[Federal Register Volume 77, Number 77 (Friday, April 20, 2012)]
[Proposed Rules]
[Pages 23638-23643]
[FR Doc No: 2012-9569]

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DEPARTMENT OF TRANSPORTATION

Federal Aviation Administration

14 CFR Part 39

[Docket No. FAA-2012-0415; Directorate Identifier 2008-SW-065-AD]
RIN 2120-AA64

Airworthiness Directives; Bell Helicopter Textron, Incorporated
Helicopters

AGENCY: Federal Aviation Administration (FAA), DOT.

ACTION: Notice of proposed rulemaking (NPRM).

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SUMMARY: We propose to supersede an existing airworthiness directive
(AD) for Bell Helicopter Textron, Inc. (BHTI) Model 204B, 205A, 205A-1,
205B, and 212 helicopters. The existing AD currently requires
conducting various inspections associated with the main rotor grip
(grip). If a crack is found, that AD requires replacing the grip before
further flight. If delamination of the buffer pad on the grip tang
inner surface is found, that AD requires inspecting the grip surface
for corrosion or other damage and repairing or replacing the grip if
corrosion or other damage is found. That AD also requires determining
and recording the hours time-in-service (TIS) and the engine start/stop
cycles for each grip on a component history card or equivalent record.
Additionally, that AD requires you to report certain inspection results
to the FAA. Since we issued that AD, additional cracks in grips have
been found. Analysis of these events has shown that a retirement life
is needed for certain grips, and the AD

applicability needs to be expanded to include additional grips similar
in design, as well as to include the Model 210 helicopter, which was
issued an FAA type certificate after the existing AD was issued. The
proposed actions are intended to prevent failure of a grip, separation
of a main rotor blade, and subsequent loss of control of the
helicopter.

DATES: We must receive comments on this proposed AD by June 19, 2012.

ADDRESSES: You may send comments by any of the following methods:
     Federal eRulemaking Docket: Go to http://www.regulations.gov. Follow the online instructions for sending your
comments electronically.
     Fax:<PHONE_NUMBER>.
     Mail: Send comments to the U.S. Department of
Transportation, Docket Operations, M-30, West Building Ground Floor,
Room W12-140, 1200 New Jersey Avenue SE., Washington, DC 20590-0001.
     Hand Delivery: Deliver to the ``Mail'' address between 9
a.m. and 5 p.m., Monday through Friday, except Federal holidays.
    Examining the AD Docket: You may examine the AD docket on the
Internet at http://www.regulations.gov or in person at the Docket
Operations Office between 9 a.m. and 5 p.m., Monday through Friday,
except Federal holidays. The AD docket contains this proposed AD, the
economic evaluation, any comments received and other information. The
street address for the Docket Operations Office (telephone 800-647-
5527) is in the ADDRESSES section. Comments will be available in the AD
docket shortly after receipt.
    For service information identified in this proposed AD, contact
BHTI, P.O. Box 482, Fort Worth, TX 76101, telephone<PHONE_NUMBER>, fax
(817) 280-6466, or at http://www.bellcustomer.com/files/. You may
review copies of service information at the FAA, Office of the Regional
Counsel, Southwest Region, 2601 Meacham Blvd., Room 663, Fort Worth,
Texas 76137.

FOR FURTHER INFORMATION CONTACT: Michael Kohner, Aviation Safety
Engineer, Rotorcraft Certification Office, 2601 Meacham Blvd., Fort
Worth, Texas 76137, telephone<PHONE_NUMBER>, fax<PHONE_NUMBER>, email
<EMAIL_ADDRESS>or<EMAIL_ADDRESS>
SUPPLEMENTARY INFORMATION:

Comments Invited

    We invite you to participate in this rulemaking by submitting
written comments, data, or views. We also invite comments relating to
the economic, environmental, energy, or federalism impacts that might
result from adopting the proposals in this document. The most helpful
comments reference a specific portion of the proposal, explain the
reason for any recommended change, and include supporting data. To
ensure the docket does not contain duplicate comments, commenters
should send only one copy of written comments, or if comments are filed
electronically, commenters should submit only one time.
    We will file in the docket all comments that we receive, as well as
a report summarizing each substantive public contact with FAA personnel
concerning this proposed rulemaking. Before acting on this proposal, we
will consider all comments we receive on or before the closing date for
comments. We will consider comments filed after the comment period has
closed if it is possible to do so without incurring expense or delay.
We may change this proposal in light of the comments we receive.

Discussion

    On December 31, 2002, we issued AD 2003-01-04, Amendment 39-13015
(68 FR 1955, January 15, 2003), for the BHTI Model 204B, 205A, 205A-1,
205B, and 212 helicopters. That AD requires the following actions:
     Within 10 hours TIS, determining and recording the hours
TIS and the engine start/stop cycles for each grip on a component
history card or equivalent record. On the single-engine model
helicopters, one ``engine start/stop cycle'' occurs when the engine is
started. On the Model 212 helicopter, one ``engine start/stop cycle''
occurs when either one or both engines are started. The intent is to
add one ``engine start/stop cycle'' each time helicopter power starts
the main rotor system turning.
     Within 10 hours TIS and thereafter at intervals not to
exceed 25 hours TIS, visually inspecting the exposed surfaces of the
upper and lower tangs of each grip for a crack, using a 10-power or
higher magnifying glass.
     At specified intervals, depending on the hours TIS or the
engine start/stop cycles, whichever occurs first, conducting initial
and repetitive ultrasonic (UT) inspections of each grip in accordance
with the Nondestructive Inspection Procedure, Log No. 00-340, Revision
E, dated April 9, 2002.
     At intervals not to exceed 1,200 hours TIS or 24 months,
whichever occurs first, inspecting each buffer pad on the tang inner
surfaces for delamination and removing the buffer pad and inspecting
the grip surface for corrosion and other damage if delamination is
found.
     Within 2,400 hours TIS or at the next overhaul of the main
rotor hub, whichever occurs first, and thereafter at intervals not to
exceed 2,400 hours TIS, inspecting the surface of each affected grip
for corrosion or other damage and conducting a fluorescent-penetrant
inspection (FPI) of the grip for a crack.
     Before further flight, replacing any grip that has a
crack, corrosion, or other damage with an airworthy grip, or repairing
a grip with corrosion or other damage if the corrosion or other damage
is within certain limits.
     Reporting certain inspection results and information to
the FAA in accordance with Appendix 1 of the AD.
    AD 2003-01-04 was prompted by three in-flight grip failures and two
cracked grips discovered during a 1,200-hour TIS inspection and on a
scheduled 2,400-hour TIS overhaul, which brought the total to 13 grips
that had cracked in the lower tang. No anomalies or damage to the
blade, blade bolt bore, or buffer pad tang surface were found in the
two cracked grips found during the inspection and overhaul. Cracking on
all of the grips has been attributed to mechanical damage from improper
blade bolt bushing installation, improper rework of the buffer pad tang
surface, or subsurface fatigue damage. All of the fatigue cracks
occurred on grip part numbers (P/N) 204-011-121-009 and -121, installed
on BHTI Model 212 helicopters. Grip P/Ns 204-011-121-005, -113, and -
117, installed on BHTI Model 204B, 205A, 205A-1, and 205B helicopters,
were also included because the grips are very similar in design and are
subjected to the same forces and loads as the grips installed on BHTI
Model 212 helicopters. We issued that AD to prevent failure of a grip,
separation of a main rotor blade, and subsequent loss of control of the
helicopter.

Actions Since Existing AD Was Issued

    Since issuing the existing AD, small cracks in three grips,
initiating in the blade bolt bore, have been detected during UT
inspections. Additionally, one crack in another grip was discovered
visually, another crack was found after an in-flight vibration
developed, and one grip fractured during flight. This brings the total
to 19 grips that have cracked in the lower tang. Based on these
additional discoveries of cracked grips and subsequent analyses
performed by the manufacturer, we have determined the following:

     The applicability of the AD needs to be expanded to
include replacement grip P/N ASI-4011-121-9, produced under a part
manufacturing approval (PMA) issued by the FAA, because the design
approval for this grip was based on its design being identical to the
original BHTI-manufactured grip, P/N 204-011-121-009;
     The applicability of the AD needs to be expanded to
include any Model 204B, 205A, 205A-1, and 212 helicopters with grip P/N
204-011-121-009, -121, or ASI-4011-121-9 installed;
     The applicability of the AD needs to be expanded to
include the Model 210 helicopter, which has a grip P/N 204-011-121-009
and -121, installed. This model was not included in AD 2003-01-04
because the FAA issued the BHTI Model 210 type certificate after AD
2003-01-04 was issued;
     A retirement life is needed for grip P/Ns 204-011-121-009,
204-011-121-121, and ASI-4011-121-9. These grips, installed on Model
205B, 210, and 212 helicopters, are currently unlimited in service life
and are only required to be replaced when damaged or corroded beyond
repair. Early failures of these grips, which have not been attributed
to mechanical damage, have indicated a need to establish retirement
lives to avoid possible cracking in the future;
     The procedure for calculating the total hours TIS for any
grip where the hours TIS cannot be determined from the helicopter
records should be modified to more accurately account for the actual
usage of the fleet and clarify the intent of the AD; and
     The reporting of inspection findings to the FAA is no
longer needed.

FAA's Determination

    We are proposing this AD because we evaluated all the relevant
information and determined the unsafe condition described previously is
likely to exist or develop in other helicopters of these same type
designs.

Related Service Information

    We reviewed the following BHTI service information:
     Operations Safety Notices 204-85-6, 205-85-9, and 212-85-
13, all dated November 14, 1985, which describe a grip with a crack in
the lower tang that was returned by an operator;
     Alert Service Bulletin (ASB) No. 212-94-92, Revision A,
dated March 13, 1995, which describes procedures for inspection and
overhaul requirements of certain grips;
     ASB No. 204-02-58, dated November 26, 2002; ASB No. 205-
02-88, dated November 26, 2002; and ASB No. 210-08-02, dated September
10, 2008. These ASBs specify a UT inspection of certain grips;
     ASB No. 205B-02-39, Revision B, dated November 22, 2002
and ASB No. 212-02-116, Revision A, dated October 30, 2002, which
specify a UT inspection of certain grips and include the Nondestructive
Inspection Procedure, Log No. 00-340, Revision E, dated April 9, 2002;
and
     Information Letter 204-08-23, 205-08-38, 205B-08-21, and
212-08-62, Revision A, dated July 23, 2008 (one letter issued with 4
identification numbers), which describes a new, improved replacement
grip, P/N 204-011-121-125, that would not require the repetitive UT
inspections and would have a retirement life of 25,000 hours TIS or a
500,000 Retirement Index Number (RIN), whichever comes first.

Proposed AD Requirements

    This proposed AD would retain certain requirements of AD 2003-01-
04, and would require additional actions. The proposed requirements are
as follows:
     Within 10 hours TIS, determining and recording the hours
TIS and the engine start/stop cycles for each grip on a component
history card or equivalent record. For each month that the hours TIS
cannot be determined, this proposed AD would assume 50 hours TIS.
     Within 10 hours TIS and thereafter at intervals not to
exceed 25 hours TIS, visually inspecting the exposed surfaces of the
upper and lower tangs of each grip for a crack, using a 10-power or
higher magnifying glass.
     At specified intervals, depending on the hours TIS or the
engine start/stop cycles, whichever occurs first, conducting initial
and repetitive UT inspections of each grip, in accordance with the
Nondestructive Inspection Procedure, Log No. 00-340, Revision E, dated
April 9, 2002.
     At intervals not to exceed 1,200 hours TIS or 24 months,
whichever occurs first, inspecting each buffer pad on the tang inner
surfaces for delamination, and removing the buffer pad and inspecting
the grip surface for corrosion and other damage if delamination is
found.
     Within 2,400 hours TIS or at the next overhaul of the main
rotor hub, whichever occurs first, and thereafter at intervals not to
exceed 2,400 hours TIS, inspecting the surface of each affected grip
for corrosion or other damage and conducting an FPI of the grip for a
crack.
     Before further flight, replacing any grip that has a
crack, corrosion, or other damage with an airworthy grip, or repairing
a grip with corrosion or other damage if the corrosion or other damage
is within certain limits.
     Before further flight, removing any grip, P/N 204-011-121-
009 or ASI-4011-121-9, that has 15,000 or more hours TIS;
     Before further flight, removing any grip, P/N 204-011-121-
121, that has 25,000 or more hours TIS;
     Revising the Airworthiness Limitations sections of the
applicable maintenance manuals or the Instructions for Continued
Airworthiness by establishing a new retirement life of 15,000 hours TIS
for grip, P/N 204-011-121-009 or ASI-4011-121-9, and 25,000 hours TIS
for grip, P/N 204-011-121-121; and
     Recording the applicable hours TIS life limits for grips,
P/N 204-011-121-009, ASI-4011-121-9, and 204-011-121 on the applicable
component history card or equivalent record.

Differences Between the Proposed AD and the Service Information

    This proposed AD requires life limits for grips, P/N 204-011-121-
009, 204-011-121-121, and ASI-4011-121-9. The manufacturer's service
bulletins do not specify a service life for these grips. Also, this
proposed AD applies to grip P/N ASI-4011-121-9, which is produced under
a parts manufacturing approval, and the manufacturer's service
bulletins do not address this particular grip.

Costs of Compliance

    We estimate that this proposed AD would affect 700 helicopters of
U.S. registry, and it would take approximately 7 work hours to create
and maintain the records, 6.25 work hours to conduct the inspections,
and 20 work hours to replace a set of grips at an average labor rate of
$85 per work hour. Required parts would cost approximately $56,385 for
a replacement set of grips. Based on these figures, we estimate the
total cost impact of the proposed AD on U.S. operators to be $6,596,875
for the entire fleet, assuming the grip set (2 grips) must be replaced
on 100 helicopters.

Authority for This Rulemaking

    Title 49 of the United States Code specifies the FAA's authority to
issue rules on aviation safety. Subtitle I, section 106, describes the
authority of the FAA Administrator. ``Subtitle VII: Aviation
Programs,'' describes in more detail the scope of the Agency's
authority.
    We are issuing this rulemaking under the authority described in
``Subtitle VII, Part A, Subpart III, Section 44701:

General requirements.'' Under that section, Congress charges the FAA
with promoting safe flight of civil aircraft in air commerce by
prescribing regulations for practices, methods, and procedures the
Administrator finds necessary for safety in air commerce. This
regulation is within the scope of that authority because it addresses
an unsafe condition that is likely to exist or develop on helicopters
identified in this rulemaking action.

Regulatory Findings

    We determined that this proposed AD would not have federalism
implications under Executive Order 13132. This proposed AD would not
have a substantial direct effect on the States, on the relationship
between the national Government and the States, or on the distribution
of power and responsibilities among the various levels of government.
    For the reasons discussed, I certify this proposed regulation:
    1. Is not a ``significant regulatory action'' under Executive Order
12866;
    2. Is not a ``significant rule'' under the DOT Regulatory Policies
and Procedures (44 FR 11034, February 26, 1979);
    3. Will not affect intrastate aviation in Alaska to the extent that
it justifies making a regulatory distinction; and
    4. Will not have a significant economic impact, positive or
negative, on a substantial number of small entities under the criteria
of the Regulatory Flexibility Act.
    We prepared an economic evaluation of the estimated costs to comply
with this proposed AD and placed it in the AD docket.

List of Subjects in 14 CFR Part 39

    Air transportation, Aircraft, Aviation safety, Incorporation by
reference, Safety.

The Proposed Amendment

    Accordingly, under the authority delegated to me by the
Administrator, the FAA proposes to amend 14 CFR part 39 as follows:

PART 39--AIRWORTHINESS DIRECTIVES

    1. The authority citation for part 39 continues to read as follows:

    Authority:  49 U.S.C. 106(g), 40113, 44701.

Sec.  39.13  [Amended]

    2. The FAA amends Sec.  39.13 by removing Amendment 39-13015 (68 FR
1955, January 15, 2003), and adding the following new airworthiness
directive (AD):

Bell Helicopter Textron, Inc. (BHTI): Docket No. FAA-2012-0415;
Directorate Identifier 2008-SW-065-AD.

(a) Applicability

    This AD applies to the following model helicopters with the
listed part-numbered main rotor grips installed, certificated in any
category:

                   Table 1--Helicopter Model and Main Rotor Grip (Grip) Part Numbers Affected
----------------------------------------------------------------------------------------------------------------
                                                                                With the following grip part
                                    Model                                                 number (P/N)
----------------------------------------------------------------------------------------------------------------
204B.........................................................................  204-011-121-005 if the grip was
                                                                                ever installed on a Model 205B
                                                                                helicopter.
204B, 205A, and 205A-1.......................................................  204-011-121-009, -117, -121 or
                                                                                ASI-4011-121-9.
205A and 205A-1..............................................................  204-011-121-005 or -113 if the
                                                                                grip was ever installed on a
                                                                                Model 205B helicopter.
205B.........................................................................  204-011-121-005, -009, -113, -
                                                                                117, or -121.
210..........................................................................  204-011-121-009 or -121.
212..........................................................................  204-011-121-009, -121, or ASI-
                                                                                4011-121-9.
----------------------------------------------------------------------------------------------------------------

(b) Unsafe Condition

    This AD defines the unsafe condition as a crack in the main
rotor grip (grip), which could result in failure of a grip,
separation of a main rotor blade, and subsequent loss of control of
the helicopter.

(c) Other Affected ADs

    This AD supersedes AD 2003-01-04, Amendment 39-13015 (68 FR
1955, January 15, 2003).

(d) Compliance

    You are responsible for performing each action required by this
AD within the specified compliance time unless it has already been
accomplished prior to that time.

(e) Required Actions

    (1) Within 10 hours time-in-service (TIS), create a component
history card or equivalent record and determine and record the total
hours TIS for each grip. If the total hours TIS cannot be determined
from the helicopter records, assume and record 50 hours TIS for each
month for which the hours cannot be determined. Continue to count
and record the hours TIS and begin to count and record the number of
times the helicopter engine(s) are started (engine start/stop
cycles).
    (2) Within 10 hours TIS, and thereafter at intervals not to
exceed 25 hours TIS, without removing the main rotor blades:
    (i) Clean the exposed surfaces of the upper and lower tangs of
each grip with denatured alcohol, and wipe dry.
    (ii) Using a 10-power or higher magnifying glass, visually
inspect the exposed surfaces of the upper and lower tangs of each
grip for a crack. Pay particular attention to the lower surface of
each lower grip tang from the main rotor blade bolt-bushing flange
to the leading and trailing edge of each grip tang. See Figure 1 of
this AD.

[GRAPHIC] [TIFF OMITTED] TP20AP12.001

                                    Table 2--Ultrasonic Inspection Intervals
----------------------------------------------------------------------------------------------------------------
                                                                               Thereafter, at intervals not to
                                                          Within 30 days,     exceed the following hours TIS or
                                                          or the following   engine start/stop cycles, whichever
                 UT inspect grip, P/N:                   hours TIS for the              occurs first:
                                                          grip, whichever  -------------------------------------
                                                           occurs later:                       Engine start/stop
                                                                                Hours TIS            cycles
----------------------------------------------------------------------------------------------------------------
204-011-121-009 or ASI-4011-121-9......................              4,000                400              1,600
204-011-121-121........................................                500                150                600
204-011-121-005 or -113, if the grip was EVER installed              4,000                400              1,600
 on a Model 205B helicopter............................
204-011-121-117, if the grip was NEVER installed on a                4,000                150                600
 Model 205B helicopter.................................
204-011-121-117, if the grip was EVER installed on a                   500                150                600
 Model 205B helicopter.................................
----------------------------------------------------------------------------------------------------------------

    The UT inspection of the grip must be performed by a Non-
Destructive Testing (NDT) UT Level I Special, Level II, or Level III
inspector who is qualified under the guidelines established by MIL-
STD-410E, ATA Specification 105, AIA-NAS-410, or an FAA-accepted
equivalent for qualification standards of NDT Inspection/Evaluation
Personnel.

    Note 1: You can find the Nondestructive Inspection Procedure
attached to BHTI Alert Service Bulletin (ASB) No. 205B-02-39,
Revision B, dated November 22, 2002, or BHTI ASB No. 212 02-116,
Revision A, dated October 30, 2002.

    (4) At intervals not to exceed 1,200 hours TIS or 24 months,
whichever occurs first:
    (i) Remove each main rotor blade, and
    (ii) Inspect each grip buffer pad on the inner surfaces of each
grip tang for delamination (see Figure 1 of this AD). If there is
any delamination, remove the buffer pad and inspect the grip surface
for corrosion or other damage.

    Note 2: This inspection interval coincides with the main rotor
tension-torsion strap replacement times.

    (5) Within 2,400 hours TIS, or at the next overhaul of the main
rotor hub, whichever occurs first, and thereafter at intervals not
to exceed 2,400 hours TIS:
    (i) Remove each main rotor blade.
    (ii) Remove each grip buffer pad (if installed) from the inner
surfaces of each grip tang.
    (iii) Visually inspect the grip tang surfaces for corrosion or
other damage.
    (iv) Fluorescent-penetrant inspect (FPI) the grip for a crack,
paying particular attention to the upper and lower grip tangs. When
inspecting any grip, P/N 204-011-121-005, -009, -113, or ASI-4011-
121-9, pay particular attention to the leading and trailing edges of
the grip barrel.

    Note 3: FPI procedures are contained in BHTI Standard Practices
Manual, BHT-ALL-SPM.

    (6) Before further flight:
    (i) Replace any cracked grip with an airworthy grip.
    (ii) Replace any grip with any corrosion or other damage with an
airworthy grip, or repair the grip if the corrosion or other damage
is within the maximum repair damage limitations found in the
applicable Component and Repair Overhaul Manual.
    (iii) Replace any grip, P/N 204-011-121-009 or ASI-4011-121-9,
which has been in service for 15,000 or more hours TIS.
    (iv) Replace any grip, P/N 204-011-121-121, which has been in
service for 25,000 or more hours TIS.
    (7) Revise the Airworthiness Limitations sections of the
applicable maintenance manuals or the Instructions for Continued
Airworthiness (ICAs) by establishing a new retirement life of 15,000
hours TIS for grip P/N 204-011-121-009 or ASI-4011-121-9, and 25,000
hours TIS for grip P/N 204-011-121-121, by making pen and ink
changes or inserting a copy of this AD into the applicable
maintenance manual or ICAs.
    (8) Record a 15,000 hours TIS life limit for each grip P/N 204-
011-121-009 or ASI-4011-121-9, and a 25,000 hours TIS life limit for
each grip P/N 204-011-121-121, on the applicable component history
card or equivalent record.

(f) Alternative Methods of Compliance (AMOCs)

    (1) The Manager, Rotorcraft Certification Office, FAA, may
approve AMOCs for this AD. Send your proposal to: Michael Kohner,
Aviation Safety Engineer, Rotorcraft Certification Office, 2601
Meacham Blvd., Fort Worth, Texas 76137, telephone<PHONE_NUMBER>,
fax<PHONE_NUMBER>, email<EMAIL_ADDRESS>or<EMAIL_ADDRESS>    (2) For operations conducted under a Part 119 operating
certificate or under Part 91, Subpart K, we suggest that you notify
your principal inspector, or lacking a principal inspector, the
manager of the local flight standards district office or certificate
holding district office before operating any aircraft complying with
this AD through an AMOC.

(g) Additional Information

    (1) BHTI ASB No. 212-94-92, Revision A, dated March 13, 1995;
BHTI ASB No. 212-02-116, Revision A, dated October 30, 2002; BHTI
205B-02-39, Revision B, dated November 22, 2002; BHTI ASB No. 204-
02-58 and ASB No. 205-02-88, both dated November 26, 2002; BHTI ASB
No. 210-08-02, dated September 10, 2008; BHTI Operations Safety
Notice 204-85-6, 205-85-9, and 212-85-13, all dated November 14,
1985; and BHTI Information Letter, 204-08-23, 205-08-38, 205B-08-21,
and 212-08-62, Revision A, dated July 23, 2008; none of which are
incorporated by reference, contain additional information about the
subject of this AD.
    (2) For service information identified in this AD, contact BHTI,
P.O. Box 482, Fort Worth, TX 76101, telephone<PHONE_NUMBER>, fax
(817) 280-6466, or at http://www.bellcustomer.com/files/. You may
review copies of this information at the FAA, Office of the Regional
Counsel, Southwest Region, 2601 Meacham Blvd., Room 663, Fort Worth,
Texas 76137.

(h) Subject

    Joint Aircraft Service Component (JASC) Code: 6220: Main Rotor
Head.

    Issued in Fort Worth, Texas, on April 11, 2012.
Lance T. Gant,
Acting Manager, Rotorcraft Directorate, Aircraft Certification Service.
[FR Doc. 2012-9569 Filed 4-19-12; 8:45 am]
BILLING CODE 4910-13-P


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Revision I Wednesday, November 06, 2002 Biodegradation Screen Study for Telomer Type Alcohols PROJECT NUMBER 3M Projects ID: E0I 0684 Pace Contract Analytical Projects ID: CA085 STUDY DIRECTOR Cleston C. Lange, Ph. D.

CLIENT James K. Lundberg, Ph. D., 3M Environmental Laboratory Bldg 2 3E 09, P. O. Box 33331, St. PauI, MN 551 33 3331 CONTRACT LABORATORY Pace Analytical Services, Inc. Bio Analytical Services Group 1700 Elm Street, Suite 200 Minneapolis, Minnesota 55414 PROJECT DATES Project Initiation: August 9, 2000 Project Completion: September 18,2001 AUTHOR Cleston C. Lange, Ph. D.

Page 1 of 38 Pace Project CAO8S: Biodegradation Screen Study for Telomer AIcohols~   Revision 1 Study Director: Cleston C. Lange. Ph. D. Pace Analytical Services, Sb Analytical Services Group Wednesday. November 06, 2002 CONTAIN NO CBI ° Ø00003 Table of Contents Title Page  1 Table of Contents 2 List of Tables & Figures 3 List of Appendices 4 Executive Summary 5 1.0 Project Personnel 6 2.0 Data Requirements and Revision Justification 6 3.0 Project Objective 6 4.0 Test Article 7 8 5.0 Reference Articles 9 6.0 Receipt/ Generation of Samples 9 7.0 Methods 9 7.1 Sample Preparation 9 7.1.1 Collection of Sludge 9 10 7.1.2 Culture Preparation 11 12 7.1.3 Solid Phase Extraction ( SPE) 12 13 7.2 Instrumental Analysis ( HPLC/ MS) 13 7.2.1 Instrument Parameters 13 14 7.2.2 Qualitative Analysis of Parents and Products 14 15 7.2.3 Quantitative Analysis 16 7,3 Data Transformations and Calculations 16 7.3.1 MolarCalculations 16 7.3.2 Conversion of ng/ mL to micromolar and nanomolar 16 7.3.3 Molar Mass Balance Calculations 17 7.4 Software Versions 17 8.0 Results 18 8.1 Sludge Characterization 18 8.2 Quality Control/ Sample Matrix Spike Results 18 8.3 Analytical Blanks Results 18 8.4 HPLC/ MS Analysis 19 8.4.1 Qualitative I IPLC/ MS and HPLC/ MS/ MS Results 19 24 8.4.2 Quantitative Analysis Results for PFOA 24 25 9.0 Conclusions 27 10.0 Literature Cited 28 11.0 Sample and Data Retention 29 Page 2 of 38 Pace Project CAO8S: Biodegradation Screen Study for Telomer Alcohol&   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06,2002 000004 List of Tables & Figures TABLES Table 1. Parent telomer alcohol expected ions 14 Table 2. Expected product fluorinated fatty acid products ions 15 Table 3. HPLC/ MS Data Table for Parent and Product Ions observed 23 Table 4. HPLC/ MS/ MS Data for n Oxidation Product Ions observed 24 FIGURES Chemical Representation of Telomer Alcohols 8 Chemical Structure of perfluorooctanoate ( PFOA) 9 Plotted HPLC/ MS Integrated Peak Areas for C8, C10 and C12 Telomer Alcohols 20 Plotted HPLC/ MS Integrated Peak Areas for C5, C14 and C15 Telomer Alcohols 20 Plotted HPLC/ MS Integrated Peak Areas for Perfluorinated Fatty Acids Detected 22 Plotted Peak Areas for Transient Polyfluorinated Fatty Acids Detected 22 Plotted PFOA Concentrations Measured in Biodegradation Samples ( SPE eluate 2) 25 Proposed Biodegradation Pathway for Telomer Alcohols 26 Figure 1. Figure 2. FIgure 3. FIgure 4. Figure 5. Figure 6. Figure 7. Figure 8.

Page 3 of 38 Pace Project CA085: 8iodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06. 2002 000005 List of Appendices Appendix A: Approval Signatures 30 Appendix B: Final Quantitative Results for PFOA ( ng/ mL) Table 1. Test Cultures Final Results 31 Table 2. Abiotic Cultures Final Results 31 Table 3. Blank Cultures Final Results 31 Appendix C: Integrated Chromatogram Peak Area Data for the Telomer Alcohols Table 1. Telorner Alcohols Peak Areas 32 Table 2. Percentage of Peak Areas For Telomer Alcohols Remaining at Time Points 32 Appendix D: Table 1. Integrated LC/ MS ( SIR) Peak Area Data for Perfluorinated Fatty Acids 33 Appendix E: Table I. Peak Area Data for Transiently Formed Polyfluorinated Fatty Acids 34 Appendix F: Table I. Total Ion Chromatogram ( TIC) for SIR Data for Biodegradation Samples 35 Appendix G: Table I. TICs for SIR Data for Day 0 and Day 16 Biodegradation Samples 36 Appendix H: Table 1. TICs for SIR Data for Day 0 and Day 16 Abiotic Control Samples 37 Appendix I: Table I. TICs for Extracted Ion Data for Transient Polyfluorinated Fatty Acids 38 Page 4 of 38 Pace Project CA085: ` Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 000006 Executive Summary A screening studywas undertaken to determine whether the fluorochemical telonier intermediate ( telomer alcohol) biodegrades when exposed to municipal wastewater treatment sludge. The study included the preparation of cultures for a six sample point comparative study, and included test cultures, blank cultures, and control cultures. The microbial inoculum for cultures was sludge obtained from the Twin Cities Metro Wastewater Treatment Facility. Telomeralcohol test substance was added to cultures at approximately 5MM concentrations. Cultures were incubated with shaking at 25 ° C. Solid phase extraction of cultures for recovery of biodegradation products and parent analytes was employed. An HPLCIMS analytical method was developed for analysis of the telomer alcohols and expected products. The I IPLC/ MS analysis of culture extracts provided strong evidence that the telamer alcohols were biodegraded. The observed loss of telomer alcohols occurred concomitant with the appearance of several expected perfluorinated carboxylic acids. Unexpectedly, transient} y formed polyfluorinated p oxidation intermediates were observed. The 3 oxidation pathway was suspected to be the major route of biodegradation resulting in the observed even numbered carbon chain length perfluorinated fatty acids. Minor end products, as odd numbered carbon chain length perfluorinated fatty acids, were also observed and were likely the products of fatty acid a oxidation, a minor pathway of fatty acid metabolism usually observed in branched chain fatty acids. After the l6daytest period, perfiourinated carboxylic acids ranging from C5 to C, 2 were observed in the test cultures. These compounds were not in controls or blank cultures,

Perfluorooctanoate ( PFOA), was the only compound quantitatively analyzed and, based on mass balance data accounted for approximately 6 7% of the total telomer alcohols initially present in the test cultures.

Page 5 of 38 Pace Project CA085: ` Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 000007 1.0 Proiect Personnel 1.1 Sponsor Company 1.2 Sponsor Representative 1.3 Contract Facility Personnel:

1.3.1 Study Director 1.3.2 Laboratory Management:

1.3.3 Sample Preparation 3M Dr. James K. Lundberg Or, Cleston C. Lange Mr. Bruce E. Warden Ms. Angela L. Schuler Dr. Cleston C. Lange Ms. Angela L. Schuler Dr. Cleston C. Lange Dr. Cleston C. Lange 2.0 Data Requirements and Revision Justification The sponsor representative desired an aerobic biodegradability screening study to be conducted using the telomer alcohol mixture as test substrate, Thestudy was initiated on August 9, 2000 as a non GLP study.

The report issued Friday, November 16, 2001 is revised to substitute " Dupont Zonyl BA type Telomer, or Zonyl type Telomer" with Telomer. The reason for the revision is to more accurately reflect the impact of results to the general class of telomer alcohols.

3.0 Project Objective This study was conducted in order to elucidate whether the fluorotelomer intermediate ( telomer alcohol) is biodegradable under aerobic conditions using a microbial rich inoculurn of municipal wastewater treatment sludge. This study was stnilar to earlier studies conducted for 3M by Pace as Pace projects CA058 and CAO9T 2 and CA1O4 ~,

CAl05 `, and CAl 32 ~. The development of an analytical method and the analysis of parent analytes and possible products were critical parameters for the determinatiofto f biodegradation.

Page 6 of 38 Pace Project CAO85: " Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Ciroup Wednesday, November 06, 2002 1.3.4 1,3.5 1,3.6 HPLC/ MS Analyst Sample Custodian:

Report Author 000008 4.0 Test Article The test article used for this study was the Zonyl BA type telomer alcohol fluorochernic&

surfactant intermediate. Two sources of test material were used for this study, The first source was provided by the sponsor company, 3M, as approximately 2.5 g of crystalline solid test material labeled Zonyl BA N telomer alcohol, 3M tracability number TN A 2186,

on July 7, 2000. The sponsor did not provide an MSDS, a chain of custody, the purity information for the test article, noran expiration date for the material. This material was given a test, control, and reference ( TCR) number at Pace as CA TCRO2 009 and was stored at 4 ° C. Material CA TCRO2 009 was used primarily for method development purposes of the study, including the first set of cultures prepared and original LC/ MS method development.

A commercial vendor, Aldrich Chemical Company, provided the second source of telomer alcohol used. Material was purchased as Zonyl BA L fluorotelomer intermediate ( 1999 Aldrich Catalog number 42,151 0), also called perfluoroalkylethanol. The material characteristics were boiling point 145 245 ° C, F( CF2)~ CH2CH2OH where n equals approximately 7 to 8, and molecular weight was reported as M~ ca. 443.70 wt. An expiration date and the purity, or percent composition, of the material were not provided.

The purchased material was received at Pace on September 22, 2000. Upon receipt at Pace, the material was given tracability number CA TNCOO 254 and was stored at 4 ° C,

or less. A representative chemical compilation of the Zonyl BA type telomer alcohol is shown in Figure 1.

Page 7 of 38 Pace Project CA085: ` Biodegradation Screen Study for Telonier Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday. November 06, 2002 00( 2009 F F F F F F F I I h__ OH F F F F F F F F F F F F J ~~ OH F F F F F F F F F F F F F F F I I I I Molecular formula = C5 H5 F130 8 Molecular Weight = 364.104 EM + CH3COO 1' = 423 Molecular formula = C10 H5 F17 0 10 Molecular Weight = 464.119 ( M + CH3COO ]` = 523 a" Molecularformula = C24 H5 F250 ` 14 Molecular Weight = 664.149 [ M + CH3COO ~ ` = 723 Figure 1. The telomer alcohols. Based on the HPLC/ MS average peak area response observed in the six abiotic control cultures, the composition of the test material was 6.4% ± 0.3% as C5 telomer alcohol, 39.3% ± 1.5% as C8, 27.7% ± 2.1%

as C10, 17.5% ± 1.2% as C12, 7.2% ± 1.0% as C54, and 2.0% ± 0.2% as C16.

Page 8 of 38 Pace Project CA085: " Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services. Bio Analytical Services Group Wednesday, November 06, 2002 ooooi~ O F F F F F Ce F F F F Molecular formula = C5 H5 F9 0 Molecular Weight = 264.069 P ( M * Cl 13C00 7 `= 323 I I I I I I F F F F F F F F F F F F F F 1FF F F F ThJ~ l C Molecular formula = C12 H5 F21 0 12 Molecular Weight = 564.134 [ M + CH3COO 1 `= 623 ` IIIflIIIIIII\

F F F F F F F F F F F F OH I £ 11 111111111 ~ ft"~   OH Molecular formula = 016 H5 F29 0 16 Molecular Weight = 764.164 [ M CH3COO] `= 823 5.0 Reference Materials The sponsor provided reference material. The neat material was stored at  80 ° C.

NS: Expiration date not specified 5.1. Perfluorooctanoate Ammonium Salt ( PFOA)

Purity: 95.2%; 3M#: TCR 99131 3?; Pace #: CA TCRO2 001; Expires: NS F F F F F F F 6 NH FLLL~ I~ IX~

Figure 2 Perfluorooctanoate ammoniurn salt 6S Receipt/ Generation of Samples Samples were not received, but were generated as an inherent part of this study. All of the experimental cultures prepared for each culture set were extracted by solid phase extraction ( SPE) methodology to generate three analytical samples per culture. The analytical samples, as SPE eluates collected for each culture, were labeled as SPE eluates 1, 2 or 3. Of the three eluates generated for each sample, eluate 1 was the 25 mL aqueous sample eluate and eluates 2 and 3 were 25 mL methanol eluates from the SPE cartridge. Qualitative and semi quantitative HPLC/ MS analysis was conducted for eluate 2 only, and the resulting data was evaluated to determine whether biodegradation occurred.

7.0 Methods 7.1 Sample Preparation 7,1.1. Collection of Sludge.

The sludge for this study was obtained from the Twin Cities Municipal waste treatment facility in a manner consistent with the sludge collected for other studies listed in section 30. The sludge used for the preparation of the first set of cultures was collected on July31, 2000 and Page 9 of 38 Pace Project cAOa5: " Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analyticat Services Group 000011 Wednesday, November 06, 2002 delivered with Pace chain of custody ( CCC) form 465254. The sludge for the second set of cultures was collected on September 18, 2000 and was accompanied with COC # 528791. Sludge used for the second set of cultures in this study was also used in preparation of cultures for other fluorochemical biodegradation studies ( CA1O5 and CA132 5), and was shown to be active for biodegradation of other fluorochemical compounds in those studies.

To prepare cultures, sludge was obtained from the primary municipal waste treatment facility in the Twin Cities area. Arrangements were made for Pace personnel to retrieve fresh mixed liquor suspended solids ( MLSS) from the aeration units at the Twin Cites Metro Wastewater Treatment Facility located in St. Paul, MN. Typically, Four liters of MLSS was collected by Pace laboratory personnel and delivered as four 1 liter Nalgene polypropylene bottles containing MLSS, and were accompanied with a corresponding chain of custody with date collected. Upon receipt at Pace Science Solutions, the individual bottles were labeled # 1 through # 4, The suspended solids in the bottles were allowed to settle at least 24 hours at 4 ° C ± 3 ° C. The settled solids " sludg& were then used to prepare MLSS plus sludge for use in preparing test cultures. A sludge characterization analysis was not conducted as pad of this screening study.

The settled sludge in each bottle constituted approximately 20% of the volume, or approximately 200 mL volume in a I liter bottle, based on visual observations and was consistent with observation of earlier sludge collections.

Page 10 of 38 Pace Project CA085: " Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 000012 7.1.2. Culture Preparation.

The first set of cultures were prepared August 9, 2000 and incubation of those cultures continued with intermittent culture harvests until the final harvest on September 13, 2000, for a total incubation time of 35 days.

The second set of cultures were prepared on September 27, 2000 and incubation of those cultures continued with intermittent culture harvests until the final harvest on October 12, 2000, for a total of 16 days incubation. The culture preparation procedure described below was used, and was documented as Pace standard operating procedure ( SOP) CAG SP 03 ~

Cultures were prepared using a mineral salts medium defined by EPA Guideline OPPTS 835.3200. The mineral salts medium pH was 7.4 and contained per liter, 0,334 g Na2HPO4 2H20, 0.005 g NH4CI, 0.2175 g K2HPO4, and 0.085 KH2PO4, 0.0275 g CaCI2 anhydrous, 0.0225 g MgSO4 7H2O, and 0.00025 g FeCl3 6H20.

Two liters of a mineral salts medium containing 100 mL of settled sludge was prepared and contained 1 mL of methanol. To each test culture was added 25 mL of this mineral medium containing sludge.

A mineral salts medium, un sterilized and without sludge, was prepared.

This mineral salts medium without sludge was used to prepare the 25 mL no sludge ( abiotic) control cultures.

All cultures were prepared by dispensing 25 mL of appr3priate mineral salts medium solution into clear sterile 125 mL Nalgene polycarbonate culture flasks containing labels with appropriate identification information.

Note: The mineral salts medium that contained sludge had to be swirled regularly during dispensing in order to keep the mixture homogenous and prevent the sludge from settling out of the solution.

The test substance, either Zonyl BA N telomer alcohol for culture set one ( Stock ID CA058 SS 004 at 10,280 ~. tg/ mLin methanol) or Zonyl BA L Page 11 of 38 Pace Project CA085: ` Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 ooool3 telomer alcohol for culture set two ( Stock ID CAO8S SW 001 at 7,288 ~ Lg/ mLin methanol), was added to the test cultures by transferring 5 . iL and 8J. LL, respectively, to the appropriate test cultures. The final test concentration in cultures was 2.056 pg/ mL of Zonyl BA N telomer alcohol for set one, and 2.332 pglmL of Zonyl BA L telomer alcohol for set two.

Blank control cultures received 25 mL of mineral medium solution, as did the test culture, but without addition of the test substrate.

All of the day zero cultures were prepared and immediately frozen at  20 ° Cfor storage until SPE preparation could be conducted. All other cultures were placed in temperature controlled shaking incubators that were maintained at 25 ° C ± 3 ° C. Cultures were removed from the incubators at designated time points. Upon removal from the incubator,

cultures were either immediately frozen, or immediately prepared for analysis by solid phase extraction.

All culture preparation information, including times, analyte additions, etc.

were recorded in sample preparation worksheets and signed and dated by the preparation analyst. All original data sheets were maintained in project specific binder labeled as Project CA085.

7.1.3. Solid Phase Extraction of Cultures The solid phase extraction procedure described below was documented as Pace standard operating procedure ( SOP) CAG SP 04 ~.

All cultures and control cultures were prepared by solid phase extraction methodology using SEP VAC C18 6cc SPE cartridges from Waters Corporation ( Part No. WAT036905). A sample label was applied to each SPE cartridge prior to use, and each cartridge was packed with plug of quartz glass wool to deter plugging. Each SPE cartridge was washed prior to use by drawing 5 niL of methanol and then 5 mL of aqueous 1%

acetic acid solution through the cartridge. These wash solution eluates were discarded to waste. All of the SPE eluates for this study were collected in clear 1 Chem vials with labels that identified them as eluate 1, 2 or 3, as defined below.

Page 12of38 Pace Project CAOB5: ` Biodegradation Screen Study for Telonier Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services. Bio Analytical Services Group Wednesday, November 06, 2002 000014 Frozen cultures were thawed at ambient room temperature before extraction. Following thawing, and prior to solid phase extraction, 0.25 mL of glacial acetic acid was added to each of the cultures yielding a final concentration of 1% acetic acid. The content of each acidified culture was swirled to mix, and then drawn by vacuum through the appropriately labeled SPE cartridge by carefully pouring the contents of the culture flask into the SPE cartridge. The aqueous eluate was collected in an 1 Chem vial labeled eluate 1, removed from the vacuum manifold, and capped. Then, 25 mL of methanol was added to the culture flask, the flask sealed, and vigorously shaken. The cap was then removed from the flask, and the methanol content ( 25 mL) drawn through the SPE cartridge, collected in an 1 Chem vial labeled eluate 2.

Eluate 2 was expected to contain a majority of the analyte that was in the original culture.

As a precaution that some analyte may be retained in the SPE cartridge or in the culture flask, a second 25 mL methanol eluate was collected in a similar fashion to that collected for eluate 2, and was labeled eluate 3.

Aliquots of eluates 2 and 3 were transferred to autovials, capped, and then quantitatively analyzed by HPLC/ MS. The remaining volume of each eluate was stored at 4 ° C ± 2 ° C.

The final SPE extractions of the day 7, 14 and 16 day cultures from set two occurred on September 24, 2001.

7.2 Instrumental Analysis ( LC/ MS)

7.2.1 Instrument Parameters The HPLC/ MS method used was a modified version of Pace method CAGORG 23 as described below.

Analysis of culture extracts from culture set one, prepared with Zonyl BA N telomer alcohol, was conducted using instrument " STING" which included an HPI100 HPLC pump with Gilson 215 liquid handling system in line with a Micromass Quattro II triple quadrapole mass spectrometer detector.

Page 13 of 38 Pace Project CAOS5: " Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06. 2002 000015 Analysis of culture extracts from culture set two, prepared with Zonyl BA L telomer alcohol, were analyzed both on instrument " STING' and instrument " 1 OLCMSO3" which consisted of a Waters 2690 HPLC system in line with a Micromass Quattro II triple quadrapole mass spectrometer detector, Typical conditions for analysis of telomer alcohols and fluorochemical acid products were as follows:

Typical injection volumes for samples and calibration standards were 50 ML. The 4.6 x 150 mm Betasil CS column used for the quantitation of extracts from culture set two had serial number 1101567H and the 4 x 35 mm NG1 column used had serial number 15104. A pressure regulated splitter was used with an approximate split ration of 2: 1 ( Waste: MS)

7.2.2 Qualitative Parent Analyte and Products Analysis.

Parent Analyte Expected tbns ( mix) I C6 telomer alcohol acetate adduct 423 C10 telomer alcohol acetate adduct 523 C12 telomer alcohol acetate adduct 623 C~ 4telomer alcohol acetate adduct 723 C16 telomer alcohol acetate adduct 823 Table 1. Parent telomer alcohols analyzed.

Page 14 of 38 Pace Project CA085: ` Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 Instrument conditions for HPLC/ MS analysis.

Mass Spectrometer HPLC Ionization mode: API ES negative Desolvation Temperature: 200 ° C Source Block Temperature: 150' C RF lens: 0.2 Extractor: 3 Cone: B Capillary: 3.50 LM Resolution: 14 HM Resolution: 14 Ion Energy: 2.5 Lens 6: 2 MS1 Multiplier: 650 MS/ MS: Collision Energy varied from MS/ MS Collision Gas: Argon MS2 Multiplier: 750 Time ( mm). ~ A 0.00 97.0 3.0 0.50 97.0 3.0 5.00 5.0 95.0 11.00 5.0 95.0 11.50 97.0 3.0 15.00 97.0 3.0 Flow: 1 mLlmin, splitter approx. 3: 1 Solvent A= 2 mM ammonium acetate Solvent6= Methanol column Temperature: ambient 10 to 40 V for individual Experiments,

000016 To conduct the analysis of parenl compounds and expected biodegradation products, the HPLC/ MS system was set up with the same chromatographic configuration as that described in method CAG ORG

23 ~. The method utilized two columns in tandem with flow to a pressure relief valve that serves as a flow through splitter to the mass spectrometer Z spray source.

Specific MS conditions for mass analysis of the telomer alcohols and acid products were developed during this study. The mass spectrometer,

Micromass triple quadrapole mass spectrometer with Z spray ion source,

was operated with electrospray ionization in either selected ion recording ( SIR) mode or with mass range ( 150 1000 m/ z) scanning on MS1.

Discrete chromatographic peaks with singly charged negative ions, ( M +

Acetate~ for parent telomer alcohols and ( M  H'~' for expected acid biodegradation products, were observed and monitored:

Expected Product Expected anion structure CF3( CF2) 2COO~ Expected anions ( mix)

Perfluorobutyrate ( C4) 213,169 Perfiuoropentanoate ( C5) CF3( CF2) 2C00' 263, 219 Perfluorohexanoate ( C6) CF3( CF2J~ COO' 313, 269 Perfluoroheptanoate ( C7) CF3( CF2) 5C00' 363, 319 Perfluorooctanoate ( C8, PFOA) CF3( CF2) sCOO~ 413, 369 Perfluorononanoate ( C9) CFa( CF2) 7C00' 463, 419 Perfluorodecanoate ( C10) CF3( CF2) 5C00' 513, 469 Perfluoroundecanoate ( C11) CF3( CF2) 9COO~ 563, 519 Perfluorododecanoate ( C12) CF3( CF2) 10C00' 613, 569 2K, 2H perfiuorooctanoate ( C3) CF3( CF2) 5CH2C00' 377 2K, 2H perfluorodecanoate ( C10) CF3( CF2) 7CH2COO' 477 2H. 2H perfluorododecanoate ( C12) CF3( CF2) 9CH2COO' 577 Table 2. Expected products, and expected anions for each.

Page 15 of 38 Pace Project CA085: " Biodegradation Screen Study for Telomer Alcohols'   Revision 1 Sludy Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 000017 7.2.3 Quantitative Analysis.

QuanUtative analysis was conducted only for perfluorooctanoate ( PFOA)

and only on extracts of culture set two. Quantitative analysis was performed by the external standard method using SPE extracted calibration standards ( eluate 2 only) and quadratic calibration curves,

The HPLCIMS analysis of culture set one extracts was used primarily for method development purposes.

Typical injection volumes for samples and calibration standards were 50 ML. The 4.6 x 150 mm Betasil C8 column used for the quantitation of extracts from culture set two had serial number 1101 567H and the 4 x 35 mm NG1 column used had serial number 15104. The pressureregulated splitter had no identifying number to distinguish it. The part number for ordering the pressure relief valve ( splitter) was Alitech catalog part number 39025.

7.3 Data Transformations and Calculations 7.3.1 Molar Calculations:

Because all data was collected on an ng/ mL basis ( part per billion, ppb),

a b'ansformation from ng/ mL to molar concentrations had to be conducted to obtain mass balance information when applicable. The mole conversion values for each analyte are as follows:

PFOA molecular weight, as ammonium salt, is 431.10 Zoriyl BA L telomer alcohol average molecular weight is 434.70.

7.3.2 Conversion of ng/ mL to micromolar ( MM) and nanomolar ( nM).

( Working Examples):

2,232 ng/ mL telomer alcohol = ( 2,232 ng/ mL) t( lnmole / 434.7 ng) =

5.135 nmole/ mL = 5.135 gmole/ liter = 5.135 MM   assuming 100% purity.

150 ng/ mL PFOA ( Nl 14+ salt) = ( 150 ng/ mL) * ( 1 nmole / 431,1 ng)

= 0.3479 nmolelmL = 0.3479 Mmole/ L = 0.3479 MM = 347.9 nM Page 16of38 Pace Project CAOBS: ` Biodegradation Screen Study for Telomer Alcohols"   Revision I Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Gb Analytical Services Group Wednesday, November 06, 2002 000018 7.3.3 Molar Mass Balance Calculations ( Theoretical Yield):

Convert all ng/ mL values to their corresponding molar concentrations as aM or nM ( see section 7.3.2, above). Divide the sum of the analyte concentrations by the known concentration of starting compound and represent the final result as a percentage of the known starting concentration.

( Working Example):

If, the starting concentration of BA L telomer alcohol was at 5.135MM And, after incubation, the following was determined:

If, PFOA was detected at 0.3479gM Then, the mass balance is as follows:

Mass balance = [( 0.3479 MM) / 5.135 MMIX 100% Mass balance = [ 0.0678] X 100% = 6.78%

Or, 6.78% of the telomer BA L alcohol was oxidized to form PFOA.

7.4 Software Versions Microsoft~ Excel 2000 was used for data processing and producing tables.

MicrosoftlM Word 2000 was used for processing the analytical report text.

Adobe Acrobat 4.0 was used for generation of the final electronic report.

Masslynx version 3.2 was used for data collection and peak integration.

ACD Chemsketch version 4.0 was used for chemical drawings.

Page 17 of 38 Pace Prcject CA085: " Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Sludy Director Cleston C. Lange, Ph. D. Pace Analytical Services, Gb Analytical Services Group Wednesday, November 06, 2002 000019 8.0 Results 8.1 Sludge Characterization A chemical analysis of the mixed liquor suspended solids used for the samples prepared on August 9, 2000 was conducted, and characterization information can be found in the final report for project CA058'. The sludge for samples prepared on September 27, 2000 was not characterized, but was obtained from the Twin Cities Municipal waste treatment facility in a manner consistent with the sludge collected and used for project CA058.

8.2 Quality Control/ Sample Matrix Spike Results,

The determination of the analyte recoveries from sample matrix spikes was not included as part of this screening study. Recoveries are only reflected in the ability to achieve molar mass balance based on expected parent and product yields from samples and by the use of a sludge extracted curve for semiquantitative analysis of PFOA.

8.3 Analytical Blanks Methanol solvent blanks were injected onto the HPLC/ MS column and quantitatively analyzed to determine the instrument background analyte concentration and carry over during the analysis.

The no sludge ( abiotic) controls containing mineral salts medium with test analyte, and culture blanks, containing mineral salts medium with test analyte and no sludge, were prepared and analyzed. Abiotic controls and blank cultures were prepared and incubated in an identical manner to biodegradation test cultures. The results were used to determine whether the sample matrix contained any of the analytes of interest and whether biodegradation could be attributed to the bioactivity of the sludge. Fluorochemical analytes were not detected in blanks and biodegradation did not occur in abiotic controls.

Page 18of38 Pace Project CA085: ` Biodegradation Screen Study for Telomer Alcohols'   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday. November 06, 2002 000020 8.4 HPLC/ MS Analysis Results Semi quantitative analysis was conducted only for the SPE eluate 2 extracts from the second set of cultures, prepared September 27, 2000, and were performed on September of 2001 as sequence CA085fi92401b. spl ( with full scan MS data), and CAOB5_ 092701a. spl ( SIR data) on instrument 1OLCMSO3 in the Bio

Analytical services group at Pace. The qualitative HPLC/ MS analyses and HPLC/ MS method development for this study were conducted in September and October of 2000 as analytical sequence runs S091500. spl, S09I800. spl,

SO92000. spl, SO92100. spl, S092600. spl, S1000200. spl, and S100400. spl on Pace instrument " STING".

8.4.2 Qualitative HPLCIMS and HPLC/ MS/ MS Results The first set of test cultures containing sludge and BA N telomer alcohol substrate provided evidence that biodegradation occurred. The HPLC/ MS data showed near total loss of all of the telomer alcohol peaks in the HPLC/ MS chromatogram, with formation of chromatographic peaks that had mass spectra consistent with perfluorinated acids. However, at that time, the analytical test method was not complete. Concerns about the integrity of the test material resulted in a second set of test cultures being prepared using commercially purchased Zonyl BA L telomer intermediate.

Although each sample, control, and blank for the second set of cultures was prepared and incubated in duplicate, analysis was conducted for SPE eluate 2 extracts from one sample, control and blank pertime point.

The data showed that the sample test cultures underwent rapid loss of the C5, C9, C10, and C12 telomer alcohols, and moderate loss of the C14 telomer alcohol over the 16 days of incubation, as determined by decreasing peak area response for each telomer alcohol ion at the sequential sampling time points ( Figures 3 and 4). The C16 telorner alcohol was degraded very slowly and showed little loss.

Page 19 of 38 Pace Project CAO85: " Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday. November 06, 2002 000021. 35000 30000 25000 20000 15000 10000 5000 0 0 5 10 15 20 ° C8 TA peak area AdO TA peak area °° C12 TA peak area Figure 3. HPLCIMS integrated peak areas for the most abundant telomer alcohols ( TA). Near complete degradation of all three was observed. Data plotted with the best manual fit of the data,

6000 5000 4000 ~ C6TA peak area 3000 ° uC14 TApeak area ° C16 TA peak area ~ 2000 0 a

1000 0 Incubaiton time ( days)

Figure 4. HPLC/ MS integrated peak areas for the less abundant telomer alcohols ( TA). Near complete degradation was observed for the C6 TA. Moderate degradation of the C1~ TAwas observed and little degradation of the C19 TA was observed. Data plotted with the best manual fit of the data.

Page 20 of 38 Pace Project CAO8S: ~ Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 000022 0 5 10 15 20 Concomitant with the loss in telomer alcohols was the formation of compounds with HPLC/ MS ions and retention times consistent with perfluorinated adds ( PFAs), inc! uding perfluorooctanoate ( PFO~ A, CrPF. A),

which was confirmed as an end product. The transformation of telomer alcohols to PFAs was rapid, with a large increase in all of the PFA peak area responses at day 1, and continuing increase through day i6 ( Figure 5). No degradation to form PFAs was observed in the abiotic no sludge controls.

Although PFOA was accurately quantified, many other perfluorinated acids were qualitatively observed and their relative concentrations determined based on the observed increasing peak area response. The observed perfluorinated fatty acid end products were: perfluoropentanoic acid ( C5 PFA), per fluorohexanoic acid ( C6 PFA), perfiuoroheptanoic acid ( CrPFA),

perfluorooctanoic acid ( C8 PFA, PFOA), perfluorononanoic acid ( CrPFA),

perfluorodecanoic acid ( C, 0 PFA), perfluoroundecanoic acid ( C11 PFA) and perfiuorododecanoic acid ( C1rPFA).

Transientlyformed intermediate compounds were also observed at early time points following the initial exposure of the sludge to the telomer alcohol substrate ( Figure 6). The transient compounds were suspected to be: 2H, 2H perfluorooctanoate; 2H, 2H   perfluorodecanoate; 2H, 2H Perfluorododecanoate and the possible f3 oxidation pathway intermediates:

21 l perfluoro 2 octenoate, 2H perfluoro 2 decenoate, and 2H perfluoro 2

dodecenoate. Table 3 shows the MS ions and retention times observed for the polyfluorinated and perfluorinated acid products and the telomer alcohol parent substrates.

HPLC/ MS/ MS Data ( Table 4) support the identification of the suspected metabolites..

Page 21 of 38 Pace Project CA085: " Biodegradation Screen Study for Telomer Alcohols'   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 000023 FIgure 5. Integrated peak area responses for the pertluorinated fatty acids ( PFAs)

observed in test culture extracts. Even numbered carbon chain length carboxylic acids were the most abundant carboxylic acid peaks and were: perfluorohexaooate ( C5 PFA),

perfluorooctanoate ( C3 PFA) and perfluorodecanoate ( Cio PFA),

Figure 6. Transient polyfluorinated fatty acid intermediates observed. The 2Hperfluorinated olefinic fatty acids are suspected ~. oxidationintermediates that formed rapidly upon initial exposure to telomer alcohols, and then were transformed to their corresponding pertluorinated carboxylic acids end products Page 22 of 38 Pace Project CA085: Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 000024 250000 200000 C,

k9 a a. 150000 100000 50000 01 0     2 4 6 8 10 12 Incubation time ( days) IA 11 14 16 18 rAnal~ esObserved Chemical Formula Retention Time LCIMS anions F observed ft~ ieiorneralcohol* CF3( CF2) 3CH2CH2OH.. .

CH3COO" 7.63 323 1 C6 telomer alcohol CF3( CF2) 6CH2CH2OH.. CH3COGY 8.13 423 010 telorner alcohol* CF3( CF2) 7CH2CH2OH. .. CH3COO~ 8.50 523 C, 2 telomer alcohol' CF3( CF2) 9CH2CI 120H... CH3COQ  8.96 623 C, 4 telomer alcohol' CF3( CF2) jiCl 42CH2OH... CH3COO~ 9.53 723 C, 6 telomer alcohol' CF3( CF2) 13CH2CH2OH.. . CH3COO 10.25 923 2H, 21 f perfluorooctanoate' CF3( CF2) 5CH2COO~ 7.17 377 2H, 2H pertluorodecanoate' CF3( CF2) 7CR2C00' 7.38 477 2H, 2H Perfluorododecanoate' CF3( CF2) gCH2COO~ 749 577 2H perfluoro 2 octenoate' CF3( CF2) 4CF= CHCOO~ 7.14 357293 2H perfluoro 2 decenoate' CF3( CF2) 6CF= CHCOO~ 735 457393 2H Perfluoro 2 dodecenoate' CF3( CF2) 8CF= CHCOO~ 752 557,493 Perfluoropentanoate CF3( CF2) 3COO~ 6.62 263, 219 Pertluorohexanoate CF3( CF2) 4C00" 6.88 313, 269 Pertluoroheptanoate CF3( CF2) 5C00 7.07 363, 319 Pertluorooctanoate CF3( CF2) 5C00  7.22 413, 369 Perfluorononanoate Pertluorodecanoate CF3( CF2) 7C00 CF3~ CF2) 6COO" 7.30 7.41 463, 419 513, 469 1 Perfluorouridecanoate CF3( CF2) 9C00 7.48 563. 519 Perfluorododecanoate QF3( CF2), oCOO~ 7.52 613, 569 Only anions corresponding to even chain length molecules were observed.

Table 3. Parent and products anions observed in by HPLC/ MS analysis of SPE eluate2 extracts from test cultures containing sludge and telomer alcohol substrate. The parent anion signals decreased in test cultures over time concomitant with increases in product signals. Many of the telomer alcohol signals decreased below detection limits in test cultures.

Page 23 of 38 Pace Project CAOB5: " Biodegradation Screen Study for Telomer Alcohols'   Revision I Study Director: Cleston C. Lange, Ph. D. Pace Anafyticat Services, Bio Analytical Services Group Wednesday, November 06, 2002 000025 Product Analytes Observed Chemical Formula Retention Time LC/ MS/ MS LC/ MS/ MS daughter Parent anion anions observed 2H ,2H~ perfiuorooctanoate* CF3( CF2) 5CH2COcY 7.17 377 333,31 3,293 21 l, 2H perfluorodecanoate CF3( CF2) 7CH2COO  7.38 477 413,393 2H, 2H Perfluorododecanoate* CF3( CF2)~ CH2COO 7,49 577 .

2H. perf1uoro~ 2~ octenoate* CF3( CF2) 4CF= CHCOO 7.14 357 313,293,243,143,119 2H. perfluoro~ 2~ decenoate* CF3( CF2) 6CFCHCOO 7.35 457 413,393.343 2H Perfluoro 2 dodecenoate CF3( CF2) 5CF= CHCOO  I ` I~

Table 4. HPLC/ MS! MS data for suspected  oxidation products. Parent and daughter anions from theHPLC/ MS/ MS analysis of the SPE eluate 2 extract from the 1 day biodegradation sample ( CA085

0927 SA 033 E2). The parent anions were observed as transiently formed anions in early sample point biodegradation samples only, with the most intense signals for the ions observed at day one. The MS/ MS data collected for each product was consistent with the predicted chemical formula. Common mass loss from the parent anion, attributed to loss of CO2 ( mass 44) and HF ( mass 20), were observed.

8.4.2 Quantitative Analysis of PFOA The HPLC/ MS multi component calibration standards used in this study for product quantitation contained the target analytes: PFOA, PFOSulfinate, PFOS,

FOSA, N MeFOSA, M556, M570, and N MeFOSE alcohol. Five calibration standards were prepared by addition of a known amount o(, perfluorinated analytes to a culture medium with sludge, and then extraction by solid phase extraction in manner identical to the treatment of samples. The PFOA was the only target analyte of that mixture that was used for this study. The exact concentrations of the standards were used in calibration curves for quantitative analysis.

Page 24 of 38 Pace Project CA085: ` Biodegradation Screen Study for Telomer Alcohols'   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 000026 All calibration standards and sanip~ eeluates were stored at 4 ° Cin refrigerator ID 0213 until analysis. Aliquots were transferred to autovials and capped for use in HPLC/ MS runs. The instrument calibrations were performed for PFOA by use of a five point calibration with quadratic fit of the data. The LLOQ was 11.0 ng/ mL PFOA, and the coefficient of determination ( r) for PFOA was 0.965. Quantitative sequence runs contained calibration standards at the beginning and end of the run.

The biodegradation of the telomer alcohols resulted in all increase in the measured levels of PFOA during the 16 day study ( Figure 7).

Figure 7. The measured perfluorooctarioate ( PFOA) in cultures containing sludge and fluorotelomer alcohol. The PFOA was not measurable ri cultures that did not contain sludge. Data plotted with the best manual fit of the data and concentrations shown were not adjusted for reference material purity of 95.2%.

Page 25 of 38 Pace Project CA085: ` Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, B( o Analytical Services Group Wednesday, November 06, 2002 180.0 160.0 140.0 j' 120.0 100.0 80.0 a. 60.0 40.0 20.0 0.0 0 2 4 6 8 10 12 14 16 15 Incubafion time ( days)

000027 F   ` j' dsMra9wlase F F F F HH F F F F 0 Oxidatlve decarboxylatlon h~ tcøan ( a oxIdation)

F F F F F F F F OH Dial F) I I! K F F F F F F F F 0 & lase n OxidatIon 0~

F F F F Ilibi ~

F F F F F F F F F F F F F F F 111a2 F 4 F F F F F P F F 0 Perftuorortonanoate ÷ Co2 F 111111! IH F F F F F F F F ,~, 4efl~ 0coq~ a~ e 4, P. ket~ c~ l FE P F F F F 0 IHb3 F________________________

F F F F F F 0 Perfi uorooctanoate +

M 3 C~~ Q Acetate Figure 8. Proposed biodegradation pathway for telomer alcohols, shown for C10 telomer alcohol as example. The p oxidation pathway branch ( Ilib) involves primary formation of a fatty acyl CoA thioester that is not shown, to clearly depict the products observed by HPLC/ MS. The fatty acyl CoA thiol bond is readily hydrolyzed to yield the corresponding fatty acids observed.

Page 26 of 38 Pace Project CA085: ~ BiodegradationScreen Study for Telorner Alcohols'   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Sb Analytical Services Group Wednesday, November 06, 2002 000028 F I I . L I I I I I ~`~ 0K C, Tetomer Aicohol diydrog&' ase ~ r r i ii ii, 9.0 Conclusions This study has demonstrated that fluorochernical telomer alcohots are biodegradable and are transformed to perfluorinated acid end products. Although the telomer alcohols are supplied only as even numbered carbon chain compounds, the end product perfluorinated acids consist of both even number and odd number carbon chains, with even numbered carbon chain length acids predominating as the major end products.

This observation suggests that, following the initial oxidation of the hydroxyl carbon to torn, the primary carboxylic acid ( telomer carboxylata), two oxidation mechanisms exist as shown in Figure 8, III. The first, less utilized route ( Figure 8, lila), involves oxidation of the a carbon concomitant with decarboxylation ( oxidative decarboxylation) to form an odd chain length perfluorinated carboxylic acid, which does not undergo further biotransformation. The second observed route of oxidation ( Figure 8, IlIb) is the more common route of fatty acid ~ oxidation, as evidenced by the formation of detectable transient p oxidation polyfluorinated fatty acid intermediates and the more abundant even number carbon chain length carboxyiic acids Although toxicity of the telomer alcohols was not addressed as part of this study, the microbial degradation observed here over the entirety of the 16 day study suggests that microorganisms present in the sludge were not significantly inhibited by the tested concentration of the test material.

Page 27 of 38 Pace Project CA085: " Biodegradation Screen Study forTelomer Alcohols'   Revision I Study Director: Cleston C. Lange, Ph. D. PaceAnalytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 000029 8.0 Literature Cited 1. Final Reports for Pace Project CA058 ( 3M ft E00 2252), " 2 Week N EtFOSE Alcohol Biodegradation Screen Study Report" and " Aerobic Biodegradation of N EtFOSE alcohol Study Report'. Author: Cleston C. Lange, Ph. D.

2. Final report for Pace Project CA097 ( 3M # E01  0415), " The 18 Day Aerobic Biodegradation Study of Perfluorooctanesulfonyl based Chemistries." Author:

Cleston C. Lange, Ph. D.

3. Final report for Pace Project CAIO4 ( 3M # E01 0444), " The 35 Day Aerobic Biodegradation Study of Perfluorooctanesulfonate ( PFOS)." Author: Cleston C.

Lange. Ph. D.

4. Final Report for Pace Project CAl 05 ( 3M ft E01 0683), " Fluorochemical Adipate ( LI 5468) Biodegradation Screen Study.' Final report issued July 10, 2001.

Study Director: Cleston C. Lange, Ph. D.

5. Final Report for Pace Project CAl 32 ( 3M ft E01 0682). " The Aerobic Biodegradation Study ot the Fluorochemical FC807 Diester". Final report issued May 29, 2001. Study Director: Cleston C. Lange, Ph. D.

6. Pace method CAG SP 03. " Culture Preparation for Assessment of Aerobic Biodegradability of Fluorochemicals Using Municipal or Industrial Sludge as Microbial Inoculum."

7. Pace method CAG SP 04. " C18 Solid Phase Extraction Procedure for Fluorochemicals Recovery from Aqueous/ Sludge Matrices."

8. Pace method CAG ORG 23 " Quantitative Analysis of Fluorochernicals by High Performance Liquid Chromatography with Mass Spectrometric Detection."

Page 28 of 38 Pace Project CA085: " Biodegradation Screen Study for Telomer Alcohols'   Revision I Study Director: Cleston C. Lange. Ph. D. Pace Analytical Services, Blo Analytical Services Group Wednesday, November 06, 2002 ooooao 9.0 Sample and Data Retention At a minimum, one copy of all pertinent raw data and one copy of the signed final report will be retained in the Pace Analytical Tier 2 data archives for a minimum period of 2 years after completion of the project. The remaining sample extracts will be retained at the Pace Analytical facility for a period of 2 years after completion of the project at 4 ° Cin the Carroll walk in cooler ( Pace ID 0140) located in the Pace Analytical Tier 2 facility.

The following will be provided to the sponsor ( 3M):

° The original signed analytical report and one copy of the signed original.

° The final scanned report ( read only) and pertinent electronic data on a CD.

° All original Data, correspondence, chromatograms, sample & standards prep sheets, etc.

* Upon request before 2 years, the stored samples may be sent to the sponsor.

All electronic copies of the instrumental raw data will be archived onto CD disks and one copy provided to 3M and one copy retained at Pace.

Facility data will be retained for a period of 10 years. Facility data is available for inspection and includes thefollowing records:

° Training records ° Controlled storage temperature logs ° Standard preparation logs ° Calibration and maintenance logs ° Chemical and solvent traceability logs ° Standard Operating Procedures ° Methods pertaining to the conduct of this project Page 29 of 38 Pace Project CA085: " Biodegradation Screen Study for Telomer Alcohols'   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Anatytical Services, Sb Analytical Services Group Wednesday, November 06, 2002 000031 APPENDIX A APPIOVAL S~ eNATuREs Project Title: Biodegradation Screen Study for Telomer Alcohols Client Project ID: EUl  0684 Contract Analytical Project Numben 0A085 Report Rev~ sedby:

MouractRahi_ 1k7 0z Date Laboratory Management:

Bruce~ j~ den   Signature Page 30 of 38 Pace Project CAO8S: " Biodegradation Screen Study for Telomer Alcohols"   Revision I Study Director: Cleston C. Lange, Ph. D. PaceAnalytical Services, BJo AnaJyticaJ Services Group Wednesday, November 06, 2002 000032 Appendix B Final Quantitative Results for PFOA in samples, controls and blanks Table 1. The measured PFOA concentrations in test cultures during the 16 day screening study for telorner alcohol aerobic biodegradability.

JConc. ntnffco, n~ 4, t~ Eluat. 21 Snh PF0A DaySb~ d~ n4a5on sacn, CA085' 0927. SA'DDt. 02 0.0 Day 0~ deVa4asonsriiyla. C4085 0027 SA. 033 02 252 Day 2 bndeo, adat, rn, aampl°, C4085 002? SA' 035, 02 650 Cay 5 bcdOnd~' onsample, CAOaSO92l'. SA.~ 1.02 I 1.4 Day 7 b~ ecradrionsample. cA085 0027. SA. 039 02 ` 39.8 Day I. scdearadaelon sample, CAOSS 0027. SA 04I 02 ` Ste Day IS S da'adaelon sample. CAOO5 0921 SA 043, 02 1600 Table 2. The measured PFOA concentrations in the abiotic control cultures during the 16 day screening study fortelomer alcohol aerobic biodegradability.

Coticentnfton OnGkaLj [ Elssat~ 2 SampM PPOA Days r~ sludge ~ mroI, CAOSS 0927. SA. 053, 22 ° 0 lay I ~ slid 0 , co, st, sl. 0A08S0921. SA. 065, 02 0.0 lay 2 Im sludge wlslrOI, CA085.0927. SA. 056 22 0.0 layS Im didse ~ mlol, CAOSSOO27. SAOSQ 22 0.0 lay? to sltdQe ~ mml, CA085' 0027.5A 060. 22 0.0 lay 14 rIO slidIm toMiol, CAOOS 0927. SA 062 22 0.0 lay IS to sludge~ r4roI. CAOOS 0027. SA ® 4, 22 0.0 Table 3. The final data results for sludge blanks during the 16 day screening study for telomer alcohol aerobic biodegradability. PFOA was not detected in any of the sludge blanks.

Concentrallon ( ngknL) IEluaO. 2J Sam~* PFOA Dayssludge Slayic. n_ A0a5.0021. SA. 045 02 0.0 Day I sludge 04*. Aoe5~ 092T SA o4a. 02 00 Day2 sludge slaM. 0A01S0027 SA 047. 02 0.0 Day S sludge 5124*. CAO85 0927 S404e. 02 00 Day? sludge slr*. CAOBS. 0927' SA DiQ, 22 0.0 Day 4 sludge bla', k, CAO5S. 0927 SA. 050. 22 00 Day II sludge blaik, CA085' 0927 SA. 051. E2 00 Page 31 of38 Pace Project CA085: " Biodegradation Screen Study for Telomer Alcohols"   Revision I Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bia Analytical Services Group Wednesday, November 06, 2002 000033 Appendix C Integrated Chromatogram Peak Area Data for telomer alcohols Table 1. Integrated LC/ MS peak area raw data for the parent telomer alcohols substrates. Data acquired from sequence run CA085_ 092701a. spl as SIR data.

Telomer Alcohol Integraled Peslc Mess SampI. PIle lb CS CS do c12 c14 Cli SUM lay_ 0_ a, 0de~ ada, FDnsampe, CA08S0927.$ A. 031, 22 CAOSS 092701a 008 3503 21704 26040 9446 5550 320 326 lay I b'od'gadau. op sampo, C4055' 0927'$ A. 033, E2 Day 2 5,03eoradat! 0l sample. CA065~ 092?~ SA. 03S, 62 cAO8S 0927Db 000 ND SI laos 0833 4766 0310 1340 CAGeS 092701a 010 ND 298 262 88fl 4403 1213 1213 Days s, sdsqradaoou ssmme Ca0e5~ 092? Sa. 037 22 CAOO5 092701 a 011 12 090 03 3070 3701 1140 1140 0807 Olsde5radalIonsampl., CA005' 012?. SA' 039, 62 ~ A005 092701a 012 ND S31 NO 1263 3240 5343 ` 363 Day 14 b'odeorsdatOl sample. CA065 092?.$ A. D41, 62 ~ A085D92701a 013 ND 594 NO 670 1124 1093 0093 Day 10b, odeoradlioy sample CAOS5 ® 2?. SA 043. 62 ~ slL,~ C0, Il0l. cAoasoO27' SA' 063. 22 CA085_ 09270l8 014 cAoes ognoia 016 ND 5072 043 30841 II?

22594 085 63734 1276 5014 lIST 1192 113?

1412 Day I to sk, Oto0074101, dAOS5' 092? SA'OSS, 22 CAO0S 092701 a 017 4890 31305 23373 1273 4712 147? 1477 Day? oo slu~ ecor4lol, cAOSS. 0927. SA. 050 22 CA005 092701a 010 5108 31002 26464 ` 3848 5379 1462 162 Days so sluOgecorIml, CA085.0927. SA. 059, 22 CAO8S 092701a 019 4303 27486 22470 12856 4941 ` 393 393 Day? no sluadgecor4ml. CA0oS.~ 7' SA'O60,22 CA065 092701a 020 3515 21400 13621 10429 4252 $ 095 1095 Day 4cm sludge 6001704. CAoa5.092? SA. 062 22 CAOOS 092701a 021 3310 21284 14368 10228 ~ 992 $ 265 0205 Day 1Gm sludge consul, C4081 092Z24.084, 62 DayS sludge blank. CA08S0027. SA'OdS. 22 CA085 D92? Ola 022 CAOOS D92701a 044 4602 ND 27329 17 27946 ND 12265 ND 5230 ND 1470 ND 5470 NO Day I slados stalk C5085.0927' SA 046 62 CAOO5 092701a 045 ND ND lID NO NO ND ND Day 2 slsoçe blank. CA08S. 0927. SA 047. 22 CAOSS D9270Ia 040 ND ND lID ND NO ND ND DayS sIsoge 5193k, cA085.0027 SA 048. 02 CA085 092701a 047 ND ND 140 ND ND ND ND Day? sliaage blank CAOSS. 003? SA 049. 02 cADSS 092701a 040 3? 102 ND ND NO ND ND Day 4 lludge blank. CA095.0927. SA. 050, 22 CAO8S 0927018 049 ND 211 ND ND ND ND ND Day lb sludge blank, CA015.0937 SA 0Sl. 62 CAO6S 0927D1a 05D I? 08 NO 269 ND ND ND Table 2. Percentage of integrated peak area remaining at different time points with respect to the peak area at time zero for their respective culture types. telomer alcohols ( TA) was observed in test cultures only. for all except C10 TA, which was 79.4%. Near complete degradation of several Near 90% remained in control cultures Sample Description . fl~ L % TA Remaining versus Day 0 c. TA C, TA C10TA c13TA C14TA c, 0TA Day 0 biodegradation sample, CAO8S 0927 SA 031, E2 0 100.0% 100.0% 100.0% 100,0% 100,0% 100.0% ) ay 1 biodegradation sanipJe. CAO8S 0927 SA 033, E2 0.0% 0.4% 4.5% 46.8% 85.8% 101.1% ) ay 2 biode~ adationsample, CA085 0927 SA 035, E2 2 0.0% 1.2% 0.9% 36.2% 80,7% 91.5% Day 5 biodegradation sample, CA055 0927 sA 037. E2 5 0.3% 3.2% 0.4% 16.6% 66.6% 86.0% Day 7 biodegradation sample, cAo8s 0927 SA 039, E2 7 0.0% 2,9% 0.0% 6.8% 58.4% 101.3% Day 14 biodegradation sample, cAO85 0927 SA 041. E2 14 0.0% 2.7% 0.0% 3.6% ` 23.8% 82.4% Day 16 biodegradation sample, CA085 0927 SA 043, E2 Day 0 no sludge control, CA085 0927 SA 053, E2 16   0 0.0%

100.0% 3.0%

100.0% 0,4%

100.0% 3,7%

100.0% 23.0%

100.0% 85.7%

100.0% Day I no sludge control, CA085 0927 SA 055, E2 1 96.4% 101.5% 90.2% 82.1% 94.0% 99.0% Day 2 no sludge control, CA065 0927 SA 056. E2 2 102.5% 103.3% 108.1% 101.0% 107.3% 98.0% DayS no sludge control, CA085 0927 SA 05g, E2 5 86.4% 89.1% 99.5% 93.6% 98.5% 93.4% Day 7 no sludge control, cA085 0927 SA 060, E2 7 69.3% 69.6% 60.3% 75.9% 84.8% 73,4% Day 14 no slud~ control. cAoss o927 SA o62 E2 14 65,4% 69.0% 63.6% 74.5% 99.6% 84.8% ) ay 16 no slud~ control. cA085 0927 SA 064, E2 16 90.7% 86.6% 79.4% 88.9% 104.3% 99.1%

Page 32 of 38 Pace Project CA085: 1Biodegradation Screen Study for Telomer Alcohols'   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday, November 06, 2002 000034 Appendix D Integrated Peak Area Data for Periluorinated Acid End Products Table 1. Integrated LC/ MS peak area raw data for the end products formed. Data acquired from sequence run CA085_ 092701a. spl as SIR data.

Inteorats Peak Arose Sample , ,

Fll'lD~'. Perflu& op ntanoata Pethooroh xanoate Pertluoro. hepunoat. Pertluorooctanost P. rfluorononasnoata Pertluorod caoo. te P. dluoro. undetafloale Pertluoro. lodocanoate SayG b, nulflbdakon oample. 0A08S0827' 5A' 031. 02 C4085 0927008 008 276 3576 12127 2715 8818 253 0589 375 Say I b. od. gradaiioo samp.. 04185.0027' 5A' 033. 02 CASeS 092701a 009 13796 46683 15484 4582$ 8388 6120 0290 505 Say? g. oaegnaoatios lamp., CA085 D921' SA' 035, 22 DAllaS 0027053 010 483$ 4 127201 25641 88508 6267 17586 1378 1192 Say S slCu900adaliOn san'p° CAO8S. 0927' SA'ODT, 22 04085 0921008 Oil 63648 204221 3574? 40809 9375 44419 1841 2605 Day? bodegraoalso nampe c4005.0921.5A. 03e. ES CAO8S 0921018 012 72810 210569 43227 170923 13751 75377 2944 * 047 Day 14 S'odadslioii lamp., C406s0927 SAo41. £ 2 05056 092701a 013 70978 221543 47751 $ 90923 15382 92188 3071 11376 ay lb Skodegnadel101 9801* cAoos. 092T'SA'oIa. 62 DAOOS 0037018 014 79828 208870 47305 692075 05069 96834 3279 10603 ay 0cm gas~ e600004. CMOSOOV. SA' 053, 92 04000 0927018 048 193 060 9244 1007 4382 134 0253 00 al Ito Sll.~ 00011110, C436S092?. SA 065, 92 ay2 cm sl,~ emood, CA085.0927. SA. 050. 62 04088 0927818 017 288 0722 16885 1695 5518 233 1692 51 04085 092701a 058 351 0437 17394 1649 5953 064 1178 20 aySm stsd9e0000,00, CA08S0927' SA' 059. 62 04085 092701a 019 478 1744 17948 1883 7120 270 1926 81 al 7cm slIme00000100, 05085' 0927' 84' 060. 62 DAllaS 092701a 020 457 0643 05212 1583 8328 173 lOIS 25 ay 08 cm sludge 01,00050. 0408S0927. SA. 06282 04085 0037018 020 580 173$ 10978 1880 7129 204 0675 98 Day 08cm lsudge6010051. cA08s0927 SA' 084. £ 2 04085 0027018 022  811 185$ 17010 1037 7203 233 1342 34 Day 0 sludge Sank. cAO8s. 1927.54445, 62 CAO85_ 092701a 044 818 525 31 II? ` 8 3 0 3 Day I nludge blank. ca00s. oo27. SA. a4~ 62 CAO8S_ 0927013_ 045 883 152 75 ` 60 262 0 0 0 Day 2 01110469 blInk. 04085.0927.54.047. 62 04085 0927018 046 686 510 51 229 722 0 0 48 Days 5111009 bllnb. C4085.0827. SA. D48. 02 04005 0927018 047 972 320 66 223 392 0 0 0 Day' sludge bla, kn, C4065,0027. SA'D40, 02 CAGeS 0027318 048 737 248 82 500 247 0 24 42 Day 14 sludge menu. 04085.0927.54.050. 62 04085 092701a 049 676 237 86 731 314 0 24 29 Day 10 sludge blank. CAOS5.0927. SA' 061. 62 04085 0927008 050 634 285 81 645 397 0 20 0 Page 33 of 36 Pace Project CAOBS: Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group 000035 Wednesday, November 06, 2002 Appendix E Integrated Peak Area Data for Transiently formed Polyfluorinated Acid Products Table 1. Integrated LC/ MS peak area data for transient products. Data from sequence CA085092401b. spl as full scan MS data from rn/ i 100 to rn/ i 1000.

 lnteQrated Peak Areas   `:.:   ~   ,  ~ ~

 Sample : FllelQ::~.: 2W porfluoro 2 octenoale 2H' p. rfluoro. 2 decenoale 2W 1H, IW IH, IHperfluoro 2' pnfluoro perfluorododecenoate octanoate decanoat, 1H, 1H.

perfluorododocanoate ayO b, od6~ adaIionsaes~ e. 04685.0927.84.031  62 ~ A085_ 09240Ib 008 1040 ND ND 3085 200 ND Day I sodooradaeonsamNe, 04685.0927 54.033 62 05085 09240W 008 3470 780 249 ISO2S 7005 958 Day Sblod, craiaoionsamøe, 0A695' 0027' SA' 025,62 Day S biodognadaoion 68,1144.. 0A085' 0837. SA' 037. 22 04005 092401b 010 223 ND ND 0020 1802 203 04086 0024006 001 ND ND ND 738 ND ND Day? biodeorldali, n 68.80. 0* 085 0927 54.039 62 0* 385 0924006 002 ND ND ND 057 ND ND Day 04 buld. 901da1000saa, pOe, C* OeSO9Rl'SA' 041 22 05086 602400b 003 ND ND ND 88 ND NP Day 1650d. gcadMloaealnpoe, C4085.0027. SA' 043. 62 04086 0824016 014 ND ND NO 128 ND ND Day acm altdge01MM. 04086 0927' SA. 063. 22 04085 0924016 008 ND ND NO ,... J~ Q,.,._.. ND ND Day Inc 906mGconlosl. C* 085.0027. SA' 065. 62 C* 085_ 092401b 017 ND ND NO ND ND ND Day 2cm slLmg0 mnl, d, 0A085' 0927. SA. 066, 62 0* 085 0924016 018 ND ND ND ND ND NO DayS nolIudgeconI, dI 04085.0037. SA. 069, 22 Day? nooludgeconInol. C* D8S0027. S*. 060. 22 0* 006 0924016 018 ND NO ND ND ND ND 0* 085 0924066 020 ND ND ND ND ND ND lay 04cm Oludge 0001001, 0* W. 0927. SA' 002 62 Say 00cm akudgemovol. 0A065' 0927' M' 064, 62 04305 0924016 020 0* 085 0924000 022 ND ND 040 20 ND NO ND ND ND NO ND ND Say0 sludge biaS 04065.0937,54045 62 0* 085 092400b 030 ND S~ NO NO ND ND Say I sludge bin cAo6a. o927. sA. ooe, 62 0* 085 0924016 031 ND ND NO ,,.. J~ 9.,.,,,,,,, ND ND Say2 sludge blank. 04085.0927 84,047,62 0* 085 0924016 032 ND ND ND ND ND ND SayS sludge blailk, C4085' 0927' SA. 04e. 62 CAO8S 092401b 033 ND ND ND ,_,, J4Q,,,,,,,,, ND ND Daylooudgeblagok. QADOS' 0927. SA 040 62 ,..., 0& 065_ 092401b 034 ND ND ND ND ND ND Say 74 lIMe blanA, 0A085.0027. SA 050. 62 Say ~ 6 ojudgeblank CAO8S 0027' SA 051. ES 05085 0920016 035 0* 065 0024016 026 ND ND ND ND ND ND ND ND ND ND ND ND Page 34 of 36 Pace Project CAOB5: " Biodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group 000036 Wednesday, November 06. 2002 Appendix F Typical Total Ion Chromatograms for SIR Data Showing Telomer Alcohols and Polyfluorinated Fatty Acid Products from Day O to Day 16 for HPLC/ MS Analysis of Biodegradation Culture SPE eluate 2.

SIR of 17 anions: rn/ i 263, 313, 363, 413, 463, 513, 563, and 613 for pm fluorinated acids; mlz 377, 477 and 577 for polyfluorinated acids; and rn/ i 323, 423, 523, 623, 723, and 823 for telomer alcohols,

Pertluorinated Fatty Acid Peaks cA385.81274* a42 22. dey II bIOde9 simple 04245 092 lOla 011 100 r ~

L ~~"` I.!, .:, lllas_ os7'/ ooa, 713 100, 91,720 742 Day 14', ~  ~

04065_ 202701a_ 617 SIR oflichann. l. ES. l. a Sloe Day 7 " 1 100 6~ 7733 1.1 9: 00 :/ LL,.,__,,__ 3___~_. 08080_ 092701a_ 211 SIR oil? Cliannoli ESNC 1.5346 0' I 722 01k ~ ) U ~

SIR oil? Cyanrslesort I 02,6 Pace Project CA085: asiodegradation Screen Study for Telomer Alcohols"   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bia Analytical Services Group Wednesday, November 06, 2002 000037 Day li" `. 8! 8,000110hannsls ES' lIT. 1.08,6 SIR 00 1105an 11 InIS 55. TIC 1.81MG 722 02 ~ J, Day~ 5~

CAo8S_ 0927011_ 000 100.

Day 2 L _.~..

lbl7oIa I lu SIR of'; Channels Es. 050 9' . rc 90 ` 747 Day i : ..... P2 ~ 79"~~ Q~,,:

III o'NIyil ` 1,8,01, ES 100 [`   I TIC Day a : J0~'~_?~~ `~ t~_' L :,..., ttr2,4 ,. ,~_._.

100 2.00 300 400 5.00 5.02 700 0.00 900 1200 11.02 12100 0300 7420 Telomer Alcohol Peaks Page 35 of 38 Appendix G SIR data Showing Ions for Telomer Alcohols at Day O and Day i 6 from Analysis of the Biodegradation Culture SPE eluate 2.

CA085 0927 84421 22. day a biodeg sample CAE 105 015.' YE 1,4 lOS ISO 0S SIR of olcnaon, 4o ES 431000001, S OdeS SIR of 17C$ oannels 6$' S23. ce 2.000a 50065 Pace Project CAOB5: " Biodegradation Screen Study for Telomer Alcohols'   Revision 1 Study Director: Cleston C. Lange, PhD. Pace Analytical Services, Blo Analytical Services Group Wednesday, November 06, 2002 SIR or Il011annOIs 53' TIC 50065 000038 7.50 913 C4500 270 a ` 108 1 SIR or IT 004,0201,05 ice ~ 23N1 I 0000 50065 44 ` 53 o  ....    ,     "~    . C4005092701a,. 006 SIR of? Channels ES. 000 523.00 1 CODa 5.0005 tan ,~ ~,

06215,. C92? Ola.. I08 SIR oil? ChofInelo ES 523.00 0, ceCa 5,0045 14 `.

05065 09270ia~ SIRoil? ChaRlEs 65, 00 4230010009 50065 S e., 2 .`   , .       .            _ . ,. ~

CROSS ( 52702a 008 SIR Oil? Channels ES 000 32300 I CODS 5.0005 S 0          r ,.,    ~~, r   CAEOS_ 0927OIafi08 SIR 0117 ChanloelS ES'

cm. 000 I .~ 1~~ nnzr~= tr   tme .00 2.00 3.00 4,00 5.00 6.00 7.00 6.00 9.00 50.00 11.00 52.00 300 14.00 cAOas 0927 sA 043 22, day it blodeg sampia CASES 0h275I~ 014 SIR 0127 0114( 100.1% ES.

100 6230015000 Scales 9.

00585 51112701a 0t4 SIR oIl? CTIOSEOOIS ES. ICE 723.00 SODa 5 00e6 S cAoos_ oo2? o'a_ o'. . SIR oil? C6, o, vlolo ES' ice 023.00 i. 000a SOOeS S CREOS_ 092701 9,014 Ice S CAII05_ 41927010_ 014 SIR all y CNan, eIs ES' 00 41220 I 0034 50045 94 SIR or? O'annos ES'

100 ` U lb 0 ~    ` 201145 1412' OIal I ice   ~

S ;,; 942 4.20 ` ""` J 4 '_._ .,___.. ._,~ ,,.   0           11150 000 2.00 3.00 ON 5.00 6.00 7,00 8.00 9.00 1000 11.00 1200 2200 1400 Page 36 of 38 Appendix H SIR data Showing Ions for Perfluorinated Fatty Acids at Day O and Day I 6 from Analysis of the Biodegradation CuLture SPE eluate 2.

C408S' 092? SA. O31 E2, day 0 biude~, ampie Cs065_ 091701a_ 008 51R of 17 CEanwIcOs ESco 813001.000S 45 2. OOeô c., ootaor7oIaj~ .  .  Sl~ 0( I? CnannOla ES' 003 500.00 CODa 2. CEleO 0'~'.`     . `"      ` _   ` .`     `    `..   `   ,   ` ..     _ r__..   ' .      '    .       t.    .,.',     , .     _ '_.'_.    ,     '   , .'   . .      ".   "           _     . '     C4088_ 002701&, 008 SIR oil? CII8nII. S 60 100  $ 13 00 I 001la aCoen .5 GA005,., J027010008 SIR III I? CEliflfdIE ES'

50 4170( 11 OSlo 2001o11 __________ __________ 6.5O   ,   _ .`    ".`_, `. In. l"'' r". l ,,._`._`   ,   _ . I     '   ' _.'__',.__.   ' _.   .      __,     _   _ ..   _   , ._   _  _,.,.,, Cs065,, 09llOia_ ce8 SIR 0117 Cr161109. ES

00 413001000a 10 2.0045 0''  ~

Ca00&,, 0021' oIa_ OoO SIR00 I? CIlaniela 25 ,00 383. OOl, 000ra 200q6 ____________ 7.07    ` `~ `"`` r''"    ~   `   `   ` ~ 1"   ~ ','   ' .' r'      , .    ~''    '.',"   ' .   '    '   n      ',     " ~ CAo05_ 092? tla_ 008 SIR 01 I? ChOl% 90I0 ES

` 00. 313.00 I, OEOa 200t0 752 1013   ` "              .   . .   ` .,   , , .   . ".,   `   . .    ,       _ ,_`~        .   1   _ _,.,     _ .   _   ' .       _ '        '      _ '   '     . ,   '    .',_.     . . 015070'. 0027000508 SIR of I? Challis ES. IOU 251100 I 0033 2 0000 S CAOSS 00270la 005 SIR oilY 011000101465. 00 210 200t0 ~~ 8090.00oan,~

.00 2.00 3.00 0.00 5.00 6.00 7,00 8.00 9.00 10.00 10,00 2.00 13.00 1400 C4085 032?' SA 043 82, day IS blodeg sample C4005_ 092? Ora_ 0I4 SIR oil? Channels ES'

1110 5i3000000g 2.0008 ~   _ .    ~".~   "       ~

CAOES. 0i12701a1314 SIR ~ I I? Cfla, rI6ls ES' 100 20000 S. 0   .   CA000.. 002l' 0l0_ 014 SIR of ITC. 1. IOnSl.; ES

100. SIII) 010004 5: 741 2110e6 01 __ CA00S., 002? 0Io_ 014 SIR 01 t I? 07405010 ES'

ISO 465.00 I 0006 2cm. s 730 O C. So, b_ 012? O 12.014 e. SIR oat? Chanoeli ES' 00 733 413.00 I. 00D~

E   `   ~` e.   "    . ,'.',..'.~

CAOOS_ 092? 0la_ 0I4 SIR oil? Chonhab ES'

ice ~ 00 I000a 2.00e6 ` 5 cRrSfr. 0E270 ca_ Ole I SIR El IC 1417,014 ES' 100 6010 3130070004 lb 1518 15127 I ( 114   01114 1 lEO 083 71', XlOa lb CAsilo 5142714` 4.514 ` SIR 0' 17 0,05,05' S 05' IOU ~ 6' F 741 2 0000 14 6.02°

L2L,... ~ ~

00 2043 3,00 4.00 500 600 TOO 8,00 0.00 16.00 11.00 12.00 1300 5400 Page 37 of 38 Pace Project CA085: " Biodegradation Screen Study for Telomer Alcohols"   Revision I Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services. Bio Analytical Services Group Wednesday, November 06, 2002 000039 Appendix I Chroniatograms of Extracted ions m/ z 377, 477, and 577 and Mass Spectra for Each Peak for Polyfluorinated Acid Intermediates Formed Transiently in Biodegradation Cultures at Day O, Day I, Day 2 and Day S. Suspected n Oxidation olefinic acid Intermediates.

cafeIlcRloaoflu, en,' I   fl   n  Se  flnj'n : 0.1 7.    EolealiIaAlo, e, nil ~ l~ l° 0l'II Day O ` 4 711 A E   z_''"'~'~"_''''''~'''''"        . .     `       .   lEO 0.00 100 050 6 CE 0.00 7EE 6.00 400 ` 700 1100 1000 13.00 10.50 Page 38 of 38  . rer,

Pace Project CA065: " Biode9radation Screen Study for Telomer Alcohols   Revision 1 Study Director: Cleston C. Lange, Ph. D. Pace Analytical Services, Bio Analytical Services Group Wednesday1 November 06, 2002 000040 " U ( a,. ,~ en.

c4r taela LEafl ` 01 `

l0e'~ o ;,,~ ~

  ~ I cA6s~ ur~ s# o33E2. I blodeg ample / / CA045,. 0124016_ OII / lio~ / 1.0001 Day 5 ~ . S~ ES' 0A065_ Oa240Ib_ 0I0 / 75757 I. 000o 000 lOOSE 7.11 Day 2 & ES 00055_ 152401b_ E00 10 357n45? 1557 10001 100 ,~ lobs  fy35 Day I ~:

a               ~ ~   ,

00245j420016,_ 008 357.4o7n557 I 0000 ISO ICOnS