PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 1 of 13 TITLE: ETHYL TERTIARY BUTYL ETHER ORAL ABSORPTION IN MALE AND FEMALE RATS SPONSOR: Section 211( b) Research Group American Petroleum Institute 1220 L Street NW Washington, DC 20005 TESTING FACILITY: Science and Engineering RTI International* Post Office Box 12194 3040 Cornwallis Road Research Triangle Park, NC 27709 RTI PROJECT NO.: 09408.007 RTI STUDY DIRECTOR: Timothy R. Fennell PROPOSED EXPERIMENTAL START DATE: April 2005 PROPOSED EXPERIMENTAL TERMINATION DATE: January 2006 SPONSOR REFERENCE NO:

AMENDMENTS:

No. Date Section Pages 1 2 3 4 Thomas E Gray, MS, DABT Sponsor's Representative Date Timothy R. Fennell, Ph. D. Study Director RTI International Date RTI International is a trade name of Research Triangle Institute. PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 2 of 13 Table of Contents Page 1.0 OBJECTIVES............................................................................................................................. 4 2.0 PERSONNEL.............................................................................................................................. 4 3.0 STUDY DESIGN......................................................................................................................... 4 4.0 JUSTIFICATIONS ...................................................................................................................... 5 4.1 Animal Species ........................................................................................................................... 5 4.2 Numbers of Animals.................................................................................................................... 5 4.3 Routes of Administration and Dose Levels .................................................................................. 5 5.0 REGULATORY COMPLIANCE .................................................................................................. 5 6.0 TEST SUBSTANCE ................................................................................................................... 5 7.0 ANIMALS.................................................................................................................................... 6 7.1 Husbandry................................................................................................................................... 6 7.1.1 Identification................................................................................................................... 7 7.1.2 Quarantine...................................................................................................................... 7 7.1.3 Feed and Water.............................................................................................................. 7 7.1.4 Environmental ................................................................................................................ 7 7.1.5 Acclimation and Housing During Studies ........................................................................ 7 7.2 Randomization and Assignment of Animals to Treatment Groups ............................................... 7 7.3 Body Weights.............................................................................................................................. 8 7.4 Found Dead/ Moribund Animals ................................................................................................... 8 7.5 Anesthesia and Euthanasia ......................................................................................................... 8 8.0 STUDY PROCEDURES.............................................................................................................. 8 8.1 Test Chemical Preparation and Analysis ..................................................................................... 8 8.2 Cannulation of Rats for Blood Collection ..................................................................................... 9 8.3 Dosing......................................................................................................................................... 9 8.4 Ante mortem Observations and Functional Assessments ............................................................ 9 8.5 Collection and Storage of Biological Samples ........................................................................... 10 8.5.1 Blood............................................................................................................................ 10 8.6 Analysis of Blood Samples. ....................................................................................................... 10 9.0 PHARMACOKINETIC ANALYSIS ............................................................................................ 10 PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 3 of 13 10.0 STATISTICAL ANALYSIS........................................................................................................ 11 11.0 DATA FROM " EXTRA" ANIMALS ........................................................................................... 11 12.0 RECORDS AND REPORT........................................................................................................ 11 13.0 MAINTENANCE OF RECORDS AND RAW DATA................................................................... 12 14.0 SAFETY PRECAUTIONS......................................................................................................... 13 15.0 PROTOCOL AMENDMENTS AND DEVIATIONS .................................................................... 13 16.0 REFERENCES ......................................................................................................................... 13 PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 4 of 13 1.0 OBJECTIVES The objectives of this study on ethyl tertiary butyl ether ( ETBE) are to:

Develop a method for the analysis of ETBE in blood by headspace GC Conduct an evaluation of oral absorption by measuring ETBE in blood in male and female Fischer 344 rats administered ETBE by gavage at one of two dose levels, 10 and 100 mg/ kg.

2.0 PERSONNEL   Timothy R. Fennell, Ph. D., Study Director   Susan Sumner, Ph. D., Chemist   Norman Gaudette, B. S.   Research Chemist   Rodney Snyder, M. S.   Research Chemist   Yan Hong, M. S Research Chemist   Jem Scott Emuakpor, DVM  Veterinarian   Melody Gower   Biologist Other personnel will be used as required. A full list of study participants will be included in the study report.

3.0 STUDY DESIGN For the pharmacokinetic analysis of ETBE, a GC method will be developed for the quantitation of ETBE in blood. To provide rat blood for development of the method, up to thirty male rats will be sacrificed for the collection of control blood. The rats will be euthanized under CO2 as needed on the study, and exanguinated by cardiac puncture.

For the analysis of blood concentrations, all time point blood samples from ETBE exposures will placed in glass crimp seal vials and sealed. Control blood samples will be used to develop a GC method for analyzing the concentration of ETBE in blood. Once the method is verified, blood samples will be used to prepare a standard curve. Aliquots of standards will be frozen at approximately  20 o C or below to provide sets of standard curve solutions for the definitive pharmacokinetic studies. Details of the method will be recorded in the raw data.

ETBE will be administered in water at a dose of 10 or 100 mg/ kg. For each dose level, six male rats and six female rats will be cannulated with jugular vein cannulas up to 4 days prior to dosing and the cannulas will be kept patent. Two additional male rats and two additional female rats will be cannulated and kept on hand in the event that a cannula fails. One additional male rat and one additional female rat will be available in the event of an accidental death prior to cannulation. ETBE will be administered by gavage in water. The syringe, needle and contents will be weighed prior to dosing, and after dosing the weight of the empty syringe and needle will be recorded. The weight of the dose administered will be calculated.

The only biological samples collected for analysis in this study will be blood. PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 5 of 13 It is anticipated that the study will be conducted with male rats administered 100 mg/ kg ETBE on day 1, female rats administered 100 mg/ kg on day 2. Analysis of the samples from this part of the study will be conducted prior to the start of the second portion of the study approximately 1 week later in which male rats will be dosed with 10 mg/ kg ETBE on day 8 and female rats will be dosed with 10 mg/ kg ETBE on day 9. Male and female rats will be ordered specifically for each dose group, and thus will not be randomly assigned to a treatment group.

4.0 JUSTIFICATIONS 4.1 Animal Species The present studies are designed to evaluate pharmacokinetics of ETBE to provide information that will be used for safety assessments to humans. No in vitro techniques are available that allow for adequate determination of uptake, distribution, and excretion of chemicals by mammals. Fischer 344 rats are an established animal species and strain for toxicological testing, and pharmacokinetic studies.

4.2 Numbers of Animals The numbers of animals used in this study are considered acceptable to develop the analytical procedures, and to evaluate the appropriateness of the exposure generation system for further study.

4.3 Routes of Administration and Dose Levels The route of administration is one of the expected exposure routes in humans ( through ground water). Oral administration is commonly used in toxicity or safety assessment studies. The dose vehicle, water, corresponds to a potential human exposure scenario. The doses are 10 and 100 mg/ kg,

and are expected to be without significant toxicity. The high dose was selected based on the concentration of ETBE that can be reasonably achieved in a dose volume of 10 ml/ kg. The solubility of ETBE in water is 1.2 %, which is expected to enable preparation of a solution of 10 mg/ ml for gavage dosing.

5.0 REGULATORY COMPLIANCE This study will be carried out in compliance with the U. S. EPA Good Laboratory Practice Standards, 40 CFR part 792.

6.0 TEST SUBSTANCE NAME: Ethyl tertiary Butyl Ether ( ETBE; tertiary butyl ethyl ether, 2 ethoxy 2 methylpropane)

CAS No.: 637 92 3 PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 6 of 13 MOLECULAR FORMULA: C6H14O MOLECULAR WEIGHT: 102.17 STRUCTURE:

CH3 C H3C H3C O H2 C CH3 SOURCE OF TEST SUBSTANCE: ETBE will be purchased from Sigma Aldrich, Milwaukee, WI ( Catalog number 253898, specified purity 99%). A certificate of analysis will obtained from the vendor.

LOT NUMBER( S): To be listed in the final report.

IDENTITY AND PURITY: The identity of the unlabeled ETBE will be confirmed at RTI by 1 H and 13 C NMR, and by mass spectrometry. The purity of the test chemical will be determined by GC MS.

STORAGE CONDITIONS: ETBE will be stored in the dark at room temperature.

.

7.0 ANIMALS 1. Species and Strains: Fischer 344 rats 2. Approximate Age: 8 9 weeks old at time of dosing. ( Male rats that are used as blood donors for analytical methods development may be significantly older than the 8 9 weeks of age, and this will not affect the study).

3. Approximate Weight: 200 g 4. Number/ Sex:

Up to 48 male rats and 18 female rats.

5. Sources: Taconic ( Germantown, NY) will be the primary source of animals. In the event that suitable animals cannot be provided from the primary source, acceptable alternate sources are Charles River Laboratories, Inc. ( Portage, MI), and Harlan ( Indianapolis, IN).

The source( s) of all animals will be included in the Study Report.

7.1 Husbandry Research Triangle Institute is accredited by AAALAC International. Animal procedures detailed in this protocol are in accordance with the Animal Welfare Act, " Guide for the Care and Use of Laboratory Animals" ( NRC, 1996), and the Office of Laboratory Animal Welfare ( NIH). All animal procedures will be reviewed by RTI's Institutional Animal Care and Use Committee ( IACUC) before PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 7 of 13 initiation of the studies. In the opinion of the Sponsor and Study Director, the study does not unnecessarily duplicate any previous work.

7.1.1 Identification Rats will be identified by individual metal ear tags. All individual animal data will be referenced to either ear tag number or to treatment group and animal number or to both.

7.1.2 Quarantine Animals will be quarantined for a minimum of five days before use on a study. Animals will be examined by a veterinarian or their representative prior to their release from quarantine, and only animals determined to be in good health as indicated by body weight gain and the absence of clinical signs will be used. During the quarantine period and prior to initiation of the experiments detailed in Section 8.0, rats will be housed ( maximum of 3 per cage) in polycarbonate cages with stainless steel bar lids accommodating a water bottle. Cage sizes are approximately 19" x 10 1/ 2" x 8" high ( ca. 143 sq. in.

floor space). Contact bedding will be Sani Chips ( P. J. Murphy Corporation, Montville, New Jersey).

7.1.3 Feed and Water Animals will be provided Certified Purina Rodent Chow ( 5002) ad libitum. Water will be provided ad libitum. The source of the water is the City of Durham, NC. The analysis of water and analysis of the rodent chow for chemical composition and possible chemical contamination will be provided by the suppliers and maintained in the Study Records. It is anticipated that contaminant levels will be below those permitted in the certified feed and will not affect the design, conduct, or conclusions of this study.

7.1.4 Environmental Air circulation will be 100% fresh air. Room temperature will be maintained at 64  79 ° F and relative humidity at 30  70% and monitored at least once a day. Light/ darkness will be cycled at 12 h intervals. Any deviations from these conditions shall be included in the study records. Environmental parameters will be recorded automatically using a computerized HVAC Monitoring and Control System.

7.1.5 Acclimation and Housing During Studies Animals will be acclimated for a minimum of 5 days prior to use on studies. Animals will be housed in polycarbonate cages with stainless steel lids. Following administration of ETBE, the cages will be placed in a hood for the duration of the post dosing period.

7.2 Randomization and Assignment of Animals to Treatment Groups Animals designated as blood donors for methods development will not be randomized. Animals designated for dosing with ETBE will be specifically ordered to be at the appropriate age and body weight at the time of dosing. To accommodate the dosing schedule, separate groups of 9 male and female rats PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 8 of 13 will be purchased for each dose group. For each dose group, a total of 9 rats will be available for cannulation prior to dosing. After cannulation, and before dosing, animals within each treatment group will also be assigned in numerical order to sequential values listed in Table VII " Random Sampling Numbers" which appears in the Appendix of The Statistical Analysis of Experimental Data, by John Mandel, Dover Publications, Inc., 1964. Assignment of an animal number from 1 9 will be conducted by ranking the random numbers in ascending order. Animals assigned numbers 1  4 in the treatment groups will be designated as " core" members of the group; animals assigned higher numbers will be designated as " extra" animals ( see Section 11.0 for an explanation of the use of the " extra" animals).

7.3 Body Weights Individual body weights will be measured during the quarantine period, the day of exposure, and at sacrifice.

7.4 Found Dead/ Moribund Animals The Study Director or the veterinarian will authorize euthanasia of animals with life threatening clinical signs that indicate that they are unlikely to survive until the next scheduled observation. The time of death will be estimated as precisely as possible and recorded.

7.5 Anesthesia and Euthanasia Anesthesia will be used to avoid undue pain or distress. Animals will be anesthetized with CO2 exposure just prior to study termination. Rats will be euthanized by exsanguination.

8.0 STUDY PROCEDURES 8.1 Test Chemical Preparation and Analysis ETBE will be prepared for gavage dosing as a solution in water. For dosing of animals, the dose solution will be prepared within 24 hours of dosing, and maintained at room temperature. The nominal concentration will be calculated from the weight of ETBE and the weight of water added to the dose preparation. Dose solutions will be prepared at two concentrations, the first at approximately 10 mg/ ml and the second at 1 mg/ ml, to ensure that the volume of dose administered per kg body weight is similar in the high and the low dose groups. Prior to dosing, a method will be prepared for the quantitative analysis of ETBE in water, by dissolving a weighed aliquot of the ETBE/ water solution in a suitable solvent such as methanol. Aliquots of this solution will be analyzed directly by GC, using a standard curve for the quantitation of ETBE. The stability of ETBE in water will be determined by analysis of aliquots of a mock dose solution of ETBE kept at room temperature for three days. PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 9 of 13 When appropriate methods have been developed for the detection of ETBE in blood, and for the quantitative analysis of ETBE in dosing solutions, and the stability of ETBE in blood and dose solution have been established, the animal study will be conducted.

8.2 Cannulation of Rats for Blood Collection Rats in the ETBE treatment groups will be prepared with an indwelling jugular cannula to facilitate painless, nontraumatic collection of serial blood samples. The surgical technique will be similar to that reported by McKenna and Bieri ( 1984). Animals will be anesthetized with ketamine/ xylazine ( 7: 1,

60 80 mg/ kg or to effect). A small incision at the level of the clavicle approximately 0.5 cm from the midline will afford access to the right, external jugular vein. The vessel will be blunt dissected and the cannula ( prefilled with sterile saline containing 20 IU/ mL of sodium heparin) will be inserted to a depth of approximately 28 mm. Placement in the atrium will be confirmed by patency. The cannula will be secured in the vessel at the insertion point by sutures to the surrounding musculature. The cannula will then be exteriorized by passing the distal end subcutaneously around the neck to exit the dorsal surface via a small incision between the scapulae. The ventral incision will be closed with sutures. The distal end of the cannula will be sutured to the base of the neck between the scapulae. Animals will be allowed to recover a minimum of two days prior to dosing. Cannula patency will be maintained with daily flushings of heparinized saline. At least six rats for each dose group will be prepared with cannulas.

8.3 Dosing Each rat will be weighed prior to dosing to determine the amount of dose to be administered.

Gavage doses will be administered using a syringe fitted with a blunt tipped gavage dosing needle. The dose administered to each animal will be determined from the weight of the full syringe minus that of the empty syringe. The dose time will be recorded. Dosing of animals will be spaced apart to allow blood collection at the appropriate times ( see section 8.4.1 below). The groups will be designated according to the dose administered and sex of the rats: Group A male 100 mg/ kg, Group B female 100 mg/ kg, Group C male 10 mg/ kg, Group D female 10 mg/ kg. At least four cannulated rats per group will be dosed with ETBE.

8.4 Ante mortem Observations and Functional Assessments Animals will be observed twice per day for mortality, morbidity, signs of toxicity, and for any acute distress that might be related to the test procedure or test substances. Animals exhibiting adverse reactions will be closely monitored. All signs of poor health or abnormal behavior will be recorded. PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 10 of 13 8.5 Collection and Storage of Biological Samples 8.5.1 Blood Blood samples will be collected prior to dosing from each rat ( time = 0), and from 4 rats of each sex and at each dose at 15 and 30 min, and at 1, 2, 4, 8, and 24 hr after dosing.

Blood samples will be drawn from the jugular vein cannula into a heparinized 1 ml syringe, and the sample will be immediately placed in a preweighed headspace vial. The amount of blood will be approximately 100 µ l. The vial will be immediately crimped and weighed, and the weight of the blood aliquot will be determined. After the last sample has been collected from each animal at 24 hr, the rats will be euthanized by exposure to CO2. Should cannulas fail between the 8 and 24 hr blood samples,

blood will be collected by cardiac puncture under CO2 anesthesia at sacrifice at 24 hr. The blood samples will be maintained at room temperature until analyzed. It is anticipated that blood samples will be analyzed by GC within 24 hours of collection.

Blood samples from unexposed rats will be used for methods development. Blood samples will be collected by cardiac puncture, under CO2 anesthesia, and the rats will be euthanized.

8.6 Analysis of Blood Samples.

A GC method will be developed for quantitating ETBE using blood from control male Fischer 344 rats. The methods will involve the development of a standard curve for analysis of ETBE in the headspace of air tight vials containing a known volume of control blood and a known concentration of ETBE. The stability of ETBE will be evaluated under storage conditions that may be used in this study will be evaluated during this method development. This will include analysis of ETBE immediately spiking in control blood, after storage at room temperature ( 8 hr) or at approximately 4 o C ( 24 hr). The concentration of ETBE in the headspace of vials containing a known volume of blood from rats administered ETBE by gavage will be determined by comparison to the standard curve.

9.0 PHARMACOKINETIC ANALYSIS Individual and mean blood concentration time data for each dose and sex will be analyzed, as appropriate, by noncompartmental ( model independent) using the least squares fitting program WinNonlin ( Statistical Consulting Inc., Cary, North Carolina). The following pharmacokinetic parameters will be determined as appropriate: terminal elimination rate constant, terminal elimination half life ( T1/ 2),

area under the blood concentration time curve extrapolated from time zero to infinity ( AUC), maximum concentration achieved ( Cmax), and time to maximum concentration ( Tmax). PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 11 of 13 10.0 STATISTICAL ANALYSIS All blood concentration data will be reported in tables as the mean ± standard deviation ( SD).

Pharmacokinetic data will be presented as mean ± SD. The methods to be used for statistical analysis will be added by amendment.

11.0 DATA FROM " EXTRA" ANIMALS To better ensure that complete data from the required number of animals in each treatment group will be obtained, one or more additional animals, predesignated as " extras", will start the study as part of each treatment group. If a " core" study animal fails to complete the study due to loss of samples,

misdosing, morbidity, or other accidents, it will be replaced with a designated " extra" animal and data will cease to be obtained from the " core" animal. The " extra" animal will then become part of the " core"

group and the original animal will be removed from the " core" group. All such substitutions will be documented as to reason and approved in writing by the Study Director. Except for animals that become part of a " core" group, samples from " extra" animals will not normally be analyzed. However, all data obtained in the study will be reported. Data used to construct group means and to obtain pharmacokinetic parameters will consist of the data obtained from the " core" study animals ( not the " extras") except in cases where an " extra" has taken the place of a " core" animal. In that case, the data from the " extra" animal will be used instead, and data from the " core" animal that was eliminated from the study will not be used in these calculations.

12.0 RECORDS AND REPORT The following will be maintained in the record:

a. Protocol and any amendments b. Animal receipt records c. Quarantine records d. Temperature and humidity records for the treatment rooms e. Animal research facility room logs f. Feed and water analysis for contaminants g. Randomization records h. Test chemical receipt, storage and use records i. Balance calibration log references j. Correspondences PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 12 of 13 k. All other raw data and documentation.

Results of the studies will be described in an audited draft report, which will be submitted to the Sponsor for approval. This report will include but not be limited to:

a. Name and address of the facility performing the study, dates of study initiation and completion, and RTI study number.

b. A copy of the signed, dated and approved protocol and all deviations and authorized amendments to the original protocol.

c. A detailed description of all methods used, including the randomization method.

d. The lot number( s) of the test substances and details of the formulation of doses.

e. Animal information to include: supply source, species, strain or substrain, sex, individual animal weights ( at randomization through sacrifice), approximate age at initiation of dosing, and procedure used for individual animal identification and assignment to the treatment group.

f. Tabulated individual results for blood concentration.

g. Tabulated mean results for blood concentration.

h. Pharmacokinetic and statistical analysis of the data.

Upon acceptance of the draft report by the Sponsor, a final report will be issued. Two bound and one unbound copies of the audited draft report and two bound and one unbound copies of the final report will be shipped to:

Matthew Todd Regulatory Analysis and Scientific Affairs American Petroleum Institute 1220 L Street NW Washington, DC 20005 p: ( 202) 682 8319 f: ( 202) 682 8031 13.0 MAINTENANCE OF RECORDS AND RAW DATA Records will be maintained in the laboratories of the study personnel while the studies are being conducted. Raw data generated while conducting the study and any transformations, calculations or operations performed on the data will be recorded in the study file. All raw data, documentation,

records, protocols, amendments, and the final report will be stored in the RTI International Archives,

under the control of the RTI Quality Assurance Unit in accordance with FDA, EPA FIFRA GLP regulations and OECD Principles of GLP. The applicable record retention requirements for this study are those of EPA TSCA GLPs, 40 CFR part 792, subpart J. Documentation and raw data will be maintained PROTOCOL RTI INTERNATIONAL POST OFFICE BOX 12194 RESEARCH TRIANGLE PARK, NC 27709 04/ 04/ 05 Draft RTI 932 Page 13 of 13 in the Archives for a period of ten years. The test substance ETBE is an ether that can generate explosive peroxides on storage. Therefore, an archived sample of test chemical will not be retained.

The samples of blood collected will not afford analysis beyond the duration of the study. Wet specimens of blood will be disposed of after quality assurance verification ( after the QAU assures that discarding the samples does not negatively impact the integrity of the study).

14.0 SAFETY PRECAUTIONS ETBE is a flammable liquid, with the potential to form explosive peroxides. The container should be tightly closed when not in use. Storage under a nitrogen atmosphere is recommended to avoid the generation of peroxides. Use personal protective equipment, including safety glasses, lab coat, and chemical resistant gloves.

15.0 PROTOCOL AMENDMENTS AND DEVIATIONS This protocol may be amended by the Study Director with agreement of the Sponsor as the study progresses. Normally, a formal amendment will be prepared and signed by the Study Director and the Sponsor's Representative prior to the change. If instances arise where a change is urgent, the change may become effective upon approval by the Study Director. A notification of the urgent change will be sent to the Sponsor's Representative ( email, facsimile, or telephone) as soon as feasible ( no more than 24 h after the Study Director's approval). Subsequently, a formal protocol amendment will be prepared for approval by the Study Director and the Sponsor's Representative.

Any deviations from the protocol that occur in the course of the conduct of the study will be documented. The cause for the deviation and its effect on the outcome of the study will be explained and the Study Director will sign the document.

16.0 REFERENCES Harms PG and Ojeda SR ( 1974) A rapid and simple procedure for chronic cannulation of the rat jugular vein. J. Appl. Physiol. 36, 391 392.

Mandel J ( 1964) The Statistical Analysis of Experimental Data. Dover Publications, Inc.

McKenna MC and Bieri JG ( 1984) Multilayer cannula for long term infusion of unrestrained rats. Lab.

Animal Sci. 34, 308 310.

National Research Council ( 1996). Guide for the Care and Use of Laboratory Animals. National Academy Press: Washington, DC.


[Federal Register: September 26, 2006 (Volume 71, Number 186)]
[Notices]
[Page 56139-56141]
[DOCID:fr26se06-61]

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

DEPARTMENT OF ENERGY

Federal Energy Regulatory Commission

[Docket No. CP06-449-000, Docket No. CP06-450-000, Docket No. CP06-451-
000, Docket No. CP06-448-000]

Kinder Morgan Louisiana Pipeline LLC, Natural Gas Pipeline
Company of America; Notice of Applications

September 19, 2006.
    Take notice that on September 8, 2006, Kinder Morgan Louisiana
Pipeline LLC (Kinder Morgan Louisiana), 747 East 22nd Street, Lombard,
Illinois 60148, filed applications in Docket Nos. CP06-451-000, CP06-
450-000 and CP06-449-000, pursuant to section 7(c) of the Natural Gas
Act (NGA) and Parts 157 and 284 of the Commission's regulations. Kinder
Morgan Louisiana is requesting certificates of public convenience and
necessity: (i) To construct and operate natural gas pipeline facilities
that will originate in Cameron Parish, Louisiana and terminate in
Evangeline Parish, Louisiana, and to lease firm capacity on a long-term
basis from Natural Gas Pipeline Company of America (Natural) [Docket
No. CP06-449-000]; (ii) for the

future construction of facilities pursuant to blanket certificate
authority under part 157, subpart F of the Commission's Regulations
[Docket No. CP06-450-000], and (iii) for the undertaking of self-
implementing interstate transportation service under part 284, subpart
G of the Commission's Regulations [Docket No. CP06-451-000]. Kinder
Morgan Louisiana seeks approval of proposed recourse rates for
transportation service, approval of its pro forma tariff and the
issuance of a preliminary determination providing for the Commission's
early determination on nonenvironmental issues.
    Also take notice that on September 8, 2006, Natural, 747 East 22nd
Street, Lombard, Illinois 60148, filed an application in Docket No.
CP06-448-000 pursuant to section 7(b) of the NGA, and part 157 of the
Commission's Regulations for approval to abandon, by lease to Kinder
Morgan Louisiana, certain firm capacity on a long-term basis. This
application is related to, and is being filed concurrently with the
applications being filed by Kinder Morgan Louisiana to construct a new
interstate natural gas pipeline system in Louisiana, as listed above.
    The details of these proposals are more fully set forth in these
applications which are on file with the Commission and open for public
inspection. These filings are available for review at the Commission in
the Public Reference Room or may be viewed on the Commission's Web site
at http://www.ferc.gov using the ``eLibrary'' link. Enter the docket

number excluding the last three digits in the docket number field to
access the document. For assistance, please contact FERC Online Support
at<EMAIL_ADDRESS>or toll free at<PHONE_NUMBER>, or for
TTY, contact<PHONE_NUMBER>.
    Any questions regarding these applications should be directed to
Bruce H. Newsome, Vice President of Certificates and Rates, Kinder
Morgan Louisiana Pipeline LLC and Vice President of Certificates and
Rates, Natural Gas Pipeline Company of America, 747 East 22nd Street,
Lombard, Illinois 60148-5072, telephone:<PHONE_NUMBER>, e-mail:
<EMAIL_ADDRESS>
    Specifically, Kinder Morgan Louisiana requests authorization to
construct its proposed Louisiana Pipeline Project which will consist
of: (1) About 132 miles of 42-inch pipeline from the Cheniere Sabine
Pass LNG Terminal (Sabine LNG Terminal) in Cameron Parish, Louisiana to
a point of interconnection with an existing Columbia Gulf Transmission
Company line in Evangeline Parish, Louisiana (Leg 1); (2) about 1 mile
of 36-inch pipeline that will extend from the Sabine LNG Terminal to a
point of interconnection with Natural's existing interstate pipeline
located north of the Terminal (Leg 2); and (3) about 2.3 miles of 24-
inch pipeline extending away from Leg 1 (at about milepost 110) to the
existing Florida Gas Transmission Company compressor station in Acadia
Parish, Louisiana.
    Kinder Morgan Louisiana has entered into precedent agreements with
Chevron U.S.A. and Total Gas Power North America providing for firm
transportation service on its Louisiana Pipeline Project away from the
Cheniere Sabine Pass LNG Terminal. To effectuate such transportation
service, in part, Kinder Morgan Louisiana proposes to lease firm
capacity from Natural. By leasing such capacity, Kinder Morgan
Louisiana will reduce the overall cost and improve the efficiency of
its Louisiana Pipeline Project, as well as lessen its environmental
impact. Thus, Kinder Morgan Louisiana also seeks authorization to lease
capacity of 200,000 Dth per day on the facilities of Natural in
southern Louisiana. The estimated cost of the Louisiana Pipeline
Project is about $517 million, including overhead, contingency, Section
2.55(a) auxiliary installations and reimbursement of facility costs to
interconnect pipelines.
    On February 17, 2006, the Commission granted Kinder Morgan
Louisiana's request to utilize the National Environmental Policy Act
(NEPA) Pre-Filing Process and assigned Docket No. PF06-16-000 to staff
activities involving its proposed Louisiana Pipeline Project. Now, with
the filing of the Louisiana Pipeline Project certificate applications,
the NEPA Pre-Filing Process for this Project has ended. From this time
forward, all proceedings involving the Louisiana Pipeline Project will
be conducted in Docket Nos. CP06-449, et al., as listed above in the
caption of this Notice.
    As its part of the proposal, Natural seeks authority to abandon
200,000 Dth/d of firm capacity on its system by lease to Kinder Morgan
Louisiana on a long-term basis pursuant to the terms of a capacity
lease agreement between the parties dated August 31, 2006. The leased
capacity will start at a point on Natural's Louisiana Line in Cameron
Parish, Louisiana where it will interconnect with the Kinder Morgan
Louisiana's Louisiana Pipeline Project. The leased capacity will extend
east about 16 miles to a 24-inch lateral of Natural that extends south
to Johnson's Bayou, where Kinder Morgan Louisiana proposes two delivery
points. The leased capacity (200,000 Dth/d) will also exist in the
interconnection facility between the Kinder Morgan Louisiana's
Louisiana Pipeline Project and Natural's Louisiana Line, and in each of
the two delivery points to be built at Johnson's Bayou.
    There are two ways to become involved in the Commission's review of
this project. First, any person wishing to obtain legal status by
becoming a party to the proceedings for this project should, on or
before the comment date, file with the Federal Energy Regulatory
Commission, 888 First Street, NE., Washington, DC 20426, a motion to
intervene in accordance with the requirements of the Commission's Rules
of Practice and Procedure (18 CFR 385.214 or 385.211) and the
Regulations under the NGA (18 CFR 157.10). A person obtaining party
status will be placed on the service list maintained by the Secretary
of the Commission and will receive copies of all documents filed by the
applicant and by all other parties. A party must submit 14 copies of
filings made with the Commission and must mail a copy to the applicant
and to every other party in the proceeding. Only parties to the
proceeding can ask for court review of Commission orders in the
proceeding. However, a person does not have to intervene in order to
have comments considered.
    The second way to participate is by filing with the Secretary of
the Commission, as soon as possible, an original and two copies of
comments in support of or in opposition to this project. The Commission
will consider these comments in determining the appropriate action to
be taken, but the filing of a comment alone will not serve to make the
filer a party to the proceeding. The Commission's rules require that
persons filing comments in opposition to the project provide copies of
their protests only to the party or parties directly involved in the
protest.
    Comments, protests and interventions may be filed electronically
via the Internet in lieu of paper. See 18 CFR 385.2001(a)(1)(iii) and
the instructions on the Commission's Web site under the ``e-Filing''
link at http://www.ferc.gov. The Commission strongly encourages

intervenors to file electronically. Persons unable to file
electronically should submit an original and 14 copies of the protest
or intervention to the Federal Energy Regulatory Commission, 888 First
Street, NE., Washington, DC 20426.
    This filing is accessible on-line at http://www.ferc.gov, using the

``eLibrary'' link and is available for

review in the Commission's Public Reference Room in Washington, DC.
There is an ``eSubscription'' link on the Web site that enables
subscribers to receive e-mail notification when a document is added to
a subscribed docket(s). For assistance with any FERC Online service,
please e-mail<EMAIL_ADDRESS>or call<PHONE_NUMBER> (toll
free). For TTY, call<PHONE_NUMBER>.
    Comment Date: 5 p.m. eastern time on October 11, 2006.

Magalie R. Salas,
Secretary.
 [FR Doc. E6-15669 Filed 9-25-06; 8:45 am]

BILLING CODE 6717-01-P