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WHO FOOD ADDITIVES SERIES: 48

SAFETY EVALUATION OF CERTAIN
FOOD ADDITIVES AND CONTAMINANTS

BENZYL DERIVATIVES

First draft prepared by Dr J. Gry1, Professor A.G. Renwick2 and Professor I.G. Sipes3
1
Institute of Food Safety and Nutrition, Danish Veterinary and Food Administration, Ministry of Food, Agriculture and Fisheries, Søborg, Denmark
2Clinical Pharmacology Group, University of Southampton, Southampton, United Kingdom
3 Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson, Arizona, USA

Evaluation

Introduction

Estimated daily intake

Metabolic considerations

Application of the Procedure for the Safety Evaluation of Flavouring Agents

Consideration of combined intakes

Conclusions

Relevant background Information

Explanation

Additional considerations on intake

Biological data

Biochemical data

Absorption, distribution, and excretion

Metabolism

Toxicological studies

Acute toxicity

Short-term studies of toxicity

Long-term studies of toxicity and carcinogenicity

Genotoxicity

Reproductive toxicity

References

1. EVALUATION

1.1 Introduction

The Committee evaluated a group of 37 flavouring agents1 that consisted of benzyl alcohol (No. 25), benzaldehyde (No. 22), benzoic acid (No. 850), and related substances (see Table 1) by the Procedure for the Safety Evaluation of Flavouring Agents (see Figure 1, Introduction). All members of this group are aromatic primary alcohols, aldehydes, carboxylic acids, or related esters or acetals. The benzene ring may be ring-substituted with alkyl substituents (Nos 863-869).

Table 1. Summary of results of safety evaluations of benzyl derivatives used as flavouring agentsa

Flavouring agent

No.

CAS no. and structure

Step A3b Does intake exceed the threshold for human intake?

Step A4 Is the substance or are its metabolites endogenous?

Comments

Conclusion based on current intake

Benzyl alcoholc

25

100-51-6

Yes
Europe: 16 000
USA: 17 000

Yes

See note 1.

No safety concern

Benzyl formate

841

104-57-4

No
Europe: 41
USA: 51

NR

See note 2.

No safety concern

Benzyl acetatec

23

140-11-4

No
Europe: 1400
USA: 850

NR

See note 2.

No safety

Benzyl propionate

842

122-63-4

No
Europe: 49
USA: 99

NR

See note 2.

No safety concern

Benzyl butyrate

843

103-37-7

No
Europe: 120
USA: 290

NR

See note 2.

No safety concern

Benzyl isobutyrate

844

103-28-6

No
Europe: 15
USA: 21

NR

See note 2.

No safety concern

Benzyl isovalerate

845

103-38-8

No
Europe: 14
USA: 19

NR

See note 2.

No safety concern

Benzyl trans-2-methyl-2- butenoate

846

37526-88-8

No
Europe: 0.01
USA: 0.03

NR

See note 2.

No safety concern

Benzyl 2,3-dimethyl- crotonate

847

7492-69-5

No
Europe: 0.01
USA: 1

NR

See note 2.

No safety concern

Benzyl acetoacetate

848

5396-89-4

No
Europe: 0.2
USA: 0.07

NR

See note 2.

No safety concern

Benzyl benzoatec

24

120-51-4

Yes
Europe: 1900
USA: 4200

Yes

See notes

No safety concern

Benzyl phenylacetate

849

102-16-9

No
Europe: 5
USA: 57

NR

See note 2.

No safety concern

Benzaldehydec

22

100-52-7

Yes
Europe: 9300
USA: 36 000

Yes

See note 3.

No safety concern

Benzaldehyde dimethyl acetal

837

1125-88-8

No
Europe: 0.2
USA: 0.3

NR

See note 4.

No safety concern

Benzaldehyde glyceryl acetal

838

1319-88-6

No
Europe: 16
USA: 300

NR

See note 4.

No safety concern

Benzaldehyde propylene glycol acetate

839

2568-25-4

No
Europe: 0.04
USA: 110

NR

See note 4.

No safety concern

Benzoic acidc,d

850

65-85-0

No
Europe: 39
USA: 340

NR

See note 5.

Evaluation not finalized

Methyl benzoate

851

93-58-3

No
Europe: 47
USA: 230

NR

See note 6.

No safety concern

Ethyl benzoate

852

93-89-0

No
Europe: 110
USA: 110

NR

See note 6.

No safety concern

Propyl benzoate

853

2315-68-6

No
Europe: 0.01
USA: 0.3

NR

See note 6.

No safety concern

Hexyl benzoate

854

6789-88-4

No
Europe: 380
USA: 1

NR

See note 6.

No safety concern

Isopropyl benzoate

855

939-48-0

No
Europe: 0.004
USA: 0.3

NR

See note 6.

No safety concern

Isobutyl benzoate

856

120-50-3

No
Europe: 0.4
USA: 1

NR

See note 6.

No safety concern

Isoamyl benzoate

857

94-46-2

No
Europe: 110
USA: 33

NR

See note 6.

No safety concern

cis-3-Hexenyl benzoate

858

25152-85-6

No
Europe: 8
USA: 0.1

NR

See note 6.

No safety concern

Linalyl benzoate

859

126-64-7

No
Europe: 10
USA: 2

NR

See note 6.

No safety concern

Geranyl benzoate

860

94-48-4

No
Europe: 4
USA: 0.03

NR

See note 6.

No safety concern

Glyceryl tribenzoated

861

614-33-5

No
Europe: ND
USA: 49

NR

See note 6.

Evaluation not finalized

Propylene glycol dibenzoated

862

19224-26-1

No
Europe: ND
USA: 14

NR

See note 6.

Evaluation not finalized

Methylbenzyl acetate (mixed ortho, meta, and para)

863

29759-11-3

No
Europe: ND
USA: 3

NR

See note 2.

No safety concern

para-Isopropylbenzyl alcohol

864

536-60-7

No
Europe: 0.3
USA: 0.3

NR

See note 1.

No safety concern

4-Ethylbenzaldehyde

865

4748-78-1

No
Europe: 0.4
USA: 6

NR

See note 3.

No safety concern

Tolualdehydes (mixed ortho, meta, and para)

866

1333-09-1

No
Europe: 260
USA: 1100

NR

See note 3.

No safety concern

Tolualdehyde glyceryl acetal

867

1333-09-1

No
Europe: 0.01
USA: 1

NR

See note 4.

No safety concern

Cuminaldehyde

868

122-03-2

No
Europe: 130
USA: 1

NR

See note 3.

No safety concern

2,4-Dimethylbenzaldehyde

869

15764-16-6

No
Europe: 0.4
USA: 0.1

NR

See note 3.

No safety concern

Flavouring agent

No.

CAS no. and structure

Step A3/B3b Does intake exceed the threshold for human intake?

Step A4 Is the substance or are its metabolites endogenous?

Step B4 NOEL for substance or structurally related agent?

Comments

Conclusion based on current intake

Benzyl 2-methoxyethyl acetal

840

7492-39-9

No
Europe: ND
USA: 1

NR

Yes

See note 7.

No safety concern

CAS, Chemical Abstracts Service; ND, no data; NR, not required for evaluation

a

Step 1: All the flavouring agents in this group are in structural class I, except for benzyl 2-methoxyethyl acetal (No. 840).

 

Step 2: All the flavouring agents in this group are predicted to be metabolized to innocuous products.

b

The threshold for human intake for class I is 1800 µg/day. All values for intake expressed in µg/day.

c.

A group ADI of 0–5 mg/kg bw was confirmed by the Committee at its forty-sixth meeting (Annex 1, reference 122) and maintained at the present meeting.

d

Further information is required to determine whether this substance is currently in use as a flavouring agent.

Notes

1.

Benzyl alcohols are oxidized to corresponding acids, which are conjugated with glycine and excreted as hippuric acid.

2.

Benzyl esters are hydrolysed to the corresponding acid and alcohol.

3.

Benzyl aldehydes are oxidized to the corresponding acids.

4.

Benzaldehyde acetals are hydrolysed to yield the aldehyde.

5.

Benzoic acid is conjugated with glycine and excreted as hippuric acid.

6.

Benzoatae esters are hydrolysed to yield corresponding alcohols and acids.

7.

Hydrolysed to acetaldehyde, benzyl alcohol, and 2-methoxyethanol. A two-generation study of reproductive toxicity with methoxyethanol in rats with a NOEL of 6 mg/kg bw per day provides a safety margin > 10 000 (Gulati et al., 1990a,b).

The Committee previously evaluated five members of the group. Benzyl alcohol (No. 25) was evaluated at the twenty-third and forty-sixth meetings (Annex 1, references 50 and 122); benzyl acetate (No. 23) was evaluated at the eleventh, twenty-seventh, twenty-ninth, thirty-first, thirty-fifth, forty-first, and forty-sixth meetings (Annex 1, references 14, 62, 70, 77, 88, 107, and 122); benzyl benzoate (No. 24) was evaluated at the fifteenth and forty-sixth meetings (Annex 1, references 26 and 122); benzaldehyde (No. 22) was evaluated at the eleventh and forty-sixth meetings (Annex 1, references 14 and 122); and benzoic acid (No. 850) was evaluated at the sixth, ninth, seventeenth, twenty-seventh, and forty-sixth meetings (Annex 1, references 6, 11, 32, 62, and 122). At its forty-sixth meeting, the Committee evaluated the five benzyl derivatives as a group and maintained the group ADI of 0–5 mg/kg bw as benzoic acid equivalents (Annex 1, reference 122).

Of the 37 substances in this group, 29 have been reported to occur naturally in foods. They have been detected in a wide variety of fruits, vegetables, meats, cheeses, and wine (CIVO-TNO, 2000).

1.2 Estimated daily intake

The total annual production of the 37 benzyl derivatives in this group is approximately 210 000 kg in Europe (International Organization of the Flavor Industry, 1995) and 460 000 kg in the USA (National Academy of Sciences, 1987; Lucas et al., 1999). Approximately 91% of the total annual production in Europe and 94% of that in the USA is accounted for by three substances in the group: benzyl alcohol (No. 25), benzaldehyde (No. 22), and benzyl benzoate (No. 24). Approximately 31% and 59% of the total annual production in Europe and the USA, respectively, is accounted for by benzaldehyde, 54% and 28% by benzyl alcohol, and 6% and 7% by benzyl benzoate. The estimated daily intake per person in Europe and the USA is 9300 µg and 36 000 µg for benzaldehyde; 16 000 µg and 17 000 µg for benzyl alcohol; and 1900 µg and 4200 µg for benzyl benzoate, respectively. The estimated daily intake of each flavouring agent in the group is reported in Table 2.

Table 2. Annual volumes of use of benzyl derivatives used as flavouring agents in Europe and the USA

Substance (No.)

Most recent annual volume (kg)

Intakea

 

Annual volume in naturally occurring foods (kg)b

Consumption ratioc

 

 

µg/day

µg/kg bw per day

 

 

Benzyl alcohol (25)

Europe

110 000

16 000

270

 

0.06

USA

130 000

17 000

290

7 100

0.05

Benzyl formate (841)

Europe

290

41

0.7

 

 

USA

390

51

0.8

12

0.03

Benzyl acetate (23)

Europe

9 500

1 400

23

 

0.4

USA

6 500

850

14

3 500

0.5

Benzyl propionate (842)

Europe

340

49

0.8

(+)

 

USA

750

99

1.6

 

NA

Benzyl butyrate (843)

Europe

850

120

2

+

 

USA

2 200

290

5

 

NA

Benzyl isobutyrate (844)

Europe

110

15

0.3

(+)

 

USA

160

21

0.4

 

NA

Benzyl isovalerate (845)

Europe

95

14

0.2

+

 

USA

150

19

0.3

 

NA

Benzyl trans-2-methyl-2-butenoate (846)

Europe

0.1

0.01

0.0002

+

 

USA

0.2

0.03

0.0005

 

NA

Benzyl 2,3-dimethylcrotonate (847)

Europe

0.1

0.01

0.002

 

USA

10

1

0.02

 

NA

Benzyl acetoacetate (848)

Europe

2

0.2

0.003

(+)

 

USA

1

0.07

0.001

 

NA

Benzyl benzoate (24)

Europe

13 000

1 900

32

 

0.008

USA

32 000

4 200

70

110

0.003

Benzyl phenylacetate (849)

Europe

35

5

0.08

 

USA

440

57

1

 

NA

Benzaldehyde (22)

Europe

65 000

9 300

160

 

0.9

USA

270 000

36 000

600

58 000

0.2

Benzaldehyde dimethyl acetal (837)

Europe

1

0.2

0.003

(+)

 

USA

2

0.3

0.005

 

NA

Benzaldehyde glyceryl acetal (838)

Europe

110

16

0.3

 

USA

2 300

300

5

 

NA

Benzaldehyde propylene glycol acetal (839)

Europe

0.3

0.04

0.0007

+

 

USA

820

110

1.8

 

NA

Benzoic acid (850)

Europe

280

39

0.6

+

 

USA

2 600

340

5.7

 

NA

Methyl benzoate (851)

Europe

330

47

0.8

 

0.08

USA

1 700

230

3.8

26

0.02

Ethyl benzoate (852)

Europe

790

110

1.8

 

1.4

USA

790

110

1.8

1 100

1.4

Propyl benzoate (853)

Europe

0.1

0.01

0.0002

(+)

 

USA

2

0.3

0.005

 

NA

Hexyl benzoate (854)

Europe

2 600

380

6.3

 

0.7

USA

6

1

0.02

1 700

280

Isopropyl benzoate (855)

Europe

0.03

0.004

0.00007

+

 

USA

2

0.3

0.005

 

NA

Isobutyl benzoate (856)

Europe

3

0.4

0.007

+

 

USA

6

1

0.02

 

NA

Isoamyl benzoate (857)

Europe

790

110

1.8

 

0.3

USA

250

33

0.6

220

0.9

cis-3-Hexenyl benzoate (858)

Europe

55

8

0.13

 

2.5

USA

1

0.1

0.002

140

140

Linalyl benzoate (859)

Europe

69

10

0.2

+

 

USA

14

2

0.03

 

NA

Geranyl benzoate (860)

Europe

28

4

0.07

 

USA

0.2

0.03

0.0005

 

NA

Glyceryl tribenzoate (861)

Europe

NR

NA

NA

 

USA

370

49

0.8

 

NA

Propylene glycol dibenzoate (862)

Europe

NR

NA

NA

 

USA

110

14

0.2

 

NA

Methylbenzyl acetate (mixed ortho, meta, para) (863)

Europe

NR

NA

NA

(+)

 

USA

20

3

0.05

 

NA

para-Isopropylbenzyl alcohol (864)

Europe

2

0.3

0.005

+

 

USA

2

0.3

0.005

 

NA

4-Ethylbenzaldehyde (865)

Europe

3

0.4

0.007

+

 

USA

45

6

0.1

 

NA

Tolualdehydes (mixed ortho, meta, para) (866)

Europe

1 900

260

4.3

+

 

USA

8 600

1 100

1.8

 

NA

Tolualdehyde glyceryl acetal (867)

Europe

0.1

0.01

0.0002

 

USA

5

1

0.02

 

NA

Cuminaldehyde (868)

Europe

940

130

2.2

+

 

USA

7

1

0.02

 

NA

2,4-Dimethylbenzaldehyde (869)

Europe

3

0.4

0.007

+

 

USA

1

0.1

0.002

 

NA

Benzyl 2-methoxyethyl acetal (840)

Europe

NR

NA

NA

 

USA

10

1

0.02

 

NA

Total

Europe

210 000

 

 

 

 

USA

460 000

 

 

 

 

NA, not applicable; NR, not reported; +, reported to occur naturally in foods (CIVO-TNO, 2000), but quantitative data were not available; -, not reported to occur naturally in foods

a

Intake expressed as µg/person per day calculated as follows: [(annual volume, kg) x (1 x 109 µg/kg)/ (population x survey correction factor x 365 days)], where population (10%, ‘eaters only’) = 32 x 106 for Europe and 26 x 106 for the USA. The correction factor = 0.6 for Europe and 0.8 for the USA, representing the assumption that only 60% and 80% of the annual volume of the flavour, respectively, was reported in the poundage surveys (International Organization of the Flavor Industry, 1995; Lucas et al., 1999). Intake expressed as µg/kg bw per day calculated as follows: [(µg/person per day)/body weight], where body weight = 60 kg. Slight variations may occur from rounding.

b

Quantitative data from Stofberg & Grundschober (1987)

c

Calculated as follows: (annual consumption in food, kg)/(most recently reported volume as a flavouring agent, kg)

In addition to its presence in food and flavours, benzoic acid is endogenous in the human body. Endogenous benzoic acid is formed through the phenylalanine-tyrosine pathway (Annex 1, reference 123).

1.3 Metabolic considerations

In general, aromatic esters are hydrolysed in vivo by the catalytic activity of carboxylesterases, which predominate in hepatocytes. The acetals in the group are anticipated to be hydrolysed readily under acidic conditions. Benzyl esters and acetals are hydrolysed to benzyl alcohol (and carboxylic acids) and to benzaldehyde (and alcohols), respectively, followed by oxidation to yield benzoic acid. Benzoate esters are hydrolysed to benzoic acid (and alcohols).

Benzyl derivatives have been shown to be absorbed rapidly through the gut, metabolized primarily in the liver, and excreted in the urine as glycine conjugates of benzoic acid derivatives. Once absorbed, benzyl derivatives are oxidized and excreted primarily as the glycine conjugate of benzoic acid (hippurate). At high doses, the formation of the glycine conjugate is limited; when glycine is depleted, free benzoic acid may sequester acetyl coenzyme A or be excreted unchanged or as the glucuronic acid conjugate. Aromatic ring substitution is anticipated to have little influence on the principal pathway of metabolism.

Oxidation of the alcohol or aldehyde group may be accompanied by oxidation of the alkyl side-chain.

1.4 Application of the Procedure for the Safety Evaluation of Flavouring Agents

Step 1. All the benzyl and benzoate esters and acetals of benzaldehyde (or acetaldehyde) are anticipated to hydrolyse readily to yield benzoic acid, benzyl alcohol, benzaldehyde, acetaldehyde, or alkyl-substituted derivatives thereof. The remaining alcohol or acid components formed by hydrolysis are simple aliphatic substances that are either oxidized to polar, excretable metabolites or metabolized in the fatty acid pathway and tricarboxylic acid cycle. In the Procedure (Annex 1, reference 131), all 37 benzyl derivatives were assigned to structural class I (Cramer et al., 1978).

Step 2. At current levels of estimated intake, 36 of the 37 substances in the group are predicted to be metabolized to innocuous products. The evaluation of these substances therefore proceeded via the left-hand side of the decision tree in the Procedure. One compound, benzyl 2-methoxyethyl acetal (No. 840), is not metabolized to innocuous products and was accordingly evaluated via the right-hand side of the scheme.

Step A3. The estimated daily per capita intakes in Europe and the USA of 33 of the flavouring agents in this group are below the human intake threshold for class I (1800 µg/person per day), and these 33 substances were considered to be of no safety concern when used at current levels. The intakes of the remaining three substances are greater than the threshold for human intake for class I: these are benzyl alcohol (16 000 µg/person per day in Europe and 17 000 µg/person per day in the USA), benzyl benzoate (1900 µg/person per day in Europe and 4200 µg/person per day in the USA), and benzaldehyde (9300 µg/person per day in Europe and 36 000 µg/person per day in the USA). Accordingly, the evaluation of these three substances proceeded to step A4.

Step A4. Benzyl alcohol, benzyl benzoate, and benzaldehyde are readily metabo-lized to benzoic acid, which is endogenous in humans.

Step B3. For benzyl 2-methoxyethyl acetal (No. 840), no data on intake were reported for Europe, and an intake of 1 µg/person per day was reported for the USA, which is below the intake threshold for substances in class I.

Step B4. The NOEL for 2-methoxyethanol, a hydrolysis product of benzyl 2-methoxyethyl acetal (No. 840), in a two-generation study of reproductive toxicity in rats (Gulati et al., 1990a,b), of 6 mg/kg bw per day, provides a safety margin greater than 10 000 in relation to the estimated intake in the USA. Therefore, benzyl 2-methoxyethyl acetal does not pose a safety concern at the estimated level of intake.

The stepwise evaluations of the 37 benzyl derivatives used as flavouring agents are summarized in Table 1.

1.5 Consideration of combined intakes

In the unlikely event that all foods containing these flavouring agents, except benzyl 2-methoxyethyl acetal (No. 840), were to be consumed simultaneously on a daily basis, the estimated combined intake would exceed the human intake threshold for class I. However, these flavouring agents are expected to be efficiently detoxicated and would not saturate the available detoxication pathways. On the basis of the evaluation of the collective data, there would be no safety concerns about combined intake. Furthermore, the total combined daily intake per kilogram of body weight of all benzyl derivatives (0.5 mg in Europe and 1 mg in the USA) is less than the group ADI of 0–5 mg/kg bw for benzoic acid, the benzoate salts (calcium, potassium, and sodium), benzaldehyde, benzyl acetate, benzyl alcohol, and benzyl benzoate, expressed as benzoic acid equivalents, which was established by the Committee at its forty-sixth meeting (Annex 1, reference 122). The three benzyl derivatives that account for more than 90% of the total intake of this group of substances in Europe and the USA are benzyl benzoate (No. 24), which is rapidly hydrolysed to benzyl alcohol and benzoic acid (Nielsen & Bundgaard, 1987), benzaldehyde (No. 22), and benzyl alcohol (No. 25), which are are all readily metabolized to benzoic acid, which is endogenous in humans. In the opinion of the Committee, the endogenous concentration of this substance would not give rise to perturbations outside the physiological range. Therefore, these three substances were considered to be of no safety concern of the current levels of intake.

1.6 Conclusions

The Committee concluded that the flavouring agents in the group of benzyl derivatives would not present safety concerns when used at estimated current levels as flavouring agents. No data on toxicity were required in application of the Procedure to 36 of the 37 benzyl derivatives in the group. However, the Committee noted that the available information was consistent with the results of the safety evaluation. The required data on toxicity were available for benzyl 2-methoxyethyl acetal (No. 840).

2. RELEVANT BACKGROUND INFORMATION

2.1 Explanation

This monograph summarizes the key data relevant to the safety evaluation of 37 derivatives of benzyl alcohol, benzaldehyde, or benzoic acid (Table 1). All members of this group are aromatic primary alcohols, aldehydes, carboxylic acids, or their corresponding esters or acetals. The structural features common to this group of substances include an oxygenated functional group bonded directly to a benzene ring which may be ring-substituted with alkyl substituents. The group contains the parent saturated alcohol, benzyl alcohol (No. 25), and related benzyl esters (Nos 23–24 and 841–849); the corresponding aldehyde, benzaldehyde (No. 22), and related acetals (Nos 837–840); and the corresponding parent carboxylic acid, benzoic acid (No. 850), and related benzoate esters (Nos 851–862). The group also includes seven additional structurally related benzyl derivatives (Nos 863–869) containing ring alkyl substituents.

A subgroup of benzyl derivatives comprising benzyl alcohol (No. 25), benzaldehyde (No. 22), benzyl acetate (No. 23), and benzoic acid (No. 850) and salts was the subject of a recent comprehensive review and safety evaluation by the Committee (Annex 1, references 122 and 123). The review and evaluation contain detailed descriptions of most of the studies on specific benzyl derivatives. The following sections give only summaries of the results of studies described in detail in that review. Other relevant studies, including studies on other benzyl derivatives not discussed in the review, are described in detail.

2.2 Additional considerations on intake

The natural occurrence and total annual production of these are discussed in sections 1.1 and 1.2, respectively, and summarized in Table 2.

Quantitative data on natural occurrence were available for 10 substances in the group (CIVO-TNO, 2000), and consumption ratios were calculated (Stofberg & Kirschman, 1985; Stofberg & Grundschober, 1987). The consumption ratio for the substance for which the reported annual volume of use in Europe is highest (benzyl alcohol, No. 25) is 0.06, and that for the substance with the highest reported annual volume in the USA (benzaldehyde, No. 22) is 0.2. The consumption ratios for benzyl formate (No. 841), benzyl benzoate (No. 24), and methyl benzoate (No. 851) are < 0.1 (Stofberg & Kirschman, 1985; Stofberg & Grundschober, 1987) (Table 2).

2.3 Biological data

2.3.1 Biochemical data

(a) Absorption, distribution, and excretion

The benzyl derivatives are rapidly absorbed through the gut, metabolized primarily in the liver, and excreted in the urine as glycine conjugates of benzoic acid derivatives (Davison, 1971; Abdo et al., 1985; Temellini, 1993), as discussed by the Committee at its forty-sixth meeting, when it evaluated benzyl alcohol (No. 25), benzyl acetate (No.23), benzaldehyde (No. 22), and benzoic acid (No. 850) (Annex 1, reference 122). After absorption, benzyl derivatives are metabolized and excreted within 24 h as polar metabolites, mainly as urinary hippuric acid (Table 3).

Table 3. Metabolism of benzyl derivatives

Flavouring agent (No.)

Species

Route

Dose

24-h urine/ faeces (% dose)

Major urinary metabolite

Minor urinary metabolites

Reference

Benzyl alcohol (25)

Humans

Oral

1.5 g

84

Hippuric acid

 

Snapper et al. (1925)

 

Humans (neonates)

Intravenous, intramuscular

0.007–0.222 mmol/kg bw

82–85

Hippuric acid

Benzoic acid

LeBel et al. (1988)

 

Rabbits

Stomach tube

0.5–16 g

52–84

Hippuric acid

Benzoyl glucuronide

Bray et al. (1952)

Benzyl acetate (23)

Humans

Oral

2.0 g

83

Hippuric acid

 

Snapper et al. (1925)

 

Rats

Oral

5, 50, or 500 mg/kg bw

90/0.3–1.3

Hippuric acid

Benzyl alcohol, benzyl mercaptic acid

Abdo et al. (1985)

 

Mice

Oral

10, 100, or 1000 mg/kg bw

90/0.3–1.3

Hippuric acid

Benzyl alcohol, benzyl mercaptic acid

Abdo et al. (1985)

 

Rats

Gavage

5, 250, or 500 mg/kg bw

70–89/4 (72 h)

Hippuric acid

Benzyl alcohol, benzoic acid, benzyl mercaptic acid

Chidgey & Caldwell (1986)

 

Rats

Subcutaneous

0.3 mg

 

Hippuric acid

Benzyl mercaptic acid

Clapp & Young (1970)

 

Rats

Oral

5 or 500 mg/kg bw

78/2

Hippuric acid

Benzyl mercaptic acid, benzyl alcohol, benzoyl glucuronide, benzoic acid

McMahon et al. (1989)

Benzyl benzoate (24)

Humans

Oral

1.0 g

85

Hippuric acid

 

Snapper et al. (1925)

Benzaldehyde (22)

Rabbits

Stomach tube

0.5–1.5 g

80–98

Hippuric acid

Benzoyl glucuronide

Bray et al. (1951)

 

Rabbits

Oral

0.35–0.75 mg/kg bw

82–83

Hippuric acid

Benzoic acid, benzoyl glucuronide, benzoyl glucuronic acid, benzyl mercaptic acid

Laham et al. (1988)

Benzoic acid (850)

Rats

Oral

0.061, 0.61, 6.1, 61, or 305 mg/kg bw

88–89/1–6

Hippuric acid

Benzoic acid, benzoyl glucuronide, 3-hydroxy-3-phenyl propionic acid

Nutley (1990)

 

Mice

Intraperitoneal

0.061, 0.61, 6.1, 61, or 305 mg/kg bw

92–98/1–10

Hippuric acid

Benzoic acid, benzoyl glucuronide, 3-hydroxy--phenyl propionic acid

Nutley (1990)

 

Various

Oral

1–400 mg/kg bw

50–100

Hippuric acid

Benzoyl glucuronide, ornithic acid

Bridges et al. (1970)

(b) Metabolism

Benzyl esters and acetals are hydrolysed to benzyl alcohol and carboxylic acids and to benzaldehyde and alcohols, respectively. Benzyl alcohol and benzaldehyde are rapidly oxidized to benzoic acid, while benzoate esters are hydrolysed to benzoic acid and alcohols. The benzoic acid derivatives are excreted primarily as glycine conjugates. Benzoic acid is readily conjugated with glycine, primarily in the liver (see Figure 1). After high doses, formation of the glycine conjugate is limited by the concentration of glycine. When glycine is depleted, free benzoic acid may sequester acetyl coenzyme A or be excreted unchanged or as the glucuronic acid conjugate (Diack & Lewis, 1928; Bray et al., 1951; Williams, 1959; Abdo & Wenk, 1995; Abdo et al., 1998). The metabolism, including hydrolysis, of flavouring agents that were not reviewed by the Committee at its forty-sixth meeting is summarized below.

FIGURE 1

Figure 1. Metabolism of benzyl derivatives

(i) Hydrolysis of esters

In general, aromatic esters are hydrolysed in vivo by the catalytic activity of carboxylesterases, which are found throughout mammalian tissues but predominate in hepatocytes (Heymann, 1980).

Benzyl and benzoate esters in the group of benzyl derivatives are anticipated to be hydrolysed rapidly to yield the corresponding alcohols and carboxylic acids, as shown in several studies in vitro and in vivo. Benzyl acetate (No. 23) is hydrolysed rapidly, both in vitro and in vivo (Heymann, 1980; Yuan et al., 1995; Annex 1, reference 122).

Methyl benzoate (No. 851), ethyl benzoate (No. 852), propyl benzoate (No. 853) and benzyl benzoate (No. 24) are all hydrolysed to benzoic acid and the corresponding alcohol after incubation with human plasma, diluted to 80% with phosphate buffer at pH 7.4 and 37 °C. The half-times were was 108 min for methyl benzoate, 210 min for ethyl benzoate, 46 min for propyl benzoate, and 19 min for benzyl benzoate (Nielsen & Bundgaard, 1987).

Benzyl phenylacetate was hydrolysed to 90% within 1 h and completely hydrolysed within 2 h of incubation with a 2% pancreatin solution (Leegwater & van Straten, 1974).

(ii) Hydrolysis of acetals

Benzaldehyde-related acetals are anticipated to hydrolyse readily to their component alcohols and benzaldehyde under acidic conditions.

Benzaldehyde propylene glycol acetal (No. 839) was 97% hydrolysed after incubation for 5 with simulated gastric juice and intestinal fluid in vitro (Morgareidge, 1962).

(iii) Metabolism of benzyl alcohol, benzoic acid, and related esters

The metabolism of benzyl alcohol (No. 25), benzyl acetate (No. 23), benzaldehyde (No. 22), and benzoic acid (No. 850) was described comprehensively by the Committee at its forty-sixth meeting (Annex 1, reference 122).

(iv) Metabolism of substituted benzyl alcohols and benzaldehydes

Aromatic ring substitution is anticipated to have little or no effect on the principal pathway of metabolism. Oxidation of the alcohol or aldehyde to the benzoic acid derivative may be accompanied by minor side-chain oxidation.

Cuminaldehyde (No. 868) was administered orally at a dose of 2 g to male rabbits, and their urine was collected for 3 days after treatment. Cuminaldehyde underwent both oxidation of the aldehyde function and oxidation of the alkyl side-chain to yield 9-hydroxycuminic acid and 8-hydroxycuminic acid as the major urinary metabolites. Cumyl alcohol and 2-(para-carboxyphenyl)propionic acid were minor metabolites (Ishida et al., 1989).

2.3.2 Toxicological studies

(a) Acute toxicity

LD50 values after oral administration have been reported for 30 of the 37 substances in this group (see Table 4). In rats, the values ranged from 1000 mg/kg bw for benzaldehyde (No. 22) to 12 000 mg/kg bw for hexyl benzoate (No. 854), and in mice they ranged from 1300 mg/kg bw for benzoic acid (No. 850) to 9400 mg/kg bw for linalyl benzoate (No. 859). The LD50 values for rabbits, guinea-pigs, and cats were in a similar range, from 1000 mg/kg bw to > 5000 mg/kg bw, indicating that the the benzyl derivatives in the group have low acute toxicity.

Table 4. Acute toxicity of benzyl derivatives

Flavouring agent (No.)

Species

Sex 

Route

LD50 (mg/kg bw)

Reference

Benzyl alcohol (25)

Rabbit
Rat

NR
NR

Oral
Oral

1000
2100

Graham & Kuizenga (1945)

Benzyl alcohol (25)

Rat

M,F

Gavage

1200

Jenner et al. (1964)

Benzyl alcohol (25)

Rat

M,F

Oral

1600

Procter & Gamble (1992)

Benzyl alcohol (25)

Rat

NR

Oral

3100

Smyth et al. (1951)

Benzyl alcohol (25)

Mouse

NR

Gavage

1600

Jenner et al. (1964)

Benzyl formate (841)

Rat

M.F

Gavage

< 5000

Shelanski (1971)

Benzyl acetate (23)

Rabbit

NR

Oral

2600

Graham & Kuizenga (1945)

Benzyl acetate (23)

Rat

M,F

Gavage

2500

Jenner et al. (1964)

Benzyl acetate (23)

Rat

NR

Oral

3700

Graham & Kuizenga (1945)

Benzyl propionate (842)

Rat

NR

Oral

3300

Moreno (1973)

Benzyl butyrate (843)

Rat

NR

Oral

1800

Moreno (1973)

Benzl butyrate (843)

Rat

M,F

Gavage

2300

Jenner et al. (1964)

Benzyl isobutyrate (844)

Rat

M,F

Oral

2800

Owen & Meyer (1971)

Benzyl isovalerate (844)

Rat

NR

Oral

> 5000

Moreno (1974)

Benzyl trans-2-methyl-2-butenoate (846)

Rat

NR

Oral

> 5000

Moreno (1979)

Benzyl benzoate (24)

Cat

NR

Oral

2200

Graham & Kuizenga (1945)

Benzyl benzoate (24)

Rabbit

NR

Oral

2000

Draize et al. (1948)

Benzyl benzoate (24)

Rabbit

NR

Oral

1700

Graham & Kuizenga (1945)

Benzyl benzoate (24)

Guinea-pig

NR

Oral

1100

Draize et al. (1948)

Benzyl benzoate (24)

Rat

NR

Oral

1900

Draize et al. (1948)

Benzyl benzoate (24)

Rat

NR

Oral

2800

Graham & Kuizenga (1945)

Benzyl benzoate (24)

Mouse

NR

Oral

1600

Draize et al. (1948)

Benzyl phenylacetate (849)

Rat

M,F

Oral

> 5000

Owen & Meyer (1971)

Benzaldehyde (22)

Guinea-pig

M,F

Gavage

1000

Jenner et al. (1964)

Benzaldehyde (22)

Rat

M,F

Gavage

1300

Jenner et al. (1964)

Benzaldehyde (22)

Rat

NR

Oral

2800

Sporn et al. (1967)

Benzaldehyde (22)

Rat

M,F

Gavage

1300

Taylor et al. (1964)

Benzaldehyde (22)

Mouse

NR

Food

1200

Schafer & Bowles (1985)

Benzalehyde dimethyl acetal (837)

Rat

NR

Oral

1200

Moreno (1977)

Benzaldehyde glyceryl acetal (838)

Rat

NR

Oral

3700

Levenstein (1974)

Benzaldehyde glyceryl acetal (838)

Rat

NR

Oral

2800

Moreno (1980)

Benzaldehyde propylene glycol acetal (839)

Rat

M,F

Gavage

3000

Lewis & Palanker

Benzoic acid (850)

Mouse

NR

Oral

1200

Schafer & Bowles (1985)

Benzoic acid (850)

Mouse

NR

Oral

2000

Sado (1973)

Benzoic acid (850)

Mouse

NR

Intragastric

2000

Shell BV (1982)

Methyl benzoate (851)

Rabbit
Rat

NR
NR

Oral
Oral

2100
2200

Graham & Kuizenga (1945)

Methyl benzoate (851)

Rat

M,F

Gavage

1400

Jenner et al. (1964)

Methyl benzoate (851)

Rat

M,F

Oral

3400

Smyth et al. (1954)

Methyl benzoate (851)

Mouse

NR

Gavage

3300

Jenner et al. (1964)

Ethyl benzoate (852)

Rabbit
Rat

NR
NR

Oral
Oral

2600
2100

Graham & Kuizenga (1945)

Ethyl benzoate (852)

Rat

M,F

Oral

6500

Smyth et al. (1954)

Hexyl benzoate (854)

Rat

NR

Oral

12 000

Smyth et al. (1951)

Isopropyl benzoate (855)

Rat

NR

Oral

3700

Smyth et al. (1951)

Isobutyl benzoate (856)

Rat

M,F

Gavage

3700

Levenstein (1973)

Isoamyl benzoate (857)

Rat

NR

Oral

6300

Wong & Weir (1971)

Linalyl benzoate (859)

Rabbit

NR

Oral

> 5000

Moreno (1973)

Linalyl benzoate (859)

Rat

NR

Oral

> 5000

Moreno (1973)

Linalyl benzoate (859)

Mouse

M

Oral

9400

Hoffman-LaRoche (1967)

Geranyl benzoate (860)

Rat

NR

Oral

> 5000

Moreno (1973)

para-Isopropylbenzyl alcohol (864)

Rat

NR

Oral

1000

Moreno (1973)

4-Ethylbenzaldehyde (865)

Rat

M,F

Oral

2000

Costello (1984)

Tolualdehydes (mixed ortho, meta, para) (866)

Rat

NR

Oral

2200

Moreno (1973)

Cuminaldehyde (868)

Rat

M,F

Gavage

1400

Jenner et al. (1964)

2,4-Dimethylbenzaldehyde (869)

Rat

M,F

Gavage

< 5000

deGroot et al. (1974)

M, male; F, female; NR, not reported

(b) Short-term studies of toxicity

The results of short-term toxicological studies have been reported for nine representative benzyl derivatives. Studies with the parent alcohol, benzyl alcohol (No. 25), the corresponding aldehyde, benzaldehyde (No. 22), and benzoic acid (No. 850) were described in detail by the Committee at its forty-sixth meeting (Annex 1, reference 122). Only the results of studies on the flavouring agents in the group that were not reported at the forty-sixth meeting are presented below. The short-term studies of toxicity on the benzyl derivatives are summarized in Table 5.

Table 5. Results of short-term studies of toxicity with benzyl derivatives

Flavouring agent (No.)

Species, sex

No. of test groupsa/ no. per groupb

Route

Length
(days)

NOEL (mg/kg bw per day)

Reference

Benzyl alcohol (25)

Rat, M,F
Rat, M,F
Mouse, M,F
Mouse, M,F

10/10
10/5
10/10
10/5

Gavage
Gavage
Gavage
Gavage

13 weeks
16
13 weeks
16

100
130
100
250

National Toxicology Program (1989)

Benzyl acetate (23)

Rat, M,F
Rat, M,F


Rat, M,F



Mouse, M,F
Mouse, M


Mouse, M,F

10/5
10/10


12/10



10/5
10/10


10/10

Gavage
Gavage


Oral



Gavage
Gavage


Gavage

14
13 weeks


13 weeks



14
13 weeks


13 weeks

500
250


460



1000
500


< 430

National Toxicology
Program (1986)
National Toxicology
Program (1993)
National Toxicology
Program (1986)
National Toxicology
Program (1993)

Benzaldehyde (22)

Rat, M,F

Rat, M,F



Rat, M,F

Mouse, M,F

Mouse, M,F

10/5

12/10



1/5

10/5

10/10

Gavage

Gavage



Oral

Gavage

Gavage

13 weeks

13 weeks



27-28 weeks

13 weeks

13 weeks

200

200



>50

300

300

Kluwe et al. (1983)
National Toxicology Program (1990)
Hagan et al. (1967)
Kluwe et al. (1983)
National Toxicology Program (1990)

Methyl benzoate (851)

Rat, NR

5/5

Intragastric

6 months

0.005

Kravets-Bekker (1970)

Glyceryl-tribenzoate (861)

Rat, M,F

8/15

Oral

90

600

Carson (1972a)

Propylene glycol dibenzoate (862)

Rat, M,F

8/15

Oral

90

2500

Carson (1972b)

Tolualdehydes (mixed ortho, meta, para) (866)

Rat, M,F

Rat, M,F

NR/15

NR/5

Oral

Gavage

90

13 weeks

36

250

Oser et al. (1965)
Brantom et al. (1972)

2,4-Dimethylbenzaldehyde (869)

Rat, M,F

NR/5

Gavage

2 weeks

0.18

deGroot et al. (1974)

M, male; F, female; NR, not reported

a

Does not include control groups

b

Both male and female animals

c

Study performed with a single or multiple doses that produced no adverse effects, and so the actual NOEL may be higher.

Benzyl butyrate (No. 843): Groups of 12 weanling rats (strain unspecified) of each sex were given a diet containing a mixture of six aromatic esters commonly used in foods for 12 weeks at concentrations calculated to provide an average daily intake of 0 or 100 times the assumed human intake, calculated to be 130 mg/kg bw per day. The agents were incorporated into the diet in the ratio of their use in foods: ethyl benzoate, 0.15 mg/kg; isobutyl benzoate, 25 mg/kg: benzyl acetate, 19 mg/kg; benzyl butyrate, 25 mg/kg; ethyl methylphenylglycidate, 25 mg/kg; and glycidate M-116, 25 mg/kg. The group receiving the ester blend had normal body-weight gain, food consumption, efficiency of food use, appearance, and behaviour. The blood haemoglobin and urine glucose concentrations did not differ significantly between test and control groups. Traces of albumin present in urine specimens from both control and test groups were regarded as not significant. At autopsy, no treatment-related abnormalities were observed, and the weights of the livers and kidneys were within normal limits for both groups. No histopathological examination was performed (Oser, 1957).

Glyceryl tribenzoate (No. 861): Groups of 15 male and 15 female weanling FDRL-weanling rats were maintained on a diet containing glyceryl tribenzoate at concentrations calculated to provide an average daily intake of 0 (control), 120, 600, or 2600 mg/kg bw, for 90 days. Weekly measurement of body weight and food consumption revealed depressed growth rate and efficiency of food use in males at the high dose. Haematological, blood chemical, and urine analyses gave normal values. No difference in the absolute or relative weights of the liver and kidney were found between test and control animals, and there was no evidence of treatment-related gross or histological lesions (Carson, 1972a).

Propylene glycol dibenzoate (No. 862): Propylene glycol dibenzoate was administered to weanling FDRL-rats at a dose of 0 (control), 130, 630, or 2500 mg/kg bw per day for 90 days in a study similar to that described above. No treatment-related effects were reported, even at the highest dose (Carson, 1972b).

Tolualdehydes (mixed ortho, meta, para) (No. 866): Groups of 15 male and 15 female weanling Sprague-Dawley rats were fed a diet containing tolualdehyde (proportions of ortho, meta, and para isomers not given) at concentrations providing an average daily intake of 36 mg/kg bw for males and 43 mg/kg bw for females, for 90 days. Weekly measurements of body weight and food consumption revealed no significant differences between test and control groups. Haematological, blood chemical, and urine analyses showed normal values. No difference in the absolute or relative weights of organs were found between test and control animals, and there was no evidence of treatment-related effects on gross or histological appearance (Oser et al., 1965).

Groups of 15 male and 15 female CFE rats were given tolualdehyde (approximately equal proportions of meta and para isomers) at a dose of 0 (control), 50, 250, or 500 mg/kg bw in corn oil daily by oral intubation for 13 weeks. Additional groups of five rats of each sex were given the compound at a dose of 0, 250, or 500 mg/kg bw per day for 2 or 6 weeks by the same route. Weekly measurements of body weights and food and water intake showed that the females at 500 mg/kg bw per day had significantly lower body weights after 2 weeks. No changes in body weight were observed in other groups at 2, 6, or 13 weeks. Haematological, blood chemical, and urine analyses performed at 2, 6, and 13 weeks showed a transient increase in erythrocyte count, erythrocyte volume fraction, and haemoglobin concentration in males, but only at 2 weeks. At necropsy at week 13, a significant decrease in the absolute or relative weight of the small intestine was found in all treated groups and decreased relative pituitary weights in females at 500 mg/kg bw per day. The magnitude of the change in the mean weight of the small intestine was not dose-related.

In a second part of the study, groups of 30 female rats were given tolualdehyde at a dose of 0 or 500 mg/kg bw per day in corn oil by gavage for 13 weeks. No significant difference in mean absolute or relative weight of the small intestine was found between the control and treated groups in this part of the study or between the control group in this part and the treated females in the first part of the study. The authors noted that the mean weight of the small intestine of control animals in the first part were abnormally high. The changes in organ weight were not associated with treatment-related gross or histological changes (Brantom et al., 1972).

2,4-Dimethylbenzaldehyde (No. 869): Groups of five male and five female rats were given 2,4-dimethylbenzaldehyde at a dose of 0 (control), 0.18, or 1.8 mg/kg bw by stomach tube daily on 6 days per week for 2 weeks. Males at the highest dose had an increased relative liver weight. Histological examination of the liver and kidneys revealed no differences from controls (deGroot et al., 1974).

(c) Long-term studies of toxicity and carcinogenicity

The results of a series of long-term studies of toxicity and carcinogenicity in mice and rats (Kieckebusch & Lang, 1960; Marquardt, 1960; Shtenberg & Ignat’ev, 1970; Toth, 1984; Abdo et al., 1985; National Toxicology Program, 1986, 1989, 1990, 1993) with four representative benzyl derivatives, benzyl alcohol (No. 25), benzaldehyde (No. 22), benzyl acetate (No. 23), and benzoic acid (No. 850) and sodium benzoate were reviewed by the Committee at its forty-sixth meeting (Annex 1, reference 122). At that meeting, the Committee commented that: "Long-term studies in which benzyl acetate, benzyl alcohol, benzaldehyde, benzoic acid and sodium benzoate were administered in the feed or by gavage to mice and rats were available for review by the Committee. No definitive conclusions could be drawn from carcinogenicity studies of sodium benzoate in mice and rats, as the information provided was insufficient for this purpose, and survival rates in the study in rats were too low to allow it to be considered as conclusive. The Committee reviewed the studies evaluated in the previous monographs and an additional study in which benzaldehyde was administered in corn oil by gavage to rats at 200 or 400 mg/kg bw per day for 103 weeks and to mice at 200 or 400 mg/kg bw per day (males), or 300 or 600 mg/kg bw per day (females) for 103 weeks. On the basis of these studies, the Committee concluded that neither benzyl acetate nor benzyl alcohol is carcinogenic. As in the studies in mice and rats given benzyl acetate in corn oil by gavage, increased incidences of pancreatic acinar cell adenomas in rats and of papillomas of the forestomach in mice were noted after administration of benzaldehyde. However, as in its previous review of benzyl acetate, the Committee concluded that the results of studies in which the compound was administered in the diet were more relevant to its safety assessment as a food additive than those in which it was given in corn oil by gavage." The Committee concluded that the data reviewed were sufficient to demonstrate lack of carcinogenic potential.

(d) Genotoxicity

The results of studies of genotoxicity in vitro and in vivo with benzyl alcohol (No. 25), benzyl acetate (No. 23), benzaldehyde (No. 22), and benzoic acid (No. 850) were reviewed by the Committee at its forty-sixth meeting (Annex 1, reference 122). The Committee concluded that: "None of the four compounds was mutagenic in the Ames test, either with or without metabolic activation. The compounds all induced gene mutations in the mouse lymphoma assay at the thymidine kinase locus (benzoic acid was not tested), although the requirement for metabolic activation varied. Some weak clastogenic activity was noted in in vitro assays, but not in in vivo assays."

Further data on these four flavouring agents and the results of studies with eight other benzyl derivatives in the group (Nos 24, 841, 842, 851, 857, 864, 867, and 868) are presented in Table 6. The results of all 20 assays in vitro with these eight benzyl derivatives were negative, except for one for DNA repair in Bacillus subtilis strains H17 and M45 with benzyl formate (Yoo, 1986).

Table 6. Results of studies of the genotoxicity of benzyl derivatives

No.

Flavouring agent

End-point

Test system

Concentration

Results

Comments

Reference

In vitro

25

Benzyl alcohol

Reverse mutation

S. typhimurium TA92, TA94, TA98, TA100, TA1535, TA1537 (preincubation)

10 000 µg/plate

Negative

Assay performed with and without S9

Ishidate et al. (1984)

 

 

Reverse mutation

S. typhimurium TA100 (plate incorporation)

1000 µg/plate

Negative

Assay performed without S9

Ball et al. (1984)

 

 

Reverse mutation

S. typhimurium TA98, TA100 (plate incorporation)

NR

Negative

Assay performed without S9

Rogan et al. (1986)

 

 

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537 (preincubation)

6700 µg/plate

Negative

Assay performed with and without S9

Mortelmans et al. (1986)

 

 

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537 (plate incorporation)

3 µmol/plate

Negative

Assay performed with and without S9

Florin et al. (1980)

 

 

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537, TA1538 (plate incorporation)

50 000 µg/plate

Negative

Assay performed with and without S9

Heck et al. (1989)

 

 

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537, TA1538 (plate incorporation)

5 µl/plate

Negative

Assay performed without S9

Milvy & Garro (1976)

 

 

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537 (preincubation)

6700 µg/plate

Negative

Assay performed with and without S9; cytotoxicity at highest concentration

National Toxicology Program (1989)

 

 

Mutation

E. coli WP2 uvrA

8 mg/plate

Negative

Japanese article, English summary; use of S9 not reported

Yoo (1986)

 

 

DNA repair

B. subtilis H17, M45

21 µg/disc

Negative

Japanese article, English summary tables

Oda et al. (1979)

 

 

DNA repair

B. subtilis H17, M45

10 µg/disk

Positive

Japanese article, English summary tables; inhibition of growth without S9

Kuroda et al.

 

 

DNA repair

B. subtilis H17, M45

20 µl/disk

Positive

Japanese article, English summary; use of S9 not reported

Yoo (1986)

 

 

Chromosomal aberration

Chinese hamster fibroblasts

1.0 mg/ml

Negative

Assay performed without S9; cells exposed for 48 h

Ishidate et al. (1984)

 

 

Chromosomal aberration

Chinese hamster ovary cells

5000 µg/ml

Equivocal

Assay performed with and without S9; positive results not reproducible; no dose– response relationship

Anderson et al. (1990)

 

 

Chromosomal aberration

Chinese hamster ovary cells

5000 µg/ml

Positive

Assay performed with and without S9; positive results reported only with S9

National Toxicology Program (1989)

 

 

Sister chromatid exchange

Chinese hamster ovary cells

5000 µg/ml

Weakly positive

Dose–response relation- ship at 500–1250 µg/ml without S9 and 500–4000 µg/ml with S9

National Toxicology Program (1989)

 

 

Sister chromatid exchange

Chinese hamster ovary cells

5000 µg/ml

Weakly positive

Assay performed with and without S9; no dose–response relationship; increase at single doses

Anderson et al. (1990)

 

 

Mutation

L5178Y mouse lymphoma cells

5000 µg/ml

Equivocal

Positive and negative responses could not be reproduced; no dose– response relationship

McGregor et al. (1988); Myhr et al. (1990)

 

 

Mutation

L5178Y mouse lympho- ma cells

4500 µg/ml

Positive

Assay performed with and without S9; positive result only without S9

National Toxicology Program (1989)

 

 

Mutation

E. coli WP2 uvrA

NR

Negative

Abstract; methods and test concentrations not reported

Kuroda et al. (1984b)

 

 

Cytotoxicity

Human alveolar tumour cells

0.5 mmol/L

Negative

 

Waters et al. (1982)

 

 

DNA damage

Human alveolar tumour cells

0.5 mmol/L

Negative

 

Waters et al. 1982)

 

 

DNA damage

Rat hepatocytes

10 mmol/L

Negative

Cytotoxicity at maximum dose

Storer et al. (1996)

 

 

DNA damage

E. coli P3478

50 µl/disc

Negative

Assay performed with and without S9

Fluck et al. (1976)

841

Benzyl formate

DNA repair

B. subtilis H17, M45

20 µl/disc

Positive

Japanese article, English summary; use of S9 not reported

Yoo (1986)

 

 

Mutation

E. coli WP2 uvrA

4.0 mg/plate

Negative

Japanese article, English summary; use of S9 not reported

Yoo (1986)

23

Benzyl acetate

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537 (preincubation)

10 mg/plate

Negative

Assay performed with and without S9

Mortelmans et al. (1986)

 

 

Reverse mutation

S. typhimurium TA98, TA100 (preincubation and plate incorporation)

5000 µg/plate

Negative

Assay performed with and without S9; cytotoxicity at three higher concentrations

Schunk et al. (1986)

 

 

Reverse mutationa

S. typhimurium TA98, TA100, TA1535, TA1537

3 µmol/plate

Negative

Assay performed with and without S9

Florin et al. (1980)

 

 

DNA repair

B. subtilis H17, M45

21 µg/disc

Negative

Assay performed without S9 Japanese article, English summary tables

Oda et al. (1979)

 

 

DNA repair

B. subtilis H17, M45

20 µl/disc

Positive

Japanese article, English summary; use of S9 not reported

Yoo (1986)

 

 

Mutation

E. coli WP2 uvrA

2.0 mg/plate

Negative

Japanese article, English summary; use of S9 not reported

Yoo (1986)

 

 

Mutation

Mouse lymphoma L5178Y cells

500 µg/ml

Positive

Assay performed with and without S9; positive results only with S9

Caspary et al. (1988)

 

 

Mutation

Human lymphoblast TK6 cells

1500 µg/ml

Positive

Assay performed with and without S9; positive results only with S9

Caspary et al. (1988)

 

 

Mutation

Mouse lymphoma L5178Y cells

1600 µL/mL

Positive

Assay performed without S9; cytotoxicity at maximum (1988) concentration

McGregor et al.

 

 

Mutation

Mouse lymphoma L5178Y cells

NR

Positive

Assay performed with and without S9; positive result only with S9

Rudd et al. (1983)

 

 

Chromosomal aberration

Chinese hamster ovary cells

5000 µg/ml

Negative

Assay performed with and without S9

Galloway et al. (1987)

 

 

Chromosomal aberration

Chinese hamster lung fibroblasts

2.4 mg/ml

Negative

Assay performed with and without S9; cytotoxicity at maximum concentration

Matsuoka et al. (1996)

 

 

Sister chromatid exchange

Chinese hamster ovary cells

5000 µg/ml

Negative

Assay performed with and without S9

Galloway et al. (1987)

 

 

Unscheduled DNA synthesis

Rat hepatocytes

NR

Negative

Abstract; methods and test concentrations not reported

Mirsalis et al. (1983)

842

Benzyl propionate

DNA repair

B. subtilis H17, M45

21 µg/disc

Negative

Assay performed without S9 Japanese article, English summary tables

Oda et al. (1979)

24

Benzyl benzoate

Reverse mutationa

S. typhimurium TA98, TA100, TA1535, TA1537

3 µmol/plate

Negative

Assay performed with and without S9

Florin et al. (1980)

 

 

Reverse mutation

S. typhimurium TA98, TA100 (preincubation and plate incorporation)

5000 µg/plate

Negative

Assay performed with and without S9; cytotoxicity at three higher concentrations

Schunk et al. (1986)

22

Benzaldehyde

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537, TA1538 (plate incorporation)

38 000 ug/plate

Negative

Assay performed with and without S9

Heck et al. (1989)

 

 

Reverse mutation

S. typhimurium TA98, TA100 (plate incorporation)

300 µl/plate

Negative

Assay of urine samples from rats given benzaldehyde by oral gavage, per- formed with and without S9

Rockwell & Raw (1979)

 

 

Reverse mutation

S. typhimurium TA98, TA100 (plate incorporation)

100 µl/plate

Negative

Assay performed with S9

Rockwell & Raw (1979)

 

 

Reverse mutationa

S. typhimurium TA98, TA100, TA2637

2 mg/plate

Negative

Japanese article, English summary; assay performed with and without S9

Nohmi et al. (1985)

 

 

Reverse mutationa

S. typhimurium TA98, TA100, TA1535, TA1537

3 µmol/plate

Negative

Assay performed with and without S9

Florin et al. (1980)

 

 

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537 (preincubation)

1000 µg/plate

Negative

Assay performed with and without S9

Haworth et al. (1983)

 

 

Reverse mutation

S. typhimurium TA100, TA102, TA104

3300 µg/plate

Negative

Assay performed with and without S9

National Toxicology Program (1990)

 

 

Reverse mutationa

S. typhimurium TA100

1 mg/plate

Negative

Use of S9 not reported

Rapson et al. (1980)

 

 

Reverse mutation

S. typhimurium TA98, TA100 (preincubation)

NR

Negative

Assay performed with and without S9

Sasaki & Endo (1978)

 

 

Reverse mutation

S. typhimurium TA100, TA102, TA104 (preincubation)

NR

Negative

Assay performed with and without S9

Dillon et al. (1992)

 

 

Reverse mutation

S. typhimurium TA100 (preincubation)

2000 nmol/plate

Negative

Assay performed with and without S9

Vamvakas et al. (1989)

 

 

Reverse mutationa

S. typhimurium TA98, TA100

500 µg/plate

Negative

Assay performed with and without S9

Kasamaki et al. (1982)

 

 

DNA repair

B. subtilis H17, M45

21 µg/disc

Negative

Japanese article, English summary; use of S9 not reported

Oda et al. (1979)

 

 

DNA repair

B. subtilis H17, M45

NR

Positive

Assay performed with and without S9; positive result only with S9

Matsui et al. (1989)

 

 

Unscheduled DNA synthesis

Rat hepatocytes

250 µg/ml

Negative

Assay performed without S9

Heck et al. (1989)

 

 

Mutation

Mouse L5178Y lymphoma cells

600 µg/ml

Positive

Positive only with S9

Heck et al. (1989)

 

 

Mutation

Mouse L5178Y lymphoma cells

800 µg/ml

Positive

Assay performed without S9; significant increase in mutant fraction at close to toxic doses

McGregor et al. (1991)

 

 

Chromosomal aberrations

Chinese hamster cells

1.2 mg/ml

Positive

Japanese article, English summary; positive results without S9; weakly positive results with S9; cytotoxicity at two higher concentrations

Sofuni et al. (1985)

22

Benzaldehyde

Chromosomal aberrations

Chinese hamster ovary cells

1600 µg/ml

Negative

Assay performed with and without S9

Galloway et al. (1987)

 

 

Chromosomal aberrations

Chinese hamster cells

50 nmol/L

Positive

Assay performed with and without S9

Kasamaki et al. (1982)

 

 

Sister chromatid exchange

Chinese hamster ovary cells

1600 µg/ml

Positive

Assay performed with and without S9

Galloway et al.

 

 

Sister chromatid exchange

Chinese hamster ovary cells

1000 µmol/L

Negative

Assay performed without S9; cytotoxicity at highest concentration

Sasaki et al. (1989)

 

 

Sister chromatid exchange

Human lymphocytes

2 mmol/L

Positive

Assay performed without S9

Jansson et al. (1988)

850

Benzoic acid

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1538 (plate incorporation)

2500 µg/plate

Negative

Assay performed with and without S9

Anderson & Styles (1978)

 

 

Reverse mutationa

S. typhimurium TA98, TA100, TA1535, TA1536

3.6 µg/plate

Negative

Assay performed with and without S9

Cotruvo et al. (1977)

 

 

Reverse mutation

S. typhimurium TA97, TA98, TA100, TA1535, TA1537 (preincubation)

10 mg/plate

Negative

Assay performed with and without S9

Zeiger et al. (1988)

 

 

Reverse mutationa

S. typhimurium TA100

1 mg/plate

Negative

Use of S9 not reported

Rapson et al. (1980)

 

 

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537 (plate incorporation)

1 mg/plate

Negative

Assay performed with S9

McCann et al. (1975)

 

 

Reverse mutation

S. typhimurium TA92, TA94, TA98, TA100, TA1535, TA1537 (preincubation)

10 mg/plate

Negative

Assay performed with and without S9

Ishidate et al. (1984)

 

 

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537, TA1538 (plate incorporation)

100 µg/plate

Negative

Assay performed without S9

Milvy & Garro (1976)

 

 

Reverse mutation

S. typhimurium TA1535, TA1537, TA1538 (plate incorporation)

0.5%

Negative

Assay performed with and without S9

Food & Drug Admin istration (1975)

 

 

Mutation (umu gene expression)

S. typhimurium TA1535/ pSK1002

1.7 mg/ml

Negative

Assay performed with and without S9

Nakamura et al. (1987)

 

 

DNA repair

B. subtilis H17, H45

NR

Positive

Abstract; methods and test concentration(s) not reported

Nonaka (1989)

 

 

Mutation

Saccharomyces cerevisiae D3

0.18%

Negative

Assay performed with and without S9

Cotruvo et al. (1977)

 

 

Mutation

S. cerevisiae D4

0.15%

Negative

Assay performed with and without S9

Food & Drug Admin- istration (1975)

 

 

Indirect DNA repair (induction of beta-gala- ctosidase)

E. coli PQ37

400 µg/ml

Negative

 

Glosnicka & Dziadziuszko (1986)

 

 

Chromosomal aberration

Chinese hamster fibroblasts

1.5 mg/ml

Weakly positive

Total incidence of cells with aberrations, 5–9%; performed without S9

Ishidate et al. (1984)

 

 

Sister chromatid exchange

Human lymphocytes

2.0 mmol/L

Negative

Assay performed without S9

Jansson et al. (1988)

851

Methyl benzoate

Reverse mutation

S. typhimurium TA97, TA98, TA100, TA1535, TA1537 (preincubation)

6700 ug/plate

Negative

Assay performed with and without S9

Zeiger et al. (1992)

 

 

Mutation

E. coli Sd-4-73

NR

Negative

Assay performed without S9

Szybalski (1958)

857

Isoamyl benzoate

Mutation

E. coli Sd-4-73

NR

Negative

Assay performed without

Szybalski (1958) S9

864

Isopropylbenzyl alcohol

Reverse mutation

S. typhimurium TA98, TA100 (plate incorporation)

100 µl/plate

Negative

Assay performed with S9

Rockwell & Raw (1979)

 

 

Reverse mutation

S. typhimurium TA98, TA100 (plate incorporation)

300 µl/plate

Negative

Assay of urine samples from rats given isopropyl- benzyl alcohol by oral gavage; performed with and without S9

Rockwell & Raw (1979)

867

Tolualdehydes (mixed ortho, meta, para)

Reverse mutation

S. typhimurium TA104 (preincubation)

0.8 µmol/plate

Negative

Assay performed with and without S9

Marnett et al. (1985)

 

 

Reverse mutationa

S. typhimurium TA98, TA100, TA1535, TA1537

3 µmol/plate

Negative

Assay performed with and without S9

Florin et al. (1980)

 

 

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537, TA1538 (plate incorporation)

19 000 ug/plate

Negative

Assay performed with and without S9

Heck et al. (1989)

 

 

Reverse mutation

S. typhimurium TA98, TA100, TA102 (plate incorporation)

0.8 mmol/plate

Negative

Assay performed with and without S9

Aeschbacher et al. (1989)

 

 

Reverse mutation

S. typhimurium TA97, TA100, TA1535, TA1537 (preincubation)

666 µg/plate

Negative

Assay performed with and without S9

Zeiger et al. (1988)

 

 

Unscheduled DNA synthesis

Rat hepatocytes

1000 µg/ml

Negative

Assay performed without S9

Heck et al. (1989)

 

 

Mutation

Mouse L5178Y lymphoma cells

300 µg/ml

Negative

Assay performed with and without S9

Heck et al. (1989)

868

Cuminaldehyde

Reverse mutation

S. typhimurium TA98, TA100 (plate incorporation)

100 µl/plate

Negative

Assay performed with S9

Rockwell & Raw (1979)

 

 

Reverse mutation

S. typhimurium TA98, TA100 (plate incorporation)

300 µl/plate

Negative

Assay of urine samples from rats given cuminaldehyde by gavage; performed with and without S9

Rockwell & Raw (1979)

 

 

Sister chromatid exchange

Chinese hamster ovary cells

333 µmol/L

Negative

Assay performed without S9; cytotoxicity at maximum concentration

Sasaki et al. (1989)

In vivo

25

Benzyl alcohol

Sex-linked recessive lethal mutation

Drosophila melanogaster

5000 mg/kg|
8000 mg/kg

Negative
Negative

In feed
By injection

Foureman et al. (1994)

 

 

Micronucleus formation

Mouse bone-marrow cells

200 mg/kg bw

Negative

By intraperitoneal injection

Hayashi et al. (1988)

 

 

Replicative DNA synthesis

Mouse hepatocytes

NR

Positive

Route of administration not reported

Yoshikawa (1996)

 

 

Sex-linked recessive lethal mutation

D. melanogaster

300 mg/kg
20 000 mg/kg

Negative
Negative

In feed
By injection

National Toxicology Program (1993); Foureman et al. (1994)

 

 

Sister chromatid exchange

Mouse bone-marrow cells

1700 mg/kg bw

Negative

By intraperitoneal injection

National Toxicology Program (1993)

23

Benzyl acetate

Chromosomal aberration

Mouse bone-marrow cells

1700 mg/kg bw

Negative

By intraperitoneal injection

National Toxicology Program (1993)

 

 

Micronucleus formation

Mouse bone-marrow cells

1300 mg/kg bw

Negative

By intraperitoneal injection

National Toxicology Program (1993); Shelby et al. (1993)

 

 

Micronucleus formation

Mouse erythrocytes

50 000 mg/kg

Negative

By intraperitoneal injection

National Toxicology Program (1993)

 

 

Unscheduled DNA synthesis

Rat hepatocytes

NR

Negative

By oral gavage; abstract; methods and doses not reported

Mirsalis et al. (1983)

 

 

Unscheduled DNA synthesis

Rat hepatocytes

1000 mg/kg bw

Negative

By oral gavage

Mirsalis et al. (1989)

 

 

Unscheduled DNA synthesis

Rat pancreatic cells

1000 mg/kg bw

Negative

By oral gavage

Steinmetz & Mirsalis (1984)

 

 

DNA damage

Rat pancreatic cells

500 mg/kg bw

Negative

By gavage

Longnecker et al. (1990)

 

 

DNA damage

Rate pancreatic cells

0.9%

Negative

In diet

Longnecker et al. (1990)

22

Benzaldehyde

Sex-linked recessive lethal mutation

D. melanogaster

1200 mg/kg
2500 mg/kg

Negative
Negative

In feed
By injection

Woodruff et al. (1985)

NR, not reported; S9, exogenous metabolic activation system consisting of 9000 x g supernatant from rodent liver

a Not reported whether plate incorporation or preincubation method used

A total of 12 benzyl derivatives in the group have been tested for genotoxicity. In view of the mainly negative results in the assays in vitro and the uniformly negative results in well-recognized assays in vivo, the Committee concluded that the group of benzyl derivatives is not genotoxic in vivo.

(e) Reproductive toxicity

At its forty-sixth meeting, the Committtee reviewed a series of studies of developmental and reproductive toxicity with benzyl alcohol (No. 25), benzyl acetate (No. 23), benzyl aldehyde (No. 22), and sodium benzoate (Annex 1, reference 122). The Committee concluded that: "Delayed development and reduced fetal and postnatal pup body weights were observed in developmental toxicity studies in rats, mice, hamsters and rabbits, but only at doses that were toxic to the mother. In a teratogenicity study with sodium benzoate, doses that induced severe maternal toxicity were associated with embryotoxic and fetotoxic effects and fetal malformations. A 4-generation study in rats showed no effect on growth, fertility, lactation or survival". The Committee concluded that the data reviewed were sufficient to demonstrate a lack of teratogenic and reproductive potential. No further studies on reproductive toxicity with benzyl derivatives in the group were available for review by the Committee at its present meeting.

3. REFERENCES

Abdo, K.M. & Wenk, M.L. (1995) Requirement for adequate glycine for the detoxification of benzyl acetate and maintenance of its detoxification pathways. Toxicologist, 15, 19.

Abdo, K.M., Huff, J.E., Haseman, J.K., Boorman, G.A., Eustis, S.L., Matthews, H.B., Burka, L.T., Prejean, J.D. & Thompson, R.B. (1985) Benzyl acetate carcinogenicity, metabolism and disposition in Fischer 344 rats and B6C3F1 mice. Toxicology, 37, 159–170.

Abdo, K.M., Wenk, M.L., Harry, G.J., Mahler, J., Goehl, T.J. & Irwin, R.D. (1998) Glycine modulates the toxicity of benzyl acetate in F344 rats. Toxicol. Pathol., 26, 395–402.

Aeschbacher, H.U., Wolleb, U., Loliger, J., Spadone, J.C. & Liardon, R. (1989) Contribution of coffee aroma constituents to the mutagenicity of coffee. Food Chem. Toxicol., 27, 227–232.

Anderson, D. & Styles, J.A. (1978) The bacterial mutation test. Br. J. Cancer, 37, 924–930.

Anderson, B.E., Zeiger, E., Shelby, M.D., Resnick, M.A., Gulati, D.K., Ivett, J.L. & Loveday, K.S. (1990) Chromosome aberration and sister chromatid exchange test results with 42 chemicals. Environ. Mol. Mutag., 16 (Suppl. 18), 55–137.

Ball, J.C., Foxall-Van Aken, S. & Jensen, T.E. (1984) Mutagenicity studies of p-substituted benzyl derivatives in the Ames Salmonella plate-incorporation assay. Mutat. Res., 138, 145–151.

Brantom, P.G., Gaunt, I.F., Grasso, P., Lansdown, A.B.G. & Gangolli, S.D. (1972) Short-term toxicity of tolualdehyde in rats. Food Cosmet. Toxicol., 10, 637–647.

Bray, H.G., Thorpe, W.V. & White, K. (1951) Kinetic studies of the metabolism of foreign organic compounds. The formation of benzoic acid from benzamide, toluene, benzyl alcohol and benzaldehyde and its conjugation with glycine and glucuronic acid in the rabbit. Biochem. J., 48, 88–96.

Bray, H.G., Thorpe, W.V. & White, K. (1952) Kinetic studies of the metabolism of foreign organic compounds. A mathematical model expressing the metabolic fate of phenols, benzoic acids and their precursors. Biochem. J., 52, 423–430.

Bridges, J.W., French, M.R., Smith, R.L. & Williams, R.T. (1970) The fate of benzoic acid in various species. Biochem. J., 118, 47–51.

Carson, S. (1972a) 90-day studies with glyceryl tribenzoate in rats. Unpublished report. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Carson, S. (1972b) 90-Day studies with propylene glycol dibenzoate in rats. Unpublished report No. 2-732 from Food and Drug Research Laboratories Inc., New York, USA. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Caspary, W.J., Langenbach, R., Penman, B.W., Crespi, C., Myhr, B.C. & Mitchell, A.D. (1988) The mutagenic activity of selected compounds at the TK locus: Rodent vs. human cells. Mutat. Res., 196, 61–81.

CIVO-TNO (2000) Volatile Compounds in Food. Database. 1996–1999. Boelens Aroma Chemical Information Service, Zeist, Netherlands.

Clapp, J.J. & Young, L. (1970) Formation of mercaptic acids in rats after the administration of aralkyl esters. Biochem. J., 118, 765–771.

Costello, B.A. (1984) Acute oral toxicity LD50 study. 4-Ethyl benzaldehyde. Unpublished report No. 84-4171A from Bioresearch, Inc., Philadelphia, USA. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Cotruvo, J.A., Simmon, V.F. & Spanggord, R.J. (1977) Investigation of mutagenic effects of products of ozonation reactions in water. Ann. N.Y. Acad. Sci., 298, 124–140.

Cramer, G.M., Ford, R.A. & Hall, R.L. (1978) Estimation of toxic hazard—A decision tree approach. Food Cosmet. Toxicol., 16, 255–276.

Davison, C. (1971) Salicylate metabolism in man. J. Pharmacol. Exp. Ther., 179, 249–268.

Diack, S.L. & Lewis, H.B. (1928) Studies in the synthesis of hippuric acid in the animal organism. J. Biol. Chem. 77, 89-95.

Dillon, D.M., McGregor, D.B., Combes, R.D. & Zeiger, E. (1992) Detection of mutagenicity in Salmonella of some aldehydes and peroxides. Environ. Mol. Mutagen., 19, 15.

Draize, J.H., Alvarez, E. & Whitesell, M.F. (1948) Toxicological investigations of compounds proposed for use as insect repellents. J. Pharmacol. Exp. Ther., 93, 26–39.

Florin, I., Rutberg, L, Curvall, M. & Enzell, C.R. (1980) Screening of tobacco smoke constituents for mutagenicity using the Ames test. Toxicology, 18, 219–232.

Fluck, E.R., Poirier, L.A. & Ruelius, H.W. (1976) Evaluation of a DNA polymerase-deficient mutant of E. coli for the rapid detection of carcinogens. Chem.–Biol. Interactions, 15, 219–231.

Food & Drug Administration (1975) Mutagenic evaluation of compound FDA 73-70, benzoic acid, certified ACS, Litton Bionetics, Inc., Washington DC.

Foureman, P., Mason, J.M., Valencia, R. & Zimmering, S. (1994) Chemical mutagenesis testing in Drosophila: X. Results of 70 coded chemicals tested for the National Toxicology Program. Environ. Mol. Mutag., 23, 208–227

Galloway, S.M., Armstrong, M.J., Reuben, C., Colman, S., Brown, B., Cannon, C., Bloom, A.D., Nakamura, F., Ahmed, M., Duk, S., Rimpo, J., Margolin, B.H., Resnick, M.A., Anderson, B. & Zeiger, E. (1987) Chromosome aberrations and sister chromatid exchange in Chinese hamster ovary cells: Evaluations of 108 chemicals. Environ. Mol. Mutag., 10 (Suppl. 10), 1–35.

Glosnicka, R. & Dziadziuszko, H. (1986) Mutagenic action of styrene and its metabolites. II. Genotoxic activity of styrene, styrene ocide, styrene glycol and benzoic acid tested with the SOS chromotest. Bull. Inst. Mar. Trop. Med. Gdynia, 37, 3–4.

Graham, B.E. & Kuizenga, M.H. (1945) Toxcity studies on benzyl benzoate and related benzyl compounds. J. Pharmacol. Exp. Ther., 84, 358–362.

deGroot, A.P., Spanjers, M.T. & van der Heijden, C.A. (1974) Acute and sub-acute oral toxicity studies in rats with five flavor compounds. Unpublished report. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Gulati, D.K., Hope E., Barnes, L.H., Russel, S. & Poonacha, K.B. (1990a) Reproductive toxicity of ethylene glycol monomethyl ether (CAS No. 109-86-4) in Sprague-Dawley rats, litter two, 1-76. Unpublished report from Environmental Health Research and Testing, Inc. (NTIS-PB 90-252313), Springfield, Virginia, USA: Department of Commerce.

Gulati, D.K., Hope, E., Christman, K.L., Barnes, L.H. & Russell, S. (1990b) Reproductive toxicity of ethylene glycol monomethyl ether (CAS No. 109-86-4) in Sprague-Dawley rats, litter two, 1-76. Unpublished report from Environmental Health Research and Testing, Inc. (NTIS-PB 90-252321), Springfield, Virginia, USA: Department of Commerce.

Hagan, E.C., Hansen, W.H., Fitzhugh, O.G., Jenner, P.M., Jones, W.I., Taylor, J.M., Long, E.L., Nelson, A.A. & Brouwer, J.B. (1967) Food flavourings and compounds of related structure. II. Subacute and chronic toxicity. Cosmet. Toxicol., 5, 141–157.

Haworth, S., Lawlor, T., Mortelmans, K., Speck, W. & Zeiger, E. (1983) Salmonella mutagenicity test results for 250 chemicals. Environ. Mutag., 5 (Suppl. 1), 3–142.

Hayashi, M., Kishi, M., Sofuni, T. & Ishidate, M., Jr (1988) Micronucleus tests in mice on 39 food additives and eight miscellaneous chemicals. Food Chem. Toxicol., 26, 487–500.

Heck, J.D., Vollmuth, T.A., Cifone, M.A., Jagannath, D.R., Myhr, B. & Curren, R.D. (1989) An evaluation of food flavoring ingredients in a genetic toxicity screening battery. Toxicology, 9, 257 (Abstract 1031).

Heymann, E. (1980) Carboxylesterases and amidases. In: Jakoby, W.B., Bend, J.R. & Caldwell, J., eds, Enzymatic Basis of Detoxication, 2nd Ed., New York: Academic Press, pp. 291–323.

Hoffmann-LaRoche (1967) Acute toxicity, eye and skin irritation tests on aromatic compound, linalyl benzoate. Unpublished report from Roche Chemical Division. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

International Organization of the Flavor Industry (1995) European inquiry on volume use. Unpublished report. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Ishida, T., Toyota, M. & Asakawa, Y. (1989) Terpenoid biotransformation in mammals. V. Metabolism of (+)-citronellal, (+–)-7-hydroxycitronellal, citral, (–)-perillaldehyde, (–)-myrtenal, cuminaldehyde, thujone, & (+–)-carvone in rabbits. Xenobiotica, 19, 843–855.

Ishidate, M., Jr, Sofuni, T., Yoshikawa, D., Hayashi, M., Nohmi, T., Sawada, M. & Marsouka, A. (1984) Primary mutagenicity screening of food additives currently used in Japan. Food Chem. Toxicol., 22, 623–636.

Jansson, T., Curvall, M., Hedin, A. & Enzell, C.R. (1988) In vitro studies of the biological effects of cigarette smoke condensate: III. Induction of SCE by some phenolic and related constituents derived from cigarette smoke: A study of structure–activity relationships. Mutat. Res., 206, 7–24.

Jenner, P.M., Hagan, E.C., Taylor, J.M., Cook, E.L. & Fitzhugh, O.G. (1964) Food flavourings and compounds of related structure. I. Acute oral toxicity. Food Cosmet. Toxicol.. 2, 327–343.

Kasamaki, A., Takahashi, H., Tsumura, N., Niwa, J., Fujita, T., & Urasawa, S. (1982) Genotoxicity of flavoring agents. Mutat. Res., 105, 387–392.

Kieckebusch, W. & Lang, K. (1960) Die Verträglichkeit die Benzoesäure im chronischen Fütterungversuch. Arzneimittel. Forsch., 10, 1001-1003 (in German).

Kluwe, W.M., Montgomery, C.A., Giles, H.D. & Prejean, J.D. (1983) Encephalopathy in rats and nephropathy in rats and mice after subchronic oral exposure to benzaldehyde. Food Chem. Toxicol., 21, 245–250.

Kuroda, K., Tanaka, S., Yoo, Y.S. & Ishibashi, T. (1984a) Rec-assay of food additives. Jpn. Soc. Public Health, 31, 277–281.

Kuroda, K., Yoo, Y.S. & Ishibashi, T. (1984b) Antimutagenic activity of food additives. Mutat. Res., 130, 369.

Laham, S., Potvin, M. & Robinet, M. (1988) Metabolism of benzaldehyde in New Zealand white rabbits. Chemosphere, 17, 517–524.

LeBel, M., Ferron, L., Masson, M., Pichette, J. & Carrier, C. (1988) Benzyl alcohol metabolism and elimination in neonates. Dev. Pharmacol. Ther., 11, 347–356.

Leegwater, D.C. & van Straten, S. (1974) In vitro study on the hydrolysis of twenty-six organic esters by pancreatin. Unpublished report No. 4319 from CIVO/TNO.

Levenstein, I. (1973) To determine the oral LD50, in fasted rats of the test material as submitted. Isobutyl benzoate. Unpublished report No. 30967 from Leberco Laboratories, Roselle Park, USA. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Levenstein, I. (1974) Acute oral toxicity (rat). Dermal toxicity (rabbit). Benzal glyceryl acetal. Unpublished report No. 41766 from Leberco Laboratories, Roselle Park, USA. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Lewis, C.A. & Palanker, A.L. (1979) Acute oral toxicity (rat). Acute dermal toxicity (rabbit). Oral LD50 (rat). Benzaldehyde propylene glycol acetal. Unpublished report No. 79104-19 from Consumer Product Testing, Fairfield, USA. Submitted to WHO by Flavor and Extract Manufacturers Association of the United States.

Longnecker, D.S., Roebuck, B.D., Curphey, T.J. & MacMillan, D.L. (1990) Evaluation of promotion of pancreatic carcinogenesis in rats by benzyl acetate. Food Chem. Toxicol., 28, 665–668.

Lucas, C.D., Putnam, J.M., Hallagan, J.B. & the Flavor and Extract Manufacturers’ Association of the United States Flavor Ingredients Committee (1999) 1995 Poundage and Technical Effects Update Survey, Washington DC: Flavor and Extract Manufacturers’ Association of the United States.

Marnett, L.J., Hurd, H.K., Hollstein, M.C., Levin, D.E., Esterbauer, H., & Ames, B.N. (1985) Naturally-occurring carbonyl compounds are mutagens in Salmonella tester strain TA104. Mutat. Res., 148, 25–34.

Marquardt, P. (1960) Zur Verträglichkeit der Benzoesäure. Arzneimittel Forsch., 10, 1033.

Matsouka, A., Yamakage, K., Kusakabe, H., Wakuri, S., Asakura, M., Noguchi, T., Sugiyama, T., Shimada, H., Nakayama, S., Kasahara, Y., Takahashi, Y., Miura, K.F., Hatanaka, M., Ishidate, M., Jr, Morita, T., Watanabe, K., Hara, M., Odawara, K., Tanaka, N., Hayashi, M. & Sofuni, T. (1996) Re-evaluation of chromosomal aberration induction on nine mouse lymphoma assay ‘unique positive’ NTP carcinogens. Mutat. Res., 369, 243–252.

Matsui, S., Yamamoto, R. & Yamada, H. (1989) The Bacillus subtilis/microsome rec-assay for the detection of DNA damaging substances which may occur in chlorinated and ozonated waters. Water Sci. Technol., 21, 857–887.

McCann, J., Choi, E., Yamasaki, E. & Ames, B.N. (1975) Detection of carcinogens as mutagens in the Salmonella/microsome test: Assay of 300 chemicals. Proc. Natl Acad. Sci. USA, 79, 5135–5139.

McGregor, D.B., Brown, A., Cattanach, P., Edwards, I., McBride, D., Riach, C. & Caspary, W.J. (1988) Responses of the L5178Y tk+/tk mouse lymphoma cell forward mutation assay: III. 72 coded chemicals. Environ. Mol. Mutag., 12, 85–154 (Erratum: Environ. Mol. Mutag., 12, 345).

McGregor, D.B., Brown, A.G., Howgate, S., McBride, D., Riach, C. & Caspary, W.J. (1991) Responses of the L5178Y mouse lymphoma cell forward mutation assay. V: 27 coded chemicals. Environ. Mol. Mutag., 17, 196–219.

McMahon, T.F., Diliberto, J.J. & Birnbaum, L.S. (1989) Age related changes in disposition of benzyl acetate (BA): A model compound for glycine conjugation. Toxicology, 9 (Abstract).

Milvy, P. & Garro, A.J. (1976) Mutagenic activity of styrene oxide (1,2 epoxyethyl benzene), a presumed styrene metabolite. Mutat. Res., 40, 15–18.

Mirsalis, J., Tyson, K., Beck, J., Loh, E., Steinmetz, K., Contreras, C., Austera, L., Martin, S. & Spalding, J. (1983) Induction of unscheduled DNA synthesis (UDS) in hepatocytes following in vitro and in vivo treatment. Environ. Mutag., 5, 482 (Abstract Ef-5).

Mirsalis, J.C., Tyson, C.K., Steinmetz, K.L., Loh, E.K., Hamilton, C.M., Bakke, J.P. & Spalding, J.W. (1989) Measurement of unscheduled DNA synthesis and S-phase synthesis in rodent hepatocytes following in vivo treatment: Testing of 24 compounds. Environ. Mol. Mutag., 14, 155–164.

Moreno, O.M. (1973) Acute oral toxicity in rats. Dermal toxicity in rabbits. Cuminyl alcohol, tolyl aldehyde, geranyl benzoate, benzyl propionate, benzyl butyrate, linalyl benzoate. Unpublished report No. MB 73-233, MB 73-204, MB 73-94, MB 73-139, MB 72-7, MB 73-98 from MB Research Laboratories, Inc., Perkasie, USA. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Moreno, O.M. (1974) Acute oral toxicity in rats. Dermal toxicity in rabbits. Benzyl isovalerate. Unpublished report No. MB 73-433 from MB Research Laboratories, Inc., Perkasie, USA. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Moreno, O.M. (1977) Acute oral toxicity in rats. Dermal toxicity in rabbits. Benzaldhyde dimethyl acetal. Unpublished report No. MB 76-1448 from MB Research Laboratories, Inc., Spinnerstown, USA. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Moreno, O.M. (1979) Acute oral toxicity in rats. Dermal toxicity in rabbits. Benzyl tiglate. Unpublished report No. MB 78-3419 from MB Research Laboratories, Inc., Spinnerstown, USA. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Moreno, O.M. (1980) Acute oral toxicity in rats. Dermal toxicity in rabbits. Benzyl glyceryl acetal. Unpublished report No. 80-4427 from MB Research Laboratories, Inc., Spinnerstown, USA. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Morgareidge, K. (1962) In vitro digestion of four acetals. Unpublished report. Submitted to WHO by Flavor and Extracts Manufacturers Association of the United States.

Mortelmans, K., Haworth, S., Lawlor, T., Speck, W., Tainer, B. & Zeiger, E. (1986) Salmonella mutagenicity tests: II. Results from the testing of 270 chemicals. Environ. Mutag., 8, 1–119.

Myhr, B., McGregor, D., Bowers, L., Riach, C., Brown, A.G., Edwards, I., McBride, D., Martin, R. & Caspary, W.J. (1990) L5178Y mouse lymphoma cell mutation assay results with 41 compounds. Environ. Mol. Mutag., 16, 138–167.

Nakamura, S., Oda, Y., Shimada, T., Oki, I. & Sugimoto, K. (1987) SOS-inducing activity of chemical carcinogens and mutagens in Salmonella typhimurium TA1535/pSK1002: Examination with 151 chemicals. Mutat. Res., 192, 239–246.

National Toxicology Program (1986) Toxicology and carcinogenesis studies of benzyl acetate in F344/N rats and B6C3F1 mice (gavage studies) (NTP-TR-250; PB-86-2506), Washington DC.

National Toxicology Program (1989) Toxicology and carcinogenesis studies of benzyl alcohol in F344/N rats and B6C3F1 mice (gavage studies) (NTP-TR-343; NIH Publication No. 89-2599), Washington DC.

National Toxicology Program (1990) Toxicology and carcinogenesis studies of benzaldehyde in F344/N rats and B6C3F1 mice (gavage studies) (NTP-TR-378; NIH Publication No. 90-2833), Washington DC.

National Toxicology Program (1993) Toxicology and carcinogenesis studies of benzyl acetate in F344/N rats and B6C3F1 mice (feed studies) (NTP-TR-431; NIH Publication No. 92-3162), Washington DC.

Nielsen, N.M. & Bundgaard, H. (1987) Prodrugs as drugs delivery systems. 68. Chemical and plasma-catalyzed hydrolysis of various esters of benzoic acid: A reference system for designing prodrug esters carboxylic acid agents. Int. J. Pharm., 39, 75–85.

Nohmi, T., Miyata, R., Yoshikawa, K. & Ishidate, M., Jr (1985) Mutagenicity tests on organic chemical contaminants in city water and related compounds. I. Bacterial mutagenicity tests. Bull. Natl Inst. Hyg. Sci., 103, 60–64.

Nonaka, M. (1989) DNA repair tests on food additives. Environ. Mol. Mutag., 14 (Suppl.15), 143 (Abstract 414).

Nutley, B.P. (1990) Investigations into the metabolism of cinnamic acid, cinnamyl alcohol and cinnamaldehyde in relation to their safety evaluation. PhD thesis, University of London, Department of Pharmacology.

Oda, Y., Hamano, Y., Inoue, K., Yamamoto, H., Nihara, T. & Kunita, N. (1979) Mutagenicity of food flavors in bacteria. Osaka-Furitsu Koshu Eisei Kenkyu Hokoku. Shokuhin Eisei Hen, 91, 325 (Chem Abstr., 1979, No. 207194r).

Oser, B.L. (1957) Toxicological screening of components of food flavors Class V. aromatic esters. Unpublished report from Food Research Laboratories, Inc.

Oser, B.L., Carson, S. & Oser, M. (1965) Toxicological tests on flavouring matters. Food Cosmet. Toxicol., 3, 563–569.

Owen, G. & Meyer, F. (1971) Acute oral toxicity investigation in rats. Benzyl phenylacetate, benzyl isobutyrate. Unpublished report No. 71-37, 71-39 from Huntingdon Research Center, Baltimore, USA. Submitted to WHO by Flavor and Extract Manufacturers Associations, USA.

Procter & Gamble (1992) Acute oral toxicity study (LD50) in the rat. Benzyl alcohol. Unpublished report No. 2131-110/233 from Hazleton, Harrogate, England. Submitted to WHO by Flavor and Extract Manufacturers Associations, USA.

Rapson, W.H., Nazar, M.A. & Butsky, V.V. (1980) Mutagenicity produced by aqueous chlorination of organic compounds. Bull. Environ. Contam. Toxicol., 24, 590–596.

Rockwell, P. & Raw, I. (1979) A mutagenic screening of various herbs, spices and food additives. Nutr. Cancer, 1, 10–15.

Rogan, E.G., Cavalieri, E.L., Walker, B.A., Balasubramanian, R., Wislocki, P.G., Roth, R.W. & Saugier, R.K. (1986) Mutagenicity of benzylic acetate, sulfates, and bromides of polycyclic aromatic hydrocarbons. Chem.–Biol. Interactions, 58, 253–275.

Rudd, C.J., Mitchell, A.D. & Spalding, J. (1983) L5178Y mouse lymphoma cell mutagenesis assay of coded chemicals incorporating analyses of the colony size distributions. Environ. Mutag., 5, 419 (Abstract Cd-19).

Sado, I. (1973) Synergistic toxicity of official permissible preservative food additives. Jpn. J. Hyg., 28, 463–476.

Sasaki, Y. & Endo, R. (1978) Mutagenicity of aldehydes in Salmonella. Mutat. Res., 54, 251–252.

Sasaki, Y.F., Imanishi, H., Ohta, T. & Shirasu, Y. (1989) Modifying effects of components of plant essence on the induction of sister chromatid exchanges in cultured Chinese hamster ovary cells. Mutat. Res., 226, 103–110.

Schafer, E.W. & Bowles, W.A. (1985) Acute oral toxicity and repellency of 933 chemicals to house and deer mice. Arch. Environ. Contam. Toxicol., 14, 111–129.

Schunk, H.H., Shibamoto, T., Tan, H.K. & Wei, C.-I. (1986) Biological and chemical studies on photochemical products obtained from euronol, benzyl acetate and benzyl benzoate. In: Lawrence, B.M., Mookherjee, B.D. & Willis, B.J., eds, Flavors and Fragrances: A World Perspective. Proceedings of the 10th International Congress of Essential Oils, Fragrance and Flavors, Amsterdam: Elsevier Science Publishers, pp. 1045–1068.

Shelanski, M.V. (1971) Acute oral toxicity study. Benzyl formate. Unpublished report No. 7114 from Food and Drug Research Laboratories, New York, USA. Submitted to WHO by Flavor and Extract Manufacturers Associations, USA.

Shelby, M.D., Erexson, G.L., Hook, G.J. & Tice, R.R. (1993) Evaluation of a three-exposure mouse bone marrow micronucleus protocol: Results with 49 chemicals. Environ. Mol. Mutag., 21, 160–179.

Shell (1950) Study of the comparative toxicity of benzoic aid and p-tertiary butyl benzoic acid. Unpublished report from School of Medicine, University of California, San Francisco, USA. Submitted to WHO by Flavor and Extract Manufacturers Associations, USA.

Shtenberg, A.J. & Ignat’ev, A.D. (1970) Toxicological evaluation of some combinations of food preservatives. Food Cosmet. Toxicol., 8, 369–380.

Smyth, H.F., Carpenter, C.P. & Weil, C.S. (1951) Range finding toxicity data: List IV. Arch. Ind. Hyg. Occup. Med., 4,119–122.

Smyth, H.F., Carpenter, C.P., Weil, C.S. & Pozzani, U.C. (1954) Range finding toxicity data: List V. Arch. Ind. Hyg. Occup. Med., 10, 61–68.

Snapper, J., Gruenbaum, A. & Sturkop, S. (1925) Uber die Spaltung und die Oxydation von Benzylalkohol und Benzylestern im Menschlichen Organismus. Biochem. Z., 155, 163–173.

Sofuni, M.D., Hayashi, M., Matsouka, A., Sawada, M., Hatanaka, M. & Ishidate, M., Jr (1985) Mutagenicity tests on organic chemical contaminants in city water and related compounds. II. Chromosome aberration tests in cultured mammalian cells. Bull. Natl Inst. Hyg. Sci., 103, 64–75.

Sporn, A., Dinu, I. & Stanciu, V. (1967) Cercetari cu privire la toxicitatea aldehidei benzoice. Igiena, 16, 23–24 (in Romanian).

Steinmetz, K.L. & Mirsalis, J.C. (1984) Measurement of DNA repair in primary cultures of rat pancreatic cells following in vivo treatment. Environ. Mutag., 6, 446.

Stofberg, J. & Grundschober, F. (1987) Consumption ratio and food predominance of flavoring materials. Perfumer Flavorist, 12, 27.

Stofberg, J. & Kirschman, J.C. (1985) The consumption ratio of flavoring materials: A mechanism for setting priorities for safety evaluation. Food Chem. Toxicol., 23, 857–860.

Storer, R.D., McKelvey, T.M., Kraynak, A.R., Elia, M.C., Barnum, J.E., Harmon, L.S., Nichols, W.W. & DeLuca, J.G. (1996) Revalidation of the in vitro alkaline elution/rat hepatocyte assay for DNA damage: Improved criteria for assessment of cytotoxicity and genotoxicity and results for 81 compounds. Mutat. Res., 368, 59–101.

Szybalski, W. (1958) Special microbial systems. II. Observations on chemical mutagenesis in microorganisms. Ann. N.Y. Acad. Sci., 76, 475–489.

Taylor, J.M., Jenner, P.M. & Jones, W.I. (1964) A comparison of the toxicity of some allyl, propenyl, and propyl compounds in the rat. Toxicol. Appl. Pharmacol., 6, 378–387.

Temellini, A. (1993) Conjugation of benzoic acid with glycine in human liver and kidney: A study on the interindividual variability. In: Fifth North American ISSX Meeting, Tucson, AZ, 17–21 October 1993. 4.

Toth, B. (1984) Lack of tumorigenicity of sodium benzoate in mice. Fund. Appl. Toxicol., 4, 494–496.

Vamvakas, S., Dekant, W. & Anders, M.W. (1989) Mutagenicity of benzyl S-haloalkyl and S-haloalkenyl sulfides in the Ames test. Biochem. Pharmacol., 38, 935–939.

Waters, R., Mirzayans, R., Meredith, J., Mallalah, G., Danford, N. & Parry, J.M. (1982) Correlations in mammalian cells between types of DNA damage, rates of DNA repair and the biological consequence. Prog. Mutat. Res., 4, 247–259.

Williams, R.T. (1959) Detoxication Mechanisms, 2nd Ed., London: Chapman & Hall.

Wong, L.C.K. & Weir, R.J. (1971) Acute oral toxicity studies—Rats. Acute dermal toxicity studies—Rabbits. Primary skin irritation—Rabbits. Unpublished report No. 2221 from Bionetics Research Laboratories, Falls Church, USA. Submitted to WHO by Flavor and Extract Manufacturers Association of the United States.

Woodruff, R.C., Mason, J.M., Valencia, R. & Zimmering, S. (1985) Chemical mutagenesis testing in Drosophila. V. Results of 53 coded compounds tested for the National Toxicology Program. Environ. Mutag., 7, 677–702.

Yoo, Y.S. (1986) Mutagenic and antimutagenic activities of flavoring agents used in foodstuffs. Osaka-Shiritsu Daigaku Igaki Zasshi, 4, 267–288.

Yoshikawa, K. (1996) Anomalous nonidentity between Salmonella genotoxicants and rodent carcinogens: Nongenotoxic carcinogens and genotoxic noncarcinogens. Environ. Health Perspectives, 104, 40–46.

Yuan, J.H, Goehl, T.J., Abdo, K., Clark, J., Espinosa, O., Bugge, C. & Garcia, D. (1995) Effects of gavage versus dosed feed administration on the toxicokinetics of benzyl acetate in rats and mice. Food Chem. Toxicol., 33, 151–158.

Zeiger, E., Anderson, B., Haworth, S., Lawlor, T. & Mortelmans, K. (1988) Salmonella mutagenicity tests: IV. Results from the testing of 300 chemicals. Environ. Mol. Mutag., 11 (Suppl.12), 1–158.

Zeiger, E., Anderson, B., Haworth, S., Lawlor, T. & Mortelmans, K. (1992) Salmonella mutagenicity tests: V. Results from the testing of 311 chemicals. Environ. Mol. Mutag., 19 (Suppl.21), 2–141.

ENDNOTES

1 During evaluation of these agents, the Committee questioned whether some substances in the group (see footnote to Table 1) were in fact used as flavouring agents and should therefore appropriately be evaluated with the Procedure. Information to address this question will be sought from the relevant manufacturers.



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       Toxicological Abbreviations