Harcros Chemicals Inc. 

5200 Speaker Road 

Kansas City, KS 66106 

Telephone : 913/621-7747 

Fax: 913/621-7746 

E-mail: OrganicsCS@Harcros.com 

Date 03/19/13 

Miff 35%M>? 

Document Control Office (DCO) (7407M) 

Office of Pollution Control and Toxics (OPPT) 

U.S. Environmental Protection Agency (EPA) 

1200 Pennsylvania Ave. NW 

Washington, D.C. 20460 (202-564-8930) 

Attn: TSCA Section 4 

40 CFR § 799.5087 (Docket # EPA-HQ-OPPT-2007-0531) 

RE: Acute Toxicity in the Rat- Up and Down Procedure CASRN 31138-65-5 

C O 

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To Whom It May Concern: 

In accordance with EPA's final rule, Testing of Certain High Production Volume Chemicals; Second 

Group of Chemicals, published in the Federal Register on January 7,2011 at 76 fed. Reg. 1067, 

please find enclosed the final report for the Acute Toxicity in the Rat - Up and Down Procedure on Dgluco-Heptonic acid, monosodium salt, (2.xi.)- [CAS# 31138-65-5]. 

This study was conducted to fulfill the requirements of testing category D as set forth in Table 2 of 40 

CFR 799.5087G). 

If any further information is required, please do not hesitate to contact me at (913) 621-7802 or at 

asavini@harcros.com. 

Regards, 

Ann Marie Savini 

Harcros Chemicals Inc. 

Regulatory Specialist 

cc: Peter Radford 

Howard Stevenson 

James Grantham 

Randy Buckner 

Don Meuschke 

John Miles 

File 

4 0 1 3000003 0 

CONTAINS NO CEfl 
PAGE 1 OF 15 PAGES 

harlan 

Sodium Glucoheptonate: 

ACUTE ORAL TOXICITY IN THE RAT 

- UP-AND-DOWN PROCEDURE 

PROJECT NUMBER: 41104687 

AUTHOR: A Sanders 

STUDY SPONSOR: 

Harcros Chemicals Inc. 

3452 Corporate Drive 

Dalton GA 30720 

UNITED STATES OF AMERICA 

STUDY MONITOR: 

John Newlove 

JN Consultancy (International) Ltd. 

1 Potterton Lane 

Barwick in Elmet 

LS15 4DU 

TEST FACILITY: 

Harlan Laboratories Ltd 

Shardlow Business Park 

Shard low 

Derbyshire' 

DE72 2GD -

UK 

Telephone: +44 (0) 1332 792896 

Facsimile: +44 (0) 1332 799018 

41104687.docx/JO 
p. 3 

PROJECT NUMBER: 41104687 PAGE 2 

QUALITY ASSURANCE REPORT 

This study type is classed as short-term. The General Study Plan for this study type was 

reviewed for compliance once only on initial production. Inspection of the routine and 

repetitive procedures that constitute the study is carried out as a continuous process 

designed to encompass the major phases of this study type at least once every three 

months. From 16t h May 2012 the frequency of these process based inspections was 

changed from a monthly to a three monthly cycle. This deviation is considered not to 

affect the purpose or integrity of the study. 

In addition, general facilities are inspected at least once a year and the results are 

reported to management. 

This report has been audited by the Quality Assurance Unit, and is considered to be an 

accurate account of the data generated and of the procedures followed. 

In each case, the outcome of QA evaluation is reported to the Study Director and 

Management on the day of evaluation. Audits of study documentation, and process 

inspections appropriate to the type and schedule of this study were as follows: 

24 August 2010 General Study Plan Compliance Audit 

18 October 2012 Test Item Preparation 

18 October 2012 Animal Preparation 

04 October 2012 Dosing 

11 October 2012 Assessment of Response 

03 October 2012 Necropsy 

§ 28 November 2012 Draft Report Audit 

§ Date of QA Signature Final Report Audit 

§ Evaluation specific to this study 

^.r.laS&a*'^ ^ DATE: 

For the Quality Assurance Unftt 

1 5 MAR 2013 

"Authorised QA Signatures: 

Senior Audit Staff: J G Riley BSc (Hons) MRQA, J M Crowther MIScT MRQA, 

G Wren ONC MRQA, S Bevan BSc (Hons) MRQA, L Blaney MRQA 
p. 4 

PROJECT NUMBER: 41104687 PAGE 3 

GLP COMPLIANCE STATEMENT 

With the exception noted below the work described was performed in compliance with 

UK GLP standards (Schedule 1, Good Laboratory Practice Regulations 1999 

(S11999/3106 as amended by SI 2004/0994)). These Regulations are in accordance 

with GLP standards published as OECD Principles on Good Laboratory Practice (revised 

1997, ENV/MC/CHEM(98)17); and are in accordance with, and implement, the 

requirements of Directives 2004/9/EC and 2004/10/EC. 

No analysis was carried out to determine the homogeneity, concentration or stability of 

the test item formulation. The test item was formulated within two hours of being applied 

to the test system; it is assumed that the formulation was stable for this duration. This 

exception is considered not to affect the purpose or integrity of the study. 

This report fully and accurately reflects the procedures used and data generated. 

DATE: 1 1 MAR 2013 

A Sanders 

Study Director 

This report may be presented in final form as a digital (pdf) document. Such documents are prepared by scanning the paper original, 

and are considered of equivalent integrity and authenticity to versions produced by optical photocopy. However, in all cases the 

hand-signed paper original, held in secure archives, is the definitive document. 
PROJECT NUMBER: 41104687 

p. 5 

PAGE 4 

CONTENTS 

QUALITY ASSURANCE REPORT 2 

GLP COMPLIANCE STATEMENT 3 

CONTENTS 4 

SUMMARY 5 

1. INTRODUCTION 6 

2. TEST ITEM AND EXPERIMENTAL PREPARATION 6 

2.1 Description, Identification and Storage Conditions 6 

2.2 Preparation of Test Item 6 

3. METHODS 7 

3.1 Animals and Animal Husbandry 7 

3.2 Procedure 8 

3.3 Evaluation of Data 9 

4. ARCHIVES 9 

5. RESULTS 10 

5.1 Mortality Data 10 

5.2 Clinical Observations 10 

5.3 Bodyweight 10 

5.4 Necropsy 10 

6. CONCLUSION 11 

Table 1 Individual Clinical Observations and Mortality Data 12 

Table 2 Individual Bodyweights and Weekly Bodyweight Changes 13 

Table 3 Individual Necropsy Findings 14 

Appendix 1 Statement of GLP Compliance in Accordance with Directive 2004/9/EC 15 
PROJECT NUMBER: 41104687 

p. 6 

PAGE 5 

Sodium Glucoheptonate: 

ACUTE ORAL TOXICITY IN THE RAT - UP-AND-DOWN PROCEDURE 

SUMMARY 

Introduction. The study was performed to assess the acute oral toxicity of the test item 

following a single oral administration in the Wistar strain rat. The method was designed 

to be compatible with the following: 

• OECD Guidelines for the Testing of Chemicals No. 425 "Acute Oral Toxicity -

Up-and-Down-Procedure (UDP)" (adopted 03 October 2008) 

Method. A total of five female animals were dosed individually in sequence with 

sufficient time (at least 48 hours) between each animal, at dose levels of 354, 1112 or 

4040 mg/kg bodyweight (equivalent to 175, 550 and 2000 mg active ingredient/kg 

bodyweight, respectively). 

The test item was administered orally undiluted at dose levels of 1112 and 4040 mg/kg 

bodyweight (equivalent to 550 and 2000 mg active ingredient/kg bodyweight, 

respectively) and as a solution in distilled water at a dose level of 354 mg/kg bodyweight 

(equivalent to 175 mg active ingredient/kg bodyweight). Clinical signs and bodyweight 

development were monitored during the study. AH animals were subjected to gross 

necropsy. 

Mortality. There were no deaths. 

Clinical Observations. Hunched posture was noted in the animal treated at a dose 

level of 1112 mg/kg (equivalent to 550 mg active ingredient/kg bodyweight). No other 

signs of systemic toxicity were noted. 

Bodyweight. Animals showed expected gains in bodyweight over the study period, 

except for one animal treated at a dose level of 4040 mg/kg (equivalent to 2000 mg 

active ingredient/kg bodyweight) which showed expected gain in bodyweight during the 

first week but no gain in bodyweight during the second week. 

Necropsy. No abnormalities were noted at necropsy. 

Conclusion. The acute oral median lethal dose (LD5o) of the test item in the female 

Wistar strain rat was found to be greater than 4040 mg/kg bodyweight (equivalent to 

2000 mg active ingredient/kg bodyweight). 
PROJECT NUMBER: 41104687 

p. 7 

PAGE 6 

Sodium Glucoheptonate: 

ACUTE ORAL TOXICITY IN THE RAT - UP-AND-DOWN PROCEDURE 

1. INTRODUCTION 

The study was performed to assess the acute oral toxicity of the test item following a 

single oral administration in the Wistar strain rat. The method was designed to be 

compatible with the following; 

• OECD Guidelines for the Testing of Chemicals No. 425 "Acute Oral Toxicity -

Up-and-Down-Procedure (UDP)" (adopted 03 October 2008) 

The rat was selected for this study as it is a readily available rodent species, historically 

used in safety evaluation studies, and is acceptable to appropriate regulatory authorities. 

The oral route was selected as the most appropriate route of exposure and the results 

are believed to be of value in predicting the likely toxicity of the test item to man. 

The study was performed between 25 September 2012 and 06 November 2012. 

2. TEST ITEM AND EXPERIMENTAL PREPARATION 

2.1 Description, Identification and Storage Conditions 

Sponsor's identification Sodium Glucoheptonate 

Description dark brown liquid 

Lot number 921000100 

Purity : 49.5% 

Date received : 09 February 2012 

Expiry date 13 January 2013 

Storage conditions room temperature in the dark 

The integrity of supplied data relating to the identity, purity and stability of the test item is 

the responsibility of the Sponsor. 

2.2 Preparation of Test Item 

For the purpose of the 1112 and 4040 mg/kg dose levels (equivalent to 550 and 

2000 mg active ingredient/kg bodyweight, respectively), the test item was used as 

supplied. The specific gravity was determined and used to calculate the appropriate 
p. 8 

PROJECT NUMBER: 41104687 PAGE 7 

dose volume for the required dose level. For the purpose of the 354 mg/kg dose level 

(equivalent to 175 mg active ingredient/kg bodyweight), the test item was freshly 

prepared, as required, as a solution at the appropriate concentration in distilled water. 

The test item was formulated within two hours of being applied to the test system. It is 

assumed that the formulation was stable for this duration. For the purpose of the study, 

the dose levels were adjusted for the purity of the test item (49.5%). 

No analysis was conducted to determine the homogeneity, concentration or stability of 

the test item formulation. This is an exception with regard to GLP and has been 

reflected in the GLP compliance statement. 

3. METHODS 

3.1 Animals and Animal Husbandry 

Female Wistar (RccHanTM:WIST) strain rats were supplied by Harlan Laboratories UK 

Ltd., Oxon, UK. On receipt the animals were randomly allocated to cages. The animals 

were nulliparous and non-pregnant. After an acclimatisation period of at least five days 

the animals were selected at random and given a number unique within the study and 

identified by a number written on a cage card. At the start of the study the animals were 

eight to twelve weeks of age. The bodyweights fell within an interval of ±20% of the 

mean weight of any previously dosed animals. 

The animals were individually housed in suspended solid-floor polypropylene cages 

furnished with woodflakes. With the exception of an overnight fast immediately before 

dosing and for approximately three to four hours after dosing, free access to mains 

drinking water and food (2014C Teklad Global Rodent diet supplied by Harlan 

Laboratories U.K. Ltd., Oxon, UK) was allowed throughout the study. The diet, drinking 

water and bedding were routinely analysed and were considered not to contain any 

contaminants that would reasonably be expected to affect the purpose or integrity of the 

study. 

The temperature and relative humidity were set to achieve limits of 19 to 25°C and 30 to 

70% respectively. Any occasional deviations from these targets were considered not to 

have affected the purpose or integrity of the study. The rate of air exchange was at least 

fifteen changes per hour and the lighting was controlled by a time switch to give twelve 

hours continuous light (06:00 to 18:00) and twelve hours darkness. 
PROJECT NUMBER: 41104687 

p. 9 

PAGE 8 

The animals were provided with environmental enrichment items which were considered 

not to contain any contaminant of a level that might have affected the purpose or integrity 

of the study. 

3.2 Procedure 

No information was available regarding the toxicity of the test item therefore the default 

values for LD5 0 and sigma were entered into AOT425 Statistical Program. The statistical 

program gave a recommended dose progression of 2000, 550, 175, 55.0, 17.5, 5.5 and 

1.75 mg/kg. 

The first animal was dosed at 354 mg/kg (equivalent to 175 mg active ingredient/kg 

bodyweight). Further animals were then treated as follows based on the short-term 

results of the previously treated animal: 

Test Sequence 

{Animal 

number) 

Dose Level 

mg/kg 

Concentration 

(mg/ml) 

Specific 

Gravity 

Dose Volume 

(ml/kg) 

Short-Term 

Result 

1 

(1-0) 3540 35.4 - 10 0 

2 

(2-0) 1112® - 1.400 0.80 0 

3 

(3-0) 4040* - 1400 2.89 0 

4 

(4-0) 4040» - 1.400 2.89 0 

5 

(5-0) 4040. - 1.400 2.89 0 

0 = Animal survived 

The test was complete after the fifth animal had been dosed as the following stopping 

criterion was met: 

• three consecutive animals survived at the maximum dose level (4040 mg/kg, 

equivalent to 2000 mg active ingredient/kg bodyweight) 

All animals were dosed once only by gavage, using a metal cannula attached to a 

graduated syringe. The volume administered to each animal was calculated according 

to the fasted bodyweight at the time of dosing. Treatment of animals was sequential. 

© = Equivalent to 175 mg active ingredient/kg bodyweight 

® = Equivalent to 550 mg active ingredient/kg bodyweight 

• = Equivalent to 2000 mg active ingredient/kg bodyweight 
PROJECT NUMBER: 41104687 

p. 10 

PAGE 9 

Sufficient time (at least 48 hours) was allowed between each individual animal to confirm 

the outcome of the previously dosed animals. 

The animals were observed for deaths or overt signs of toxicity K, 1,2 and 4 hours after 

dosing and subsequently once daily for fourteen days. 

Individual bodyweights were recorded prior to dosing and seven and fourteen days after 

treatment. 

At the end of the observation period the animals were killed by cervical dislocation. All 

animals were subjected to gross pathological examination. This consisted of an external 

examination and opening of the abdominal and thoracic cavities for examination of major 

organs. The appearance of any macroscopic abnormalities was recorded. No tissues 

were retained. 

3.3 Evaluation of Data 

The oral LD5o will be calculated by the maximum likelihood method. Data evaluations 

also included the relationship, if any, between the exposure of the animal to the test item 

and the incidence and severity of all abnormalities including behavioural and clinical 

observations, gross lesions, bodyweight changes, mortality and any other toxicological 

effects. 

Using the mortality data obtained, an estimate of the acute oral median lethal dose 

(LD5 0 ) of the test item was calculated by the statistical program. 

4. ARCHIVES 

Unless instructed otherwise by the Sponsor, all original data and the final report will be 

retained in the Harlan Laboratories Ltd, Shardlow, UK archives for five years, after which 

instructions will be sought as to further retention or disposal. 
p. 11 

PROJECT NUMBER: 41104687 PAGE 10 

5. RESULTS 

Individual mortality data and individual clinical observations are given in Table 1. 

5.1 Mortality Data 

The mortality data are summarised as follows: 

Dose Level 

mq/kq 

Number of Animals Survived Number of Animals Died Total Number of Animals 

354© 1 0 1 

1112® 1 0 1 

4040* 3 0 3 

All doses 5 0 5 

5.2 Clinical Observations 

Hunched posture was noted in the animal treated at a dose level of 1112 mg/kg 

(equivalent to 550 mg active ingredient/kg bodyweight). No other signs of systemic 

toxicity were noted. 

5.3 Bodyweight 

Individual bodyweights and weekly bodyweight changes are given in Table 2. 

Animals showed expected gains in bodyweight over the study period, except for one 

animal treated at a dose level of 4040 mg/kg (equivalent to 2000 mg active ingredient/kg 

bodyweight) which showed expected gain in bodyweight during the first week but no gain 

in bodyweight during the second week. 

5.4 Necropsy 

Individual necropsy findings are given in Table 3. 

No abnormalities were noted at necropsy. 

© = Equivalent to 175 mg active ingredient/kg bodyweight 

® = Equivalent to 550 mg active ingredient/kg bodyweight 

• = Equivalent to 2000 mg active ingredient/kg bodyweight 
p. 12 

PROJECT NUMBER: 41104687 PAGE 11 

6. CONCLUSION 

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat 

was found to be greater than 4040 mg/kg bodyweight (equivalent to 2000 mg active 

ingredient/kg bodyweight). 
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PROJECT NUMBER: 41104687 PAGE 15 

Appendix 1 Statement of GLP Compliance in Accordance with Directive 

2004/9/EC 

THE DEPARTMENT OF HEALTH O f TH E GOVERNMENT 

OF TH E UNITED KINGDOM 

GOOD LABORATORY PRACTICE 

STATEMENT OF COMPLIANCE 

IN ACCORDANCE WITH DIRECTIVE 2004/9/EC 

TEST FACILITY TEST TYP£(S} 

Harlan Laboratories Ltd Analytical/Clinical 

DATE OF INSPECTION 

10 July 2012 

An inepection for compliance with tha Principles of Good Laboratory Practice was 

carried out at the above test facBfty as part of the UK Good Laboratory Practice 

Compliance Monitoring Programme. 

This statement confirms that> on tha date of issue, the UK Good Laboratory 

Practice Monitoring Authority were satisfied that the above test facility was 

operating in compliance with the OECD Principles of Good Laboratory Practice. 

This statement constitutes a Good Laboratory Practice instrument (as defined in the 

UK Good Laboratory Practice Regulations 1999). 

Shardlow Business Park 

London Road 

Shard low 

Darby, 0E27 2GD 

Chemistry 

Environmental Toxicity 

Environmental Fate 

Mutagenicity 

Phya/Chem. Tests 

Toxicology 

Dr. Andrew J. Gray 

Head, UK GLP Monitoring Authority 
p. 17 

ORIGIN ID: KCKA (913) 621-7733 

JAKE BOOREtl 

HARCROS CHEMICALS INC 

5200 SPEAKER RD 

KANSAS CITY. KS 66106 

UNITED STATES US 

SHIP DATE: 19HAR13 

ACTWGT: O.S LB MAN 

CAD: 0978415/CAFE2606 

BILL THIRD PARTY 

TO(DCO)(7407M)(OPPT) 

US EPA TSCA SECTION 4 

1200 PENNSYLVANIA AVE NW 

WASHINGTON DC 20460 

(202)664 - 6930 

REF: POSTAGE 

FedEx 

Express 

WED - 20 MAR 3:00P 

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10201 

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