DR.U.NOACK-LABORATORIEN 

Report Page 4 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Personnel Involved 

Study Director: Silke Fiebig 

 (Engineer, Biotechnologist) 

Deputy: Gunda Winkelmann 

 (Engineer of Horticulture) 

Technical Staff: Karin Ruthenberg 

 Doris Streichhardt 

 Barbara Wienker 

 Henriette Winkler 

 

Quality Assurance Unit: Gudrun Möhrmann-Kalabokidis 

 (Biologist) 

Deputy: Ute Kutzner 

 

Head of Testing Facility: Dr. Udo Noack 

 (Biologist) 

 

Sponsor representative Dr. Dieter Lämmermann 
DR.U.NOACK-LABORATORIEN 

Report Page 5 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table of Content 

 Page 

 Title Page 1 

 Statement of GLP Compliance 2 

 Statement of the Quality Assurance Unit 3 

 Personnel Involved 4 

 Table of Content 5 

 List of Tables 6 

1 Summary 7

2 Characterisation Data of the Test Item 8

3 Method 9

4 Results 16

4.1 Test of the Reference Items 16

4.2 Results of the Test Item 17

4.2.1 Definitive Tests 17

5 Validity Criteria 32

6 Conclusions 32

7 Literature / References 32

8 GLP-Certificate of DR.U.NOACK-LABORATORIEN 33
DR.U.NOACK-LABORATORIEN 

Report Page 6 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

List of Tables 

 Page 

 

Table 1: Mutagenic and Cytotoxic Effects of the Test Item 7 

Table 2: Induction Rates of Reference Items (Positive Controls) 16 

Table 3: Spontaneous Revertants with DMSO (Plate Incorporation Method) 18 

Table 4: Spontaneous Revertants with Double Distilled Water (Plate Incorporation 

 Method) 19 

Table 5: Spontaneous Revertants with DMSO (Preincubation Method) 20 

Table 6: Spontaneous Revertants with Double Distilled Water (Preincubation 

 Method) 21 

Table 7: Induction Rates TA 1537 - S9 (Plate Incorporation Method) 22 

Table 8: Induction Rates TA 1537 - S9 (Preincubation Method) 22 

Table 9: Induction Rates TA 1537 + S9 (Plate Incorporation Method) 23 

Table 10: Induction Rates TA 1537 + S9 (Preincubation Method) 23 

Table 11: Induction Rates TA 98 - S9 (Plate Incorporation Method) 24 

Table 12: Induction Rates TA 98 - S9 (Preincubation Method) 24 

Table 13: Induction Rates TA 98 + S9 (Plate Incorporation Method) 25 

Table 14: Induction Rates TA 98 + S9 (Preincubation Method) 25 

Table 15: Induction Rates TA 100 - S9 (Plate Incorporation Method) 26 

Table 16: Induction Rates TA 100 - S9 (Preincubation Method) 26 

Table 17: Induction Rates TA 100 + S9 (Plate Incorporation Method) 27 

Table 18: Induction Rates TA 100 + S9 (Preincubation Method) 27 

Table 19: Induction Rates TA 1535 - S9 (Plate Incorporation Method) 28 

Table 20: Induction Rates TA 1535 - S9 (Preincubation Method) 28 

Table 21: Induction Rates TA 1535 + S9 (Plate Incorporation Method) 29 

Table 22: Induction Rates TA 1535 + S9 (Preincubation Method) 29 

Table 23: Induction Rates E. coli WP2 (uvrA) - S9 (Plate Incorporation Method) 30 

Table 24: Induction Rates E. coli WP2 (uvrA) - S9 (Preincubation Method) 30 

Table 25: Induction Rates E. coli WP2 (uvrA) + S9 (Plate Incorporation Method) 31 

Table 26: Induction Rates E. coli WP2 (uvrA) + S9 (Preincubation Method) 31 
DR.U.NOACK-LABORATORIEN 

Report Page 7 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

1 Summary 

The mutagenic effects of the test item DOBA were determined in a bacterial reverse mutation 

test according to OECD Guideline No. 471 for Testing of Chemicals (1997) / Directive 

2000/32/EC Method B. 13/14 (2000) in two independent studies (plate incorporation method 

and preincubation method) from June 21, 2006 to July 06, 2006 at DR.U.NOACK-LABORATORIEN

in 31157 Sarstedt, Germany. Test systems were the Salmonella typhimurium strains TA 1537, 

TA 98, TA 100 and TA 1535 and Escherichia coli WP2 (uvrA) with (+) and without (-) the 

metabolic activation system S9 (from male Wistar rats), respectively. Positive and negative 

controls were included in each study. The test item was dissolved in DMSO and applied once at 

test initiation with the concentration ranges as given in Table 1. Duration of each study was 

48 - 72 h. Mutagenic and cytotoxic effects are summarized in Table 1. 

Table 1: Mutagenic and Cytotoxic Effects of the Test Item

Tested Concentration Range 

 (factor √10) 

[mg/plate] 

Lowest Cytotoxic 

Concentration [mg/plate] 

Strain S9 

Plate 

Incorporation 

Method 

Preincubation 

Method 

Lowest 

Mutagenic 

Concentration 

[mg/plate] 

Plate 

Incorporation 

Method 

Preincubation 

Method 

- 1.6 0.5 

TA 1537 

+ 1.6 – 0.016 1.6 1.6 

- > 5.0 0.5 

TA 98 

+ > 5.0 > 5.0 

- 5.0 0.5 

TA 100 

+ 5.0 5.0 

- 1.6 0.5 

TA 1535 

+ > 5.0 > 5.0 

E. coli - > 5.0 > 5.0 

WP2 

(uvrA) + 

5.0 - 0.05 5.0 - 0.05 none 

> 5.0 > 5.0 

The test item is regarded to be not mutagenic under test conditions. 
DR.U.NOACK-LABORATORIEN 

Report Page 8 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

2 Characterisation Data of the Test Item 

TEST ITEM DOBA

Batch Number BES06024 

Purity 98.1 % 

Appearance Colourless solid 

Water solubility < 1 g/L 

Stability Not specified 

Density 0.55 g/L (20 °C) 

Melting point 143 – 145 °C 

 

Expiry date 2007-03-06 

Recommended storage Room temperature (20 °C) 

Storage at testing facility Room temperature, protected from moisture and light 

Retention At least 1 g has been retained. 

Identification parameter Name, batch number, state, consistency, colour and melting point 

at testing facility 

The test item and the information concerning the test item were provided by the sponsor. 
DR.U.NOACK-LABORATORIEN 

Report Page 9 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

3 Method 

TEST GUIDELINES OECD Guideline No. 471 for Testing of Chemicals (1997) / 

 Directive 2000/32/EC Method B. 13/14 (2000) 

 

TYPE AND PURPOSE Determination of the mutagenic effects of the test item to four 

OF THE STUDY different Salmonella typhimurium strains and one Escherichia coli

strain over 48 / 72 h. 

TEST SYSTEM Salmonella typhimurium strains TA 1537, TA 98, TA 100 and TA 

1535 and Escherichia coli WP2 (uvrA) with (+) and without (-) 

metabolic activation system (S9). 

Reason for the selection Salmonella typhimurium strains and Escherichia coli WP2 

of the test system were selected according to the guidelines. 

 

Origin University of California, Berkeley Divison of Biochemistry & 

Molecular Biology, CA 94720 USA (TA 98, TA 100, TA 1535) 

 TRINOVA BIOCHEM GMBH, Kerkrader Straße 10, 35394 Gießen 

(E. coli WP2 (uvrA), TA 1537 and TA 100 derived from master 

culture obtained directly from University of California) 

Storage at test facility Under liquid nitrogen 

Inoculum An overnight culture was prepared. Incubation was done for 18 h 

at 37 °C. Titer of the overnight culture was ≥ 1 × 108

 cells/mL. 

Metabolic activation As metabolic activation system a certified post-mitochondrial (S9) 

system (S9) fraction from livers of male Wistar rats, which were induced with 

Phenobarbital intraperitoneally and β-Naphtoflavone orally was 

used. The S9 fraction was received from CCR (In den 

Leppsteinwiesen 19, D-64380 Roßdorf). At test start an aliquot of 

the S9-fraction was thawed and enriched with cofactors according 

to DIN Guideline 38415 part 4 (S9-Mix). Batch number of S9-

fraction was 230905 and 020606 (31.2 mg protein/mL S9), 

respectively. 
DR.U.NOACK-LABORATORIEN 

Report Page 10 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

TEST ITEM DOBA

Stock solution 50 g/L, prepared with DMSO 

Dispersion treatment Agitation 

Test concentrations 

Test strain [mg/plate] 

TA 98, TA 100, TA 1535, 

E. coli WP2 (uvrA) 

5.0 – 1.6 – 0.5 – 0.16 – 0.05 

TA 1537 

Plate Incorporation Method: 

5.0 – 1.6 – 0.5 – 0.16 – 0.05 

Preincubation Method: 

1.6 – 0.5 – 0.16 – 0.05 – 0.016 

CONTROLS Untreated controls with double distilled water were tested. 

SOLVENT CONTROLS Controls with DMSO were tested. 

REFERENCE ITEMS 

Reference item Batch No. Purity 

[%] 

Salmonella / 

E. coli strain 

Test conc. 

[μg/plate] 

ICR 191 026K1228 99.6 TA 1537 - S9 2.0 

4-Nitro-ophenylendiamine 

10625LB355 98 TA 98 - S9 0.5 

Benzo(a)pyrene 1224946 97.2 (rel.) TA 98 + S9 10 

Nitrofurantoine 014K1284 100 TA 100 - S9 0.2 

Sodium azide 448503/1 

10405368 99.9 TA 1535 - S9 0.25 

4-Nitroquinolin 1-

oxide 084K0988 100 E. coli WP2 (uvrA) -S9 1 

TA 1537, 

TA 100, 

TA 1535 + S9 

2 

2-Aminoanthracen S29093-

215 99.8 

E. coli WP2 

(uvrA) + S9 25 
DR.U.NOACK-LABORATORIEN 

Report Page 11 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

TEST METHOD 

Media According to DIN Guideline 38415 Part 4 (1999) 

Test container Petri dishes with cams (ø 94 mm, 16 mm high) with about 25 mL 

VOGEL-BONNER minimal medium 

Application Application was carried out once at test start. 

Composition of the test Plate Incorporation Method

media 2.0 mL top agar with 0.5 mM Histidin-/Biotin solution and 

L-Tryptophan solution (for E. coli WP (uvrA)), respectively, 

for plates without metabolic activation system 

 1.5 mL top agar with 0.5 mM Histidin-/Biotin solution and 

L-Tryptophan solution (for E. coli WP (uvrA)), respectively, 

for plates with metabolic activation system 

 0.5 mL S9-Mix for plates with metabolic activation system 

0.1 mL test / reference item solution, double distilled water 

and DMSO, respectively 

0.1 mL bacteria (overnight culture) 

were mixed and poured over the surface of a Vogel-Bonner 

minimal agar plate 

Preincubation Method

 0.35 mL test / reference item solution, double distilled water and 

DMSO, respectively 

1.75 mL buffer solution or S9-Mix 

0.35 mL bacteria (overnight culture) 

were preincubated in a microtiter plate for at least 20 min at 35 

± 2 °C on a shaker. 

0.7 mL of the preincubated solution 

 1.5 mL top agar with 0.5 mM Histidin-/Biotin- and L-Tryptophansolution (for E. coli WP (uvrA)), respectively, 

were mixed and poured over the surface of a Vogel-Bonner 

minimal agar plate 

Titer control Plate Incorporation Method

 2.0 mL top agar with 5 mM Histidin- / 0.5 mM Biotin-solution 

and 5 mM L-Tryptophan-solution (for E. coli WP (uvrA)), 

respectively 

 0.5 mL S9-Mix 

0.1 mL bacteria (10-5 dilution from overnight culture) 
DR.U.NOACK-LABORATORIEN 

Report Page 12 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

were mixed and poured over the surface of a Vogel-Bonner 

minimal agar plate 

Preincubation Method

 0.35 mL double distilled water 

 1.75 mL S9-Mix 

 0.35 mL bacteria (overnight culture) 

 were preincubated in a microtiter plate for at least 20 min at 35 

± 2 °C on a shaker. 

 0.7 mL of the preincubated solution was mixed with 

1.5 mL top agar with 0.5 mM Histidin-/Biotin- and L-Tryptophansolution (for E. coli WP (uvrA)), respectively, 

were mixed and poured over the surface of a Vogel-Bonner 

minimal agar plate 

Replicates Three replicates per concentration level and control. 

 Titer control replicates were prepared 2-fold. 

Confirmation of genotypes The genotypes of the tested strains were checked within each 

study: histidine and tryptophan auxotrophy, respectively, 

ampicillin resistance, UV-sensitivity and growth inhibition with 

crystal violet. 

Temperature (Min-Max) 36.0 - 36.9 °C 

 Preincubation temperature (Min-Max): 34.0 – 36.9 °C 

Test duration 48 - 72 h 

KIND AND FREQUENCY Genotypes were evaluated for each study. Plates of the 

OF MEASUREMENT AND mutagenicity test were inspected for present and reduced 

OBSERVATION background lawn after an incubation time of 48 h. 

Plates with colonies which correspond not with the typical shape 

and colour of Salmonella typhimurium and Escherichia coli, 

respectively, were regarded as contaminated and were not 

included in calculations. 

Colonies per plate (revertants) were counted. Plates with reduced 

background were considered carefully for evaluation as 

appropriate. 

Equipment Microflow biological safety cabinet (MDH) 

 Colony counter (manual counting), BZG 30 (WTW) 

 Thermometer, Oregon Scientific 

 Analysis balance, LA230-S, SARTORIUS

 
DR.U.NOACK-LABORATORIEN 

Report Page 13 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Balance, PJ Precisa junior 2000 C, DIGITANA

 Laboratory incubators, B 5042 E and B 6060 (HERAEUS) 

 Water bath, WB 22 (MEMMERT) 

COURSE OF THE STUDY Each study was conducted with 5 different concentration levels of 

the test item in a geometrical series of factor √10. Application was 

carried out as specified above. Plates for confirmation of the 

genotypes were incubated for 24 h and plates of the mutagenicity 

test for 48 – 72 h, respectively, at 37 ± 1 °C. The results were 

confirmed in a second independent study (preincubation method). 

Evaluations were performed as described below. 

EVALUATIONS AND Since a reduced background lawn is regarded to be a cytotoxic 

STATISTICS effect, plates with reduced background lawn normally were not 

included into evaluation procedures.

 Arithmetic mean values and standard deviations were calculated 

out of colonies per plate of three replicates. 

 For evaluation of the results the induction rate of the mean values 

was calculated (1): 

 revertant colonies of test item 

 Induction rate =  (1) 

 revertant colonies of the 

 corresponding control 

 Corresponding control for the reference item was double distilled 

water and DMSO for the test item. 

The test item is to be classified mutagenic if there is a 

concentration effect relationship over at least three 

concentrations and / or the induction rate is ≥ 2. 

Software The data presented in the tables are computer generated and 

rounded for presentation. Thus manual calculation of results 

based on the data in this report may yield in minor deviations 

from these figures.

 

 Calculations were carried out using software 

- Excel 2000 (2000), MICROSOFT CORPORATION
DR.U.NOACK-LABORATORIEN 

Report Page 14 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

VALIDITY CRITERIA Following genotypes of the tested strains have to be confirmed: 

 - Histidine- and tryptophan auxotrophy, respectively 

 - Ampicillin resistance 

 - UV-sensitivity 

 - growth inhibition with crystal violet (rfa-mutation) 

 Titer of the overnight culture has to be ≥ 1 ⋅ 108

 cells/mL 

Spontaneous revertants (negative controls) should be within the 

following ranges: 

Test strain Metabolic 

system 

MV ± SD Valid range 

(± S9) 

- S9 8 ± 4 

TA 1537 

+ S9 10 ± 7 

2 - 40 

- S9 27 ± 8 

TA 98 

+ S9 32 ± 14 

15 - 50 

- S9 118 ± 34 

TA 100 

+ S9 101 ± 33 

60 - 200 

- S9 25 ± 15 

TA 1535 

+ S9 16 ± 14 

5 - 40 

- S9 45 ± 12 

E. coli WP2 (uvrA) + S9 57 ± 17 30 - 100 

MV = Mean of all reference tests until January 2006 SD = standard deviation 

 The induction rates of the positve controls had to be ≥ 2.

DATES 

Study initiation 2006-06-20 

Experimental starting 2006-06-21 

Experimental completion 2006-07-06 

Study completion Please refer to page 1 

DEVIATIONS FROM Validity criteria were taken from the literature and DIN guideline, 

THE GUIDELINE since in OECD and EC guideline no validity criteria are described. 
DR.U.NOACK-LABORATORIEN 

Report Page 15 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

DEVIATIONS FROM None 

THE STUDY PLAN 

ARCHIVING The following will be retained in the archive of the testing facility 

for the period as specified in the operative national GLP 

regulations: 

• all raw data 

• study plan 

• final report 

• all records performed by the quality assurance programme 

 including master schedules 

• sample of test and reference items 

 Additionally microfilms will be retained in a safe-deposit by 

Volksbank Sarstedt, D-31157 Sarstedt. 
DR.U.NOACK-LABORATORIEN 

Report Page 16 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

4 Results 

4.1 Test of the Reference Items 

Positive controls were tested parallel to the test item. 

Table 2: Induction Rates of Reference Items (Positive Controls) 

Strain Reference item [μg/plate] S9 Induction Rate* 

 Plate 

Incorporation 

Method 

Preincubation 

Method 

ICR 191 2.0 - 112.1 > 240.0 

 TA 1537 

2-Aminoanthracene 2.0 + 55.4 26.9 

4-Nitro-o-phenylenediamine 0.5 - 3.9 4.0 

 TA 98 

Benzo(a)pyrene 10 + 22.4 10.9 

Nitrofurantoine 0.2 - 2.7 4.6 

 TA 100 

2-Aminoanthracene 2.0 + > 6.9 > 6.5 

Sodium azide 0.25 - 5.7 6.8 

 TA 1535 

2-Aminoanthracene 2.0 + 10.6 13.0 

4-Nitroquinolin-1-oxid 1.0 - 8.3 > 22.2 

 

E. coli

WP2 

(uvrA) 2-Aminoanthracene 25.0 + 9.3 13.3 

Plates with number of colonies higher than 1000/ 1200 colonies were not counted, 

induction rates were calculated with this number and given as > values. 

*) Induction Rates were calculated with the double distilled water control 
DR.U.NOACK-LABORATORIEN 

Report Page 17 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

4.2 Results of the Test Item 

4.2.1 Definitive Tests 

The test item was clearly dissolved in DMSO. During the application the test item precipitated, 

fine white particles occured in the petridish. To distinguish the particles from revertant colonies, 

the incubation was extended to 72 h. Nevertheless cytotoxic effects (reduced background lawn) 

were not unequivocal determinable for all strains. 

Revertant colonies of the test item plates are given in Table 7 to Table 26. Spontaneous 

revertants are listed in Table 3 to Table 6. 

The arithmetic mean value and the standard deviation were calculated out of the three 

replicates of the revertant colony plates. 

For evaluation of the results the induction rates of the mean values were calculated with the 

DMSO control (Table 7 to Table 26). Mutagenic and cytotoxic effects are summarized in 

Table 1. 
DR.U.NOACK-LABORATORIEN 

Report Page 18 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 3: Spontaneous Revertants with DMSO (Plate Incorporation Method)

 colonies / plate standard 

strain S9 valid range counted mean value deviation 

 7 

 - 4 7 2.5 

 9 

TA 1537 2 - 40 

 4 

 + 6 7 3.1 

 10 

 17 

 - 15 17 2.5 

 20 

TA 98 15 - 50 

 24 

 + 18 25 7.0 

 32 

 103 

 - 121 103 18.0 

 85 

TA 100 60 - 200 

 157 

 + 172 165 10.6 

 contaminated 

 31 

 - 39 32 6.1 

 27 

TA 1535 5 - 40 

 14 

 + 10 14 3.5 

 17 

 49 

 - 52 45 10.2 

 33 

EC WP2 30 - 100 

(uvrA) 36 

 + 43 46 12.3 

 60 
DR.U.NOACK-LABORATORIEN 

Report Page 19 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 4: Spontaneous Revertants with Double Distilled Water (Plate Incorporation Method)

 colonies / plate standard 

strain S9 valid range counted mean value deviation 

 3 

 - 4 5 2.6 

 8 

TA 1537 2 - 40 

 8 

 + 7 8 0.6 

 8 

 23 

 - 20 22 1.5 

 22 

TA 98 15 - 50 

 23 

 + 17 18 4.2 

 15 

 91 

 - 89 99 15.6 

 117 

TA 100 60 - 200 

 156 

 + 139 145 9.3 

 141 

 25 

 - 36 32 5.9 

 34 

TA 1535 5 - 40 

 18 

 + 10 17 7.0 

 24 

 47 

 - 52 46 6.6 

 39 

EC WP2 30 - 100 

(uvrA) 48 

 + 61 53 7.2 

 49 
DR.U.NOACK-LABORATORIEN 

Report Page 20 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 5: Spontaneous Revertants with DMSO (Preincubation Method)

 colonies / plate standard 

strain S9 valid range counted mean value deviation 

 8 

 - 3 6 2.6 

 7 

TA 1537 2 - 40 

 2 

 + 5 5 3.0 

 8 

 31 

 - 31 30 2.3 

 27 

TA 98 15 - 50 

 34 

 + 21 25 7.5 

 21 

 122 

 - 128 124 3.8 

 121 

TA 100 60 - 200 

 188 

 + 118 142 39.6 

 121 

 31 

 - 33 35 5.9 

 42 

TA 1535 5 - 40 

 14 

 + 21 16 4.4 

 13 

 37 

 - 31 34 3.1 

 33 

EC WP2 30 - 100 

(uvrA) 41 

 + 58 51 9.1 

 55 

 
DR.U.NOACK-LABORATORIEN 

Report Page 21 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 6: Spontaneous Revertants with Double Distilled Water (Preincubation Method)

 colonies / plate standard 

strain S9 valid range counted mean value deviation 

 7 

 - 4 5 1.7 

 4 

TA 1537 2 - 40 

 11 

 + 11 10 1.7 

 8 

 38 

 - 43 41 2.9 

 43 

TA 98 15 - 50 

 42 

 + 42 42 0.6 

 41 

 151 

 - 161 191 61.4 

 262 

TA 100 60 - 200 

 130 

 + 145 184 81.5 

 278 

 22 

 - 42 33 10.1 

 34 

TA 1535 5 - 40 

 19 

 + 17 15 5.3 

 9 

 52 

 - 35 45 8.9 

 48 

EC WP2 30 - 100 

(uvrA) 51 

 + 52 52 1.5 

 54 
DR.U.NOACK-LABORATORIEN 

Report Page 22 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 7: Induction Rates TA 1537 - S9 (Plate Incorporation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 7 

control + 4 7 2.5 - 

 + 9 

 - 0 

5 - 0 1 1.2 0.1 

 - 2 

 - 2 

1.6 - 0 1 1.2 0.1 

 - 0 

 + 4 

0.5 + 8 8 3.5 1.2 

 + 11 

 + 5 

0.16 + 6 6 0.6 0.9 

 + 6 

 + 3 

0.05 + 4 4 1.5 0.7 

 + 6 

 

Table 8: Induction Rates TA 1537 - S9 (Preincubation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 8 

control + 3 6 2.6 - 

 + 7 

 - 0 

1.6 - 0 0 0.0 0.0 

 - 0 

 - 0 

0.5 - 4 2 2.0 0.3 

 - 2 

 + 3 

0.16 + 4 3 1.0 0.5 

 + 2 

 + 6 

0.05 + 3 4 1.7 0.7 

 + 3 

 + 9 

0.016 + 6 8 1.5 1.3 

 + 8 

+ = present - = reduced 
DR.U.NOACK-LABORATORIEN 

Report Page 23 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

 Table 9: Induction Rates TA 1537 + S9 (Plate Incorporation Method) 

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 4 

control + 6 7 3.1 - 

 + 10 

 - 6 

5 - 4 6 2.0 0.9 

 - 8 

 - 1 

1.6 - n.d. 1 n.d. 0.2 

 - n.d. 

 + 5 

0.5 + 5 5 0.6 0.7 

 + 4 

 + 1 

0.16 + 2 3 2.6 0.5 

 + 6 

 + 5 

0.05 + 5 5 0.0 0.8 

 + 5 

Table 10: Induction Rates TA 1537 + S9 (Preincubation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 2 

control + 5 5 3.0 - 

 + 8 

 - n.d. 

1.6 - n.d. n.d. n.d. n.d. 

 - n.d. 

 + 9 

0.5 + 4 8 3.6 1.6 

 + 11 

 + 8 

0.16 + 8 7 2.3 1.3 

 + 4 

 + 8 

0.05 + 7 8 0.6 1.5 

 + 8 

 + 10 

0.016 + 7 8 1.7 1.6 

 + 7 

+ = present - = reduced n.d. = not determinable 
DR.U.NOACK-LABORATORIEN 

Report Page 24 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 11: Induction Rates TA 98 - S9 (Plate Incorporation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 17 

control + 15 17 2.5 - 

 + 20 

 + 3 

5 + 6 7 4.0 0.4 

 + 11 

 + 3 

1.6 + 6 3 2.5 0.2 

 + 1 

 + 11 

0.5 + 19 18 7.0 1.1 

 + 25 

 + 10 

0.16 + 12 13 3.6 0.8 

 + 17 

 + 23 

0.05 + 26 23 3.0 1.3 

 + 20 

Table 12: Induction Rates TA 98 - S9 (Preincubation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 31 

control + 31 30 2.3 - 

 + 27 

 - n.d. 

5 - n.d. n.d. n.d. n.d. 

 - n.d. 

 - 3 

1.6 - 4 4 1.5 0.1 

 - 6 

 - 11 

0.5 - 6 7 3.2 0.2 

 - 5 

 + 24 

0.16 + 19 22 2.6 0.7 

 + 23 

 + 23 

0.05 + 24 21 3.8 0.7 

 + 17 

+ = present - = reduced n.d. = not determinable
DR.U.NOACK-LABORATORIEN 

Report Page 25 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 13: Induction Rates TA 98 + S9 (Plate Incorporation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 24 

control + 18 25 7.0 - 

 + 32 

 + 7 

5 + 8 6 2.6 0.2 

 + 3 

 + 18 

1.6 + 15 17 1.7 0.7 

 + 18 

 + 14 

0.5 + 20 18 3.2 0.7 

 + 19 

 + 17 

0.16 + 19 20 3.6 0.8 

 + 24 

 + 19 

0.05 + 18 20 2.6 0.8 

 + 23 

Table 14: Induction Rates TA 98 + S9 (Preincubation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 34 

control + 21 25 7.5 - 

 + 21 

 + 3 

5 + 6 6 3.0 0.2 

 + 9 

 + 27 

1.6 + 17 21 5.1 0.8 

 + 20 

 + 36 

0.5 + 25 28 7.4 1.1 

 + 22 

 + 33 

0.16 + 33 31 3.5 1.2 

 + 27 

 + 20 

0.05 + 34 27 7.0 1.1 

 + 27 

+ = present 
DR.U.NOACK-LABORATORIEN 

Report Page 26 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 15: Induction Rates TA 100 - S9 (Plate Incorporation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 103 

control + 121 103 18.0 - 

 + 85 

 - n.d. 

5 - n.d. n.d. n.d. n.d. 

 - n.d. 

 + 23 

1.6 + 28 25 2.9 0.2 

 + 23 

 + 80 

0.5 + 84 85 5.6 0.8 

 + 91 

 + 49 

0.16 + 31 38 9.9 0.4 

 + 33 

 + 132 

0.05 + 91 101 27.4 1.0 

 + 80 

Table 16: Induction Rates TA 100 - S9 (Preincubation Method) 

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 122 

control + 128 124 3.8 - 

 + 121 

 - 54 

5 - 44 52 6.8 0.4 

 - 57 

 -/+ 72 

1.6 -/+ 61 69 7.0 0.6 

 -/+ 74 

 - n.d. 

0.5 - n.d. n.d. n.d. n.d. 

 - n.d. 

 + 99 

0.16 + 108 95 14.8 0.8 

 + 79 

 + 136 

0.05 + 111 118 15.4 1.0 

 + 108 

+ = present - = reduced n.d. = not determinable + / - = slight cytotoxicity 
DR.U.NOACK-LABORATORIEN 

Report Page 27 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 17: Induction Rates TA 100 + S9 (Plate Incorporation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 157 

control + 172 165 10.6 - 

 + contaminated 

 - 18 

5 - n.d. n.d. n.d. n.d. 

 - n.d. 

 + 81 

1.6 + 76 77 4.0 0.5 

 + 73 

 + 106 

0.5 + 86 97 10.1 0.6 

 + 98 

 + 107 

0.16 + 115 109 5.3 0.7 

 + 105 

 + 166 

0.05 + 137 138 27.0 0.8 

 + 112 

 

Table 18: Induction Rates TA 100 + S9 (Preincubation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 188 

control + 118 142 39.6 - 

 + 121 

 - n.d. 

5 - n.d. n.d. n.d. n.d. 

 - n.d. 

 + 36 

1.6 + 42 43 8.1 0.3 

 + 52 

 + 97 

0.5 + 167 121 39.6 0.9 

 + 100 

 + 121 

0.16 + 100 109 10.8 0.8 

 + 106 

 + 116 

0.05 + 110 110 5.5 0.8 

 + 105 

+ = present - = reduced n.d. = not determinable 
DR.U.NOACK-LABORATORIEN 

Report Page 28 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 19: Induction Rates TA 1535 - S9 (Plate Incorporation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 31 

control + 39 32 6.1 - 

 + 27 

 - 3 

5 - 6 4 1.5 0.1 

 - 4 

 +/- 13 

1.6 +/- 13 12 2.3 0.4 

 +/- 9 

 + 12 

0.5 + 21 18 5.2 0.6 

 + 21 

 + 30 

0.16 + 59 40 16.7 1.2 

 + 30 

 + 26 

0.05 + 30 29 3.1 0.9 

 + 32 

Table 20: Induction Rates TA 1535 - S9 (Preincubation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 31 

control + 33 35 5.9 - 

 + 42 

 - n.d. 

5 - n.d. n.d. n.d. n.d. 

 - n.d. 

 - n.d. 

1.6 - n.d. n.d. n.d. n.d. 

 - n.d. 

 - 1 

0.5 - 2 2 0.7 0.0 

 - n.d. 

 + 19 

0.16 + 21 19 2.5 0.5 

 + 16 

 + 30 

0.05 + 42 37 6.4 1.1 

 + 40 

+ = present - = reduced n.d. = not determinable + / - = slight cytotoxicity 
DR.U.NOACK-LABORATORIEN 

Report Page 29 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 21: Induction Rates TA 1535 + S9 (Plate Incorporation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 14 

control + 10 14 3.5 - 

 + 17 

 + 2 

5 + 4 5 3.1 0.3 

 + 8 

 + 11 

1.6 + 3 7 4.0 0.5 

 + 6 

 + 16 

0.5 + 15 16 1.5 1.2 

 + 18 

 + 10 

0.16 + 8 10 1.5 0.7 

 + 11 

 + 13 

0.05 + 20 13 6.5 1.0 

 + 7 

Table 22: Induction Rates TA 1535 + S9 (Preincubation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 14 

control + 21 16 4.4 - 

 + 13 

 + 1 

5 + 2 1 0.6 0.1 

 + 1 

 + 7 

1.6 + 9 8 1.2 0.5 

 + 9 

 + 9 

0.5 + 8 10 2.1 0.6 

 + 12 

 + 7 

0.16 + 17 14 5.8 0.9 

 + 17 

 + 20 

0.05 + 12 15 4.4 0.9 

 + 13 

+ = present 
DR.U.NOACK-LABORATORIEN 

Report Page 30 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 23: Induction Rates E. coli WP2 (uvrA) - S9 (Plate Incorporation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 49 

control + 52 45 10.2 - 

 + 33 

 + 51 

5 + 42 45 4.9 1.0 

 + 43 

 + 44 

1.6 + 48 44 4.0 1.0 

 + 40 

 + 44 

0.5 + 42 45 3.6 1.0 

 + 49 

 + 44 

0.16 + 54 48 5.1 1.1 

 + 47 

 + 38 

0.05 + 51 41 8.5 0.9 

 + 35 

Table 24: Induction Rates E. coli WP2 (uvrA) - S9 (Preincubation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 37 

control + 31 34 3.1 - 

 + 33 

 + 33 

5 + 20 30 9.3 0.9 

 + 38 

 + 38 

1.6 + 37 36 3.2 1.1 

 + 32 

 + 43 

0.5 + 49 43 6.5 1.3 

 + 36 

 + 36 

0.16 + 24 29 6.1 0.9 

 + 28 

 + 41 

0.05 + 44 41 2.5 1.2 

 + 39 

+ = present 
DR.U.NOACK-LABORATORIEN 

Report Page 31 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

Table 25: Induction Rates E. coli WP2 (uvrA) + S9 (Plate Incorporation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 36 

control + 43 46 12.3 - 

 + 60 

 + 49 

5 + 50 52 3.8 1.1 

 + 56 

 + 58 

1.6 + 63 53 12.7 1.2 

 + 39 

 + 46 

0.5 + 46 50 7.5 1.1 

 + 59 

 + 63 

0.16 + 55 59 4.0 1.3 

 + 58 

 + 62 

0.05 + 61 61 1.5 1.3 

 + 59 

Table 26: Induction Rates E. coli WP2 (uvrA) + S9 (Preincubation Method)

test item background colonies / mean value standard induction rate 

concentration lawn plate deviation 

[mg/plate] 

 + 41 

control + 58 51 9.1 - 

 + 55 

 + 43 

5 + 33 39 5.5 0.8 

 + 42 

 + 47 

1.6 + 51 47 3.5 0.9 

 + 44 

 + 42 

0.5 + 67 50 15.0 1.0 

 + 40 

 + 48 

0.16 + 42 41 8.1 0.8 

 + 32 

 + 44 

0.05 + 35 39 4.7 0.8 

 + 37 

+ = present 
DR.U.NOACK-LABORATORIEN 

Report Page 32 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

5 Validity Criteria 

• The genotypes of the tested strains have been confirmed: 

 - Histidine- and tryptophan auxotrophy, respectively 

 - Ampicillin resistance 

 - UV-sensitivity 

 - growth inhibition with crystal violet (rfa-mutation) 

• Titer of the overnight cultures was > 1⋅108

 cells/mL. 

• Spontaneous mutation rates (negative controls) met the requirements. 

• The induction rates of the positive controls was ≥ 2. 

6 Conclusions 

In this study DOBA was found to have no mutagenic effects on Salmonella typhimurium strains 

TA 1537, TA 98, TA 100, TA 1535 and Escherichia coli WP2 (uvrA) with (+) and without (-) the 

metabolic activation system S9 from male Wistar rats. 

7 Literature / References 

OECD Guideline 471 for testing of chemicals: Salmonella typhimurium. Reverse Mutation Assay 

(adopted July 21. 1997) 

Richtlinie 2000/32/EC Methode B. 13/14.: Rückmutationsversuch unter Verwendung von 

Bakterien (Juni 2000) 

DIN-Richtlinie 38415 Teil 4 (Dezember 1999)

LEVIN et al. (1982): A new Salmonella tester strain (TA 102) with A⋅T base pairs at the site of 

mutation detects oxidative mutagens. Proc. Natl. Acad. Sci.. USA. Vol. 79. pp 7445-7449 

MARON. D.M. and AMES. B.N. (1983): Revised methods for the Salmonella mutagenicity test. 

Mutation Research 113. 173-215 

 
DR.U.NOACK-LABORATORIEN 

Report Page 33 of 33 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / Directive 2000/32/EC Method B. 13/14 Study-No. USO108573 

8 GLP-Certificate of DR.U.NOACK-LABORATORIEN 
DR.U.NOACK-LABORATORIEN 

Robust Summary Page 1 of 3 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / EC Directive 2000/32/EC Method B. 13/14 Study-No. USO108573

Robust Summary 

Test Item DOBA 

 (batch number: BES06024) 

Remarks none 

Testing Facility DR.U.NOACK-LABORATORIEN

 Käthe-Paulus-Str. 1, D-31157 Sarstedt 

 Tel. +49(0)5066 70670, email: info@noack-lab.de 

Author Silke Fiebig 

Report issued 2006 - 08 - 11 

Method/Guideline followed OECD Guideline No. 471 for Testing of Chemicals (1997) / 

 EC Directive 2000/32/EC Method B. 13/14 (2000) 

Type Bacterial reverse mutation test 

GLP Yes [X] No [ ] 

Year 2006 

Test system Salmonella typhimurium strains TA 1537, TA 98, TA 100, TA 

1535, and Escherichia coli WP2 (uvrA) with (+) and without (-) 

metabolic activation system (S9) 

Exposure period 48 - 72 h 

Metabolic activation As metabolic activation system a post-mitochondrial (S9) 

system (S9) fraction from livers of male Wistar rats, which were induced with 

Phenobarbital intraperitoneally and β-Naphtoflavone orally was 

used. The S9 fraction was received from CCR (In den 

Leppsteinwiesen 19, D-64380 Roßdorf). At test start an aliquot of 

the S9-fraction was thawed and enriched with cofactors according 

to DIN Guideline 38415 part 4 (S9-Mix). Batch number of S9-

fraction was 230905 and 020606 (31.2 mg protein/mL S9), 

respectively. 

Concentrations tested 

Test strain [mg/plate] 

TA 98, TA 100, TA 1535, 

E. coli WP2 (uvrA) 

5.0 – 1.6 – 0.5 – 0.16 – 0.05 

TA 1537 

Plate Incorporation Method: 

5.0 – 1.6 – 0.5 – 0.16 – 0.05 

Preincubation Method: 

1.6 – 0.5 – 0.16 – 0.05 – 0.016 

Statistical Methods Arithmetic mean values and standard deviations were calculated 

out of colonies per plate of three replicates. Induction rates of the 

mean values were calculated out of the relation of revertant 

colonies of control and test item plates. 
DR.U.NOACK-LABORATORIEN 

Robust Summary Page 2 of 3 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / EC Directive 2000/32/EC Method B. 13/14 Study-No. USO108573

Test Conditions / 

Test Design Three replicates per concentration level and control. Titer control 

replicates were prepared 2-fold. 

 Frequency of dosing: Once at test start. 

 Composition of test media: 

 Plate Incorporation Method

 2.0 mL top agar with 0.5 mM Histidin-/Biotin solution and 

L-Tryptophan solution (for E. coli WP (uvrA)), respectively, 

for plates without metabolic activation system 

 1.5 mL top agar with 0.5 mM Histidin-/Biotin solution and 

L-Tryptophan solution (for E. coli WP (uvrA)), respectively, 

for plates with metabolic activation system 

 0.5 mL S9-Mix for plates with metabolic activation system 

0.1 mL test / reference item solution, double distilled water and 

DMSO, respectively 

0.1 mL bacteria (overnight culture) 

were mixed and poured over the surface of a Vogel-Bonner 

minimal agar plate 

Preincubation Method

 0.35 mL test / reference item solution, double distilled water and 

DMSO, respectively 

1.75 mL buffer solution or S9-Mix 

0.35 mL bacteria (overnight culture) 

were preincubated in a microtiter plate for at least 20 min at 35 

± 2 °C on a shaker. 

0.7 mL of the preincubated solution 

 1.5 mL top agar with 0.5 mM Histidin-/Biotin- and L-Tryptophansolution (for E. coli WP (uvrA)), respectively, 

were mixed and poured over the surface of a Vogel-Bonner 

minimal agar plate 

Description of follow up The results were confirmed in two independent studies. 

repeat study Plate incorporation method was carried out in the first study and 

preincubation method in the second study.
DR.U.NOACK-LABORATORIEN 

Robust Summary Page 3 of 3 

DOBA 

Bacterial Reverse Mutation Test (Amestest) Project-No. 060601CL 

acc. OECD 471 / EC Directive 2000/32/EC Method B. 13/14 Study-No. USO108573

Criteria for evaluating Genotypes were evaluated for each study. 

Plates of the mutagenicity test results were inspected for present 

and 

 reduced background lawn after incubation. Plates with colonies 

which correspond not with the typical shape and colour of 

Salmonella typhimurium or Escherichia coli, respectively, were 

regarded as contaminated and were not included in calculations. 

Colonies per plate (revertants) were counted. Plates with reduced 

background were considered carefully for evaluation as 

appropriate. 

Results 

Table 1: Mutagenic and Cytotoxic Effects of the Test Item

Tested Concentration Range 

 (factor √10) 

[mg/plate] 

Lowest Cytotoxic 

Concentration [mg/plate] 

Strain S9 

Plate 

Incorporation 

Method 

Preincubation 

Method 

Lowest 

Mutagenic 

Concentration 

[mg/plate] 

Plate 

Incorporation 

Method 

Preincubation 

Method 

- 1.6 0.5 

TA 1537 

+ 1.6 – 0.016 1.6 1.6 

- > 5.0 0.5 

TA 98 

+ > 5.0 > 5.0 

- 5.0 0.5 

TA 100 

+ 5.0 5.0 

- 1.6 0.5 

TA 1535 

+ > 5.0 > 5.0 

E. coli - > 5.0 > 5.0 

WP2 

(uvrA) + 

5.0 - 0.05 5.0 - 0.05 none 

> 5.0 > 5.0 

Conclusions

The test item is regarded to be not mutagenic under test conditions.