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WASHINGTON, D .C. 20460 ' 

DEC 11 2008 

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OFFICE QP 

PREVENTION, PES'~I'~iDES AND 

TOXIC SUBST)6~iCES 

Ms . Simone A. Miller, Corporate Director 

Environmental, Health, Safety & Regulatory Affairs 

Aceto Corporation 

One Hollow Lane, Suite 201 

Lake Success, NY 11042 

IlllllllllllllllllllllllllllllllllllllllllllllllfIII 

RE : Repeat of Study entitled "Human Percutaneous Absorption and Cutaneous Disposition of 

[r4C]-Nitroaniline In Vitro" 

Dear Ms. Miller : 

On January 24, 2005, Aceto Corporation notified EPA of its intent to conduct a test of pnitroaniline (CAS No. 100-01-6) required by a TSCA section 4 test rule in part 799.5115 (69 FR 

22402, April 26, 2004). The test protocol and data were submitted to EPA by Aceto on 

December 22, 2005 . In an e-mail on August 28, 2007 and again on December 4, 2007, EPA,__ 

informed Aceto that the study did not meet certain requirements of the test standard and that it 

must be repeated (Attachment 1) . Your response on January 16, 2008, included a letter from _ 

Celsis In Vitro Technologies, which performed the study and said it was prepared to repeat the 

study (also Attachment 1) . In a phone conversation on August 7, 2008, you informed a member 

of my staff, Catherine Roman, that the laboratory expects to be paid again to repeat the study -

with only a 10% discount (Attachment 2) . During that same conversation, you also relayed that 

Aceto Corporation no longer imports or handles p-nitroaniline and does not want to spend 

$70,000 to have the study repeated, especially because Aceto is not confident that another Tier 1 

company (which was not named) and customer will share in the test costs, although the unnamed 

Tier 1 company committed to pay 50% of the costs when the test was initially performed. 

I want to advise you that, per 40 CFR 790.59(a), Aceto will be in violation of the test rule 

if it fails to conduct the test in accordance with the test standard . EPA has identified how the 

study did not meet the requirements of the test standard and the laboratory states that it can 

perform the test and that it successfully performed the test with another chemical in the same test 

rule. To avoid being found in violation of the test rule and subject to penalties, Aceto should 

move toward having the test repeated. 

Although Aceto submitted the letter of intent to test, it might not bear the full burden of 

the cost of testing or repeat testing if there are companies from whom Aceto can seek 

CONTAINS NO C0 

Internet Address (URL) " http :l/www .epa .gov 

Recycled/Recyclable -Printed with Vegetable Oil Based Inks on Recycled Paper (Minimum 50% Postconsumer content) 

-3161 
reimbursement under 40 CFR 791 . Per 799.5115(b)(1), the persons subject to the rule include 

those who manufacture, import, or process p-nitroaniline during the reimbursement period, i.e ., 

any time after the effective date of the rule (May 26, 2004) until the end of the reimbursement 

period (December 22, 2010) . Under 799.5115(c)(3), Tier 2 companies are granted an automatic 

conditional exemption from testing. TSCA section 4(b)(3) provides for reimbursement by 

exemption holders of persons who perform the required testing, if that testing is the basis for the 

exemption. Therefore, in addition to the other Tier 1 company and customer/processor that you 

have already identified, other companies that begin or began manufacturing, importing, or 

processing p-nitroaniline during the reimbursement period are subject to the rule and may 

ultimately have reimbursement obligations. Under 791 .50(a)(3), eligible costs include "[t]he cost 

of repeating or repairing tests where failure was demonstrably due to some cause other than 

negligence of the tester ." EPA is not expressing an opinion regarding the applicability of 

791 .50(a)(3) in this case, a determination of this issue could be made under the hearing 

procedures in part 791, subpart B. 

2006 TSCA Inventory Update Rule reporting, which is based on 2005 data, only includes 

one company, other than Aceto, as an importer of p-nitroaniline . That company is Flexsys 

America L.P ., which reported importing 3 .6 million pounds in 2005 . Flexsys has not requested 

an exemption from testing. 2006 TSCA Inventory Update reporting did not include any 

manufacturers in the U.S ., and processors are not required to report . Under 40 CFR 791 .40(b) 

and 791 .48, a company's fair share of the test costs is in proportion to that company's share of 

the total production/import volume of the test chemical during the period that begins one 

calendar year before the publication of the final test rule in the Federal Register [i .e . 2003] and 

continues up to the latest data available upon resolution of a dispute. If Aceto has identified 

companies from whom it seeks reimbursement and with whom Aceto has been unable to agree 

on reimbursement, Aceto may initiate reimbursement hearings by following the procedures in 40 

CFR 791 .20. 

It is unfortunate that Aceto will be bearing the cost of testing a second time. In the future, 

I suggest stipulating in the laboratory contract that a test must meet the requirements of the test 

standard, or be repeated at no additional charge. 

I would appreciate a response indicating your intentions by January 15, 2009. If 

additional testing is conducted, a new report should be prepared. Copies of the new report 

should be provided to Catherine Roman and the EPA docket indicating that they are a revision of 

the original submission. 

0 

illis, Director 

mical Control Division 

Attachments (2) 
cc Catherine Roman (7405M) 

Renee Kearney/Docket No . EPA-HQ-OPPT-2003-0006 

Ms. Jennifer Fleischer, Celsis In Vitro Technologies, 1450 South Rolling Road, 

Baltimore, MD 21277 

Ms. Sharen Breyer, Flexsys America L.P ., Americas Headquarters, Akron Marketing & 

Technical Center, 260 Springside Drive, Akron, OH 44333-2433 
"Simone A. Miller" 

<SMiller@aceto .com> 

01/16/2008 10:51 AM 

Happy New Year! 

To Catherine Roman/DC/USEPA/US@EPA 

cc "Fred Trudwig" <FTrudwigQa aceto .com> 

bcc 

Subject RE: In Vitro Dermal Absorption Rate study of p-nitroaniline 

I just wanted to give you an update about Aceto's status with the below request. 

Aceto has been in contact with the testing lab which informed us that the study director that originally 

performed the test is no longer with the company. Thus, Aceto has been assigned a new study director 

who has provided a response (see below in red) . 

In response to our phone conversation of December 18, 2007, 1 am providing the following response to 

~ the questions raised by the EPA in regard to the skin permeability testing of p-nitroaniline. 

Based on the results from our initial study with this compound, we have determined that follow-up studies 

are necessary in order to address adequately the EPA's concerns and to ensure regulatory compliance, 

particularly as specified in 40 CFR Part 799.5115 . We therefore plan to conduct two studies . 

We will first conduct a pilot study for two purposes: 1 . to determine the solubility of p-nitroaniline in 

isopropyl myristate (IPM) and 2. to determine the time required to achieve steady state absorption of 

p-nitroaniline when dissolved in IPM and applied to cadaver skin. 

Conduct of this short pilot study will enable us to select the appropriate concentration of p-nitroaniline 

required to maintain receptor fluid p-nitroaniline levels at less than 10% of the initial level applied to the 

donor side of the skin. Results from the pilot study will also allow for appropriate selection of 4 steady state 

timepoints from which the permeability coefficient (Kp) can be correctly calculated in a statistically sound 

manner. 

A full study will then be conducted to follow 40 CFR Part 799.5115, including GLP compliance . We will use 

the protocol generated for our study of isophorone as a guide in developing the protocol for the full study 

of p-nitroaniline. 

We are formulating a Statement of Work for these two studies, which will be available to you shortly . 

Please don't hesitate to contact me if the EPA requires further details of our plan or has additional 

questions or concerns. Of course I am always available to you for discussion of study plans and protocols 

as well . 

Sincerely, 

Jennifer Fleischer 

Study Director, ADMEITox 

Celsis In Vitro Technologies 

jfleischer@celsis .com 

Best regards, 

Simone A. Miller 

Corporate Director, EHS & Regulatory Affairs 

Aceto Corporation 

One Hollow Lane, Suite 201 
Lake Success, NY 11042 

Tel : (516) 627-6000 ext . 515 

Fax: (516) 627-6093 

-----Original Message----- 

From : Roman.Catherine@epamail .epa.gov fmailto : Roman.Catherine(cr~epamail .epa.ctovl 

Sent: Tuesday, December 04, 2007 3:34 PM 

To: Simone A. Miller 

Subject: Fw: In Vitro Dermal Absorption Rate study of p-nitroaniline 

Sending again . 

Catherine Roman 

USEPA 

1200 Pennsylvania Ave., NW, 7405M 

Washington, DC 20460 

(202) 564-8172 

(202)564-4765(Fax) 

----- Forwarded by Catherine Roman/DC/USEPA/US on 12/04/2007 03:33 PM 

Catherine 

Roman/DC/USEPA/U 

S To 

smiller@aceto .com 

08/28/2007 02:59 cc 

PM Greg Schweer/DC/USEPA/US@EPA, 

Ward Penberthy/DC/USEPA/US@EPA 

Subject 

In Vitro Dermal Absorption Rate 

study of p-nitroaniline 

Simone Miller : 

On December 22, 2005, you submitted subject study (attached) on behalf of Aceto Corp. to comply 

with a TSCA section 4 test rule (69 FR 22402, April 26, 2004). A recent review of the study (attached) and 

an e-mail message (attached) by Dr. Val Schaeffer of the Occupational Safety and Health Administration 

identified some concerns which you should forward to the laboratory that conducted the study, In Vitro 

Technologies, Inc., for resolution/explanation . Following are Dr. 
Schaeffer's concerns: 

The technical data sheets were not provided that verify that the 

source of radiolabeled test compound used in the study was the 

purified para isomer of nitroaniline as specified in the final 

rule . 

In several instances, EPA's test standard (799.5115(h)) for measuring 

the permeability constant (Kp) was not followed : 

1 . Paragraph 799.5115 (h)(iii) of the test standard states 

the testing laboratory must verify that the concentration of 

test substance in the receptor fluid is less than 10 percent 

of the initial concentration in the donor chamber. This 

specification ensures that there is an undepletable , 

reservoir of test compound at the surface of the skin and 

that a strong concentration gradient is maintained across 

the epidermal membrane (i .e. infinite dosing conditions) 

during the course of the experiments. However, the study 

data show that p-nitroaniline concentration in the receptor 

fluid for all six skin experiments exceeded the 10 percent 

value (range from 25 to 68 percent of the initial 

concentration by the end of the 24 hour study period - table 

6). Since appropriate dosing conditions may not have 

occurred, it is conceivable that a maximum steady state flux 

was not achieved in any of these experiments and the true Kp 

underestimated . 

2 . Paragraph 799.5115 (h)(viii) of the test standard states 

that the in vitro dermal absorption must be performed until 

at least four absorption measurements per diffusion cell 

experiment are obtained during the steady state absorption 

portion of the experiment. Two of the samples achieved a 

marginally steady flux for a roughly four-hour period (4 to 

8 hours) covering only two absorption measurements per 

diffusion cell experiment. For the sake of consistency 

across all the experiments, the testing laboratory chose to 

calculate the permeability coefficient (Kp) from a single 

eight-hour absorption measurement per diffusion cell 

experiment rather than from a larger and more statistically 

robust number of measurements . 

3 . Paragraph 799.5115 (h)(vi) of the test standard states 

that if the concentration of a hydrophobic test substance in 

water is not high enough so that a steady state absorption 

can be obtained, the test substance must be dissolved in 
isopropyl myristate (IPM). Based on the problems with 

consistently achieving infinite dosing conditions due to the 

limited p-nitroaniline solubility in water, In Vitro 

Laboratories should have carried out the permeability 

experiments using IPM as the vehicle . 

Dr. Schaeffer also added that it is useful to compare the rather questionable infinite dosing study data 

used to determine the Kp for p-nitroaniline with the more rigorous and more acceptable (in my view) study 

data generated for isophorone by the same testing laboratory. In the case of isophorone, a 30-fold higher 

concentration (25 mg/ml) of radiolabeled test substance in IPM was applied to the skin that generated 

receptor fluid concentrations following the 24-hour exposure period that were less than 5 percent of the 

initial concentration in the donor chamber. In all experiments steady state flux was achieved and 

maintained for over 16 hours (8 to 24 hours). This data allowed the Kp to be determined from an average 

of six absorption measurements for each of the six diffusion cell experiments. 

Failing to conduct a test in accordance with the test standard adopted in the test rule is considered a 

violation of the test rule under 40 CFR 790.59(a) . To avoid possibly being found in violation of the test 

rule, you should address the concerns identified by Dr. 

Schaeffer . 

Please provide a written response to me that addresses these concerns by September 30, 2007. 

Catherine Roman 

USEPA 

1200 Pennsylvania Ave., NW, 7405M 

Washington, DC 20460 

(202) 564-8172 

(202) 564-4765 (Fax) 

(See attached file : p-Nitroaniline .pdf) (See attached file : 

nitroaniline test review.doc) (See attached file : 

VaISchaefferE-mail .pdf) 
p. 9 

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