Quaternary ammonium compounds, dicoco alkyldimethyl, nitrites

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Read across data is presented on the "fragments" from which this substance is manufactured :-

Quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides, CAS Number 68391-05-9, EC Number 269-924-1 and from sodium nitrite, CAS Number 7632-00-0, EC Number 231-555-9.

The justification in taking this approach is as follows :-

In the stomach the gastic juice is acidic, made p of acids and enzymes. In such an evironment it is highly unlikely that the quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites substance (s) will remain ionically bound to each other and thus are prone to dissociation in which case the released cation(s) will associate with other anions and the released anion will associate with cations.re, it is suggested read-across data from the corresponding quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides is considered approriate in that such substances are likely to dissociate in a similar manner.

Furthermore, in 1988, the US EPA, Office of Pesticides and Toxic Substances issued a Notice to producers, Formulators, Distributors and Registrants regarding quaternary ammonium compounds with regard to "Clustering" of such quaternary ammonium compounds.

Prior to this, EPA had required each quat compound to be individually coded and registered as a new chemical, even when the chemical structure of individual compounds differed only slightly in alkyl distribution and chain lengths. This procedure was continued with the new generations of quats having two, three, and four chains. As a result, EPA records showed that some 211 registered technical grade active ingredient products containing varying concentrations of Quats, each coded separately on the basis of alkyl chain length and percentage carbon distribution within the chain. At this time, there are approximately eight to ten thousands (8-10,000) registered end-use formulations.

However, questions were raised regarding whether the EPA could cluster or group the quats and pick one or more representative members of each cluster to be used in toxicity studies, instead of requiring separate studies on each quat. These same questions were raised when the EPA issued its March 4, 1987 Data Call-In Notice requiring all registrants of antimicrobial active ingredients to submit subchronic and chronic toxicological data to support the continued registration of their products.

In response to these questions, EPA solicited information from industry, the public, academia, industry cooperative work groups, the state of California, and Canada. EPA then reviewed all of the assembled information along with the chemical structure of most of the quats. Based on the results of this review, EPA developed the following four groupings of currently registered quat compounds:

Group I. The alkyl or hydroxyalkyl (straight chain) substituted Quats

Group II. The non-halogenated benzyl substituted Quats (includes hydroxybenzyl, ethylbenzyl, hydroxyethybenzyl, napthylmethyl, dodecylbenzyl, and alkyl benzyl)

Group III. The di-and tri-chlorobenzyl substituted

Group IV. Quats with unusual substituents (charged heterocyclic ammonium compounds).

Fundamental to this discussion EPA determined that "X-" in all of these structures would be attributed to "any anionic species". Therefore, this would mean in terms of toxicological evaluation the coutner anion in such quaternary ammonium compounds could be regarded as; e.g  halogen (Cl-, Br-, I-,), saccharinate or cyclohexylsulphamate. It is therefore suggested here that nitrite (NO2-) could also be regarded as a pertinent anion.

Since the US EPA deem that such a clustering of structures for toxicological evaluation is well founded then it would seem that to consider read-across data from quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides to the closely structurally analogous quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites to be equally justifiable.

Similarly since the US EPA deem that the counter anion could be regarded as "any anionic species" then it would seem that to consider available toxicological data on sodium nitrite, in order to evaluate any health effects that may be incurred from exposure to the nitrite anion (NO2-), is equally justifiable.

Furthermore, in certain organic solvents it has been reported that the exchange constants between nitrite and chloride in quaternary ammonium salts (QAS) are approximately equal. [Zhurnal Analiticheskoi Khimii, 2010, Vol. 65, No. 6, pp. 579–584. (E.M. Rakhman’ko, M.S. Markovskaya, L.S. Stanishevskii, Yu.S. Zubenko, A.R. Tsyganov)]

To that end one study is presented from quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides, CAS Number 68391-05-9, EC Number 269-924-1 and three from sodium nitrite,

CAS Number 7632-00-0, EC Number 231-555-9.

Quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides, CAS Number 68391-05-9, EC Number 269-924-1

A reproduction study was conducted in Sprague-Dawley rats by dietary administration of test material (containing DDAC) at target dosage levels of 0, 300, 750, or 1500 ppm (0, 22.52, 56.3 and 112.6 mg/kg bw/day ) for two generations.
Under the test conditions, continuous exposure to test material (containing DDAC) in the diet for two generations resulted in no adverse reproductive effects. Parental toxicity was observed at 1500 ppm (112.6 mg/kg/day), limited to body weight reduction, weight gain depression, and decreased food consumption. Postnatal toxicity at 1500 ppm was indicated by reduced pup body weights. The NOEL for both adults and offspring was 750 ppm (56.3 mg/kg bw/day), indicating no increased risk to offspring in the absence of indications of adult toxicity.

Sodium nitrite, CAS Number 7632-00-0, EC Number 231-555-9 (1)

In a publication entitled "Effects of imipramine, nitrite and dimethylnitrosamine on reproduction in mice" (Anderson, L.M., Goner-Sorolla, A., Ebeling, D., Research Communications in Chemical Pathology and Pharmacology Vo. 19 (2), 311 ff, 1978), the fertility of CD-1 mice (females) upon exposure to sodium nitrite was investigated.

Under the conditions of the study no NOAEL could be determined.

Sodium nitrite, CAS Number 7632-00-0, EC Number 231-555-9 (2)

In a study conducted in 1990 the fertility of CD-1 mice (females) upon exposure to sodium nitrite was investigated. No guidle;ine is available although the study was conducted to GLP.

Under the conditions of this study the NOAEL (first parental animals) was determined to be 370 mg/kg bw/day.

However, at concentrations that reduced water consumption slightly but did not reduce body weight, sodium nitrite had no adverse effect on reproductive performance or necropsy end points in Swiss CD-1 mice.

Sodium nitrite, CAS Number 7632-00-0, EC Number 231-555-9 (3)

In a publication entitled "The Relationships among Reproductive Endpoints in Swiss Mice, Using the Reproductive Assessment by Continuous Breeding Database" ( Chapin, R.E., Sloane, R.A., Haseman, J.K., Fund. Appl. Toxicol. 38, 129 - 142, 1997) the reproductive toxicity effects of sodium nitrite was investigated in a two generation study (primary source).

Under the conditions of the study the NOAEL for parental animals were determined to be 425 mg/kg bw/day (reproductive toxicity) and 260 mg/kg bw/day (general toxicity).

Link to relevant study records

Reference

Reference 1

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Report peer-reviewed by governmenatal competent authority. Well documented study, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Read across justification is presented from the structurally analogous quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides to the quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites.

In the stomach the gastic juice is acidic, made up of acids and enzymes. In such an evironment it is highly unlikely that the quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites substance (s) will remain ionically bound to each other and thus are prone to dissociation in which case the released cation(s) will associate with other anions and the released anion will associate with cations. Thererfore, it is suggested read-across that data from the corresponding quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides is considered approriate in that such substances are likely to dissociate in a similar manner.

Furthermore, in 1988, the US EPA, Office of Pesticides and Toxic Substances issued a Notice to producers, Formulators, Distributors and Registrants regarding quaternary ammonium compounds with regard to "Clustering" of such quaternary ammonium compounds.

Prior to this, EPA had required each quat compound to be individually coded and registered as a new chemical, even when the chemical structure of individual compounds differed only slightly in alkyl distribution and chain lengths. This procedure was continued with the new generations of quats having two, three, and four chains. As a result, EPA records showed that some 211 registered technical grade active ingredient products containing varying concentrations of Quats, each coded separately on the basis of alkyl chain length and percentage carbon distribution within the chain. At this time, there are approximately eight to ten thousands (8-10,000) registered end-use formulations.

However, questions were raised regarding whether the EPA could cluster or group the quats and pick one or more representative members of each cluster to be used in toxicity studies, instead of requiring separate studies on each quat. These same questions were raised when the EPA issued its March 4, 1987 Data Call-In Notice requiring all registrants of antimicrobial active ingredients to submit subchronic and chronic toxicological data to support the continued registration of their products.

In response to these questions, EPA·solicited information from industry, the public, academia, industry cooperative work groups, the state of California, and Canada. EPA then reviewed all of the assembled information along with the chemical structure of most of the quats. Based on the results of this review, EPA developed the following four groupings of currently registered quat compounds:

Group I. The alkyl or hydroxyalkyl (straight chain) substituted Quats
Group II. The non-halogenated benzyl substituted Quats (includes hydroxybenzyl, ethylbenzyl, hydroxyethybenzyl, napthylmethyl, dodecylbenzyl, and alkyl benzyl)
Group III. The di-and tri-chlorobenzyl substituted
Group IV. Quats with unusual substituents (charged heterocyclic ammonium compounds).

Fundamental to this discussion EPA determined that "X-" in all of these structures would be attributed to "any anionic species". Therefore, this would mean in terms of toxicological evaluation the coutner anion in such quaternary ammonium compounds could be regarded as; e.g halogen (Cl-, Br-, I-,), saccharinate or cyclohexylsulphamate. It is therefore suggested here that nitrite (NO2-) could also be regarded as a pertinent anion.

Since the US EPA deem that such a clustering of structures for toxicological evaluation is well founded then it would seem that to consider read-across data from quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides to the closely structurally analogous quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites to be equally justifiable.

Furthermore, in certain organic solvents it has been reported that the exchange constants between nitrite and chloride in quaternary ammonium salts (QAS) are approximately equal. [Zhurnal Analiticheskoi Khimii, 2010, Vol. 65, No. 6, pp. 579–584. (E.M. Rakhman’ko, M.S. Markovskaya, L.S. Stanishevskii, Yu.S. Zubenko, A.R. Tsyganov)]

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Deviations:

not specified

Principles of method if other than guideline:

Not applicable

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): Didecyldimethylammonium chloride (DDAC) CAS [7173-51-5], EC 269-924-1

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Not reported

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
No data

Details on mating procedure:

No data

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

Not applicable

Duration of treatment / exposure:

Exposure period: Continuous
Premating exposure period (males): 10 wk F1A, and approx. 19 wk F1B
Premating exposure period (females): 10 wk F1A, and approx. 19 wk F1B
Duration of test: Weaning second generation

Details on study schedule:

Exposures continued through mating, gestation, parturition and lactation
- At least 10 d after weaning the F1A litters, FO parents were mated in different male-female pairings, within dose groups, to produce the F1B generation.

Dose / conc.:

0 ppm (nominal)

Remarks:

Basis : nominal in diet

Dose / conc.:

300 ppm (nominal)

Remarks:

Basis : nominal in diet

Dose / conc.:

750 ppm (nominal)

Remarks:

Basis : nominal in diet

Dose / conc.:

1 500 ppm (nominal)

Remarks:

Basis : nominal in diet

No. of animals per sex per dose:

28/sex/dose

Control animals:

yes, concurrent no treatment

Details on study design:

No data

Positive control:

Not used in the study

Parental animals: Observations and examinations:

DETAILED CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data

Oestrous cyclicity (parental animals):

No data

Sperm parameters (parental animals):

No data

Litter observations:

No data

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals
- Maternal animals: All surviving animals

GROSS NECROPSY
- High dose and control animals

Postmortem examinations (offspring):

No data

Statistics:

Not reported

Reproductive indices:

No data

Offspring viability indices:

No data

Clinical signs:

not specified

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Body weight reduction

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Body weight reduction

Histopathological findings: non-neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS): Body weight reduction, weight gain depression and decreased food consumption at 1500 ppm

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS): No adverse effects

Key result

Dose descriptor:

NOAEL

Effect level:

750 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Body weight reduction, weight gain depression and decreased food consumption at 1500 ppm

Clinical signs:

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

BODY WEIGHT (OFFSPRING): Reduced pup body weights at 1500 ppm

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

750 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Reduced pup body weights at 1500 ppm

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

750 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Reduced pup body weights at 1500 ppm

Reproductive effects observed:

not specified

The details of the study were reproduced from the original report of Bradley NTL (1991). [Union Carbide, Bushy Run Research Center, Export, PA, USA. Report No. 52-648, (unpublished)]

Conclusions:

Under the test conditions, continuous exposure to test material (containing DDAC) in the diet for two generations resulted in no adverse reproductive effects. Parental toxicity was observed at 1500 ppm (112.6 mg/kg/day), limited to body weight reduction, weight gain depression, and decreased food consumption. Postnatal toxicity at 1500 ppm was indicated by reduced pup body weights. The NOEL for both adults and offspring was 750 ppm, indicating no increased risk to offspring in the absence of indications of adult toxicity.

Executive summary:

A reproduction study was conducted in Sprague-Dawley rats by dietary administration of test material (containing DDAC) at target dosage levels of 0, 300, 750, or 1500 ppm (0, 22.52, 56.3 and 112.6 mg/kg bw/day ) for two generations.

This study is assigned a reliability score of 2 in accordance with the criteria for assessing data quality set forth by Klimisch et al (1997).

Dose levels in terms of mg/kg/day decreased throughout the study as animal body weight increased and varied within and between dose groups and generations, so are not reported here. A total of 28 males and 28 females were evaluated at each dose level. Animals were exposed to test material for 10 wk prior to mating, and each of the two generations produced two litters. The original rats, called the F0 generation, were randomly paired within dose groups and mated over a three wk period to produce the F1A generation. Exposures continued through mating, gestation, parturition, and lactation. At least 10 d after weaning the F1A litters, FO parents were mated in different male-female pairings, within dose groups, to produce the F1B generation.  Exposures to test material again occurred from mating to lactation. After the FIB animals were weaned, F0 parents were necropsied and high dose and control animals were examined for histopathologic lesions.

Selected F1 parents were exposed to the same concentrations of test material as their parents for at least 10 wk, and were then paired as described above to produce F2A and F2B generations. Mating, gestation, lactation, and necropsy of the  F1  parents  and  selected  F2A  and  F2B  pups  were  performed as outlined above, except that no F2 animals were selected as parents.

Under the test conditions, continuous exposure to test material (containing DDAC) in the diet for two generations resulted in no adverse reproductive
effects. Parental toxicity was observed at 1500 ppm (112.6 mg/kg/day), limited to body weight reduction, weight gain depression, and decreased food consumption. Postnatal toxicity at 1500 ppm was indicated by reduced pup body weights. The NOEL for both adults and offspring was 750 ppm (56.3 mg/kg bw/day), indicating no increased risk to offspring in the absence of indications of adult toxicity.

Reference 2

Endpoint:

fertility, other

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1978

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: basic information given

Justification for type of information:

Since in the stomach the gastic juice is acidic, made up of acids and enzymes, it is suggested that in this evironment it is highly unlikely that the quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites substance (s) will remain ionically bound to each other and thus are prone to dissociation in which case the released cation(s) will associate with other anions and the released anion will associate with cations. Thererfore, it is suggested that read-across data from sodium nitrite is considered appropriate in order to arrive at a conclusion in terms of any health effects that may be incurred from exposure to "nitrite" whichever cation it may be associated with.

In addition, in 1988, the US EPA, Office of Pesticides and Toxic Substances issued a Notice to producers, Formulators, Distributors and Registrants regarding quaternary ammonium compounds with regard to "Clustering" of such quaternary ammonium compounds.

Prior to this, EPA had required each quat compound to be individually coded and registered as a new chemical, even when the chemical structure of individual compounds differed only slightly in alkyl distribution and chain lengths. This procedure was continued with the new generations of quats having two, three, and four chains. As a result, EPA records showed that some 211 registered technical grade active ingredient products containing varying concentrations of quats, each coded separately on the basis of alkyl chain length and percentage carbon distribution within the chain. At this time, there are approximately eight to ten thousands (8-10,000) registered end-use formulations.

However, questions were raised regarding whether the EPA could cluster or group the quats and pick one or more representative members of each cluster to be used in toxicity studies, instead of requiring separate studies on each quat. These same questions were raised when the EPA issued its March 4, 1987 Data Call-In Notice requiring all registrants of antimicrobial active ingredients to submit subchronic and chronic toxicological data to support the continued registration of their products.

In response to these questions, EPA solicited information from industry, the public, academia, industry cooperative work groups, the state of California, and Canada. EPA then reviewed all of the assembled information along with the chemical structure of most of the quats. Based on the results of this review, EPA developed the following four groupings of currently registered quat compounds:

Group I. The alkyl or hydroxyalkyl (straight chain) substituted Quats
Group II. The non-halogenated benzyl substituted Quats (includes hydroxybenzyl, ethylbenzyl, hydroxyethybenzyl, napthylmethyl, dodecylbenzyl, and alkyl benzyl)
Group III. The di-and tri-chlorobenzyl substituted
Group IV. Quats with unusual substituents (charged heterocyclic ammonium compounds).

Fundamental to this discussion EPA determined that "X-" in all of these structures would be attributed to "any anionic species". Therefore, this would mean in terms of toxicological evaluation the coutner anion in such quaternary ammonium compounds could be regarded as; e.g halogen (Cl-, Br-, I-,), saccharinate or cyclohexylsulphamate. It is therefore suggested here that nitrite (NO2-) could also be regarded as a pertinent anion.

Since the US EPA deem that such a clustering of structures for toxicological evaluation is well founded and that the counter anion could be regarded as "any anionic species" then it would seem that to consider available toxicological data on sodium nitrite, in order to evaluate any health effects that may be incurred from exposure to the nitrite anion (NO2-), is justifiable.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

no guideline available

Principles of method if other than guideline:

Determination of effects on reproductive parameters, including rates of fertility and perinatal survival, litter sizes, and weanling weights

GLP compliance:

no

Specific details on test material used for the study:

Sodium nitrite

Species:

mouse

Strain:

CD-1

Sex:

female

Details on test animals or test system and environmental conditions:

CD-1 female mice (5-6 weeks old, Charles River Breeding Farms) were housed in groups of 10 and treated as follows. Group 1 received imipramine (Bio-Craft Laboratories) thoroughly mixed (100 mg/kg) with powdered Purina Laboratory Chow. Group 2 received 1 g/l NaNO2 in their drinking water. Group 3 received both imipramine and nitrite. Group 4 received in their water 0.1 ppm DMN diluted twice weekly from a stock solution kept cold and dark and prepared fresh monthly . All mice were housed in plastic cages with hardwood shavings and filter bonnets, in a room at 24 ± 2°C, 40% relativehumidity and a 14/10 hr light/dark cycle.

Route of administration:

oral: drinking water

Vehicle:

water

Details on mating procedure:

After 10 wk males were added. The females were separated to individual cages when a vaginal plug was found and treatment was continued. Near term they were inspected for births once daily, except weekends . Females showing no signs of pregnancy were returned to the males for a second fertilization.The reproductive success of each group was measured by the number of females with surviving litters and the total number of offspring at weaning. Only in the control group did each of the 10 females have surviving offspring. Fewer offspring were weaned in the DMN-, imipramine-, and nitrite-treatment groups, compared with the controls. This effect was of statistical significance for the imipramine group and the nitrite group. However, imipramine and nitrite administered together did not have an additive adverse effect of reproduction; indeed the progeny yield in this group was not significantly differentfrom that in the control group, even though 1 female had no litter.After Exp . 1 indicated potentially interesting effects of the chemicals on reproductive success, Exp. 2 was carried out to study these effects in more detail.

Duration of treatment / exposure:

Exposure period: During mating and pregnancyPremating exposure period (males): no dataPremating exposure period (females): 20 weanling female mice were given tap water for 1 week and then drinking water with NaNO2 at a concentration of 1000 mg/l for the next 75 days (pretreatment period).Duration of test: about 100 days

Frequency of treatment:

Continuously

Dose / conc.:

190 mg/kg bw/day (nominal)

Remarks:

Basis : nominal in water

No. of animals per sex per dose:

20

Control animals:

yes, concurrent no treatment

Parental animals: Observations and examinations:

Preconception water consumption and weight gain, conception time, litter size, rates of stillbirth and neonatal death, histopathology of the major organs of dead pups

Litter observations:

Histopathology of the major organs of dead pups, weaning weights and sex ratios of the offspring

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

Compared to control values, average conception time was 5 days longer, and more females littered more than 30 days after the males were added. Litters were on the average slightly smaller than control.

Perinatal death was more common than among the controls; most of the dead young were stillborn. At weaning the offspring were somewhat smaller than controls, a difference which was statistically significant for males.

Key result

Dose descriptor:

NOAEL

Basis for effect level:

reproductive performance

Remarks on result:

not determinable

Remarks:

no NOAEL identified. Generation not specified (migrated information)

Key result

Reproductive effects observed:

not specified

Conclusions:

No NOAEL could be determined.

Executive summary:

In a publication entitled "Effects of imipramine, nitrite and dimethylnitrosamine on reproduction in mice" (Anderson, L.M., Goner-Sorolla, A., Ebeling, D., Research Communications in Chemical Pathology and Pharmacology Vo. 19 (2), 311 ff, 1978), the fertility of CD-1 mice (females) upon exposure to sodium nitrite was investigated.

This study is assigned a reliability score of 2 in accordance with the criteria for assessing data quality set forth by Klimisch et al (1997).

Under the conditions of the study no NOAEL could be determined.

Reference 3

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Remarks:

based on test type (migrated information)

Adequacy of study:

weight of evidence

Study period:

1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Basic information given

Justification for type of information:

Since in the stomach the gastic juice is acidic, made up of acids and enzymes, it is suggested that in this evironment it is highly unlikely that the quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites substance (s) will remain ionically bound to each other and thus are prone to dissociation in which case the released cation(s) will associate with other anions and the released anion will associate with cations. Thererfore, it is suggested that read-across data from sodium nitrite is considered appropriate in order to arrive at a conclusion in terms of any health effects that may be incurred from exposure to "nitrite" whichever cation it may be associated with.

In addition, in 1988, the US EPA, Office of Pesticides and Toxic Substances issued a Notice to producers, Formulators, Distributors and Registrants regarding quaternary ammonium compounds with regard to "Clustering" of such quaternary ammonium compounds.

Prior to this, EPA had required each quat compound to be individually coded and registered as a new chemical, even when the chemical structure of individual compounds differed only slightly in alkyl distribution and chain lengths. This procedure was continued with the new generations of quats having two, three, and four chains. As a result, EPA records showed that some 211 registered technical grade active ingredient products containing varying concentrations of quats, each coded separately on the basis of alkyl chain length and percentage carbon distribution within the chain. At this time, there are approximately eight to ten thousands (8-10,000) registered end-use formulations.

However, questions were raised regarding whether the EPA could cluster or group the quats and pick one or more representative members of each cluster to be used in toxicity studies, instead of requiring separate studies on each quat. These same questions were raised when the EPA issued its March 4, 1987 Data Call-In Notice requiring all registrants of antimicrobial active ingredients to submit subchronic and chronic toxicological data to support the continued registration of their products.

In response to these questions, EPA solicited information from industry, the public, academia, industry cooperative work groups, the state of California, and Canada. EPA then reviewed all of the assembled information along with the chemical structure of most of the quats. Based on the results of this review, EPA developed the following four groupings of currently registered quat compounds:

Group I. The alkyl or hydroxyalkyl (straight chain) substituted Quats
Group II. The non-halogenated benzyl substituted Quats (includes hydroxybenzyl, ethylbenzyl, hydroxyethybenzyl, napthylmethyl, dodecylbenzyl, and alkyl benzyl)
Group III. The di-and tri-chlorobenzyl substituted
Group IV. Quats with unusual substituents (charged heterocyclic ammonium compounds).

Fundamental to this discussion EPA determined that "X-" in all of these structures would be attributed to "any anionic species". Therefore, this would mean in terms of toxicological evaluation the coutner anion in such quaternary ammonium compounds could be regarded as; e.g halogen (Cl-, Br-, I-,), saccharinate or cyclohexylsulphamate. It is therefore suggested here that nitrite (NO2-) could also be regarded as a pertinent anion.

Since the US EPA deem that such a clustering of structures for toxicological evaluation is well founded and that the counter anion could be regarded as "any anionic species" then it would seem that to consider available toxicological data on sodium nitrite, in order to evaluate any health effects that may be incurred from exposure to the nitrite anion (NO2-), is justifiable.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

no guideline available

Principles of method if other than guideline:

The Reproductive Assessment by Continuous Breeding (RACB) study design has been used by the National Toxicology Program since approx 1981 as a definitive test of reproductive toxicity.

GLP compliance:

not specified

Specific details on test material used for the study:

Sodium nitrite

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

The animals were purchased from Kingston, New York, or Portage, Michigan, and were 11 weeks of age at the start of exposure.Without exception, sentinel mice were negative for murine viruses or viral antibodies. Deionized or filtered water was provided ad lib, and food was either pelleted or ground Purina rodent chow, available ad lib. Photoperiods were 14 :10 light :dark, temperature was maintained at 23 +/- 3°C and relativehumidity was 50 ± 20% .

Route of administration:

oral: drinking water

Vehicle:

water

Details on exposure:

Only the high-dose and control group pups were carried through to breed a second generation. Following weaning, these (F1) animals were exposed to sodium nitrite in their drinking water, at the same dose their parents had received.

Frequency of treatment:

Daily

Dose / conc.:

0 other: %

Remarks:

Basis : nominal in water

Dose / conc.:

0.06 other: %

Remarks:

Equivalent to ca.125 mg/kg bw/day
Basis : nominal in water

Dose / conc.:

0.12 other: %

Remarks:

Equivalent to ca. 260 mg/kg bw/day
Basis : nominal in water

Dose / conc.:

0.24 other: %

Remarks:

Equivalent to ca. 425 mg/kg bw/day
Basis : nominal in water

No. of animals per sex per dose:

20 mated pairs/dose group; 40 mated pairs as controls

Control animals:

yes, concurrent vehicle

Details on study design:

High dose and control pups were continued on to breed a 2nd generation.

Key result

Dose descriptor:

NOAEL

Effect level:

425 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

other: Reproductive toxicity

Key result

Dose descriptor:

NOAEL

Effect level:

260 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

other: General toxicity

Key result

Remarks on result:

other: No information reported on the offspring generation

Key result

Reproductive effects observed:

not specified

No treatment-related effects on: mean number of litters/pair, cumulative days to deliver each litter, mean litter size, birth weight, or pup viability.

No effect on postnatal mortality of pups exposed during lactation.

Reduced body weights of nursing pups.

No effects on reproduction of high-dose F1 animals, or on viability and weight of F2 offspring.

No effects on post-delivery estrous cycle or sperm parameters.

Nine deaths during the study period were not considered to be treatment related.

No effect on F1 terminal body and organ weights.

Conclusions:

The NOAEL for parental animals were determined to be 425 mg/kg bw/day (reproductive toxicity) and 260 mg/kg bw/day (general toxicity).

Executive summary:

In a publication entitled "The Relationships among Reproductive Endpoints in Swiss Mice, Using the Reproductive Assessment by Continuous Breeding Database" ( Chapin, R.E., Sloane, R.A., Haseman, J.K., Fund. Appl. Toxicol. 38, 129 - 142, 1997) the reproductive toxicity effects of sodium nitrite was investigated in a two generation study (primary source).

This publication is also referenced in :-

Toxicology and Carcinogenesis Studies of Sodium Nitrite in F344/N Rats and B6C3f1 Mice (Drinking Water Studies), National Toxicology Program (NTP) Technical Report, (2001; NIH Publication No. 01-3954)

The Screenimg Information DataSet (SIDS) for sodium nitrite as a secondary source and in the disseminated REACH regiatration dossier for sodium nitrite.

This study is assigned a reliability score of 2 in accordance with the criteria for assessing data quality set forth by Klimisch et al (1997).

Under the conditions of the study the NOAEL for parental animals were determined to be 425 mg/kg bw/day (reproductive toxicity) and 260 mg/kg bw/day (general toxicity).

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

56.3 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Effects on developmental toxicity

Description of key information

Read across data is presented on the "fragments" from which this substance is manufactured :-

Quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides, CAS Number 68391-05-9, EC Number 269-924-1 and from sodium nitrite, CAS Number 7632-00-0, EC Number 231-555-9.

The justification in taking this approach is as follows :-

In the stomach the gastic juice is acidic, made up of acids and enzymes. In such an evironment it is highly unlikely that the quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites substance (s) will remain ionically bound to each other and thus are prone to dissociation in which case the released cation(s) will associate with other anions and the released anion will associate with cations.re, it is suggested read-across data from the corresponding quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides is considered approriate in that such substances are likely to dissociate in a similar manner.

Furthermore, in 1988, the US EPA, Office of Pesticides and Toxic Substances issued a Notice to producers, Formulators, Distributors and Registrants regarding quaternary ammonium compounds with regard to "Clustering" of such quaternary ammonium compounds.

Prior to this, EPA had required each quat compound to be individually coded and registered as a new chemical, even when the chemical structure of individual compounds differed only slightly in alkyl distribution and chain lengths. This procedure was continued with the new generations of quats having two, three, and four chains. As a result, EPA records showed that some 211 registered technical grade active ingredient products containing varying concentrations of Quats, each coded separately on the basis of alkyl chain length and percentage carbon distribution within the chain. At this time, there are approximately eight to ten thousands (8-10,000) registered end-use formulations.

However, questions were raised regarding whether the EPA could cluster or group the quats and pick one or more representative members of each cluster to be used in toxicity studies, instead of requiring separate studies on each quat. These same questions were raised when the EPA issued its March 4, 1987 Data Call-In Notice requiring all registrants of antimicrobial active ingredients to submit subchronic and chronic toxicological data to support the continued registration of their products.

In response to these questions, EPA solicited information from industry, the public, academia, industry cooperative work groups, the state of California, and Canada. EPA then reviewed all of the assembled information along with the chemical structure of most of the quats. Based on the results of this review, EPA developed the following four groupings of currently registered quat compounds:

Group I. The alkyl or hydroxyalkyl (straight chain) substituted Quats

Group II. The non-halogenated benzyl substituted Quats (includes hydroxybenzyl, ethylbenzyl, hydroxyethybenzyl, napthylmethyl, dodecylbenzyl, and alkyl benzyl)

Group III. The di-and tri-chlorobenzyl substituted

Group IV. Quats with unusual substituents (charged heterocyclic ammonium compounds).

Fundamental to this discussion EPA determined that "X-" in all of these structures would be attributed to "any anionic species". Therefore, this would mean in terms of toxicological evaluation the coutner anion in such quaternary ammonium compounds could be regarded as; e.g  halogen (Cl-, Br-, I-,), saccharinate or cyclohexylsulphamate. It is therefore suggested here that nitrite (NO2-) could also be regarded as a pertinent anion.

Since the US EPA deem that such a clustering of structures for toxicological evaluation is well founded then it would seem that to consider read-across data from quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides to the closely structurally analogous quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites to be equally justifiable.

Similarly since the US EPA deem that the counter anion could be regarded as "any anionic species" then it would seem that to consider available toxicological data on sodium nitrite, in order to evaluate any health effects that may be incurred from exposure to the nitrite anion (NO2-), is equally justifiable.

To that end two studies are presented from quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides, CAS Number 68391-05-9, EC Number 269-924-1 and two from sodium nitrite,

CAS Number 7632-00-0, EC Number 231-555-9.

Quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides, CAS Number 68391-05-9, EC Number 269-924-1 (1)

A study was conducted to evaluate the prenatal developmental toxicity of the test material (40% didecyldimethylammonium chloride (DDAC) in water) in rabbits by oral gavage route. The study was conducted according to OECD guideline 414 and EPA OPPTS 870.3700 and in compliance with GLP.

Based on the results of this study, there are no specific concerns for developmental toxicity by test material. NOAEL for maternal toxicity and embryotoxic/teratogenic effects of the test material were determined to be 4 and 12 mg/kg/d, respectively.

Quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides, CAS Number 68391-05-9, EC Number 269-924-1 (2)

A teratogenicity study was performed with test material (containing 90.1 % of dialkyldimethylammoniurn chloride) in rats. The study was performed similar or equivalent to OECD guideline 414. The test material was applied topicall to rats at the dose levels of 8, 16.5, 33 mg/rats on Day 6 -15 of the pregnancy.

Under the test conditions, the NOAEL of the test material (containing 90.1 % of dialkyldimethylammoniurn chloride) for the maternal toxicity and

teratogenicity was determined to be 132 mg/kg bw/day in rats.

Sodium nitrite, CAS Number 7632-00-0, EC Number 231-555-9 (1)

In a publication entitled "Effect of Maternally Administered Sodium Nitrite on Hepatic Erythropoiesis in Fetal CD-1 Mice" ( Globus, M., Samuel, D., Teratology 18, 367 - 378, 1978) the developmental toxicity effects of sodium nitrite was investigated. (primary source).

With regard to fetal effects under the conditions of this study, no significant differences between groups for: litter size, weight, resorptions, fetal death, or skeletal abnormalities or variations.Significant changes in hepatic erythropoiesis.No sustained increase in hepatic mature red blood cells on day 18.

However, a NOAEL could not be determined.

Sodium nitrite, CAS Number 7632-00-0, EC Number 231-555-9 (2)

In a publication entitled "Evaluation of the Developmental Toxicity of Sodium Nitrite in Long-Evan Rats." (Roth, A.C., Herkert, G.E., Bercz, J.P., Smith, M.K, Fund. Appl. Toxicol. 9, 668 - 677, 1987) the developmental toxicity effects of sodium nitrite was investigated. (primary source).

Under the conditions of this study administration of 1g NaNO2/Liter resulted in hematological effects but did not affect growth or mortality. NaNO2 (0.5 g/liter) was at or near the no observed effect level.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

4 mg/kg bw/day

Study duration:

subacute

Species:

rat