-or ported as intact complexes. However, studies with B. subtilis
Zn-citrate-containing medium transported both complexes,in- using radioisotopes of Mg and Mn and [1,5-14C]citric acid
dicating that the adaptation was reciprocal.These results are in indicated that intact complexes were transported inside the cell
good agreement with the degradation of Ni- and Zn-citrate (6).It is also not known whether a cation efflux system operates
complexes by cells grown in medium containing Ni- or Zn- on the Ni and Zn released from the complexes after the bio-
citrate (Fig.2). degradation of citric acid,similar to the plasmid-encoded efflux
Degradation of metal-citrate complexes by cell extract.The system that confers resistance to Cd,Co,Ni,and Zn in Alcali-
specific activity of aconitase in the cell extract of P.fluorescens genes eutrophus(31,34).The partial degradation of Ni-citrate
grown in medium containing citric acid was 26 U/mg of pro- observed may be due to either toxicity of the released nickel
tein.Table 3 shows the degradation of metal-citrate complexes ion or a change in the nature of the Ni-citrate complex at
by the cell extract. Citric acid and the bidentate Fe(III)-, Ni- higher pH values. o
and Zn-citrate complexes were degraded, and the released Tridentate Cd- and Cu-citrate complexes that involve theo
metals were associated with the particulates recovered after hydroxyl group in complex formation were neither transported 0
centrifugation.Tridentate Cd-and Cu-citrate complexes were nor biodegraded by the bacteria or the cell extract.It has been m
not degraded by the cell extract,and the metals were detected proposed that the hydroxyl group of citric acid is the key factor a
in solution. In contrast,U-citrate was degraded at the rate of determining biodegradability of the metal-citrate complexes 5
12 ± 1 µmol min-1 pg of protein'.Almost all the released (16, 17). Aconitase catalyzes the removal of both a hydrogen d
uranium(90%)was detected in the particulate fraction recov- and hydroxyl group from citric acid (19). Electron paramag- b
ered after centrifugation. These findings were similar to the netic resonance and Mossbauer spectroscopy studies with ac- 3
results of growth studies except for the degradation of U- onitase showed that the hydroxyl group of isocitric acid is o
citrate. critical in the mechanism of enzyme action (13, 14, 25), be- v
cause it is required for binding the iron on the aconitase mol- o
DISCUSSION ecule(27).If the hydroxyl group of citric acid is bound tightly0
to a metal in the complex, the isomerization of citrate to o
o
isocitrate may be inhibited because of steric hindrance. a-
Citric acid is metabolized intracellularly in bacteria by the CD
enzymes, aconitase and citrate lyase. Aconitase catalyzes the Speciation calculations with Cu-citrate indicate that most of V
isomerization of citrate to isocitrate, a key reaction in the the metal and citric acid are complexed,and Cu-citrate was not N
biodegraded by growing or resting cells or the cell extract.Lack o
tricarboxylic acid cycle in aerobic metabolism,whereas citrate of biodegradation of the Cu-citrate complex was not due to 0
lyase is involved in the anaerobic metabolism of citric acid(4, toxicity of the copper, as any excess citric acid added to the
36). The degradation of the metal-citrate complexes by intact Cu-citrate was readily biodegraded(17).
cells did not correlate with the stability constants of the com- Speciation calculations of Cd-citrate predict that only 56.5%
plexes but rather with the nature of the complex formed be of the total cadmium and citric acid is present as a (CdCit)-
tween the metal and citric acid. However, the degradation of complex, and of the citric acid and the metal remains
the complexes by the cell extract correlated inversely with the uncomplexed.The presence of free citric acid was not evident
stability constants. Fe(III)-citrate, which is predominantly in from experiments with growing or resting cells or the cell
the [Fe(OH)ZCit] form with a stability constant of 2.6, Nde- extract. Also, transport studies do not corroborate the pres-
citrate (log K = 5.4), and Zn citrate (log K = 4.9)were de- ence of free citric acid,as it would be readily transported inside
to
graded faster than U-citrate.U-citrate is believed to exist as a the cells.Studies with resting cells,in which the toxic effects of
the metal,if any, should be minimal or eliminated because of
the high cell density did not show biodegradation of Cd-citrate,
TABLE 3. Degradation of metal-citrate complexes by cell extract indicating the absence of free citric acid. Addition of excess
Complex Degradation rate citric acid did not result in metabolism of the complex. For
(µmol min-1 µg of protein 1)° example, the addition of 1:1 Cd-citrate complex to culture
Citric acid 23±2 medium containing 0.52 mM uncomplexed citric acid at the
Fe(III)-citrate 20±3 beginning of the experiment or after 10 h of growth of the
Ni-citrate 17±2 bacterium on citric acid showed no degradation of the excess
Zn-citrate 17±3 citric acid or the complex.Formation of multiligand complexes
U-citrate 12±1 of Cd and citric acid has been reported (7), but the stability
Cd-citrate ND constants for these complexes are not known. These results
Cu-citrate ND suggest that the free Cd2+ ion,if present,is toxic to the bac-
Results are means-- standard errors of the means for three independent teria and inhibits citrate transport as well as aconitase activity
experiments.ND,not degraded. in the cell extract.Alternatively,formation of multiligand com-

VoL.177,1995 BIODEGRADATION OF METAL-CITRATE COMPLEXES 1993
plexes may render citric acid in the Cd-citrate complex unavail- erlchia colt.Eur.J.Biochem.45:555-559.
able for biodegradation. 12. Dodge,C.J.,and A.J.Francis.1994.Photodegradation of uranium-citrate
complex with uranium recovery.Environ.Sci.Technol.28:1300-1306.
U-citrate was neither transported nor biodegraded by the 13. Emptage,M.H.,J.-L.Dreyer,M.C.Kennedy,and H.Beinert.1983.Optical
bacteria,including resting cells.However,the cell extract corn- and EPR characterization of different species of active and inactive aconi-
pletely degraded the complex. Speciation calculations of the Lase.J.Biol.Chem.258:11106-11111.
U-citrate complex in the defined medium predict that 76% of 14. Emptage,M.H.,T.A.Kent,M.C.Kennedy,H.Beinert,and K Munck.1983.
Mossbauer and EPR studies of activated aconitase:development of a local-
the uranium and citric acid is complexed and 24%of the citric ized valence state at a subsite of the[4Fe-4S]cluster on binding of citrate.
acid is free in solution.The presence of the free citric acid was Proc.Natl.Acad.Sci.USA 80:4674-4678.
not evident from biodegradation and transport studies. Spe- 15. Francis,A.J.,and C.J.Dodge.1992.Reclamation and recovery of radio-
ciation of uranium and citric acid as a function of pH predicts nuclides and toxic metals from contaminated materials,soils,and wastes,p.
109-117.Technology 2002;NASA conference publication no.3189,vol.1.
that the amount of the U-citrate complex will decrease rapidly National Aeronautical and Space Administration Washington,D.C.
above pH 6.0.However,the existence of a cyclic 3:2 U-citrate 16. Francis,A.J.,and C.J.Dodge.1993.Influence of complex structure on the
complex involving hydroxo and oxo bridges above pH 6.5 has biodegradation of iron-citrate complexes.Appl.Environ.Microbiol.59:109-
been postulated,although its stability has not been determined 113.
(32)32 Potentiometric and s ectro hotometric analyses also 17. Francis,A. J., C. J. Dodge, and J. B. Gillow. 1992. Biodegradation of
. p lJ y metal-citrate complexes and implication for toxic metal mobility.Nature
showed the existence of U-citrate complexes above pH 6.5 (London)356:140-142.
(12).Lack of biodegradation of the U-citrate complex was due 18. Fu,C.,and R.J.Maier.1991.Competitive inhibition of an energy-dependent
to lack of transport of the complex inside the cell and not due nickel transport system by divalent cations in Bradyrhizobium japonicun:JR.
to toxicity (17).For example, excess citric acid in the mineral
Appl.Environ.Microbiol.57:3511-3516.
P 19. Glusker,J.P.1980.Citrate conformation and chelation:enzymatic implica-
salts medium (17) and in citric acid extracts of uranium con- tions.Acc.Chem.Res.13:345-352.
taminated soils and wastes(12,15)showed degradation of the 20. Good,N.E.,G.D.Wmget,W.Winter,T.N.Connolly,S.Izawa,and R M.M.
excess citric acid and several metal-citrate complexes but not Singh. 1966. Hydrogen ion buffers for biological research. Biochemistry
the U-citrate complex. Nevertheless, additional studies are 5:467-477.
PGuerinot,uerinot,M.L.,E.J.Meld!,and O.Plessner.1990.Citrate as a siderophore
needed to elucidate the structure-function relationships in the in Bradyrhizobium japonicwn.J.Bacteriol.172:3298-3303. o
metabolism of metal-citrate complexes. 22. Hamm,R.E.,C.M.Schull,and D.M.Grant.1954.Citrate complexes with
o
iron(II)and iron(III).I.Am.Chem.Soc.76:2111-2114. 5
23. Hughes,M.N.,and R.K.Poole.1989.Metal mimicry and metal limitation in o
ACKNOWLEDGMENTS studies of metal-microbe interactions.In R.K.Poole and G.M.Gadd(ed.), a
Metal-microbe interactions.IRL Press,Oxford. -.,
We thank C. J.Dodge for assistance and F.J.Wobber, Program 24. Johnson,D.A.,and T.M.Florence.1971.Spectrophotometric determination 3
Manager,for continued support. of uranium (VI) with 2-(-5-bromo-2-pyridylazo)-5-diethylaminophenol.
This research was performed under the auspices of the Environmen- Anal.Chim.Acta 53:73-79. er
tal Sciences Division's Subsurface Science Program,Office of Health 25. Kaltwasser,H.,and W.Frings.1980.Transport and metabolism of nickel in y
and Environmental Research, Office of Energy Research, U. S.De- microorganisms.In J.Nriagu(ed.),Nickel in the environment.John Wiley& 3
partment of Energy,under contract no.DE-ACO2-76CH00016. Sons.Inc.,New York. to
26. Kent,T.A.,M.H.Emptage,H.Merkle,M.C.Kennedy,H.Beinert,and E.
Munck.1987.Mossbauer studies of aconitase.J.Biol.Chem.260:6871-6881. .-e
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1. Allison,J. D.,D. S. Brown, and K.J.Voto•Gradac. 1991. MINTEQA2/ structures of aconitase with isocitrate and nitroisocitrate bound.Biochem- O
PRODEFA2:a geochemical assessment model for environmental systems. istry 31:2735-2748. n
EPA-600/3-91-021.U.S.Environmental Protection Agency,Athens,Ga. 28. Lohmeyer,M.,and C.G.Friedrich. 1987.Nickel transport in Alcaligenes a
2. American Public Health Association.1975.Standard methods for the exam- eutrophus.Arch.Microbiol.149:130-135. m
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Permit No.G-10
9590 9402 3665 7335 1548 97
United States •Sender:Please print your name,address,and ZIP+4®in this box'
Postal Service
NCDEQ-DWR
Water Quality Regional Operations Section
ATTN: SHRISTI SHRESTHA
1636 Mail Service Center
Raleigh,NC 27699-1636

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