[Federal Register Volume 75, Number 127 (Friday, July 2, 2010)]
[Notices]
[Pages 38566-38569]
[FR Doc No: 2010-16141]

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SECURITIES AND EXCHANGE COMMISSION

[Release No. 34-62386; File No. SR-CBOE-2010-060]

Self-Regulatory Organizations; Chicago Board Options Exchange,
Incorporated; Notice of Filing and Immediate Effectiveness of Proposed
Rule Change Relating to the Establishment of the Initial Fees for Post-
Demutualization Trading Permits, Tier Appointment and Bandwidth Packets

June 25, 2010.
    Pursuant to Section 19(b)(1) of the Securities Exchange Act of 1934
(``Act''),\1\ and Rule 19b-4 thereunder,\2\ notice is hereby given that
on June 21, 2010, the Chicago Board Options Exchange, Incorporated
(``CBOE'' or the ``Exchange'') filed with the Securities and Exchange
Commission (``Commission'') the proposed rule change as described in
Items I, II, and III below, which Items have been prepared by CBOE. The
Commission is publishing this notice to solicit comments on the
proposed rule change from interested persons.
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    \1\ 15 U.S.C. 78s(b)(1).
    \2\ 17 CFR 240.19b-4.
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I. Self-Regulatory Organization's Statement of the Terms of Substance
of the Proposed Rule Change

    CBOE proposes to establish the initial fees for CBOE's initial
post-demutualization Trading Permits, tier appointment and bandwidth
packets. The text of the proposed rule change is available on the
Exchange's Web site (http://www.cboe.org/Legal/), at the Exchange's
Office of the Secretary, at the Commission's Public Reference Room, and
on the Commission's Web site at http://www.sec.gov.

II. Self-Regulatory Organization's Statement of the Purpose of, and
Statutory Basis for, the Proposed Rule Change

    In its filing with the Commission, CBOE included statements
concerning the purpose of and basis for the proposed rule change and
discussed any comments it received on the proposed rule change. The
text of these statements may be examined at the places specified in
Item IV below. CBOE has prepared summaries, set forth in Sections A, B,
and C below, of the most significant aspects of such statements.

A. Self-Regulatory Organization's Statement of the Purpose of, and
Statutory Basis for, the Proposed Rule Change

1. Purpose
    In connection with the demutualization of the Exchange through
which the Exchange is restructuring from a non-stock corporation to a
stock corporation and a wholly-owned subsidiary of CBOE Holdings, Inc.,
the Exchange has amended its Rules to provide for the use of Trading
Permits, instead of memberships, to access the Exchange.
    CBOE Rule 2.20 grants the Exchange the authority to, from time to
time, fix the fees and charges payable by Trading Permit Holders. The
purpose of this proposed rule change is to establish the initial fees
for CBOE's initial post-demutualization Trading Permits, tier
appointment and bandwidth packets. These post-demutualization Trading
Permits, tier appointment and bandwidth packets will become effective
immediately following the close of trading on the date of the closing
of the Exchange's demutualization transaction, and CBOE members on the
date of the closing of the demutualization transaction will retain
their then current access to the Exchange until the close of trading on
that date. The Exchange also proposes to amend the CBOE Stock Exchange,
LLC (``CBSX'') \3\ Fees Schedule to cross-reference that CBSX Trading
Permit access fees are set forth in the CBOE

Fees Schedule. As further described below, there is no access fee
proposed for CBSX-only Trading Permit Holders.
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    \3\ CBSX is a facility of CBOE for the trading of non-option
securities, and trading on CBSX is governed by CBOE Rules.
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    Trading Permit Fees:
    CBOE Rule 1.1(ggg) defines a Trading Permit as a license issued by
the Exchange that grants the holder or the holder's nominee the right
to access one or more of the facilities of the Exchange for the purpose
of effecting transactions in securities traded on the Exchange without
the services of another person acting as broker, and otherwise to
access the facilities of the Exchange for purposes of trading or
reporting transactions or transmitting orders or quotations in
securities traded on the Exchange, or to engage in other activities
that, under CBOE Rules, may only be engaged in by Trading Permit
Holders, provided that the holder or the holder's nominee, as
applicable, satisfies any applicable qualification requirements to
exercise those rights. A Trading Permit does not convey any ownership
interest in the Exchange, is only available through the Exchange, and
is subject to the terms and conditions set forth in CBOE Rule 3.1.
Holders of Trading Permits fall within the definition of ``member'' in
Section 3(a)(3)(A) of the Securities Exchange of 1934, as amended
(``Act'').\4\
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    \4\ 15 U.S.C. 78c(a)(3)(A).
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    CBOE Rule 3.1 provides, among other things, that the Exchange may
issue different types of Trading Permits and determine the fees for
those Trading Permits. Specifically, under Rule 3.1(a)(iv), the
Exchange may issue different types of Trading Permits that allow
holders to trade one or more products authorized for trading on the
Exchange and to act in one or more trading functions authorized by CBOE
Rules. The Exchange will have four initial types of Trading Permits
immediately following its demutualization. These Trading Permits will
be issued in accordance with the provisions of CBOE Rule 3.1A which
addresses the initial issuance of post-demutualization Trading Permits
and Rule 3.1 which sets forth the general provisions that are
applicable to post-demutualization Trading Permits. In addition, Rule
3.1(a)(v) provides, in relevant part, that Trading Permits will be
subject to such fees and charges as are established by the Exchange
from time to time pursuant to CBOE Rule 2.20 and the Exchange Fees
Schedule.
    The first type of Trading Permit is a Market-Maker Trading Permit
that entitles the holder to act as a Market-Maker (including a Market-
Maker trading remotely), DPM, eDPM or LMM. This permit provides an
appointment credit of 1.0, a quoting and order entry bandwidth
allowance, up to three logins, trading floor access and Trading Permit
Holder status. A Market-Maker Trading Permit also provides trading
access to CBSX. The Exchange is proposing to establish the initial fee
for a Market-Maker Trading Permit at $7,500 per month, commencing July
1, 2010. However, for the remainder of 2010, CBOE will provide a 20%
discount on this fee, such that the fee for a Market-Maker Trading
Permit will be $6,000 per month between July 2010 and December 2010.
    The quoting bandwidth allowance for a Market-Maker Trading Permit
is equivalent to a maximum of 31,200,000 quotes over the course of a
trading day. The quoting bandwidth allowance for a Market-Maker Trading
Permit in which the holder has a Market-Maker appointment in a Hybrid
3.0 option class shall be proportionately reduced by the appointment
cost of the class. To the extent that a Market-Maker is able to submit
electronic quotes in a Hybrid 3.0 class (such as an LMM that streams
quotes in the class), the Market-Maker shall receive the quoting
bandwidth allowance attributable to that Hybrid 3.0 class to quote in,
and only in, that class. For example, the appointment cost for SPX,
which is a Hybrid 3.0 class, is .95. Accordingly, the quoting bandwidth
for a Market-Maker Trading Permit in which the holder has a Market-
Maker appointment in SPX would be .05 of the quoting bandwidth for non-
Hybrid 3.0 classes, unless the Market-Maker is an LMM and is using a
Market-Maker Trading Permit to stream quotes in SPX. With the exception
of LMMs, Market-Makers do not quote electronically in Hybrid 3.0
classes so they are not allocated quoting bandwidth with respect to
Market-Maker appointments in those classes. This is consistent with
CBOE's pre-demutualization quoting bandwidth allocations to SPX Market-
Makers.
    The second type of Trading Permit is a Floor Broker Trading Permit
that entitles the holder to act as a Floor Broker. This permit provides
an order entry bandwidth allowance, up to three logins, trading floor
access and Trading Permit Holder Status. A Floor Broker Trading Permit
also provides access to CBSX. The Exchange is proposing to establish
the initial fee for a Floor Broker Trading Permit at $7,500 per month,
commencing July 1, 2010. However, for the remainder of 2010, CBOE will
provide a 20% discount on this fee, such that the fee for a Floor
Broker Trading Permit will be $6,000 per month between July 2010 and
December 2010.
    The third type of Trading Permit is the Electronic Access Permit
that entitles the holder to electronic access to the Exchange.
Electronic Access Permit holders must be broker-dealers registered with
the Exchange in one or more of the following capacities: (a) Clearing
Trading Permit Holder; (b) TPH organization approved to transact
business with the public; (c) Proprietary Trading Permit Holder;\5\ and
(d) order service firm. The Electronic Access Permit does not grant
access to the trading floor. This permit provides an order entry
bandwidth allowance, up to three logins and Trading Permit Holder
status. An Electronic Access Permit also provides access to CBSX. The
Exchange is proposing to establish the initial fee for an Electronic
Access Permit at $2,000 per month, commencing July 1, 2010. However,
for the remainder of 2010, CBOE will provide a 20% discount on this
fee, such that the fee for an Electronic Access Permit will be $1,600
per month between July 2010 and December 2010.
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    \5\ A Proprietary Trading Permit Holder is a proprietary broker-
dealer engaged in principal trading. A proprietary broker-dealer may
not use an Electronic Access Permit to submit Market-Maker orders
(i.e., M orders) for its own account or an affiliated Market-Maker
account.
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    The fourth type of Trading Permit is a CBSX Trading Permit that
entitles the holder to perform any of the trading functions of a
Trading Permit Holder on CBSX and to receive the CBSX login and
bandwidth allowances. The CBSX Trading Permit provides trading access
only to CBSX. The proposed initial CBSX Trading Permit fee is the same
as the access fee that applied with respect to the CBSX trading permits
that granted trading access to CBSX prior to CBOE's demutualization in
that there was no access fee charged for pre-demutualization CBSX
trading permits (just as there is no access fee initially proposed to
be charged with respect to post-demutualization CBSX Trading Permits).
Because CBSX is a relatively new trading venue, CBSX has sought to
encourage broker-dealers to become participants in CBSX's market in
order to build volume and market share by, among other things, not
assessing an access fee. This proposed rule change continues that
approach with respect to the initial CBSX Trading Permit access fee
following CBOE's demutualization.
    Trading Permit fees will be assessed by the Exchange commencing on
July 1, 2010. The Exchange is delaying the commencement of the fee
until then because the leases for the transferable memberships that
existed prior to CBOE's demutualization generally provided for monthly
lease payments

and the Exchange assessed temporary members and Interim Trading Permit
holders monthly access fees. Therefore, most post-demutualization
Trading Permit holders have already paid to access the Exchange for the
month of June 2010 in some form. As a result, the Exchange will not
charge an additional amount for access to the Exchange during that
month. Trading Permit fees are non-refundable and will be assessed
through the integrated billing system during the first week of the
following month.\6\ If a Trading Permit is issued during a calendar
month after the first trading day of the month, the access fee for the
Trading Permit for that calendar month is prorated based on the
remaining trading days in the calendar month. Trading Permits will be
renewed automatically for the next month unless the Trading Permit
Holder submits by the 25th day of the prior month (or the preceding
business day if the 25th is not a business day) a written notification
\7\ to cancel the Trading Permit effective at or prior to the end of
the applicable month.\8\
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    \6\ Thus, Trading Permit Fees for access during July 2010 will
be assessed through the integrated billing system during the first
week of August 2010.
    \7\ Written notification may be submitted to the Registration
Services Department by e-mail to<EMAIL_ADDRESS>or by other
means of written notification, including, but not limited to, a
hand-delivered letter or facsimile to the Registration Services
Department.
    \8\ Thus, if a Trading Permit Holder has a Trading Permit in
July 2010, notice must be provided by July 23, 2010 (the preceding
business day prior to July 25, 2010 since July 25, 2010 is not a
business day) if the Trading Permit Holder would like to terminate
the Trading Permit by the end of July 2010 and not be assessed the
applicable Trading Permit Fee for August 2010.
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    The Exchange proposes to amend the CBOE Fees Schedule effective on
June 21, 2010 to set forth the initial access fees for these four
initial Trading Permit types by including in the Fees Schedule the
initial access fee applicable to each Trading Permit type, the
description of each Trading Permit type included above, and the
procedural provisions included above describing the manner in which
Trading Permit access fees will be assessed by the Exchange.
    Tier Appointment Fee:
    CBOE Rule 8.3(e) provides that the Exchange may establish one or
more types of tier appointments. In accordance with CBOE Rule 8.3(e), a
tier appointment is an appointment to trade one or more options classes
that must be held by a Market-Maker to be eligible to act as a Market-
Maker in the options class or options classes subject to that
appointment. The Exchange will have one type of tier appointment
immediately following its demutualization, the SPX Tier Appointment. A
Market-Maker Trading Permit Holder must obtain an SPX Tier Appointment
to act as a Market-Maker in SPX. Further, consistent with the
provisions of Rule 8.3(e), each SPX Tier Appointment may only be used
with one designated Market-Maker Trading Permit. The Exchange is
proposing that the initial fee for an SPX Tier Appointment be set at
$3,000 per month, commencing July 1, 2010, the same date as the
commencement of the Trading Permit fees. The SPX Tier Appointment fee
is not eligible for the 20% discount for the remainder of 2010 that is
applicable to the Trading Permit fees.
    SPX Tier Appointment fees are non-refundable and will be assessed
through the integrated billing system during the first week of the
following month. The SPX Tier Appointment fee will be assessed to any
Market-Maker Trading Permit Holder, registered with the Exchange to
conduct business on the Exchange as a Market-Maker, that either (a) has
an SPX Tier Appointment at any time during a calendar month; or (b)
conducts any open outcry transactions in SPX at any time during a
calendar month. SPX Tier Appointments will be renewed automatically for
the next month unless the Trading Permit Holder submits by the 25th day
of the prior month (or the preceding business day if the 25th is not a
business day) a written notification to cancel the SPX Tier Appointment
effective at or prior to the end of the applicable month.
    Bandwidth Packet Fees:
    CBOE is also proposing to establish fees for bandwidth packets.
Bandwidth packets provide Trading Permit Holders with additional
bandwidth to use to electronically access the Exchange. Market-Makers
will be offered the opportunity to purchase one or more Quoting and
Order Entry Bandwidth Packets. Each Quoting and Order Entry Bandwidth
Packet will entitle the Trading Permit Holder to up to three additional
logins and contain the standard Market-Maker quoting and order entry
bandwidth allowance, which may then be added onto the total bandwidth
pool for a Market-Maker's acronym(s) and Trading Permit(s) without the
Market-Maker having to obtain additional Trading Permits. All Trading
Permit Holders will have the opportunity to purchase one or more Order
Entry Bandwidth Packets. Each Order Entry Bandwidth Packet will entitle
the Trading Permit Holder to up to three additional logins and an order
entry bandwidth allowance.
    The Exchange is proposing that the initial fee for a Quoting and
Order Entry Bandwidth Packet be set at $3,750 per month. In addition,
the Exchange is proposing that the initial fee for an Order Entry
Bandwidth Packet be set at $2,000 per month. Bandwidth packet fees will
be assessed by the Exchange commencing on July 1, 2010, the same date
as the commencement of the Trading Permit and SPX Tier Appointment
fees. However, for the remainder of 2010, CBOE will provide a 20%
discount on these fees, such that, between July 2010 and December 2010,
the fee for a Quoting and Order Entry Bandwidth Packet will be $3,000
per month and the fee for an Order Entry Bandwidth Packet will be
$1,600 per month. Bandwidth packet fees are non-refundable and will be
assessed through the integrated billing system during the first week of
the following month. If a bandwidth packet is issued during a calendar
month after the first trading day of the month, the bandwidth packet
fee for that calendar month is prorated based on the remaining trading
days in the calendar month. Bandwidth packets will be renewed
automatically for the next month unless the Trading Permit Holder
submits by the 25th day of the prior month (or the preceding business
day if the 25th is not a business day) a written notification to cancel
the bandwidth packet effective at or prior to the end of the applicable
month.
    The same quoting and order entry bandwidth allowance will be
provided for each Market-Maker Trading Permit and each Quoting and
Order Entry Bandwidth Packet (except to the extent described above with
respect to each Market-Maker Trading Permit in which the holder has a
Market-Maker appointment in a Hybrid 3.0 option class). Similarly, the
same order entry bandwidth allowance will be provided for each Floor
Broker Trading Permit, Electronic Access Permit, and Order Entry
Bandwidth Packet and the same bandwidth allowance will be made
available for each CBSX Trading Permit. Accordingly, bandwidth will be
available to all Trading Permit Holders on an equal basis. The Exchange
has provided the Commission with a detailed description of the
foregoing bandwidth allowances pursuant to a Freedom of Information Act
confidential treatment request. To the extent that the Exchange changes
these bandwidth allowances in the future, the Exchange will comply with
the rule filing requirements of Section 19 of the Act.\9\
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    \9\ 15 U.S.C. 78s.
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    In addition to the proposed changes to the Fees Schedule described
above, CBOE is proposing to revise its regulatory circular that sets
forth the existing Trading Permit Holder

application and other related fees. The Exchange proposes to revise
this circular to incorporate the Trading Permit, tier appointment and
bandwidth packet fees. The proposed changes to the circular are
included as Exhibit 2 to the Form 19b-4.
    CBOE believes that the proposed fees are reasonable when compared
to the average access fee previously charged to CBOE temporary members
and interim trading permit holders by the Exchange over the last twelve
months. Specifically, the average of these access fees between July
2009 and June 2010 was $8,967. Additionally, these access fees were
above $10,000 between July 2009 and November 2009, peaking at $11,900
in October 2009.
    The Exchange may adjust the proposed Trading Permit, tier
appointment and bandwidth packet fees in the future if the Exchange
determines that it would be appropriate to do so based upon the
circumstances at the time. The Exchange may also make future additions
or changes to the types of Trading Permits, tier appointments or
bandwidth packets in accordance with Exchange Rules. Any future Trading
Permit, tier appointment or bandwidth packet fee changes and the fees
for any new or modified types of Trading Permits, tier appointments or
bandwidth packets will be reflected in amendments to the CBOE Fees
Schedule that will be submitted to the Commission through further rule
filings pursuant to Section 19(b)(3)(A)(ii) of the Act \10\ (provided
that the fee changes will only apply to Trading Permit Holders and
their associated persons). Any other such fee changes, including those
that will be applicable to persons that are not Trading Permit Holders
or associated persons of Trading Permit Holders, will be submitted to
the Commission through further rule filings pursuant to Section
19(b)(2) of the Act.\11\
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    \10\ 15 U.S.C. 78s(b)(3)(A)(ii).
    \11\ 15 U.S.C. 78s(b)(2).
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2. Statutory Basis
    The proposed rule change will treat similarly situated Trading
Permit Holders in the same manner by assessing the same Trading Permit,
tier appointment and bandwidth packet fees (and applying the same
discount to the trading permit and bandwidth packet fees) to all
Trading Permit Holders based on the type of Trading Permit(s), tier
appointment and bandwidth packet(s) requested and by assessing no
Trading Permit fee to all Trading Permit Holders with access solely to
CBSX. Accordingly, the Exchange believes that the proposed rule change
is consistent with Section 6(b) of the Act,\12\ in general, and
furthers the objectives of Section 6(b)(4) of the Act \13\ in
particular, in that it is designed to provide for the equitable
allocation of reasonable dues, fees, and other charges among persons
using its facilities for the reasons described above.\14\
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    \12\ 15 U.S.C. 78f(b).
    \13\ 15 U.S.C. 78f(b)(4).
    \14\ The Exchange also believes that it is equitable to assess
different access fees for trading permits that provide differential
access as long as the same access fee is assessed to all Holders of
the same type of Trading Permit.
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B. Self-Regulatory Organization's Statement on Burden on Competition

    CBOE does not believe that the proposed rule change will impose any
burden on competition that is not necessary or appropriate in
furtherance of the purposes of the Act.

C. Self-Regulatory Organization's Statement on Comments on the Proposed
Rule Change Received From Members, Participants or Others

    No written comments were solicited or received with respect to the
proposed rule change.

III. Date of Effectiveness of the Proposed Rule Change and Timing for
Commission Action

    Because the foregoing rule change establishes or changes a due,
fee, or other charge imposed by the Exchange, it has become effective
pursuant to Section 19(b)(3)(A) of the Act \15\ and subparagraph (f)(2)
of Rule 19b-4 \16\ thereunder. At any time within 60 days of the filing
of the proposed rule change, the Commission may summarily abrogate such
rule change if it appears to the Commission that such action is
necessary or appropriate in the public interest, for the protection of
investors, or otherwise in furtherance of the purposes of the Act.
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    \15\ 15 U.S.C. 78s(b)(3)(A).
    \16\ 17 CFR 240.19b-4(f)(2).
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IV. Solicitation of Comments

    Interested persons are invited to submit written data, views, and
arguments concerning the foregoing, including whether the proposed rule
change is consistent with the Act. Comments may be submitted by any of
the following methods:

Electronic Comments

     Use the Commission's Internet comment form (http://www.sec.gov/rules/sro.shtml); or
     Send an e-mail to<EMAIL_ADDRESS>Please include
File Number SR-CBOE-2010-060 on the subject line.

Paper Comments

     Send paper comments in triplicate to Elizabeth M. Murphy,
Secretary, Securities and Exchange Commission, 100 F Street, NE.,
Washington DC 20549-1090.

All submissions should refer to File Number SR-CBOE-2010-060. This file
number should be included on the subject line if e-mail is used. To
help the Commission process and review your comments more efficiently,
please use only one method. The Commission will post all comments on
the Commission's Internet Web site (http://www.sec.gov/rules/sro.shtml). Copies of the submission, all subsequent amendments, all
written statements with respect to the proposed rule change that are
filed with the Commission, and all written communications relating to
the proposed rule change between the Commission and any person, other
than those that may be withheld from the public in accordance with the
provisions of 5 U.S.C. 552, will be available for Web site viewing and
printing in the Commission's Public Reference Room, 100 F Street, NE.,
Washington, DC 20549, on official business days between the hours of 10
a.m. and 3 p.m. Copies of such filing also will be available for
inspection and copying at the principal office of the Exchange. All
comments received will be posted without change; the Commission does
not edit personal identifying information from submissions. You should
submit only information that you wish to make available publicly. All
submissions should refer to File No. SR-CBOE-2010-060 and should be
submitted on or before July 23, 2010.

    For the Commission, by the Division of Trading and Markets,
pursuant to delegated authority.\17\
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    \17\ 17 CFR 200.30-3(a)(12).
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Florence E. Harmon,
Deputy Secretary.
[FR Doc. 2010-16141 Filed 7-1-10; 8:45 am]
BILLING CODE 8011-01-P


[Federal Register: January 25, 2008 (Volume 73, Number 17)]
[Rules
and Regulations]
[Page 4449-4450]
[DOCID:fr25ja08-20]

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DEPARTMENT OF TRANSPORTATION

Federal Aviation Administration

14 CFR Part 71

[Docket No. FAA-2007-29264; Airspace Docket No. 07-AEA-04]

Establishment of Class E Airspace; Tappahannock, VA

AGENCY: Federal Aviation Administration (FAA), DOT.

ACTION: Direct final rule; confirmation of effective date.

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SUMMARY: This action confirms the effective date of a direct final rule
that establishes a Class E airspace area to support Area Navigation
(RNAV) Global Positioning System (GPS) Standard Instrument Approach
Procedures (SIAPs) that serve Tappahannock-Essex County Airport,
Tappahannock, VA.

DATES: Effective 0901 UTC, December 20, 2007. The Director of the
Federal Register approves this incorporation by reference action under
Title 1, Code of Federal Regulations, part 51, subject to the annual
revision of FAA Order 7400.9 and publication of conforming amendments.

FOR FURTHER INFORMATION CONTACT: Daryl Daniels, Airspace Specialist,
System Support, AJO2-E2B.12, FAA Eastern Service Center, 1701 Columbia
Ave., College Park, GA 30337; telephone<PHONE_NUMBER>; fax (404) 305-
5572.

SUPPLEMENTARY INFORMATION:

Confirmation of Effective Date

    The FAA published this direct final rule with a request for
comments in the Federal Register on October 30, 2007 (72 61294-61296).
The FAA uses the direct final rulemaking procedure for a non
controversial rule where the FAA believes that there will be no adverse
public comment. This direct final rule advised the public that no
adverse comments were anticipated, and that unless a written adverse
comment, or a written notice of intent to submit such an adverse
comment, were received within the comment period, the regulation would
become effective on December 20, 2007. No adverse comments were
received, and thus this notice confirms the effective date.

    Issued in College Park, GA on December 17, 2007.
Mark D. Ward,
Manager, System Support Group, Eastern Service Center.
[FR Doc. 08-207 Filed 1-24-08; 8:45 am]

BILLING CODE 4910-13-M


[Federal Register Volume 81, Number 117 (Friday, June 17, 2016)]
[Notices]
[Pages 39637-39638]
[FR Doc No: 2016-14340]

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DEPARTMENT OF ENERGY

Federal Energy Regulatory Commission

Combined Notice of Filings #1

    Take notice that the Commission received the following electric
rate filings:

    Docket Numbers: ER12-637-005; ER11-2370-005; ER10-2211-005.
    Applicants: Calhoun Power Company, LLC, Cambria CoGen Company,
Vandolah Power Company L.L.C.
    Description: Non-Material Change in Status Filing of Calhoun Power
Company, LLC, et al.
    Filed Date: 6/10/16.
    Accession Number: 20160610-5313.
    Comments Due: 5 p.m. ET 7/1/16.

    Docket Numbers: ER16-228-002.
    Applicants: Southwest Power Pool, Inc.
    Description: Compliance filing: 3113 Basin & MDU Attachment AO
Amended Compliance Filing to be effective 10/1/2015.
    Filed Date: 6/13/16.
    Accession Number: 20160613-5033.
    Comments Due: 5 p.m. ET 7/5/16.

    Docket Numbers: ER16-1689-001.
    Applicants: ArcelorMittal Cleveland LLC.
    Description: Tariff Amendment: Supplement to Petition for
Acceptance of Market-Base Rate Tariff to be effective 6/30/2016.
    Filed Date: 6/13/16.
    Accession Number: 20160613-5154.
    Comments Due: 5 p.m. ET 7/5/16.

    Docket Numbers: ER16-1912-000.
    Applicants: Southwest Power Pool, Inc.
    Description: Sec.  205(d) Rate Filing: Out-of-Merit Energy
Clarification to be effective 7/1/2016.
    Filed Date: 6/10/16.
    Accession Number: 20160610-5274.
    Comments Due: 5 p.m. ET 7/1/16.

    Docket Numbers: ER16-1913-000.

    Applicants: River Bend Solar, LLC.
    Description: Baseline eTariff Filing: River Bend Solar, LLC
Application for Market-Based Rates to be effective 9/1/2016.
    Filed Date: 6/10/16.
    Accession Number: 20160610-5281.
    Comments Due: 5 p.m. ET 7/1/16.

    Docket Numbers: ER16-1914-000.
    Applicants: Patua Project LLC.
    Description: Sec.  205(d) Rate Filing: Notice of Succession to be
effective 5/11/2016.
    Filed Date: 6/10/16.
    Accession Number: 20160610-5283.
    Comments Due: 5 p.m. ET 7/1/16.

    Docket Numbers: ER16-1915-000.
    Applicants: Oildale Energy LLC.
    Description: Tariff Cancellation: Notice of Cancellation to be
effective 6/11/2016.
    Filed Date: 6/10/16.
    Accession Number: 20160610-5284.
    Comments Due: 5 p.m. ET 7/1/16.

    Docket Numbers: ER16-1916-000.
    Applicants: Public Service Company of Colorado.
    Description: Sec.  205(d) Rate Filing: 2016-6-13 Wind Integration
Correction Filing to be effective 1/1/2015.
    Filed Date: 6/13/16.
    Accession Number: 20160613-5108.
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    Dated: June 13, 2016.
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[FR Doc. 2016-14340 Filed 6-16-16; 8:45 am]
 BILLING CODE 6717-01-P


CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 2 of 52 STATEMENT OF CONFIDENTIALITY OR NO CONFIDENTIALITY CLAIMS ( To be completed by the Sponsor) CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 5 of 52 PAGE RESERVED FOR 40 CFR 158.34 ( c) ( 1) CERTIFICATION ( To be completed by the Sponsor) CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 6 of 52 CONTENTS STATEMENT OF CONFIDENTIALITY OR NO CONFIDENTIALITY CLAIMS 2 STATEMENT OF THE STUDY DIRECTOR AND CIT SCIENTIFIC MANAGEMENT 3 OTHER SCIENTIST INVOLVED IN THIS STUDY 3 STATEMENT OF QUALITY ASSURANCE UNIT 4 PAGE RESERVED FOR 40 CFR 158.34 ( c) ( 1) CERTIFICATION 5 SUMMARY 8 1. INTRODUCTION 10 2. MATERIALS AND METHODS 10 2.1 TEST ITEM 10 2.1.1 Identification 10 2.1.2 Preparation of the test solutions 10 2.2 TEST SYSTEM 11 2.2.1 Animals 11 2.2.2 Environmental conditions during culture 11 2.2.3 Environmental conditions during the test 12 2.2.4 Food 12 2.3 TREATMENT 12 2.3.1 Administration of the test item 12 2.3.2 Time schedule 12 2.3.3 Definitions 13 2.3.4 Observations 13 2.4 CHEMICAL ANALYSIS 13 2.5 DATA EVALUATION 13 2.5.1 Determination of the EC50 14 2.5.2 Determination of the No Observed Effect Concentration ( NOEC) 14 2.6 VALIDITY CRITERIA 14 2.7 ARCHIVING 14 2.8 STUDY PLAN ADHERENCE 15 CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 7 of 52 3. RESULTS 16 3.1 VALIDITY CRITERIA 16 3.2 CHEMICAL ANALYSIS 16 3.3 WATER QUALITY 16 3.4 RESULTS 16 4. CONCLUSION 18 Figure 1: Total number of living neonates per parent alive after 21 days versus concentration 19 Table 1: Physico chemical parameters over the 21 day period 20 Table 2: Number of immobilized parents 21 Table 3a: Living, dead and aborted eggs produced at each brood release  Control 22 Table 3b: Living, dead and aborted eggs produced at each brood release   4.31 mg/ L 23 Table 3c: Living, dead and aborted eggs produced at each brood release   9.16 mg/ L 24 Table 3d: Living, dead and aborted eggs produced at each brood release   20.0 mg/ L 25 Table 3e: Living, dead and aborted eggs produced at each brood release   44.2 mg/ L 26 Table 3f: Living, dead and aborted eggs produced at each brood release   88.6 mg/ L 27 Table 4: Living neonates per parent alive at the end of the test versus concentration of the test item 28 Table 5: Length of parent alive at the end of the test 29 APPENDICES 30 1. Analytical certificate and composition of the test item 31 2. Preparation of M4 reconstituted water 34 3. Chemical analysis of test solutions 37 4. EC50 estimation 43 5. 21 day NOEC determination based on reproduction capacity 45 6. Statistical analysis of Daphnia length data 48 to 52 CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 8 of 52 SUMMARY At the request of APME, Brussels, Belgium, the toxicity to reproduction of the test item AMMONIUM PERFLUOROOCTANOATE ( APFO) was evaluated in the Cladoceran, Daphnia magna STRAUS ( clone 5) using a 21 day semi static test according to OECD guideline No. 211 ( 21st September 1998). The criterion of toxicity measured was the effect of the test item in solution on the reproductive efficacy. This is expressed as:

  the EC50, the concentration required to reduce the number of living neonates per adult alive at the end of the test to 50% of the control value,

  the No Observed Effect Concentration ( NOEC), obtained by analysis of variance from these data.

Methods The test item was dissolved in reconstituted water ( M4) prepared from deionized water with a conductivity < 10 µ S/ cm. The total hardness of the reconstituted water is generally between 238 and 289 mg/ L ( as CaCO3) and the pH is 8.0 ± 0.5.

Reproduction test Five concentrations of AMMONIUM PERFLUOROOCTANOATE ( APFO) were used together with a control at 0, 6.25, 12.5, 25.0, 50.0 and 100 mg/ L expressed as the APFO ( solids) content of the test item.

Test solutions were changed every 2 or 3 days.

Ten daphnids ( ten individual replicates) were exposed to each concentration for 21 days and immobilization of adults and number of dead and living neonates were recorded daily throughout the test.

Environmental parameters were:

  pH: 7.56 to 8.26,

  temperature: 18.4 ° C to 20.4 ° C,

  dissolved oxygen: 7.8 to 9.2 mg/ L (> 3 mg/ L throughout the test),

  water hardness: 255 to 289 mg/ L as CaCO3.

Chemical analysis Chemical analysis was undertaken to measure the concentration of the test item in each test solution, except for the control, at the beginning of the test, before and after the medium renewals and at the end of the test. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 9 of 52 Results Chemical analysis Some measured concentrations were not within ± 20% of the corresponding nominal values. However, the maximum CV of the measured concentrations ( at the beginning of the test and before medium renewals) at each of the different nominal target values was only 8.50% ( at 6.25 mg/ L nominal), indicating that the test item was sufficiently stable between medium renewals. All study results were therefore based on harmonic means of measured concentrations in test solutions throughout the test, i. e. 21 days. These harmonic means were calculated as 4.31, 9.16, 20.0, 44.2 and 88.6 mg/ L expressed as the APFO ( solids) content of the test item, for nominal values of 6.25, 12.5, 25.0, 50.0 and 100 mg/ L, respectively.

Mortalities The test item did not induce significant mortality in parent animals at concentrations tested. At the end of the test, the cumulative mortality in parent animals was 20% ( 2/ 10) in the control, 10% ( 1/ 10) at 4.31 mg/ L, 0% ( 0/ 10) at 9.16 mg/ L, 0% ( 0/ 10) at 20.0 mg/ L, 10% ( 1/ 10) at 44.2 mg/ L and 10% ( 1/ 10) at 88.6 mg/ L.

Reproduction A significant inhibition of the reproduction capacity ( expressed as the number of living neonates produced per parent alive at the end of the test) was obtained at 44.2 and 88.6 mg/ L, compared to the control.

The reproduction capacity values were as follows: 21 day EC50: 39.6 mg/ L Confidence limits: 36.7 to 42.5 ( p = 0.05) No Observed Effect Concentration ( 21 day NOEC): 20.0 mg/ L ( p = 0.05)

Length A significant inhibition of the length of parent animals ( alive at the end of the test) was obtained at 88.6 mg/ L, compared to the control.

The length values were as follows: 21 day EC50: > 88.6 mg/ L No Observed Effect Concentration ( 21 day NOEC): 44.2 mg/ L ( p = 0.05)

Conclusion Under our experimental conditions, the 21 day EC50 based on reproduction capacity of AMMONIUM PERFLUOROOCTANOATE ( APFO) in a semi static test system was 39.6 mg/ L for Daphnia magna. The NOEC after 21 days based on survival of the parent animals was 88.6 mg/ L ( the maximum concentration tested). The NOEC at 21days based on reproduction capacity was 20.0 mg/ L. The NOEC at 21 days based on length of the parent animals was 44.2 mg/ L.

These results are expressed as the APFO ( solids) content of the test item. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 10 of 52 1. INTRODUCTION The objective of this study was to assess the reproductive toxicity of AMMONIUM PERFLUOROOCTANOATE ( APFO) to Daphnia magna ( clone 5) using a 21 day semi static test according to OECD guideline No. 211 ( 21st September 1998).

In this test, the criterion of toxicity measured is the effect of the test item in solution on the reproductive efficacy, expressed as the EC50, the concentration required to reduce the number of living neonates per adult alive at the end of the test to 50% of the control value. The No Observed Effect Concentration ( NOEC) is derived statistically from the data using analysis of variance.

2. MATERIALS AND METHODS 2.1 TEST ITEM 2.1.1 Identification   supplier: 3M   name:  Study plan: AMMONIUM PERFLUOROOCTANOATE ( APFO)  labeling: FLUORAD TM Both names correspond to the same test item.

  batch number: 10004   description at receipt: colorless liquid   description on the analytical certificate: 3M( TM) Fluorad( TM) Fluorochemical Emulsifier   containers: three drums   date of receipt: 28 January 2002   storage conditions: at room temperature   purity: 19.6% ( expressed as solids) in purified water.

Data relating to the characterisation of the test item are documented in an analytical certificate and in the composition of the test item ( presented in appendix 1) provided by the Sponsor.

2.1.2 Preparation of the test solutions The stock solutions, for the test, were prepared by dissolving the test item directly in test water ( see § 2.2.2).

The conditions of preparation of the stock solutions are reported in the following table:

Day of addition Quantity of test item ( as received) ( mg/ L) Quantity of APFO ( solids) ( mg) ( 1) Volume of water ( mL) Concentration of APFO ( solids) ( mg/ L) ( 1) Duration of the agitation ( minutes) 0, 2, 5, 7, 9, 12, 14, 16 and 19 1020.4 200 2000 100 5 ( 1) expressed as APFO ( solids) based on stated test item composition of 19.6% APFO in purified water The agitation was continued until the stock solutions were used to prepare the test solutions.

Test solutions were prepared by further dilution of the stock solution with test water to provide a geometric series of concentrations: 0, 6.25, 12.5, 25.0, 50.0 and 100 mg/ L. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 11 of 52 Test vessels containing daphnids were filled directly from the test solution containers immediately after preparation. Solutions were changed every 2 or 3 days ( every Monday, Wednesday and Friday).

The pH of the test solutions remained within acceptable limits ( between 6.5 and 9 and in the range ± 0.5 unit of the test water) after preparation and there was no adjustment of pH before addition of the animals.

2.2 TEST SYSTEM 2.2.1 Animals Species: Daphnia magna STRAUS clone 5. Reason for this choice: a species commonly used in Europe for aquatic toxicity testing and recommended in OECD guideline. Breeder: CIT. Origin: Centre Technique du Bois et de l'Ameublement, Paris, France. Parentage: the parents of the daphnids to be used in the test were born within the same 24 hour period. They had produced more than three broods and the first before day 12, prior to their use as breeding stock to produce neonates for this test. Life stage: at the beginning of the test, first instar daphnids between 6 and 24 hours old and produced from parthenogenically reproducing brood female population. Acclimation: animals were held in water of the same quality as the test water from birth, until use in the test. Allocation of the animals: animals were randomly placed first into one beaker containing water of the same quality and conditions as the culture water and then 6 hours later, the remaining daphnids were randomly assigned to the test item solutions using a pipette. Number of animals: Ten animals per concentration and in the control with one animal per vessel.

2.2.2 Environmental conditions during culture During the culture period, the stock aquarium conditions are as follows: Water: reconstituted ( M4). The pH of this solution is 8.0 ± 0.5; the sum of Ca and Mg ions in the solution is 2.5 mmol/ L ( molar ratio Ca: Mg is approximately 4: 1 and Na: K approximately 10: 1); total hardness is between 238 and 289 mg/ L as CaCO3. Details of its preparation are presented in appendix 2. Temperature in water: between 18 ° C and 22 ° C. Temperature is recorded continuously and checked generally twice a week. Light/ dark cycle: 16/ 8 hours. Forced aeration: none. Oxygen concentrations remain above 80% of the saturation value at the selected temperature. Loading: the animals are loaded at one animal/ 20 mL of culture water. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 12 of 52 The medium is replaced generally each Monday, Wednesday and Friday. On each of these occasions, neonates are separated from the adults and either removed or, once monthly, used to start a new culture. During holding, pH, water hardness and dissolved oxygen concentrations are checked in the " new" and " old" media when solutions are replaced generally twice a week.

2.2.3 Environmental conditions during the test The test conditions were as follows: Test water: see § 2.2.2. Water was changed thrice weekly during the 21 day test period. Temperature: between 18 ° C and 22 ° C throughout the test. The temperature was controlled using a water bath into which the beakers were placed and the maximum and minimum temperature were recorded daily using a max min thermometer. Light/ dark cycle: 16/ 8 hours. Light intensity: less than 1200 lux. Duration of test: 21 days. Test vessels: glass beakers. Loading rate: one daphnid per vessel containing 50 mL of solution. Forced aeration: was not used during the test. Dilution water was aerated prior to the addition of the test item. Oxygen concentrations: were maintained > 3 mg/ L and > 60% of the air saturation value throughout the test.

Temperature, pH, dissolved oxygen concentration and total hardness were measured in the control solution and the highest concentration ( where animals were still alive) at the beginning and the end of the test as well as at least once a week before and after the solution change. Temperature was also measured continuously in a vessel placed near the test vessels.

2.2.4 Food During culture and the test, animals were fed daily ( until day 20) with a diet of approximately 6.106 cells of Chlorella vulgaris and 6.106 cells of Pseudokirchneriella subcapitata ( both strains are cultured at CIT) per daphnid. The carbon content of this diet corresponded to 0.16 mg of carbon per daphnid per day ( until day 20).

2.3 TREATMENT 2.3.1 Administration of the test item Ten animals were used for each concentration with five concentrations and one control. The nominal concentrations used were as follows: 0, 6.25, 12.5, 25.0, 50.0 and 100 mg/ L expressed as the APFO ( solids) content of the test item.

2.3.2 Time schedule Experimental starting date ( first day of treatment): 6 November 2002, Experimental completion date: 27 November 2002. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 13 of 52 2.3.3 Definitions Living progeny ( or living neonates): offspring found alive in solution after its birth. Dead progeny ( or dead neonates): offspring found dead in solution after its birth. Aborted egg: egg ( or embryo) found in solution after an abortion.

The death is defined as the inability of a daphnid to swim within 15 seconds after gentle agitation of the test vessel.

2.3.4 Observations Observations were made to: 1) determine the number of immobilized or dead parent daphnids in each test solution during the test and the day of death. Immobilization is defined as the inability of a daphnid to swim within 15 seconds after gentle agitation of the test vessel, 2) count the number of neonates ( alive and dead progeny) for each parent animal throughout the test and per day of test and record the dates of release, 3) count the number of aborted eggs and record the dates of observation, 4) establish the length of all surviving parent daphnids at the end of the test ( to the nearest 0.1 mm).

Observations were undertaken everyday. Neonates ( alive and dead progeny) and aborted eggs were removed from the test vessels everyday.

2.4 CHEMICAL ANALYSIS Chemical analysis was undertaken to measure the concentration of the test item in each test solution, except for the control, throughout the test.

Samples of 5 mL were taken from each test solution container ( except for the control) at the beginning of the test. Samples totaling 5 mL were also taken before and after the medium renewals and at the end of the test. Samples were stored at  20 ° C until analysis.

The analytical procedure is presented in appendix 3.

2.5 DATA EVALUATION The following measurements and calculations were performed when possible:

  the number of parent animals which were immobilized or died during the test and day of death,

  the number of neonates ( live and dead progeny) for each parent animal and dates of release,

  the number of aborted broods and dates of observation,

  the number of neonates per parent throughout the test and per day of test,

  the coefficient of variation for control fecundity ( based on the number of neonates per parent animal alive at the end of the test per day and based on the total number of living neonates per parent animal alive at the end of the test),

  a plot of total number of living neonates per parent animal for each replicate alive at the end of the test versus concentration of the test item,

  the results of tests of homogeneity of data,

  the results of analysis of variance ( Dunnett test or Bonferroni t test) on the number of living neonates per surviving parent at the end of the test and the length of these surviving parents, used to calculate the No Observed Effect Concentration ( NOEC) and also the Lowest Observed Effect Concentration ( LOEC) after 21 days for each parameter,

  where applicable, an EC50 based on reproduction capacity ( or mortality or length) of parent animals and confidence limits, at the end of the test. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 14 of 52 2.5.1 Determination of the EC50 When for at least two concentrations, effect ( mortality/ inhibition of reproduction capacity reduction of length) is > 0% and < 100%, the EC50 is calculated according to Probit analysis ( i. e. Finney's method, published by E. Weber, combined with Bliss's method). The confidence interval limits are calculated statistically according to Fieller's method. When only one partial effect is observed, the EC50 is estimated ( using Probit analysis or an interpolation procedure, depending on the data obtained) and the highest concentration causing no effect and the lowest concentration producing 100% effect are used as confidence limits. If at all concentrations effect is 0% or 100%, a EC50 can not be calculated. It is estimated using the geometric mean of the highest concentration causing no effect and the lowest concentration producing 100% effect.

2.5.2 Determination of the No Observed Effect Concentration ( NOEC) The following method was applied to determine the No Observed Effect Concentration after 21 days ( 21 day NOEC) based on reproduction capacity. After checking of the normality of the data with Chi square and Shapiro Wilks tests ( normal at p = 0.01) as well as the variance homogeneity ( Bartlett test); ( p = 0.01), the NOEC was determined by ANOVA ( Bonferroni t test; p = 0.05).

Another statistical procedure, described in appendix 5 was used to determine the 21 day NOEC ( based on length of parent animals at the end of the test) since length data, although variances were homogeneous, were not normally distributed.

2.6 VALIDITY CRITERIA For the test to be valid the following criteria should be fulfilled:

  the mortality in the controls ( parent females) should not exceed 20% at the end of the test,

  the average cumulative number of living young produced per surviving parent female should be   60 in the controls at the end of the test.

2.7 ARCHIVING The following study materials are archived by CIT ( BP 563, 27005 Evreux, France) for 5 years after the end of the in vivo phase of the study:

  Study plan and possible amendments,

  raw data,

  correspondence,

  final report and possible amendments.

On completion of this period, the archived study materials will be returned to the Sponsor, or may be archived at CIT for a further period ( at additional cost). The total duration of archiving ( depending on regulations) will be the responsibility of the Sponsor. In addition, raw data not specific to the study including, but not limited to, records of environmental data and equipment calibration, will also be archived by CIT and retained for at least 30 years. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 15 of 52 2.8 STUDY PLAN ADHERENCE The study was performed in accordance with the Study plan No. 22658 ECD and subsequent amendments, with the following deviation from the agreed Study plan:

  temperature in test solutions was between 18 ° C and 22 ° C but varied slightly by more than ± 1 ° C during the test ( 1.3 ° C).

This minor deviation was not considered to have compromised the validity or integrity of the study. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 16 of 52 3. RESULTS 3.1 VALIDITY CRITERIA The two validity criteria were fulfilled:   the mortality in the controls was   20% ( 2/ 10 parents) at the end of the test,   the average cumulative number of living young produced per surviving parent was   60 in the controls at the end of the test.

3.2 CHEMICAL ANALYSIS Results are presented in table A, appendix 3.

Some measured concentrations were not within ± 20% of the corresponding nominal values. However, the maximum CV of the measured concentrations ( at the beginning of the test and before medium renewals) at each of the different nominal target values was only 8.50% ( at 6.25 mg/ L nominal), indicating that the test item was sufficiently stable between medium renewals. All study results were therefore based on harmonic means of measured concentrations in test solutions throughout the test, i. e 21 days. These harmonic means were calculated as 4.31, 9.16, 20.0, 44.2 and 88.6 mg/ L expressed as the APFO ( solids) content of the test item, for nominal values of 6.25, 12.5, 25.0, 50.0 and 100 mg/ L, respectively.

3.3 WATER QUALITY A translucent colorless solution was obtained at all test item concentrations.

The minimum and maximum values of parameters measured during the test were:

  pH: 7.56 and 8.26,

  temperature: 18.4 ° C and 20.4 ° C,

  dissolved oxygen: 7.8 and 9.2 mg/ L (> 3 mg/ L throughout the test),

  water hardness: 255 and 289 mg/ L as CaCO3.

Results of water quality measurements are included in table 1.

3.4 RESULTS   Number of parent animals which were immobilized or died during the test and day of death ( table 2).

The test item did not induce significant mortality in parent animals at concentrations tested. At the end of the test, the cumulative mortality in parent animals was 20% ( 2/ 10) in the control, 10% ( 1/ 10) at 4.31 mg/ L, 0% ( 0/ 10) at 9.16 mg/ L, 0% ( 0/ 10) at 20.0 mg/ L, 10% ( 1/ 10) at 44.2 mg/ L and 10% ( 1/ 10) at 88.6 mg/ L.

The EC50 value based on mortality of parent animals after 21 days was as follows:

  21 day EC50: > 88.6 mg/ L. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 17 of 52   Coefficient of variation for control fecundity ( tables 3a and 4).

The coefficient of variation for control fecundity was 16.4% based on the total number of living neonates per parent animal alive at the end of the test ( see table 4). The same coefficient was obtained using the total number of neonates ( alive and dead progeny) per parent animal alive at the end of the test per day ( see table 3a) since no dead progeny was observed in the control group during the study.

  Numbers of broods, neonates ( alive and dead progeny), aborted eggs and dates of observation ( tables 3a to 3f, table 4).

For parent animals of the control and the groups at 4.31 and 9.16 mg/ L, the first brood occurred between day 8 and day 10. Four to five broods were obtained in the test solution replicates where the parent animal was alive after 21 days. Between 9.16 and 88.6 mg/ L, the test item induced a dose related effect on occurrence of the first brood ( delay) and also, on the number of broods obtained in the test solution replicates where the parent animal was alive after 21 days ( decrease). The average number of brood per parent animal alive at the end of the test and per test solution was 4.75, 4.67, 4.80, 4.50, 2.25 and 1.22 at 0, 4.31, 9.16, 20.0, 44.2 and 88.6 mg/ L, respectively. Consequently, the total number of neonates ( alive and dead progeny) and aborted eggs produced per parent animal alive at the end of the test decreased quickly between 9.16 and 88.6 mg/ L.

After 21 days, the average number of neonates ( alive and dead progeny) produced per parent alive at the end of the test and per test solution reached 63.3, 70.7, 64.2, 62.9, 27.3 and 0.9 at 0, 4.31, 9.16, 20.0, 44.2 and 88.6 mg/ L, respectively. Furthermore, the number of aborted eggs produced per parent alive at the end of the test increased with the test item concentration from 20.0 mg/ L, corresponding to 93% of total progeny at 88.6 mg/ L. Hence, the production of neonates was highly affected by the test item. Dead progeny and aborted eggs in parent animals alive at the end of the test represented 0%, 0.2%, 0.2%, 1.4%, 12.1% and 99.2% of the total progeny ( including aborted eggs) for test solutions at 0, 4.31, 9.16, 20.0, 44.2 and 88.6 mg/ L, respectively.

  Number of living neonates per parent alive at the end of the test versus concentration of the test item ( table 4 and figure 1).

After 21 days, the average number of living neonates produced per parent alive at the end of the test and per test solution reached 63.3, 70.6, 64.1, 62.3, 24.0 and 0.1 at 0, 4.31, 9.16, 20.0, 44.2 and 88.6 mg/ L, respectively. The level of reproduction in the control ( 0 mg/ L) was within the range previously observed under these conditions in this laboratory. This average, as % of control value, corresponded to 112% at 4.31 mg/ L, 101% at 9.16 mg/ L, 98% at 20.0 mg/ L, 38% at 44.2 mg/ L and 0% at 88.6 mg/ L. Hence, as for the total production of neonates, that of living neonates was highly affected by the test item and this effect was statistically significant from 44.2 mg/ L, compared to the control.

It was therefore concluded that the test item, when tested at concentrations of 44.2 and 88.6 mg/ L, induced adverse effect on the reproduction capacity of daphnids.

The reproduction capacity values, based on production of living neonates per parent alive at the end of the test, were as follows: 21 day EC50: 39.6 mg/ L Confidence limits: 36.7 to 42.5 ( p = 0.05) No Observed Effect Concentration ( 21 day NOEC): 20.0 mg/ L ( p = 0.05)

The statistical results for the determination of the 21 day EC50 and 21 day NOEC based on reproductive capacity of parent animals are reported in appendices 4 and 5, respectively. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 18 of 52   Length of parent alive at the end of the test ( table 5)

The average length of parent animals alive at the end of the test, as % of control value, corresponded to 99% at 4.31 mg/ L, 94% at 9.16 mg/ L, 95% at 20.0 mg/ L, 94% at 44.2 mg/ L and 85% at 88.6 mg/ L. A significant inhibition of the length of parent animals ( alive at the end of the test) was obtained at 88.6 mg/ L, compared to the control.

The length values were as follows: 21 day EC50: > 88.6 mg/ L No Observed Effect Concentration ( 21 day NOEC): 44.2 mg/ L ( p = 0.05)

The statistical result for the determination of the 21 day NOEC based on length of parent animals is presented in appendix 6.

4. CONCLUSION Under our experimental conditions, the 21 day EC50 based on reproduction capacity of AMMONIUM PERFLUOROOCTANOATE ( APFO) in a semi static test system was 39.6 mg/ L for Daphnia magna. The NOEC after 21 days based on survival of the parent animals was 88.6 mg/ L ( the maximum concentration tested). The NOEC at 21days based on reproduction capacity was 20.0 mg/ L. The NOEC at 21 days based on length of the parent animals was 44.2 mg/ L.

These results are expressed as the APFO ( solids) content of the test item. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 19 of 52 Figure 1: Total number of living neonates per parent alive after 21 days versus concentration

: replicate;

: mean 0 10 20 30 40 50 60 70 80 90 100 0 10 20 30 40 50 60 70 80 90 100 Concentration ( mg/ L) Number of neonates CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 20 of 52 Table 1: Physico chemical parameters over the 21 day period Control Renewal 0 1 3 Day Solution new old new old new Beaker 1 2 1 2 1 2 3 4 3 4 Oxygen ( mg/ L) 8.9 8.8 8.1 8.1 8.4 8.3 8.0 7.9 8.1 8.1 Oxygen (%) 94 94 93 92 96 96 94 92 95 94 pH 8.06 8.12 8.03 8.19 7.56 7.63 7.76 7.96 7.79 7.81 Total Hardness ( mg/ L as Ca CO3) 255 255 255 272 255 255 289 272 255 255 Temperature ( ° C) 19.9 19.9 19.8 19.7 19.7 19.7 18.5 18.6 18.7 18.7 Renewal 8 Day Solution old new old new old Beaker 5 6 5 6 8 9 8 9 9 10 Oxygen ( mg/ L) 7.9 7.8 8.3 8.4 7.9 7.8 8.1 8.1 8.0 7.9 Oxygen (%) 93 93 94 99 91 90 94 93 94 94 pH 8.22 8.26 7.81 7.92 8.22 8.16 8.08 8.07 7.86 7.89 Total Hardness ( mg/ L as Ca CO3) 289 272 255 272 272 272 255 255 289 272 Temperature ( ° C) 19.4 19.2 19.0 19.0 19.3 19.2 19.1 19.3 19.8 19.8 Highest concentration ( where animals were still alive)

Renewal 0 1 3 Day 9 Solution new old new old new Concentration ( mg/ L) Beaker 51 52 51 52 51 52 53 54 53 54 Oxygen ( mg/ L) 9.2 9 8.1 8.2 8.4 8.4 8.1 8.1 8.3 8.3 Oxygen (%) 96 95 93 94 96 97 93 93 97 97 pH 8.09 8.13 8.24 8.26 7.89 7.86 8.02 8.03 7.73 7.74 Total Hardness ( mg/ L as Ca CO3) 255 255 272 255 255 255 272 289 255 255 Temperature ( ° C) 19.8 19.7 18.9 19.3 19.7 19.5 18.4 18.4 19.0 19.1 Renewal 8 Day 21 Solution old new old new old Concentration ( mg/ L) Beaker 55 56 55 56 57 58 57 58 59 60 Oxygen ( mg/ L) 7.8 7.8 8.8 8.8 8.0 8.0 8.4 8.5 7.9 7.9 Oxygen (%) 90 90 99 99 91 91 95 96 94 94 pH 8.25 8.22 7.88 7.89 8.22 8.25 8.09 8.11 7.96 8.00 Total Hardness ( mg/ L as Ca CO3) 289 272 272 255 289 272 255 255 272 272 Temperature ( ° C) 19.1 18.9 18.9 19.0 19.1 19.1 19.2 19.3 19.6 19.6 new: new test solution; old: old test solution Temperature in test water throughout the test: minimum 18.4 ° C / maximum 20.4 ° C 0 2 9 0 2 16 6 19 88.6 16 6 88.6 19 CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 21 of 52 Table 2: Number of immobilized parents Concentration ( mg/ L) 1st imm. 2nd imm. 3rd imm. 4th imm. 5th imm. 6th imm. 7th imm. 8th imm. 9th imm. 10th imm. Number of daphnids immobilized at the end of the test ( d 21)

control d 09 d 15 2 4.31 d 14 1 9.16 20.0 44.2 d 19 1 88.6 d 21 1 d : day imm.: immobilization CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 22 of 52 Table 3a: Living, dead and aborted eggs produced at each brood release  Control NEONATES PER ADULT  Group 1: 0 mg/ L Parent Brood 1 Brood 2 Brood 3 Brood 4 Brood 5 Total Total Total number L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. L. P.+ D. P. L. P.+ D. P. ( 1) ( 1) per day ( 1) 1 6   9 19   12 18   15 12   19 3   21 58 58 2.76                        2 4   10 14   13 18   17 4   20     40 40 1.90                        3    + D9                                           4 8   9 22   13 21   16 11   19     63 63 3.00 1   10                    5 10   9 17   12 17   15 17   19 6   21 72 72 3.43         5   16            6 8   8 16   11 19   15 17   18 10   21 70 70 3.33                        7 8   9 15   12 4   + D15                                   8 7   9 16   12 21   15 15   18 7   21 69 69 3.29             3   19        9 7   8 4   11 20   15 14   18 8   21 66 66 3.14 1   9 12   12                10 7   8 18   11 18   14 16   18 9   21 68 68 3.24                        Total 67 0 0  153 0 0  161 0 0  109 0 0  43 0 0  506 506 Mean 63.3 63.3 3.01 CV 16.4%

L. P.: Living Progeny D. P.: Dead Progeny A. E.: Aborted Eggs + = mortality of the adult at the corresponding study day ( 1) = for parent animals alive at the end of the test CV: Cefficient of Variation based on the number of living and dead progeny per parent alive at the end of the test and per day CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 23 of 52 Table 3b: Living, dead and aborted eggs produced at each brood release   4.31 mg/ L NEONATES PER ADULT  Group 2: 4.31 mg/ L Parent Brood 1 Brood 2 Brood 3 Brood 4 Brood 5 Total Total number L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. L. P.+ D. P. ( 1) ( 1) 1 4   9 7 2 3 12   + D14                                 2 8   8 25   11 22   15 20   18 7   21 82 82                       3 10   8 17   11 7   14 12   18 4   21 50 50                       4 9   9 18 1  12 19   15 18   18     64 65                       5 11   8 25   12 23   15 15   18 10   21 84 84                       6 12   8 18   11 19   14 18   18 10   21 77 77                       7 9   9 21   12 13   15 15   19     58 58                       8 12   8 21   11 25   14 18   18 11   21 87 87                       9 10   9 17   12 18   15 17   18 8   21 70 70                       10 11   8 12   12 23   15 16   18     63 63             1   19       Total 96 0 0  181 3 3  169 0 0  150 0 0  50 0 0  635 636 Mean 70.6 70.7 L. P.: Living Progeny D. P.: Dead Progeny A. E.: Aborted Eggs + = mortality of the adult at the corresponding study day ( 1) = for parent animals alive at the end of the test CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 24 of 52 Table 3c: Living, dead and aborted eggs produced at each brood release   9.16 mg/ L NEONATES PER ADULT  Group 3: 9.16 mg/ L Parent Brood 1 Brood 2 Brood 3 Brood 4 Brood 5 Total Total number L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. L. P.+ D. P. ( 1) ( 1) 1 8   8 17   11 11   15 18   18 9   21 63 63                       2 10   9 16   13 19   16 6   19     53 53 2   10                   3 5   8 12   11 11   14 18   18 5   21 52 52             1   19       4 9   9 14   11 17   13 21   16 1   21 64 65      1  12     2   19       5 11   8 14   11 14   14 13   17 10   21 69 69             7   18       6 8   9 17   12 16   15 12   18     58 58         1   16 4   19       7 9   8 9   11 18   15 16   18 4   21 56 56                       8 9   8 12   11 10   14 20   17 12   20 71 71 1   9 6   13     1   18       9 14   8 15   11 18   14 16   18 9   21 72 72                       10 12   8 23   11 19   14 19   18 10   21 83 83                       Total 98 0 0  155 1 0  154 0 0  174 0 0  60 0 0  641 642 Mean 64.1 64.2 L. P.: Living Progeny D. P.: Dead Progeny A. E.: Aborted Eggs + = mortality of the adult at the corresponding study day ( 1) = for parent animals alive at the end of the test CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 25 of 52 Table 3d: Living, dead and aborted eggs produced at each brood release   20.0 mg/ L NEONATES PER ADULT  Group 4: 20.0 mg/ L Parent Brood 1 Brood 2 Brood 3 Brood 4 Brood 5 Total Total number L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. L. P.+ D. P. ( 1) ( 1) 1 5   8 17   11 13   14 25   18     60 60                       2 12   8 19   11 20   14  1  17 7   21 80 81             22   18       3 9   8 11   11 10   14 19   18 2   21 51 51                       4 7   8 15   11 26   14 18   18     66 66                       5 13   8 18   11 22   14 13   17 9   21 78 79     1   12     2 1  18       6 10   10 4 1  13 8  3 17 4   21     26 27                       7 13   8 16   11 16   14 19   18 9   21 73 73                       8 20   15 7 2  18             27 29                       9 15   8 18   11 23   14 21   18 11   21 88 88                       10 12   8 15   11 20   14 19 1  18 8   21 74 75                       Total 116 0 0  141 3 0  158 0 3  162 3 0  46 0 0  623 629 Mean 62.3 62.9 L. P.: Living Progeny D. P.: Dead Progeny A. E.: Aborted Eggs + = mortality of the adult at the corresponding study day ( 1) = for parent animals alive at the end of the test CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 26 of 52 Table 3e: Living, dead and aborted eggs produced at each brood release   44.2 mg/ L NEONATES PER ADULT  Group 5: 44.2 mg/ L Parent Brood 1 Brood 2 Brood 3 Brood 4 Brood 5 Total Total number L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. L. P.+ D. P. ( 1) ( 1) 1                     0 0                       2 8   8 13   11 16 2  14 18 2  18    + D19                         3 3 2  14                 3 5                       4                     0 0                       5 2   14                 2 2                       6 5 3  9 10   12 15   15 18   19     50 53         2   16           7 1 4  9 18   12 10 5  15 15   19     44 53                       8  2  8 15   11 20 2  14 16   17 4 2  21 61 67             6   18       9 2 6  9 15   12 17 1  15 17   19     51 58                       10 4 2  12 1 1  14   2 21         5 8                       Total 21 19 0  72 1 0  80 10 2  90 2 0  4 2 0  216 246 Mean 24.0 27.3 L. P.: Living Progeny D. P.: Dead Progeny A. E.: Aborted Eggs + = mortality of the adult at the corresponding study day ( 1) = for parent animals alive at the end of the test CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 27 of 52 Table 3f: Living, dead and aborted eggs produced at each brood release   88.6 mg/ L NEONATES PER ADULT  Group 6: 88.6 mg/ L Parent Brood 1 Brood 2 Brood 3 Brood 4 Brood 5 Total Total number L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. D. P. A. E. Day L. P. L. P.+ D. P. ( 1) ( 1) 1  5  14                 1 7 1 1  16                   2                     0 0                       3                     0 0                       4   3 12    + D21             0 0                       5   5 12   8 21             0 0                       6                     0 0                       7   18 16   18 19             0 0                       8  1 7 12                 0 1   3 16                   9  1 10 12   15 16   7 21         0 1   9 14                   10   9 12                 0 0                       Total 1 8 64  0 0 41  0 0 7  0 0 0  0 0 0  1 9 Mean 0.1 0.9 L. P.: Living Progeny D. P.: Dead Progeny A. E.: Aborted Eggs + = mortality of the adult at the corresponding study day ( 1) = for parent animals alive at the end of the test CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 28 of 52 Table 4: Living neonates per parent alive at the end of the test versus concentration of the test item For parents alive at the end of the test Test item concentrations ( mg/ L) Parent number 0 4.31 9.16 20.0 44.2 88.6 1 58  63 60 0 1 2 40 82 53 80  0 3  50 52 51 3 0 4 63 64 64 66 0  5 72 84 69 78 2 0 6 70 77 58 26 50 0 7  58 56 73 44 0 8 69 87 71 27 61 0 9 66 70 72 88 51 0 10 68 63 83 74 5 0 Mean 63.3 70.6 64.1 62.3 24.0 0.1 ( as % Control)  ( 112) ( 101) ( 98) ( 38) ( 0) CV 16.4% 18.1% 15.3% 34.6% 110% 300%

CV: Coefficient of Variation based on the number of living progeny per parent alive at the end of the test CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 29 of 52 Table 5: Length of parent alive at the end of the test Length ( mm) of parents alive at the end of the test Test item concentrations ( mg/ L) Parent number 0 4.31 9.16 20.0 44.2 88.6 1 3.3  3.1 3.1 3.1 3.0 2 3.4 3.4 3.3 3.1  2.8 3  3.3 3.1 3.1 3.3 2.8 4 3.3 3.4 3.4 3.4 3.1  5 3.4 3.4 3.1 3.1 3.1 2.6 6 3.4 3.3 3.1 3.4 3.0 3.0 7  3.3 3.3 3.3 3.3 2.6 8 3.4 3.4 3.1 3.1 3.1 2.8 9 3.3 3.3 3.1 3.1 3.1 3.0 10 3.4 3.1 3.1 3.3 3.3 3.0 Mean 3.4 3.3 3.2 3.2 3.2 2.8 ( as % Control)  ( 99) ( 94) ( 95) ( 94) ( 85) CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 30 of 52 APPENDICES CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 31 of 52 1. Analytical certificate and composition of the test item CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 32 of 52 CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 33 of 52 CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 34 of 52 2. Preparation of M4 reconstituted water CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 35 of 52 PREPARATION OF MEDIUM M4 RECONSTITUTED WATER Trace elements Amount of trace elements for the preparation of stock solutions I, which contain only one salt per solution. From the different stock solutions I a second stock solution, stock solution II is prepared, which contains all trace elements ( combined solution I) 20 fold concentrated ( related to M4):

Stock solution( s): I _____________________________________________________________________________ ( single substance) to prepare the combined 20 fold amount concentration concentrated stock added to ( in relation to solution II add the deionized water medium M4) following amount of ( mg/ L) stock solution I to deionized water ( mL/ L) _____________________________________________________________________________ M4 H3BO3 57190 20000 fold 1.0 MnCl2, 4 H2O 7210 20000 fold 1.0 LiCl 6120 20000 fold 1.0 RbCl 1420 20000 fold 1.0 SrCl2, 6 H2O 3040 20000 fold 1.0 NaBr 320 20000 fold 1.0 Na2MoO4, 2 H2O 1260 20000 fold 1.0 CuCl2, 2 H2O 335 20000 fold 1.0 ZnCl2 260 20000 fold 1.0 CoCl2, 6 H2O 200 20000 fold 1.0 KI 65 20000 fold 1.0 Na2SeO3 43.8 20000 fold 1.0 NH4VO3 11.5 20000 fold 1.0 _____________________________________________________________________________

Na2EDTA, 2 H2O 5000 2000 fold  FeSO4, 7 H2O 1991 2000 fold

Both solutions ( 1 L) are prepared individually, poured together and autoclaved immediately.

This gives: Fe EDTA solution ( 2 L) 1000 fold 20.0 _____________________________________________________________________________

: not applicable CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 36 of 52 With the 20 fold concentrated stock solution II ( which contains all trace elements), the macro nutrients and vitamins, M4 is prepared as follows: _____________________________________________________________________________ Amount Concentration Amount of stock added to ( related to solution added deionized medium M4) to prepare medium water ( mg/ L) ( mL/ L) _____________________________________________________________________________ M4 stock solution II ( combined trace elements) 20 fold 50 Macro nutrients stock solution( s) single substance CaCl2, 2 H2O 293800 1000 fold 1.0 MgSO4, 7 H2O 246600 2000 fold 0.5 KCl 58000 10000 fold 0.1 NaHCO3 64800 1000 fold 1.0 Na2SiO3, 9 H2O 50000 5000 fold 0.2 NaNO3 2740 10000 fold 0.1 KH2PO4 1430 10000 fold 0.1 K2HPO4 1840 10000 fold 0.1 Vitamins A vitamin stock solution is prepared by adding the three vitamins to one liter deionized water:

mg/ L M4 thiamine hydrochloride 750 10000 fold  cyanocobalamine ( B12) 10 10000 fold  biotine 7.5 10000 fold

combined vitamin stock  10000 fold 0.1 _____________________________________________________________________________ : not applicable The vitamin stock is stored frozen in small aliquots. Vitamins are added to the media shortly before use.

To avoid precipitation of salts when preparing the complete media add the aliquots of stock solutions to approximately 500 800 mL deionized water and then complete to 1 liter. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 37 of 52 3. Chemical analysis of test solutions CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 38 of 52 CHEMICAL ANALYSIS Principle An aliquot of each sample was diluted and analyzed by Ion Chromatography with Electrochemical Detection ( Conductimetry). The concentrations of AMMONIUM PERFLUOROOCTANOATE ( APFO) were determined from a calibration curve of peak area against concentration of AMMONIUM PERFLUOROOCTANOATE ( APFO) in standard solutions.

Sample preparation All samples were mixed. If necessary, samples were then diluted with Milli Q to achieve concentrations in the range 1 100 mg/ L of AMMONIUM PERFLUOROOCTANOATE ( APFO).

Chromatographic conditions Pump : GP 50 Gradient Pump ( Dionex)

Mobile phase : phase A: aqueous sodium hydroxide solution 50 mM* phase B: Milli Q water phase C: acetonitrile * Aqueous sodium hydroxide solution 50 mM: 2.6 mL of sodium hydroxide solution at 46/ 48% ( Fisher Chemicals, Ref.: S/ 4930/ 05) was added to 1 L of Milli Q Water.

Time ( min) phase A (%) phase B (%) phase C (%) curve 0 10 85 5 6 5 40 0 60 6 12 40 0 60  13 10 85 5  25 10 85 5

Flow rate : 1 mL/ min Precolumn : Ionpac ATC 3, 4 mm ( Dionex)

Column : Ionpac AS16 ( Dionex) length = 250 mm, inner diameter = 4 mm Temperature : 30 ° C Detector : ED 50 Electrochemical Detector ( Dionex) Conductimetry after eluent neutralization CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 39 of 52 Neutralization : ASRS Ultra ( Dionex), electric current 70 mA or AMMS ( Dionex)

Injector : AS50 ( Dionex), at 10 ° C Injected volume : 25 µ L Data acquisition software : Multichrom 2 ( Fisons Instruments)

Retention time : APFO, approx. 9 min Analysis time : 30 min Calibration curve Peak areas were determined for standard solutions ranging from 1 to 100 mg/ L of AMMONIUM PERFLUOROOCTANOATE ( APFO) ( six levels). A calibration curve was obtained by plotting test item peak areas against concentrations.

The regression analysis of the calibration data gave an equation of the following form: Y = aX + b where Y = test item peak area ( µ Vs) X = concentration of test item ( mg/ L) a = slope value b = intercept Assay Samples of AMMONIUM PERFLUOROOCTANOATE ( APFO) were analyzed by Ion Chromatography with Electrochemical Detection ( Conductimetry).

One dilution was prepared for each sample and one injection ( of 25 µ L aliquots) was performed for each final dilution.

The peak area was determined for each sample. The concentration of AMMONIUM PERFLUOROOCTANOATE ( APFO) in each sample was calculated using the equation obtained from the calibration data.

All the results are expressed as mg/ L of APFO. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 40 of 52 Table A: Concentration of AMMONIUM PERFLUOROOCTANOATE ( APFO) in the test solutions during the study ( mg/ L)

Nominal day 0 day 2 day 5 day 7 old fresh old fresh old fresh 6.25 3.94 ( 63) 4.22 ( 68) 4.02 ( 64) 3.87 ( 62) 4.10 ( 66) 3.88 ( 62) 4.28 ( 68)

12.5 8.97 ( 72) 8.50 ( 68) 8.57 ( 69) 8.68 ( 69) 9.81 ( 78)

25 19.0 ( 76) 19.9 ( 80) 19.0 ( 76) 19.0 ( 76) 18.9 ( 76) 18.8 ( 75) 21.1 ( 84)

50 43.3 ( 87) 46.4 ( 93) 42.0 ( 84) 44.4 ( 89) 42.5 ( 85) 45.1 ( 90) 42.2 ( 84)

100 82.6 ( 83) 86.1 ( 86) 82.0 ( 82) 91.5 ( 92) 80.0 ( 80) 83.5 ( 84) 87.9 ( 88)

Nominal day 12 day 19 old fresh old fresh old fresh old 6.25 4.63 ( 74) 4.50 ( 72) 4.45 ( 71) 4.83 ( 77) 5.24 ( 84) 4.34 ( 69) 4.41 ( 71)

12.5 9.65 ( 77) 9.26 ( 74) 9.30 ( 74) 9.19 ( 74) 9.48 ( 76) 9.10 ( 73) 9.63 ( 77)

25.0 21.2 ( 85) 20.9 ( 84) 19.8 ( 79) 20.0 ( 80) 22.4 ( 90) 19.5 ( 78) 21.7 ( 87)

50.0 43.9 ( 88) 45.1 ( 90) 41.2 ( 82) 43.8 ( 88) 45.3 ( 91) 44.2 ( 88) 50.6 ( 101)

100 98.0 ( 98) 94.8 ( 95) 90.8 ( 91) 80.7 ( 81) 98.8 ( 99) 93.6 ( 94) 103 ( 103)

Nominal Harmonic mean ( days 0 21)

6.25 12.5 25.0 50.0 100 vd: value discarded ( aberrant value) CV: Coefficient of Variation ( 2): based on measured concentrations at the beginning of the test and before medium renewals 4.59 4.52 2.20 7.25 day 16 Measured ( 1)

Measured ( 1) day 21 vd vd ( 1): numbers in brackets represent percentages of the nominal concentration Measured CV (%) ( 2)

4.31 9.16 20.0 44.2 89.0 8.50 CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 41 of 52 VALIDATION OF THE ANALYTICAL METHOD The validation of the analytical method was performed according to CIT Standard Operating Procedures. The specificity, limit of quantification, linearity, repeatability of injections, accuracy and precision ( Coefficient of Variation: CV %) of the analytical method were determined.

Specificity The specificity of the analytical method was demonstrated as follows:

  analysis of a standard solution of AMMONIUM PERFLUOROOCTANOATE ( APFO) in Milli Q water,

  analysis of Milli Q water or test water ( LC, M4 or dechlorinated; desionized water) without dilution, No relevant interference between the test item peak and Milli Q water or test water was observed on chromatograms.

Limit of quantification The limit of quantification of the analytical method was established as 1 mg/ L for a standard solution of APFO. This limit corresponds to a limit of quantification of 1 mg/ L for the test item in aqueous phase.

Linearity Linearity was checked by analysis of three different sets of six standard solutions containing 1, 5, 10, 20, 50 and 100 mg/ L of AMMONIUM PERFLUOROOCTANOATE ( APFO) in Milli Q water. Satisfactory linearity was demonstrated in the range 1 to 100 mg/ L since the coefficients of determination obtained were higher than 0.999.

Repeatability of injections Replicate analysis ( n = 10) of a solution containing 50 mg/ L of the test item gave satisfactory results since the coefficients of variation values obtained were as follows:

  1% based on peak height,

  1% based on peak area. Based on these results, the repeatability of injections of the analytical method was validated taking into account peak area. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 42 of 52 Accuracy and precision Six analyses of solutions containing 2.03 and 50.7 mg/ L of the test item in Milli Q water were carried out.

The accuracy and the precision ( CV%) obtained were as follows:

Concentration ( mg/ L) Dilution CV Accuracy Nominal Mean measured* factor (%) (%) 2.03 2.02 1 6 99 50 .7 53.7 1 2 106 *: mean values of six determinations Conclusion The analytical method was validated and considered to be suitable for the analysis of the samples of the study. CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 43 of 52 4. EC50 estimation CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 44 of 52 21 day EC50 ESTIMATION BASED ON REPRODUCTIVE CAPACITY _____________________________________________________________________________ Concentration Percentage inhibition mg/ L _____________________________________________________________________________ 0 4.31 0 9.16 0 20.0 2 44.2 62 88.6 100 _____________________________________________________________________________

Slope: 3.183 Equation: Y = 3.183 x X   21.369 21 day EC50 LOWER LIMIT UPPER LIMIT ______________________________________________

39.6 ( 36.7  42.5 ) mg/ L Significance: 95% CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 45 of 52 5. 21 day NOEC determination based on reproduction capacity CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 46 of 52 21 day NOEC determination based on reproduction capacity CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 47 of 52 21 day NOEC determination based on reproduction capacity ( continued) CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 48 of 52 6. Statistical analysis of Daphnia length data CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 49 of 52 CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 50 of 52 CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 51 of 52 CIT/ Study No. 22658 ECD/ AMMONIUM PERFLUOROOCTANOATE ( APFO)/ APME 52 of 52


[Federal Register Volume 75, Number 220 (Tuesday, November 16, 2010)]
[Rules and Regulations]
[Pages 69861-69862]
[FR Doc No: 2010-28579]

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DEPARTMENT OF TRANSPORTATION

Federal Aviation Administration

14 CFR Part 39

[Docket No. FAA-2008-1328; Directorate Identifier 2008-CE-066-AD;
Amendment 39-15776; AD 2008-26-10]
RIN 2120-AA64

Airworthiness Directives; Cessna Aircraft Company (Cessna) 172,
175, 177, 180, 182, 185, 206, 207, 208, 210, 303, 336, and 337 Series
Airplanes

AGENCY: Federal Aviation Administration (FAA), DOT.

ACTION: Final rule; correction.

-----------------------------------------------------------------------

SUMMARY: The FAA is correcting an airworthiness directive (AD) that was
published in the Federal Register. That AD applies to the products
listed above. In the Information Heading and in the SUMMARY section of
the published AD, we incorrectly included Cessna 188 series airplanes.
In the Unsafe Condition section, we incorrectly designated that
paragraph as (e) instead of (d). Also in the Compliance section,
paragraph (f)(2), and in Figure 1, we incorrectly stated the mailing
address for the report. We are issuing this document to help eliminate
any confusion that this AD may have created in the Information Heading
and in the SUMMARY and Unsafe Condition sections. This document
corrects those errors. In all other respects, the original document
remains the same.

DATES: This final rule is effective November 16, 2010. The effective
date for AD 2008-26-10 remains January 5, 2009.

ADDRESSES: You may examine the AD docket on the Internet at http://www.regulations.gov; or in person at the

Docket Management Facility between 9 a.m. and 5 p.m., Monday through
Friday, except Federal holidays. The AD docket contains this AD, the
regulatory evaluation, any comments received, and other information.
The address for the Docket Office (phone:<PHONE_NUMBER>) is Document
Management Facility, U.S. Department of Transportation, Docket
Operations, M-30, West Building Ground Floor, Room W12-140, 1200 New
Jersey Avenue, SE., Washington, DC 20590.

FOR FURTHER INFORMATION CONTACT: Ann Johnson, Aerospace Engineer, FAA,
Wichita Aircraft Certification Office, 1801 Airport Road, Room 100,
Wichita, Kansas 67209; telephone:<PHONE_NUMBER>; fax:<PHONE_NUMBER>; e-
mail address<EMAIL_ADDRESS>
SUPPLEMENTARY INFORMATION: Airworthiness Directive 2008-26-10,
Amendment 39-15776 (73 FR 78939, December 24, 2008), currently requires
inspecting the alternate static air source selector valve to assure
that the part number identification placard does not obstruct the
alternate static air source selector valve port. If the part number
identification placard obstructs the port, this AD also requires
removing the placard, assuring that the port is unobstructed, and
reporting to the FAA if obstruction is found for certain Cessna 172,
175, 177, 180, 182, 185, 206, 207, 208, 210, 303, 336, and 337 series
airplanes.
    As published, the Information Heading and the Summary sections of
the AD incorrectly included Cessna 188 series airplanes. The Unsafe
Condition section is incorrectly designated as paragraph (e) instead of
paragraph (d). Also, the mailing address for the report specified in
the Compliance section, paragraph (f)(2), and in Figure 1 is
incorrectly stated as 1804 instead of 1801.
    No other part of the preamble or regulatory information has been
changed; therefore, only the changed portion of the final rule is being
published in the Federal Register.
    The effective date of AD 2008-26-10 remains January 5, 2009.

Correction of Non-Regulatory Text

    In the Federal Register of December 24, 2008, AD 2008-26-10;
Amendment 39-15776 is corrected as follows:
    On page 78939, in the second column, on line 10, under the heading
DEPARTMENT OF TRANSPORTATION, remove 188 from affected series
airplanes.
    On page 78939, in the second column, on line 19, under the heading
DEPARTMENT OF TRANSPORTATION, in the SUMMARY section, remove 188 from
affected series airplanes.

Correction of Regulatory Text

Sec.  39.13  [Corrected]

    In the Federal Register of December 24, 2008, AD 2008-26-10;
Amendment 39-15776 is corrected as follows:
    On page 78942, in the first column, under the Unsafe Condition
section, change paragraph (e) to (d).
    On page 78943, in the second column, in paragraph (f)(2), on line
3, change 1804 to 1801.
    On page 78943, in Figure 1, in the address for the Wichita
Manufacturing Inspection District Office, change 1804 to 1801.

    Issued in Kansas City, Missouri, on November 4, 2010.
James E. Jackson,
Acting Manager, Small Airplane Directorate, Aircraft Certification
Service.
[FR Doc. 2010-28579 Filed 11-15-10; 8:45 am]
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