Harcros Chemicals Inc. k/\ 0 ^^*2/ ( i L 

5200 Speaker Road

 J (V ^ V? / /fc> 

p.1 

Kansas City, KS 66106 

Telephone : 913/621-7747 

Fax: 913/621-7746 

E-maif: OrganicsCS@Harcros.com 

Date 03/07/13 

Document Control Office (DCO) (7407M) 

Office of Pollution Control and Toxics (OPPT) 

U.S. Environmental Protection Agency (EPA) 

1200 Pennsylvania Ave. NW r-» 

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

Attn: TSCA Section 4 

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

. ~o<r> i> 

RE: Micronucleous Test in the Mouse CASRN 31138-65-5 £pj ^ 

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 Micronucleous Test in the Mouse on D-gluco-Heptonic 

acid, monosodium salt, (2.xi.)- [CAS# 31138-65-5]. 

This study was conducted to fulfill the requirements of testing category E 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 300000 2 

ISO ON SNIV1NOO 
PAGE 1 OF 24 PAGES 

harlan 

Sodium Glucoheptonate: 

MICRONUCLEUS TEST IN THE MOUSE 

PROJECT NUMBER: 41104690 

AUTHOR: L Flanders 

STUDY SPONSOR: 

Harcros Chemicals Inc. 

3452 Corporate Drive 

Dalton GA 30720 

UNITED STATES OF AMERICA 

TEST FACILITY: 

Harlan Laboratories Ltd 

Shardlow Business Park 

Shard low 

Derbyshire 

DE72 2GD 

UK 

Telephone: +44 (0) 1332 792896 

41104690/LF 

Facsimile: +44 (0) 1332 799018 
PROJECT NUMBER: 41104690 PAGE 2 

QUALITY ASSURANCE REPORT 

The Genera] Study P/an for this study type was reviewed for compliance once only on 

initial production. The conduct of this study has been subject to periodic inspection by 

the Quality Assurance Unit. 

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 inspections 

appropriate to this study were as follows: 

01 July 2011 General Study Plan Compliance Audit 

§ 28 November 2012 Assessment of Response 

§ 28 November 2012 Harvest 

09 January 2013 Draft Report Audit 

§ Date of QA Signature Final Report Audit 

§ Evaluation specific to this study 

For the Quality Assurance Unit* 

DATE: 

1 8 JAN 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: 41104690 PAGE 3 

GLP COMPLIANCE STATEMENT 

With the exception noted beiow, the work described was performed in compliance with 

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

1999/3106 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 it 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. 

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, 

DATE: 

1 6 JAN 2013 

L Flanders 

Study Director 
p. 5 

PROJECT NUMBER: 41104690 PAGE 4 

CONTENTS 

QUALITY ASSURANCE REPORT 2 

GLP COMPLIANCE STATEMENT 3 

CONTENTS 4 

SUMMARY 5 

1. INTRODUCTION 7 

2. PRINCIPLES OF INVESTIGATION 7 

3. TEST AND CONTROL ITEMS AND EXPERIMENTAL PREPARATION 8 

3.1 Test Item 8 

3.2 Positive Control Item 9 

3.3 Vehicle Control Item 9 

4. METHODS 9 

4.1 Animals and Animal Husbandry 9 

4.2 Procedure 10 

5. ARCHIVES 13 

6. RESULTS 14 

6.1 Range-finding Toxicity Test 14 

6.2 Micronucleus Test 15 

7. CONCLUSION 15 

Table 1 Micronucleus Test - Summary of Group Mean Data 16 

Table 2 Micronucleus Test - Individual Data and Group Means and Standard 

Deviations: Vehicle Control (10 ml/kg) 24-Hour Sampling Time 17 

Table 3 Micronucleus Test - Individual Data and Group Means and Standard 

Deviations: Cyclophosphamide (50 mg/kg) 24-Hour Sampling Time 18 

Table 4 Micronucleus Test - Individual Data and Group Means and Standard 

Deviations: Test Item (2000 mg/kg) 48-Hour Sampling Time 19 

Table 5 Micronucleus Test - Individual Data and Group Means and Standard 

Deviations: Test Item (2000 mg/kg) 24-Hour Sampling Time 20 

Table 6 Micronucleus Test - Individual Data and Group Means and Standard 

Deviations: Test Item (1000 mg/kg) 24-Hour Sampling Time 21 

Table 7 Micronucleus Test - Individual Data and Group Means and Standard 

Deviations: Test Item (500 mg/kg) 24-Hour Sampling Time 22 

Appendix 1 Historical Control Data (2009 and 2010) 23 

Appendix 2 Statement of GLP Compliance in Accordance with Directive 2004/9/EC 24 
PROJECT NUMBER: 41104690 PAGE 5 

Sodium Glucoheptonate: 

MICRONUCLEUS TEST IN THE MOUSE 

SUMMARY 

Introduction. The study was performed to assess the potential of the test item to 

produce damage to chromosomes or aneuploidy when administered to mice. The 

method was designed to be compatible with the 1997 OECD Guidelines for Testing of 

Chemicals No.474 "Mammalian Erythrocyte Micronucleus Test", Method 812 of 

Commission Regulation (EC) No. 440/2008 of 30 May 2008, the US EPA (TSCA) 

OPPTS 870.5395, EPA 712-C-98-226, August 1998 guidelines, and be acceptable to the 

Japanese METI/MHLW/MAFF guidelines for testing of new chemical substances. 

Methods. A range-finding test was performed to find suitable dose levels of the test 

item, route of administration, and to investigate if there was a marked difference in toxic 

response between the sexes. There was no marked difference in toxicity of the test item 

between the sexes; therefore the main test was performed using only male mice. 

Following discussions with the Sponsor, the micronucleus test was conducted using the 

intraperitoneal route in groups of seven mice (males) at the maximum recommended 

dose (MRD) of 2000 mg/kg and with 1000 and 500 mg/kg as the two lower dose levels. 

Animals were killed 24 or 48 hours later, the bone marrow extracted, and smear 

preparations made and stained. Polychromatic (PCE) and normochromatic (NCE) 

erythrocytes were scored for the presence of micronuclei. 

Additional groups of mice were given a single intraperitoneal dose of phosphate buffered 

saline (PBS) (7 male mice) or dosed orally with cyclophosphamide (5 male mice), to 

serve as vehicle and positive controls respectively. Vehicle and positive control animals 

were killed after 24 hours. 

Results. There were no premature deaths seen in any of the dose groups in the main 

test. Clinical signs were observed in animals dosed with the test item at and above 

1000 mg/kg in both the 24 and 48-hour dose groups, where applicable, and included 

hunched posture and ptosis. 
PROJECT NUMBER: 41104690 PAGE 6 

Statistically significant decreases in the PCE/NCE ratio were not observed in any of the 

test item dose groups when compared to the vehicle control group. However, the 

observation of clinical signs was taken to indicate that systemic absorption had occurred 

and exposure to the target tissue had been achieved. 

There was no evidence of any statistically significant increases in the incidence of 

micronucleated polychromatic erythrocytes in animals dosed with the test item when 

compared to the vehicle control group. 

The positive control group showed a marked increase in the incidence of micronucleated 

polychromatic erythrocytes hence confirming the sensitivity of the system to the known 

mutagenic activity of cyclophosphamide under the conditions of the test. 

The test item was found not to produce a toxicologlcally significant increase in the 

frequency of micronuclei in polychromatic erythrocytes of mice under the conditions of 

the test. 

Conclusion. The test item was considered to be non-mutagenic under the conditions of 

the test. 
p. 8 

PROJECT NUMBER: 41104690 PAGE 7 

Sodium Glucoheptonate: 

MICRONUCLEUS TEST IN THE MOUSE 

1. INTRODUCTION 

The micronucleus test is a mammalian in vivo test that detects damage to the 

chromosomes induced by chemicals. In addition, numerical changes due to 

chromosome loss during cell division can be detected in this test. 

The study was performed according to a method that was designed to be compatible 

with the 1997 OECD Guidelines for Testing of Chemicals No.474 "Mammalian 

Erythrocyte Micronucleus Test", Method B12 of Commission Regulation (EC) No. 

440/2008 of 30 May 2008, the US EPA (TSCA) OPPTS 870.5395, EPA 712-C-98-226, 

August 1998 guidelines, and be acceptable to the Japanese METI/MHLW/MAFF 

guidelines for testing of new chemical substances. The results of the test are believed to 

be of value in predicting the mutagenic potential of the test item to man. The test system 

was chosen because the mouse has been shown to be a suitable model for this type of 

study and is the recommended species in the test method. 

The experimental phases of the study were performed between 28 February 2012 and 

11 December 2012. 

2. PRINCIPLES OF INVESTIGATION 

In mitotic cells in which chromosome damage has been caused by the test item or its 

metabolites, fragments (centric or acentric) or whole chromosomes tend to lag behind in 

the anaphase stage of cell division. After telophase a large proportion of the fragments 

are not included in the nuclei of the daughter cells and hence form a single or multiple 

micronuclei (Howell-Jolly bodies) in the cytoplasm of these cells. These micronuclei are 

seen in a wide variety of cell types, but erythrocytes are chosen since micronuclei are 

easily detected in these cells. 

A few hours after the last mitosis is completed, erythrocytes expel their nuclei. Immature 

erythrocytes, less than 24 hours old, stain blue with May-Grunwald/Giemsa due to the 

presence of minute fragments of nuclear material in the cytoplasm. This material is 

mainly ribonucleic acid (RNA), which gradually disappears so that more mature 
PROJECT NUMBER: 41104690 PAGE 8 

erythrocytes (normochromatic erythrocytes) stain pink with May-Grunwald/Giemsa. The 

immature blue staining cells are known as polychromatic erythrocytes and mauve 

stained micronuclei are easily detected in this cell type. If scoring is restricted to 

polychromatic erythrocytes, all the chromosomal damage detected will have been 

caused during the final cell cycle of the nucleated precursor cells. Thus by examining 

polychromatic cells at various periods after administration, the effect of the test item over 

the previous 30 hours can be monitored. 

Any toxic effects of the test item on the immature nucleated cells may lead to a reduction 

in cell division and cell death. This in turn leads to a reduction in cell volume and to 

compensate for this, peripheral blood is shunted into the bone marrow. If the ratio of 

polychromatic to normochromatic erythrocytes is scored and found to be significantly 

lower than the control value, this is taken as being indicative of cytotoxicity. 

3. TEST AND CONTROL ITEMS AND EXPERIMENTAL PREPARATION 

3.1 Test Item 

Sponsor's identification : Sodium Glucoheptonate 

Description Dark brown liquid 

Batch number 921000100 

Purity 50.5% active ingredient (accounted for in principle) 

Date received 09 February 2012 

Expiry date : 09 February 2015 

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. 

For the purpose of this study the test item was freshly prepared as required as a solution 

at the appropriate concentration in phosphate buffered saline (PBS). An adjustment was 

made for the water content of the test item (49.5%) when preparing dose concentrations. 

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 it being 

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

This is an exception with regard to GLP and has been reflected in the GLP compliance 

statement. 
p. 10 

PROJECT NUMBER: 41104690 PAGE 9 

3.2 Positive Control Item 

The positive control item was supplied by Acros Organics, as follows: 

Supplier's identification 

Supplier's lot number 

In-house serial number 

Date received 

Expiry date 

Storage conditions 

Cyclophosphamide 

A0302605 

R-5359 

27 April 2012 

27 April 2014 

Approximately 4°C in the dark 

For the purpose of this study the positive control item was freshly prepared as required 

as a solution at the appropriate concentration in distilled water (Aguettant Ltd batch 

no. 3008937 01). 

3.3 Vehicle Control Item 

The vehicle was supplied as a 10x concentrated solution by Gibco, as follows: 

Supplier's identification 

Supplier's lot number 

In-house serial number 

Date received 

Expiry date 

Storage conditions 

Dulbecco's Phosphate Buffered Saline Solution 

938281 

V-5241 

02 November 2011 

01 May 2013 

Room temperature 

For the purpose of this study the vehicle control item was freshly prepared as required as 

a solution at the appropriate concentration in distilled water (Aguettant Ltd batch 

no. 3008937 01). 

4. METHODS 

4.1 Animals and Animal Husbandry 

Sufficient albino Hsd: ICR (CD-1®) strain mice were obtained from Harlan Laboratories 

UK Ltd., Oxon, UK. At the start of the main test the mice weighed 21 to 30g and were 

approximately six to ten weeks old. After a minimum acclimatisation period of five days 
p. 11 

PROJECT NUMBER: 41104690 PAGE 10 

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

marking and a number written on a colour coded cage card. 

The animals were housed in groups of up to seven in solid-floor polypropylene cages 

with wood-flake bedding. Free access to mains drinking water and food (Harlan Teklad 

2014C Global Certified Rodent Diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) 

was allowed throughout 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 

approximately fifteen changes per hour and the lighting was controlled by a time switch 

to give twelve hours light and twelve hours darkness. 

4.2 Procedure 

4.2.1 Range-finding Toxicity Test 

A range-finding toxicity test was performed to determine a suitable dose level and route 

of administration for the micronucleus test. The dose level selected should ideally be the 

maximum tolerated dose level or that which produces some evidence of toxicity up to a 

maximum recommended dose of 2000 mg/kg. The range-finding toxicity test was also 

used to determine if the main test was to be performed using both sexes or males only. 

With no evidence of toxicity via the oral route, and following discussions with the 

Sponsor, it was considered necessary to investigate the intraperitoneal route of 

administration to maximise exposure of the test item. 
p. 12 

PROJECT NUMBER: 41104690 PAGE 11 

Groups of mice were dosed as follows: 

Dose Level Concentration Dose Volume Number of Mice 

(mg/kg) (mg/ml) (ml/kg) Male Female 

2000 (oral) 200 10 1 1 

1000 (ip) 100 10 1 1 

1500 (ip) 150 10 1 1 

2000 (ip) 200 10 1 1 

2000 (ip) 200 10 2 0 

All animals were dosed once only at the appropriate dose level by gavage using a metal 

cannula or a hypodermic needle attached to a graduated syringe. The volume 

administered to each animal was calculated according to its bodyweight at the time of 

dosing. 

Animals were observed one hour after dosing and subsequently once daily for up to two 

days. Any deaths and evidence of overt toxicity were recorded at each observation. No 

necropsies were performed. 

4.2.2 Micronucleus Test 

Groups, each of seven male mice, were dosed once only Via the intraperitoneal route 

with the test item at 2000, 1000 or 500 mg/kg. One group of mice from each dose level 

was killed by cervical dislocation 24 hours following treatment and a second group dosed 

with test item at 2000 mg/kg was killed after 48 hours. Two additional groups of male 

mice were included in the study; one group (seven mice) was dosed via the 

intraperitoneal route with the vehicle alone (PBS) and a second group (five mice) was 

dosed orally with cyclophosphamide. Cyclophosphamide is a positive control item 

known to produce micronuclei under the conditions of the test. The vehicle and positive 

control group animals were killed 24 hours following dosing. 

ip = Intraperitoneal 
p. 13 

PROJECT NUMBER: 41104690 PAGE 12 

The experimental design is summarised in the following table: 

Dose Group 

Dose Level 

(mg/kg) 

Concentration 

(mg/ml) 

Dose 

Volume 

(ml/kg) 

Kill Time 

(Hours 

Arter 

Dosing) 

Animal 

Numbers 

1. Vehicle Control (PBS) 0 0 10 24 1 -7 

2. Positive Control 

(Cyclophosphamide) 50 5 10 24 8-12 

3. Sodium Glucoheptonate 2000 200 10 48 13-19 

4. Sodium Glucoheptonate 2000 200 10 24 20-26 

5. Sodium Glucoheptonate 1000 100 10 24 27-33 

6. Sodium Glucoheptonate 500 50 10 24 34-40 

Animals were observed for signs of overt toxicity and death one hour after dosing and 

then once daily as applicable and immediately prior to termination. 

4.2.3 Slide Preparation 

Immediately following termination (i.e. 24 or 48 hours following dosing), both femurs 

were dissected from each surviving animal, aspirated with foetal bovine serum and bone 

marrow smears prepared following centrifugation and re-suspension. The smears were 

air-dried, fixed in absolute methanol, stained in May-Grunwald/Giemsa, allowed to air-dry 

and a cover slip applied using mounting medium. 

4.2.4 Slide Evaluation 

Stained bone marrow smears were coded and examined blind using light microscopy at 

x1000 magnification. Where possible, the incidence of micronucleated cells per 2000 

polychromatic erythrocytes (PCE-blue stained immature cells) per animal was scored. 

Micronuclei are normally circular in shape, although occasionally they may be oval or 

half-moon shaped, and have a sharp contour with even staining. In addition, the number 

of normochromatic erythrocytes (NCE-pink stained mature cells) associated with 1000 

erythrocytes was counted; these cells were also scored for incidence of micronuclei. 
p. 14 

PROJECT NUMBER: 41104690 PAGE 13 

The ratio of polychromatic to normochromatic erythrocytes was calculated together with 

appropriate group mean values and standard deviations. 

4.2.5 Interpretation of Results 

A comparison was made between the number of micronucleated polychromatic 

erythrocytes occurring in each of the test item groups and the number occurring in the 

vehicle control group. 

A positive mutagenic response would be demonstrated when a statistically significant, 

dose-responsive, toxicologically relevant increase in the number of micronucleated 

polychromatic erythrocytes was observed for either the 24 or 48-hour kill times when 

compared to the vehicle control group. 

If these criteria were not fulfilled, then the test item would be considered non-genotoxic 

under the conditions of the test. 

A positive response for bone marrow toxicity would be demonstrated when the dose 

group mean polychromatic to normochromatic ratio was shown to be statistically 

significantly lower than the vehicle control group. 

All data were statistically analysed using appropriate statistical methods as 

recommended by the UKEMS Sub-committee on Guidelines for Mutagenicity Testing 

Report, Part III (1989). The data was analysed following a TJ(X +1) transformation using 

Student's t-test (two tailed) and any significant results were confirmed using the one way 

analysis of variance. 

5. 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. 15 

PROJECT NUMBER: 41104690 PAGE 14 

6. RESULTS 

6.1 Range-finding Toxicity Test 

The mortality data are summarised as follows: 

Dose Level 

(mg/kg) Sex 

Number of 

Animals 

Treated 

Route 

Deaths on Day 

Total Deaths Dose Level 

(mg/kg) Sex 

Number of 

Animals 

Treated 

Route 

0 1 2 

Total Deaths 

2000 

Male 1 

oral 

0 0 0 

2000 0/2 

Female 1 

oral 

0 0 0 

0/2 

1000 

Male 1 

iP 

0 0 0 

1000 0/2 

Female 1 

iP 

0 0 0 

0/2 

1500 

Male 1 

ip 

0 0 0 

1500 0/2 

Female 1 

ip 

0 0 0 

0/2 

2000 

Male 1 

iP 

0 0 0 

2000 0/2 

Female 1 

iP 

0 0 0 

0/2 

2000 Male 2 ip 0 0 0 0/2 

In animals dosed with the test item at 2000 mg/kg via the oral route clinical signs were 

not observed in any of the animals. Therefore, with no evidence of toxicity the 

intraperitoneal route of administration was investigated to maximise exposure of the test 

item. 

Using the intraperitoneal route, the clinical signs hunched posture, ptosis, increased 

activity, and irritation and over grooming of the injection site were observed in animals 

dosed with the test item at and above 1000 mg/kg. Therefore, with evidence of modest 

toxicity, the maximum dose level in the main test was set at the maximum recommended 

dose of 2000 mg/kg with 1000 and 500 mg/kg as the two lower dose levels. The test 

item showed no marked difference in its toxicity to male or female mice; therefore the 

main test was performed using male mice only. 

ip - Intraperitoneal 
p. 16 

PROJECT NUMBER: 41104690 PAGE 15 

6.2 Micronucleus Test 

6.2.1 Mortality Data and Clinical Observations 

There were no premature deaths seen in any of the dose groups in the main test. 

Clinical signs were observed in animals dosed with the test item at and above 

1000 mg/kg in both the 24 and 48-hour dose groups, where applicable, and included 

hunched posture and ptosis. 

6.2.2 Evaluation of Bone Marrow Slides 

A summary of the results of the micronucleus test is given in Table 1. Individual and 

group mean data are presented in Tables 2 to 7. 

Statistically significant decreases in the PCE/NCE ratio were not observed in any of the 

test item dose groups when compared to the vehicle control group. However, the 

observation of clinical signs was taken to indicate that systemic absorption had occurred 

and exposure to the target tissue had been achieved. 

There was no evidence of any statistically significant increases in the incidence of 

micronucleated polychromatic erythrocytes in animals dosed with the test item when 

compared to the vehicle control group. 

The positive control group showed a marked increase in the incidence of micronucleated 

polychromatic erythrocytes hence confirming the sensitivity of the system to the known 

mutagenic activity of cyclophosphamide under the conditions of the test. 

The test item was found not to produce a toxicologically significant increase in the 

frequency of micronuclei in polychromatic erythrocytes of mice under the conditions of 

the test. . 

7. CONCLUSION 

The test item was considered to be non-mutagenic under the conditions of the test. 
p. 17 

PROJECT NUMBER: 41104690 PAGE 16 

Sodium Glucoheptonate: MICRONUCLEUS TEST IN THE MOUSE 

Table 1 Micronucleus Tes t - Summar y of Group Mean Data 

Treatment Group 

Number of PCE with M/cronuclet 

per 2000 PCE PCE/NCE Ratio 

Treatment Group 

Group Mean SD Group Mean SD 

1. Vehicle Control (PBS) 

10 ml/kg 

24-hour Sampling Time 

1.0 1.2 0.84 0.27 

2, Positive Control 

(Cyclophosphamide) 

50 mg/kg 

24-hour Sampling Time 

23.0*** 8.4 0.95 0.20 

3. Sodium Glucoheptonate 

2000 mg/kg 

48-hour Sampling Time 

2.0 1.6 1.27 0.47 

4. Sodium Glucoheptonate 

2000 mg/kg 

24-hour Sampling Time 

2.1 2.9 1.56 0.43 

5. Sodium Glucoheptonate 

1000 mg/kg 

24-hour Sampling Time 

0.9 1.6 1.91 0.94 

6. Sodium Glucoheptonate 

500 mg/kg 

24-hour Sampling Time 

0.6 0.5 1.47 0.31 

PCE = Polychromatic erythrocytes 

NCE = Normochromatic erythrocytes 

SD = Standard deviation 

*** = p < 0.001 
J*: 

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PROJECT NUMBER: 41104690 PAGE 23 

Sodium Glucoheptonate: MICRONUCLEUS TEST IN THE MOUSE 

Appendix 1 Historical Control Data (2009 and 2010) 

24-Hour Vehicle Control Group Mean Data 

PCE + MN Per 2000 PCE PCE/NCE 1000 Erythrocytes 

Mean 1.23 0.90 

SD 0.72 0.19 

Minimum 0.10 0.61 

Maximum 3.40 1.22 

24-Hour Positive Control Group Mean Data 

PCE + MN Per 2000 PCE PCE/NCE 1000 Erythrocytes 

Mean 31.59 0.92 

SD 8.81 0.24 

Minimum 18.00 0.55 

Maximum 60.40 1.53 

PCE = Polychromatic erythrocytes 

NCE = Normochromatic erythrocytes 

SD = Standard deviation 
p. 25 

PROJECT NUMBER: 41104690 PAGE 24 

Appendix 2 Statement of GLP Compliance in Accordance with Directive 

2004/9/EC 

LOH 7 Department 

' of Health 

THE DEPARTMENT OF HEALTH OF THE GOVERNMENT 

OF THE UNITED KINGDOM 

GOOD LABORATORY PRACTICE 

STATEMENT OF COMPLIANCE 

1H ACCORDANCE WITH DIRECTIVE 2Q434&JEC 

TEST FACILITY 

Harlan Laboratory* Ltd 

Sbafdlow BusirwwB Park 

London Road 

StUttdlQW 

0»rby 

wnua> 

T£STTYP£(S) 

Annlytie»l/Ctitt>eal 

Chwnittry 

£nvE«mrri«rtlal Toxicity 

Envirenowntal Fato 

Mutagenicity 

Phys/Gtwro. Tists 

ToxteoJosy 

DATE OF INSPECTION 

10 July 2012 

An inspection tor cwnpflanc« wtth the Pfinoptes of Good Laboratory PnacUcewas 

carried otwt at lha abewfl las* facwlfty as part or the UK Good laboratory Practice 

Corr^fence Monftorinfj programme 

This stabameni confirms thai, on mo #rt» of taeua, the UK Good Laboratory 

Practice Monitoring Authority wow satisfied that the above lest feei% was 

aparaling «n compliance with the OECD PrincipIsG of Good Laboratory PmMtofc 

Tftss statement constitutes a Good laboratexy PreicticB Instrument (as ifcfinod 3n the 

UK Good Laboratory Practice RcguWIom 198B). 

Or, Andrew J, Gray 

HW<t„ UK GU» Monltorino Authority 
ORIGIN ID: KCKA (913) 621-7733 

JAKE BOOREM 

HARCROS CHEMICALS INC 

5200 SPEAKER RD 

KANSAS CITY, KS 66106 

UNITED STATES US 

SHIP DATE: 0?MAR1SI 

ACTWGT: 0.6 LB MAN 

CAD: 0378415/CAFE2606 

BILL THIRD PARTY 

,o

(DCO)(7407M)(OPPT) 

US EPA TSCA SECTION 4 

1200 PENNSYLVANIA AVE NW 

WASHINGTON DC 20460 

REF: POSTAGE 

FedEx) 

Express ^ 

TRK# 

I0201I 5408 5999 7899 

XC RDVA 

FRI - 08 MAR 10:30A 

PRIORITY OVERNIGHT 

20460 

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