Reaction mass of 1-methyl-4-(1-methylethylidene)cyclohexyl acetate and p-menth-1-en-8-yl acetate

Endpoint summary

Description of key information

Oral-gavage: 28 - 42 days: Terpinyl Acetate multi has a NOAEL of >=250 mg/kg bw based on read across from Terpineol multi which was tested in an OECD 422 study for which effects were seen at 750 mg/kg bw but considered due to gavage dosing (see text for discussion).

Oal-diet: 20 weeks: For support: Terpinyl Acetate multi is expected to have a NOAEL ≥400 mg/kg bw based on read across from Terpinyl Acetate alpha, which was tested in a 20-week study using 10000 ppm in diet.

Inhalation: 90 days: Terpinyl Acetate multi has a NOAEL ≥2.23 mg/L (2230 mg/m3) based on read across from an OECD 413 study with Terpineol multi.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

Terpinyl Acetate multi information is derived from Terpineol multi with Terpinyl Acetate as a bridging study. The adverse effects are considered due to overloading because these were only seen during gavage dosing and not via dietary doses.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

An OECD 413 study performed under GLP.

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

An OECD 413 study performed under GLP.

Additional information

Introduction

For Terpinyl Acetate multi insufficient information is available for covering the repeated dose toxicity. Two tests from Terpineol multi and one from Terpinyl Acetate alpha are used to cover this endpoint. First the executive summaries of the source substances are presented and thereafter the read-across justification.

Terpineol multi: oral -> 28day - Repeated dose toxicity study (OECD TG 422) - First key study because lowest NOAEL is derived from this study

The repeated dose information from Terpineol multi is used as key here because it covers repeated dose as well as fertility and developmental screening.

Method

In the combined repeated dose toxicity study with reproduction/developmental toxicity screening test, conducted following GLP guidelines and OECD guideline 422, three groups, each comprising of ten male and ten female rats for the Main (reproductive) phase and five female rats for the Toxicity phase received Terpineol multi by gavage at doses of 60, 250 or 750 mg/kg bw/day at a dose volume of 5 mL/kg bw/day (Thacker 2010a). Main phase males and Toxicity phase females were dosed daily for a minimum of five consecutive weeks. A similarly constituted Control group received the vehicle, corn oil, at the same volume-dose. An additional ten males and ten females were dosed with the vehicle or at 750 mg/kg/day for five weeks and then given two weeks of recovery before termination. Main phase females were dosed daily for two weeks before pairing, throughout mating, gestation and until Day 6 of lactation. During the study, data was recorded on clinical condition, performance under detailed physical and arena examination, sensory reactivity, grip strength, motor activity, bodyweight, food consumption, water consumption (visual), haematology, blood chemistry, oestrous cycles, mating performance and fertility and gestation length. Organ weight, macroscopic and microscopic pathology investigations were undertaken in the adults. The clinical condition of offspring, litter size and survival, sex ratio and offspring bodyweight were assessed and macroscopic pathology investigations were undertaken.

Results, Mortality, clinical signs and body weight: In the 60 mg/kg/day dose group, one male was found dead on Day 31 of study and one female was killed because of parturition difficulties. In the absence of any other death in the intermediate and high dose groups these deaths are not attributed to the test material. No significant findings were recorded for clinical signs, detailed physical examination and arena observations. Underactive behaviour and unsteady reactions, in males and females were observed briefly during Week 1 in animals receiving 750 mg/kg/day and dose‑related increases in post‑dosing salivation and chin rubbing were seen. There were no clear effects on bodyweight in males or unmated females receiving up to 750 mg/kg/day. Males receiving 750 mg/kg/day showed lower overall weight gain (Week 0-5) compared with Control. Bodyweight during the recovery phase was similar to Controls. Bodyweight and bodyweight gain were unaffected during gestation. During lactation females receiving 250 mg/kg/day showed lower weight gain than Controls. There were no adverse effects on food consumption in males, unmated females or females during gestation and lactation but visual assessment of water consumption indicated that males and females receiving 750 mg/kg/day were consuming more water than the Controls during the dosing period.

Results organ weights: At 750 mg/kg/day, relative liver weights were significantly higher than Control in males and females and relative kidney weights were significantly higher than Control in males. Testis weight was markedly low in males receiving 750 mg/kg/day and there was also an indication of low epididymal weights at this dose. Liver and kidney weights returned to normal after two weeks when the animals did not receive Terpineol multi but testis and epididymal weights showed no evidence of recovery.

Results histopathology on liver and kidney: Adaptive centrilobular hepatocyte hypertrophy in the liver of females dosed with Terpineol multi at 750 mg/kg/day was not present after 2 weeks recovery and histopathological findings in the kidneys of males receiving 250 and 750 mg/kg/day also resolved after the end of dosing.

Results male reproductive organs: At 750 mg/kg/day, reduced numbers or complete absence of spermatozoa, accompanied by the presence of degenerate spermatogenic cells in duct(s) were observed in the epididymidis and were still present following the 2‑week recovery period. Spermatocele granuloma(ta) that were seen in two males receiving 750 mg/kg/day and one receiving 60 mg/kg/day were not seen at the end of the recovery period. The significance of this change in the single male receiving 60 mg/kg/day is uncertain as spermatocele granuloma(ta) can occur spontaneously in rats of this age and there was no other degenerative change in the testes or epididymidis of this animal. Moderate to severe seminiferous tubular atrophy/degeneration was seen in the testes of all animals dosed with Terpineol multi at 750 mg/kg/day, accompanied by minimal to moderate spermatid giant cells and minimal to slight seminiferous tubular vacuolation. Similar findings were still evident following the 2‑week recovery period but at a lower incidence and severity suggesting a degree of recovery. However, gavage is considered to be a worst case for this effect (confirmed by diet administration).

Conclusion: The No-Observed-Adverse–Effect-Level (NOAEL) for males is considered to be >=250 mg/kg bw because of the testicular effects seen at 750 mg/kg bw. These effects are considered to be caused by gavage type dosing and no adverse effects of other organs were seen. This means that the maximum tolerable dose in this study is 250 mg/kg bw. The NOAEL of the females is considered to be 750 mg/k/g/day.

Terpineol multi: inhalation - 90 day - Inhalation repeated dose toxicity - Second key study because it covers the required 90 -day repeated dose toxicity needed for this Annex

Introduction and method: In a repeated dose toxicity study conducted according to OECD Guideline 413 and in compliance with GLP, terpineol multi was administered by inhalation-aerosol to groups of Crl:CD(SD) rats (10 rats/sex/ group) by snout-only inhalation exposure at target exposure levels of 0.2, 0.6 and 2 mg/L for 6 hours per day, 5 days per week for 13 weeks. Control animals received air only. Recovery animals were similarly treated for 13 weeks followed by a 4 week off dose period. Control and high dose recovery groups were included (10/sex/group). During the study, clinical condition, body weight, food consumption, ophthalmoscopy, haematology (peripheral blood), blood chemistry, organ weight, macropathology and histopathology investigations were undertaken.

Results exposure analysis: The achieved levels were 101, 95 and 112% of the target concentrations of 0.2, 0.6 and 2 mg/L, respectively (achieved concentrations 0.202, 0.572 and 2.23 mg/L).MMAD: <0.52, 0.7 and 1.6 µm for achieved concentrations of 0.202, 0.572 and 2.23 mg/L, respectively. GSD: 2.99 and 1.75 for achieved concentrations of 0.572 and 2.23 mg/L, respectively. MMAD showed a general increase with increasing aerosol concentration. The MMAD for the low dose group could not be calculated, as virtually all the measurable test material was captured on the final filter stage, and the value presented is based on the cut point of the penultimate impactor stage. The mid dose groups particle size distribution values showed a bi-modal distribution with an average of 49% of the captured droplet having a MMAD below 0.52 µm. The MMAD value for the high dose group was within the ideal range (1 to 3 µm), indicating that the aerosol was respirable to the rats. The MMADs for the low and mid dose groups were below the ideal range of 1 to 3 µm. However, since the delivered aerosol was a liquid, it is likely that those inhaled droplets with an aerodynamic diameter below 1 µm would still have impacted on airway surfaces and not been exhaled.

Results repeated dose: There were no treatment related deaths or effects on food consumption, blood chemistry, ophthalmoscopy, organ weights or macropathology findings. Group mean body weight gains were lower than control for males exposed to 0.202 mg/L and for both sexes exposed to 0.572 and 2.23 mg/L. In both sexes, no relationship between exposure concentration and body weight gain was observed but the decrease in mean body weight gain was statistically significant for males exposed to 2.23 mg/L. Body weights showed full recovery for animals previously exposed to 2.23 mg/L. Clinical pathology measurements following 13 weeks of exposure revealed statistically significantly lower group mean reticulocyte percentages and absolute counts for males exposed to 0.572 or 2.23 mg/L, compared to control (as low as 0.82X control). A similar effect was observed for females exposed to 2.23 mg/L (as low as 0.87X control) but this did not attain statistical significance. During Recovery Week 4, values for both sexes previously exposed to 2.23 mg/L were similar to controls. Histopathological changes related to treatment were observed in the nasal turbinates for the majority of animals given the test substance and nasal pharynx for a limited number of animals given 0.572 or 2.23 mg/L. In the nasal turbinates, minimal to slight hyperplasia of the mucous cells in the respiratory epithelium was present at all exposure levels and did not exhibit a clear dose response in terms of incidence or severity in males, although there were slightly higher incidences in females at 0.572 mg/L or 2.23 mg/L compared with females exposed to 0.202 mg/L. After 4 weeks of recovery, partial recovery was evident, in terms of the severity observed. After 13 weeks of exposures, these changes were generally not associated with an inflammatory cell infiltrate or cellular degeneration; such changes were observed in a minority of animals and were of minimal severity. After 4 weeks of recovery, degeneration and inflammation of the respiratory epithelium showed completed recovery. However, degeneration of the olfactory epithelium showed only partial recovery in terms of the incidence observed. In the nasal pharynx, minimal hyperplasia of the mucous cells was also observed but at a lower incident and severity and was only evident for animals exposed to 0.572 or 2.23 mg/L. Complete recovery from this effect was observed following the 4 week recovery period. The aforementioned changes were typically associated with chronic exposure to an irritant material and are not considered adverse at the incidence and severity seen. Therefore, the No Observed Adverse Effect Concentration (NOAEC) was considered to be ≥2.23 mg/L (2230 mg/m3).

Terpinyl Acetate Alpha: oral - 20 weeks repeated dose toxicity study - Supporting and bridging study between Terpineol multi and Terpinyl Acetate multi

In a 20 -weeks oral exposure study Osborne-Mendel rats (10/dose/sex) were administered alpha-Terpinyl Acetate via diet at concentrations of 0 (control), 10000, 2500 and 1000 ppm. Animals were then observed for mortality, weight, food intake and general condition (Hagan 1967). Haematological examinations were made at termination. At the termination of the experiments the rats were sacrificed and exsanguinated. The tissues of all the rats were examined macroscopically at the time of sacrifice. The viscera were removed and the liver, kidneys, spleen, heart, and testes were weighed. Detailed microscopic examinations were done on the animals from the high dose group and the control group. No effect on growth or haematology, and no macroscopic or microscopic change in the tissues in the 10000 ppm exposure group were observed. No effect on growth or haematology, and no macroscopic change in the tissues in the 2500 and 1000 ppm exposure groups. No microscopic examination was performed on rats exposed to 2500 and 1000 ppm. These values were not converted into mg/kg bw in the publication. Therefore the ECHA guidance was used, according to R8 table R 8 -17, rats eat 40 to 50 mg/kg bw/day. 10000 ppm is equivalent to 1% in the diet. Consequently, rats exposed to 10000 ppm in the diet consumed between 400 and 500 mg alpha-Terpinyl Acetate per kg bodyweight per day. Therefore, the NOAEL was calculated to be 400 mg/kg bw.

The repeated dose toxicity of Terpinyl Acetate multi using read across from Terpineol multi

Introduction and hypothesis for the analogue approach

The constituents of Terpinyl Acetate multi have a cyclohexyl ring with an unsaturated bond with a methyl group and a tertiary acetate attached on the para-position. For Terpinyl Acetate multi insufficient repeated dose toxicity information is available.

In accordance with Article 13 of REACH, lacking information should be generated whenever possible by means other than vertebrate animal tests, i.e. applying alternative methods such as in vitro tests, QSARs, grouping and read-across. For assessing the repeated dose toxicity of Terpinyl Acetate multi, the analogue approach is selected because for the key metabolite repeated dose toxicity information is available which can be used for read across. This type of analogue approach reflects the RAAF-1 approach: (Bio)transformation to a common metabolite.

Hypothesis considering the outcome of the read across: Terpinyl Acetate multi has the same repeated dose toxicity as Terpineol multi resulting in the same effects and NOAELbecause Terpinyl acetate will not reach the systemic circulation. The acetate will be metabolized into the alcohol and/or its ring will become similarly oxidised/hydroxylatedas Terpineol multi. Conversion considering molecular weight is not needed because Terpinyl Acetate multi has a higher molecular weight and the results for Terpineol multi can be considered conservative.

The hypothesis considering the adverse outcome pathway and key events are: 1) Terpinyl Acetate multi will be metabolised into Terpineol multi and other hydroxylated products; 2) Metabolic overload can occur at > 600 mg/kg bw Terpineol and; 3) The toxicity can be seen by liver effects and testicular atrophy.

Available kinetic experimental information: The Terpinyl Acetate multi plasma DT50 of ca 50 minutes and its DT50 in acidic media of ca 10 minutes are used to predict the absence of Terpinyl Acetate in the systemic circulation (key event 1, See IUCLID Toxico-kinetic section). No metabolites or degradation products were seen in the chromatograms including Terpineol, which is likely a limitation in the detection methodology. This study receives Klimisch score 2 because it is not a GLP study and information on degradation products is missing. Therefore, for further reasoning of this absence of the acetate and formation of the alcohol, metabolism data on Linalyl acetate is used for support.

Available repeated dose toxicity information: The repeated dose toxicity of Terpineol multi is determined according to OECD TG 422 and receives reliability 1. The substance caused non-adverse liver and adverse testicular toxicity at 750 mg/kg bw and the NOAEL for repeated and reproductive toxicity was set at >=250 mg/kg bw based on additional studies showing that gavage dosing caused the testicular effect. In the 90-day repeated dose inhalation toxicity test no adverse effects were seen at 2230 mg/m3 (OECD TG 413). The NOAEL of this inhalation study can be converted to ca 400 mg/kg bw Terpineol (1 mg/kg bw is 0.0052 mg/l air). The bridging study between Terpineol multi and Terpinyl Acetate multi is the 20-wk dietary study from Hagan (1967) with Terpinyl Acetate alpha, the key constituent of Terpinyl Acetate Multi, in which liver and testes were weighed and no effects were seen at the highest dose tested of 400 mg/kg bw (Klimisch 2, non OECD guideline and non-GLP, conversion from 10000 ppm to 400 mg/kg bw (R8, Table R8-17 ECHA guidance).

2. Target chemical and source chemical(s)

Chemical structures of the target chemical and the source chemicals are shown in the data matrix.

3. Purity / Impurities

In table 1 the constituents of Terpinyl Acetate multi are presented as well as Terpineol multi to show the overlap in concentrations between constituents in the substances.

Table 1: The constituents of Terpineol multi and Terpinyl Acetate multi

Terpinoids in %	Alpha-Terpineol	Gamma-Terpineol	cis-beta-Terpineol	trans-beta-Terpineol	Alpha-Terpinyl Acetate	Gamma-Terpinyl Acetate	cis-beta- Terpinyl Acetate	trans-beta Terpinyl Acetate
Cas nu	98-55-5	586-81-2	138-87-4,generic	138-87-4, generic	80-26-2	10235-63-9	2777-47-3	59632-85-8
Terpineol multi	60	29	9	3	0	0	0	0
Terpinyl Acetate multi	0	0	0	0	64	20	7	4

The target substance Terpinyl Acetate multi consists of Terpinyl Acetate alpha and Terpinyl Acetate gamma, with Terpinyl Acetate beta (trans and cis) as impurities. In Terpineol multi a very similar distribution is present for the alcohols. This means that once the Terpinyl Acetate multi conversion to the alcohols are sufficiently characterised it can be assumed that all constituents are sufficiently addressed.

4. Analogue approach justification

According to Annex XI 1.5 read across can be used to replace testing when the similarity can be based on a common backbone and a common functional group or when there is a common metabolite. When using read across the result derived should be applicable for C&L and/or risk assessment and it should be presented with adequate and reliable documentation, which is presented below.

Analogue justification: For Terpinyl Acetate multi, Terpineol multi is selected as analogue because the constituents of Terpineol multi are the key metabolitesof the constituents of Terpinyl Acetate multi.For Terpineol multi the required information is available. The biotransformation of the alkene terpinoid Linalyl acetate is used to support the read-across justification from the acetate to the alcohol. The key constituent of Terpinyl Acetate multi is Terpinyl Acetate alpha for which a 20-week repeated dose study will be used as bridging study to support the read across, because the study as such is insufficient documented (and some current parameters may be missed) to be used as a stand-alone study.

Structural similarities and differences: The constituents of Terpinyl Acetate multi and Terpineol multi have the same non-saturated cyclohexyl ring, with a methyl group attached to the unsaturated bond. At the opposite side (C4 position) two methyl group and a tertiary acetate or tertiary alcohol is attached, respectively.

Toxico-kinetic, Absorption: The similarities considering absorption between the acetate and the alcohol are less relevant because the read across approach is applicable after the absorption step. For completeness both the acetate and alcohol are fully absorbed based on their physico-chemical characteristics with MW < 200 and log Kow values between 2-7 (see Toxico-kinetic section, and data matrix).

Metabolism: Terpineol Acetate multi will be hydrolysed and/or cleaved by carboxyl esterases in the gastro-intestinal tract, the liver and blood as presented by e.g. Belsito et al. (2008) for cyclic acetates and by Wu et al. (2010) for Terpinyl acetate. To verify this degradation of the acetate experimental tests were performed in which 50% of the Terpinyl acetate multi was cleaved ca. 10 minutes in aqueous solutions with pH 2, 4 and 7. In plasma the half-life was somewhat longer ca. 50 minutes. In the plasma experiment both Terpinyl Acetate alpha and gamma decreased in a similar manner (see toxico-kinetic section for details).

Though the decrease of the Terpinyl Acetate was seen, no formation of any metabolite was seen including the alcohol. This is considered a limitation of the GC-MS MS method used or how the method was applied. To support the acetate-alcohol conversion there is experimental metabolism information on Linalyl Acetate in which Linalool is detected. Terpinyl Acetate is very similar to Linalyl acetate. The only difference is that Terpinyl Acetate has a closed ring and Linalyl acetate has a straight alkene chain to with a tertiary ester is attached (Table 2). The formation of Linalool as identified by Hall (1979) supports the formation of Terpineol from Terpinyl Acetate(EFSA (2011) and kinetic section). Therefore, the read across of Terpinyl Acetate multi from Terpineol can be pursued. Further metabolisation work is ongoing to detect the metabolic products of Terpinyl Acetate multi.

After the cleavage of the ester and/or hydroxylation of the ring, in Phase 2 of the metabolisation, the alcohol will be glucuronidated for excretion (WHO, 1999, WHO, 2000,RIFM, 2016)

Table 2Half-life (DT50 or T ½) are shown for Terpinyl Acetate multi and Linalyl Acetate based on experimental information

	Terpinyl Acetate multi; T1/2 minutes (k elimination in brackets)	Linalyl acetate; T1/2 minutes (k elimination in brackets)
Cas no.	8000-41-7 (generic)	115-95-7
Structural features of metabolism
MW	196	196
Acidic artificial gastric juice	8.8 (water, pH2)	5.5
Pancreatic fluids	7.5 (water, pH4)	52.5
Neutral gastric fluids	10.3 (water, pH7)	121
Intestinal mucosa, blood and liver	51 (plasma, k=0.0137)	T1/2 not reported (k=0.01 to 0.0055)
Reference	IFF, 2020 (CRL)	Hall, 1979 Unpublished report; As sited in EFSA, 2011.

Toxico-dynamics: Terpinyl Acetate multi may have the same liver effects and testicular atrophy effects as Terpineolin case overloading of the metabolic pathway occurs.

For Terpineol it was shown that at gavage doses up to 600 mg/kg bw the metabolic pathway can keep up with increases in gavage doses but at higher doses (750 mg/kg bw) the AUC increased disproportionally, which is a sign of overloading of this metabolic pathway.As a result, toxic metabolite(s) become available in the systemic circulation (ECHA Terpineol multi dossier).That 600 mg/kg bw is indeed a cut off is supported with the Terpineol multi 90-day inhalation study. Absence of testicular toxicity were found at ca 2230 mg/m3, which can be converted to 400 mg/kg bw using a respiration rate for rat of 0.38 and taking into account 6/24 hours and 5/7 days weekly exposureand 100% absorption.

The 20-week dietary Terpinyl Acetate alpha of Hagan (1967) can be a bridging study between Terpinyl Acetate multi and Terpineol multi. It supports the absence of overloading the metabolic pathway up to 400 mg/kg bw(the highest dietary dose in this Hagan study, 10000 ppm)and absence of testicular effects.

Similarities in results for toxicological endpoints other than repeated dose: There are no acute systemic toxicity differences and no genotoxic differences between Terpinyl Acetate multi and Terpineol multi: these substances are non-genotoxic, while they show cytotoxicity at fairly low concentrations.

Uncertainty of the prediction: There is uncertainty on the exact quantitative metabolic pathway of Terpinyl Acetate multi to Terpineol multi. The absence of Terpinyl Acetate multi in the systemic circulation has been experimentally shown in plasma and acidic conditions. However, no degradation products could be identified due to methodological limitations of the test set-up. Therefore, information on Linalyl acetate is used, which is also a tertiary terpinyl acetate and metabolises into Linalool. Further study is currently initiated to find the metabolites of Terpinyl Acetate multi.

The available information on in vitro and in vivo metabolism studies of Terpineol (alpha and multi) indicated that at high gavage dosing an overload mechanism can occur (RIFM (2016) and Terpineol multi dossier on ECHA dissemination site, respectively). This is confirmed with additional dietary dosing at the same doses. When testing Terpinyl Acetate multi at similar high gavage doses, this may occur as well. The dose for Terpinyl Acetate multi would be 955 mg/kg bw when using a conversion from Terpineol (750 mg/kg bw effect dose Terpineol with a MW of 154, result in 955 mg/kg bw Terpinyl Acetate with a MW of 196). No testicular effects were seen for Terpinyl Acetate alpha in the 20-week dietary study at ca 400 mg/kg bw, which is below the converted critical value of 955 mg/kg bw Terpinyl Acetate when using LOAEL conversion from Terpineol. There is uncertainty considering the exact NOAEL for testicular effects, however, it is likely that the NOAEL of >=250 mg/kg bw from Terpineol can be considered conservative for Terpinyl Acetate multi.

5. Data matrix

The relevant information on physico-chemical properties and toxicological characteristics are presented in the Data Matrix.

6. Conclusions for repeated dose toxicity

For Terpinyl Acetate multi insufficient repeated dose information is available. For its key metabolites (constituents of Terpineol multi) such data is present, which can be used for read across. This analogue approach is adequate and reliably presented in the present document and the result can be used for C&L and/or risk assessment.

For assessing the repeated dose toxicity of Terpinyl Acetate multi, the absence of Terpinyl Acetate in the systemic circulation and the formation of Terpineol multi are the basis this assessment. Terpinyl Acetate multi has half-lives in plasma of ca 50 minutes and in aqueous environments of ca 10 minutes. This means that after passage of stomach, gastro-intestinal tract and liver no Terpinyl Acetate multi will be present in the systemic circulation. In absence of any degradation product, additional metabolism information from Linalyl acetate (the open version (alkene tertiary acetate) of Terpinyl acetate) is used in which the formation of Linalool is shown. Therefore, the constituents of Terpinyl Acetate multi are expected to turn into the constituents of Terpineol multi.

For Terpineol there is repeated dose toxicity data available: Repeated dose Reproscreen study according to OECD TG 422 and a 90-day inhalation study according to OECD TG 413. The read across between these two is bridged with repeated dose toxicity data from Terpinyl Acetate alpha (20-week dietary study including histopathology of key organs a.o. testes). The available information on Terpineol multi indicates liver effect and testicular atrophy at high gavage doses (750 mg/kg bw). Available kinetic information on Terpineol multi indicates overload of the metabolic pathway at doses >=600 mg/kg bw. Using all this information for Terpineol multi and Terpinyl Acetate multi this would result in a NOAEL >= 600 mg/kg bw because toxicity caused by saturation of the enzymes in the metabolic pathway do not need to be considered for toxicity. In view of a non adverse liver toxicity and testicular toxicity derived in the OECD TG 422 the NOAEL is considered to be >=250 mg/kg bw, this result will be used for the risk assessment being the lowest and therefore conservative value.

Final conclusion on hazard:Terpineol Acetate multi has a NOAEL for repeated dose toxicity of >=250 mg/kg bw.

Data matrix for the read across to Terpinyl acetate multi from Terpineol multiwith support from Terpinyl Acetate alpha.

Common names	Terpinyl Acetate multi	Terpineol multi	Terpinyl Acetate alpha
	Target	Source	Source supporting
Chemical structures		See Terpineol alpha as key const.	See Terpinyl Acetate multi
CAS no	8000-41-7 (generic)	(8007-35-0, generic)	80-26-2
ECHA registration	904-693-9	701-188-3	201-265-7
Molecular weight	196	154	196
Physico-chemical data		ECHA site	ECHA site
Appearance	Liquid	Liquid	Liquid
Water solubility, mg/l	36	2540	36
Log Kow	4.4	2.6	4.4
Human health endpoints
Toxico-kinetic, metabolism	Read across from Linalyl Acetate	Key metabolite of Terpinyl Acetate multi constituents
Acute oral tox in mg/kg bw	Read across from Terpinyl Acetate alpha	>2000 (OECD TG 401)	5075 (Similar to OECD 401)
Genotoxicity – Ames test	Negative	Negative	Negative
Genotoxicity-cytogenicity	Negative	Negative
Genotoxicity_MLA	Negative	Negative
Repeated dose toxicity
28-day Repeated dose/ Reprosceen study via gavage mg/kg bw	Read across from Terpineol multi	NOAEL >=250 (OECD TG 422)
90-day Inhalation study	Read across from Terpineol multi	2230 mg/m3 Ca 400 mg/kg bw (OECD TG 413)
20-week dietary study			NOAEL >=400 mg/kg bw (eqv OECDTG 408)
Fertility
28-day Repeated dose/ Reprosceen study via gavage mg/kg bw	Read across from Terpineol multi	NOAEL >=250 (OECD TG 422)
20-wk dietary study mg/kg bw			No effects on gonads at 400 mg/kg bw (eqv (OECD TG 408)
90-day inhalation study	Read across from Terpineol multi	No effects on gonads (OECD TG 413)
Developmental toxicity
Developmental toxicity in mg/kg bw:	Read across from Terpineol multi	600 (OECD TG 414)

References

Belsito, D., Bickers, D., Bruze, M., Calow, P., Greim, H., Hanifin, J.M., Rogers, A.E., Saurat, J.H., Sipes, I.G., Tagami, H., 2008, A toxicologic and dermatologic assessment of cyclic acetates when used as fragrance ingredients, Food and Chemical Toxicology 46, Suppl 12:S1-27.

EFSA, 2011, Aliphatic, alicyclic and aromatic and unsaturated tertiary alcohols, aromatic tertiary alcohols and their esters from chemical group 6 and 8,https://efsa.onlinelibrary.wiley.com/doi/epdf/10.2903/j.efsa.2011.1847 

RIFM, 2016, Alpha Terpineol: Metabolism in rats after single oral (gavage) exposure, unpublished report, RIFM no 70825.

WHO, 1999, Food additive series 42, 1999, Evaluation of certain food additives, Aliphatic acyclic and alicyclic terpenoid tertiary alcohols and structurally related compounds,http://www.inchem.org/documents/jecfa/jecmono/v042je17.htm

WHO, 2000, Evaluation of certain food additives, Technical Report Series 891, page 51-54,http://whqlibdoc.who.int/trs/WHO_TRS_891.pdf.

Wu, S., Blackburn, K., Amburgery, J., Jaworska, J., and Federle, T., 2010, A framework for using structural, reactivity, metabolic and physico-chemical similarity to evaluate the suitability of analogs for SAR-based toxicological assessments, Regul. Toxicol. Pharmacol., 56, 67-81.

 

Justification for classification or non-classification

Based on all available data Terpinyl Acetate multi does not need to be classified and labelled for its repeated dose toxicity according to EU CLP (EC No. 1272/2008 and its amendments).