Isooctadecyl isooctadecanoate

Endpoint summary

Additional information

There are no experimental studies available in which the toxicokinetic behaviour of isooctadecyl isooctadecanoate (CAS 41669-30-1) has been assessed. In accordance with Annex VIII, Column 1, Item 8.8.1, of Regulation (EC) 1907/2006 and with Guidance on information requirements and chemical safety assessment Chapter R.7c: Endpoint specific guidance (ECHA, 2012), assessment of the toxicokinetic behaviour of the substance is conducted to the extent that can be derived from the relevant available information. This comprises a qualitative assessment of the available substance specific data on physicochemical and toxicological properties according to the relevant Guidance (ECHA, 2012) and taking into account available information on the analogue substances from which data was used for read-across to cover data gaps. 

The substance isooctadecyl isooctadecanoate is an UVCB with an aliphatic, but branched C16-18 -alcohol moiety and a mostly C18iso (isostearic)-acid moiety.

 

It is a liquid at 20 °C (Mir, 2014), with a water solubility of 0.044 mg/L (Frischmann, 2014). The log Pow was calculated to be > 10 (Müller, 2014) and the vapour pressure is < 0.0001 Pa at 20 °C (Nagel, 2014).

 

Absorption

 

Absorption is a function of the potential for a substance to diffuse across biological membrane. The most useful parameters to provide information on this potential are the molecular weight, octanol/water coefficient (log Pow) value and water solubility (ECHA, 2012). The log Pow value provides information on the relative solubility of the substance in water and lipids (ECHA, 2012).

 

Oral

 

The molecular weight of isooctadecyl isooctadecanoate is 508.90 – 536.96 g/mol, which is only moderately in favour of absorption, generally the smaller the molecule (<500 g/mol) the more easily it may be taken up (ECHA, 2008). The high log Pow >10 and low water solubility (0.044 mg/L) of the substance suggests that any absorption will likely be via micecullar solubilisation (ECHA, 2012). The available acute oral toxicity on isooctadecyl isooctadecanoate resulted in LD50 value > 2000 mg/kg bw/d and no systemic effects were seen (Saboureau, D). However, in an OECD 422 study conducted with a structural analogue substance, isodecyl oleate (CAS 59231-34-4), revealed a test item-related decrease in body weight on lactaion day 4 and a decrease in food consumption during the lactation/gestation period in the female animals of the high dose group (1000 mg/kg bw/d) (Hansen, 2013). The effects were considered adverse and leading to maternal toxicity; thus, the NOAEL for systemic toxicity was determined to be 300 mg/kg bw/d in this study. This indicates that isooctadecyl isooctadecanoate does have the potential to be absorbed.

 

The potential of a substance to be absorbed in the gastrointestinal (GI) tract may be influenced by chemical changes taking place in GI fluids as a result of metabolism by GI flora, by enzymes released into the GI fluids tract or by hydrolysis. These changes will alter the physicochemical characteristics of the substance and hence predictions based upon the physio-chemical characteristics of the parent substance may no longer apply (ECHA, 2012).

 

In general, alkyl esters are readily hydrolysed in the gastrointestinal tract, blood and liver to the corresponding alcohol and fatty acid by the enzymatic activity of ubiquitous carboxylesterases. There are indications that the hydrolysis rate in the intestine by action of pancreatic lipase is lower for alkyl esters than for triglycerides, the natural substrate for this substance. The hydrolysis rate of linear esters increases with increasing chain length of either the alcohol or acid. Branching reduces the ester hydrolysis rate, compared with linear esters (Mattson and Volpenhein, 1969, 1972; WHO, 1999). The substance isooctadecyl isooctadecanoate is therefore anticipated to be enzymatically hydrolysed to mostly, isohexadecanol (C16), isooctadecanol (C18) and 16-methylheptadecanoic acid.

 

Free fatty acids and alcohols are readily absorbed by the intestinal mucosa. Within the epithelial cells, fatty acids are (re-)esterified with glycerol to triglycerides. In general, short-chain or unsaturated fatty acids are more readily absorbed than long-chain, saturated fatty acids. In rats given a single dose of radiolabelled octadecanol via duodenal cannula, 56.6 ± 14% of the administered material was absorbed within 24 h. As for fatty acids, the rate of absorption is likely to increase with decreasing chain length (Greenberger et al., 1966; IOM, 2005; Mattson and Volpenhein, 1962, 1964; OECD, 2006; Sieber, 1974).

In conclusion, based on the available information, the physiochemical properties and molecular weight of isooctadecyl isooctadecanoate suggest oral absorption. However, the substance is anticipated to undergo enzymatic hydrolysis in the gastrointestinal tract and absorption of the ester hydrolysis product is also relevant. The absorption rate of the hydrolysis products is considered to be high.

 

Dermal

 

The dermal uptake of liquids and substances in solution is higher than that of dry particulates, since dry particulates need to dissolve into the surface moisture of the skin before uptake can begin. Molecular weights below 100 favour dermal uptake, while for those above 500 the molecule may be too large. Dermal uptake is anticipated to be low, if the water solubility is < 1 mg/L; low to moderate if it is between 1-100 mg/L; and moderate to high if it is between 100-10000 mg/L. Dermal uptake of substances with a water solubility > 10000 mg/L (and log Pow < 0) will be low, as the substance may be too hydrophilic to cross the stratum corneum. Log Pow values in the range of 1 to 4 (values between 2 and 3 are optimal) are favourable for dermal absorption, in particular if water solubility is high. For substances with a log Pow above 4, the rate of penetration may be limited by the rate of transfer between the stratum corneum and the epidermis, but uptake into the stratum corneum will be high. Log Pow values above 6 reduce the uptake into the stratum corneum and decrease the rate of transfer from the stratum corneum to the epidermis, thus limiting dermal absorption (ECHA, 2012).

 

Isooctadecyl isooctadecanoate is almost insoluble in water; indicating a low dermal absorption potential (ECHA, 2012). The molecular weight of 508.90 – 536.96 g/mol is above the 500 g/mol limit above which dermal absorption is low. The log Pow is > 10, which means that the uptake into the stratum corneum is likely to be slow and the rate of transfer between the stratum corneum and the epidermis will also be slow (ECHA, 2012).

If the substance is a skin irritant or corrosive, damage to the skin surface may enhance penetration (ECHA, 2012).

The experimental animal data onisooctadecyl isooctadecanoateshows that no significant skin irritation occurred, which excludes enhanced penetration of the substance due to local skin damage (Dufour, 1991).

Overall, based on the available information, the dermal absorption potential of isooctadecyl isooctadecanoateis predicted to be low.

 

 

Inhalation

 

As the vapour pressure of isooctadecyl isooctadecanoate is very low (< 0.0001 Pa at 20 °C), the volatility is also low. Therefore, the potential for exposure and subsequent absorption via inhalation during normal use and handling is considered to be negligible.

If the substance is available as an aerosol, the potential for absorption via the inhalation route is increased. While droplets with an aerodynamic diameter < 100 μm can be inhaled, in principle, only droplets with an aerodynamic diameter < 50 μm can reach the bronchi and droplets < 15 μm may enter the alveolar region of the respiratory tract (ECHA, 2012).

As for oral absorption, the molecular weight, log Pow and water solubility are suggestive of absorption across the respiratory tract epithelium by micellar solubilisation.

Esterase’s present in the lung lining fluid may also hydrolyse the substance, hence making the resulting alcohol and acid available for inhalative absorption.

An acute inhalation toxicity study was performed with the read-across substance 2-ethylhexyl oleate (CAS 26399-02-0), in which rats were exposed nose-only to 5.7 mg/L of an aerosol for 4 hours (Van Huygevoort, 2010). No mortality occurred and no toxicologically relevant effects were observed. Thus, the test substance is not acutely toxic by the inhalation route, but no firm conclusion can be drawn on respiratory absorption. Due to the limited information available, absorption via inhalation is assumed to be as high as via the oral route in a worst case approach.

 

Distribution and Accumulation

 

Distribution of a compound within the body depends on the physicochemical properties of the substance; especially the molecular weight, the lipophilic character and the water solubility. In general, the smaller the molecule, the wider is the distribution. If the molecule is lipophilic, it is likely to distribute into cells and the intracellular concentration may be higher than extracellular concentration, particularly in fatty tissues (ECHA, 2012).

The substance isooctadecyl isooctadecanoate will mainly be absorbed in the form of the hydrolysis products. The fraction of ester absorbed unchanged will undergo enzymatic hydrolysis by ubiquitous esterases, primarily in the liver (Fukami and Yokoi, 2012). Consequently, the hydrolysis products are the most relevant components to assess. Both hydrolysis products are expected to be distributed widely in the body.

After being absorbed, fatty acids are (re-)esterified along with other fatty acids into triglycerides and released in chylomicrons. Fatty acids of carbon chain length ≤ 12 may be transported as the free acid bound to albumin directly to the liver via the portal vein, instead of being re-esterified. Chylomicrons are transported in the lymph to the thoracic duct and eventually to the venous system. Upon contact with the capillaries, enzymatic hydrolysis of chylomicron triacylglycerol fatty acids by lipoprotein lipase takes place. Most of the resulting fatty acids are taken up by adipose tissue and re-esterified into triglycerides for storage. Triacylglycerol fatty acids are likewise taken up by muscle and oxidised for energy or they are released into the systemic circulation and returned to the liver (IOM, 2005; Johnson, 1990; Johnson, 2001; Lehninger, 1993; Stryer, 1996).

Absorbed alcohols are likewise transported via the lymphatic system. Twenty-four hours after intraduodenal administration of a single dose of radiolabelled octadecanol to rats, the percent absorbed radioactivity in the lymph was 56.6 ± 14. Thereof, more than half (52-73%) was found in the triglyceride fraction, 6-13% as phospholipids, 2-3% as cholesterol esters and 4-10% as unchanged octadecanol. Almost the entire radioactivity recovered in the lymph was localized in the chylomicron fraction. Thus, the alcohol is oxidised to the corresponding fatty acid and esterified in the intestine as described above (Sieber, 1974).

Taken together, the hydrolysis products of isooctadecyl isooctadecanoate are anticipated to distribute systemically. Long-chain alcohols are rapidly converted into the corresponding fatty acids by oxidation and distributed in form of triglycerides, which can be used as energy source or stored in adipose tissue. Stored fatty acids underlie a continuous turnover as they are permanently metabolised for energy and excreted as CO2. Bioaccumulation of fatty acids takes place, if their intake exceeds the caloric requirements of the organism.

 

 

Metabolism

 

The metabolism of isooctadecyl isooctadecanoate initially occurs via enzymatic hydrolysis of the ester resulting in 16-methylheptadecanoic acid and branched C16-18 alcohols.The esterases catalysing the reaction are present in most tissues and organs, with particularly high concentrations in the GI tract and the liver (Fukami and Yokoi, 2012). Depending on the route of exposure, esterase-catalysed hydrolysis takes place at different places in the body. After oral ingestion, esters of alcohols and fatty acids undergo enzymatic hydrolysis already in the gastrointestinal tract. In contrast, substances which are absorbed through the pulmonary alveolar membrane or through the skin may enter the systemic circulation directly before entering the liver where hydrolysis will generally take place.

The C16-18 alcohols will mainly be metabolised to the corresponding carboxylic acid via the aldehyde as a transient intermediate (Lehninger, 1993). The stepwise process starts with the oxidation of the alcohol by alcohol dehydrogenase to the corresponding aldehyde, where the rate of oxidation increases with increased chain-length. Subsequently, the aldehyde is oxidised to carboxylic acid, catalysed by aldehyde dehydrogenase. Both the alcohol and the aldehyde may also be conjugated with e.g. glutathione and excreted directly, by passing further metabolism steps (WHO, 1999).

A major metabolic pathway for linear and branched fatty acids is the beta-oxidation for energy generation. In this multi-step process, the fatty acids are at first esterified into acyl-CoA derivatives and subsequently transported into cells and mitochondria by specific transport systems. In the next step, the acyl-CoA derivatives are broken down into acetyl-CoA molecules by sequential removal of 2-carbon units from the aliphatic acyl-CoA molecule. Further oxidation via the citric acid cycle leads to the formation of H2O and CO2 (Lehninger, 1993). Branched-chain acids can be metabolised via the same beta-oxidation pathway as linear, depending on the steric position of the branch, but at lower rates (WHO, 1999). The alpha-oxidation pathway is a major metabolic pathway for branched-chain fatty acids where a methyl substituent at the beta-position blocks certain steps in the beta-oxidation (Mukherji, 2003). Generally, a single carbon unit is cleaved off the branched acid in an additional step before the removal of 2-carbon units continues. Alternative pathways for long-chain fatty acids include the omega-oxidation at high dose levels (WHO, 1999). The fatty acid can also be conjugated (by e.g. glucuronides, sulfates) to more polar products that are excreted in the urine.

The potential metabolites following enzymatic metabolism of the two main constituents of the substance were predicted using the QSAR OECD toolbox (OECD, 2011). This QSAR tool predicts which metabolites may result from enzymatic activity in the liver and in the skin, and by intestinal bacteria in the gastrointestinal tract. Nine hepatic metabolites and 9 dermal metabolites were predicted for isooctadecyl isooctadecanoate. Primarily, the ester bond is broken both in the liver and in the skin and the hydrolysis products may be further metabolised. Besides hydrolysis, the resulting liver and skin metabolites are all the product of alpha-, beta- or omega-oxidation (= addition of hydroxyl group). In the case of omega-oxidation, it is followed by further oxidation to the aldehyde, which is then oxidised to the corresponding carboxylic acid. In a few cases the ester bond remains intact, and only fatty acid oxidation products are found, which result in the addition of one hydroxyl group to the molecule. In general, the hydroxyl groups make the substances more water-soluble and susceptible to metabolism by phase II-enzymes. The metabolites formed in the skin are expected to enter the blood circulation and have the same fate as the hepatic metabolites. Between 114 and 122 metabolites were predicted to result from all kinds of microbiological metabolism for the ester with C16 alcohol and C18 alcohol, respectively. Most of the metabolites were found to be a consequence of fatty acid oxidation and associated chain degradation of the molecule. The results of the OECD Toolbox simulation support the information retrieved in the literature.

There is no indication that the substance isooctadecyl isooctadecanoate is activated to reactive intermediates under the relevant test conditions. The experimental studies performed on genotoxicity (Ames test, gene mutation in mammalian cells in vitro, chromosome aberration assay in mammalian cells in vitro) using isooctadecyl isooctadecanoate and the read-across substances were negative, with and without metabolic activation (Gattefosse, 2002; Croda, 1998; Emery, 1994). The result of the skin sensitisation study performed with the structural analogue substance, 2-octyldodecyl isooctadecanoate was likewise negative (Croda, 1998).

 

 

 

Excretion

The C16 and C18-fatty acids from the oxidation of the corresponding alcohols will be metabolised for energy generation or stored as lipid in adipose tissue or used for further physiological functions e.g. incorporation into cell membranes (Lehninger, 1993). Therefore, the fatty acid metabolites are not expected to be excreted to a significant degree via the urine or faeces but excreted via exhaled air as CO2 or stored as described above. Experimental data with ethyl oleate (CAS 111-62-6, ethyl ester of oleic acid) support this principle. The absorption, distribution, and excretion of 14C-labelled ethyl oleate was studied in Sprague Dawley rats after a single, oral dose of 1.7 or 3.4 g/kg bw. At sacrifice (72 h post-dose), mesenteric fat was the tissue with the highest concentration of radioactivity. The other organs and tissues had very low concentrations of test material-derived radioactivity. The main route of excretion of radioactivity in the groups was via expired air as CO2. 12 h after dosing, 40-70% of the administered dose was excreted in expired air (consistent with β -oxidation of fatty acids). 7-20% of the radioactivity was eliminated via the faeces, and approximately 2% via the urine (Bookstaff et al., 2003).

In addition, the alcohol components may also be conjugated to form a more water-soluble molecule and excreted via the urine (WHO, 1999). In an alternative pathway, the alcohol may be conjugated with e.g. glutathione and excreted directly, bypassing further metabolism steps.

In contrast, the isononanoic acid is unlikely to be used for energy generation and storage, since saturated aliphatic, branched-chain acids are described to be subjected to omega-oxidation due to steric hindrance by the methyl groups at uneven position, which results in the formation of various diols, hydroxyl acids, ketoacids or dicarbonic acids. In contrast to the products of beta-oxidation, these metabolites may be conjugated to glucuronides or sulphates, which subsequently can be excreted via urine or bile or cleaved in the gut with the possibility of reabsorption (entero-hepatic circulation) (WHO, 1998).

 

A detailed reference list is provided in the technical dossier (see IUCLID, section 13) and within CSR.