The relative rates of enzymatic hydrolysis of Benzyl cinnamate were investigated using a preparation of pancreatin. Incubations were performed in 0.5 M phosphate buffer at pH 7.5 and 37°C. The extent of hydrolysis was determined after 2 hours by gas chromatography of the ester. When 18 mg/L Benzyl cinnamate was incubated with simulated intestinal fluid, 80% hydrolysis was measured. [1]

A spot test for mutagenicity based on the Ames test was conducted using Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 with and without S9 from Aroclor-induced rats. A dose of 3 μmol/plate (equivalent to 715 μg/plate) of Benzyl cinnamate in ethanol was applied. The material precipitated under the test conditions. [1]

A rec assay was performed using Bacillus subtilis strains H17 (rec+) and M45 (rec-). A dose of 1000 μg/disk of Benzyl cinnamate in dimethyl sulfoxide produced no effects. [1]

Oral acute toxicity: Benzyl cinnamate dissolved in corn oil was administered via gavage to Charles River rats (5/sex/dose) at dose levels of 2.0, 2.25, 3.0, and 5.0 g/kg body weight. Observations were made for 14 days. [1]

Oral acute toxicity (different strain): Osborne-Mendel rats (5/sex) were dosed orally with Benzyl cinnamate and observed for 14 days. [1]

Oral acute toxicity in guinea pigs: Benzyl cinnamate was tested for toxicity in guinea pigs (number not reported) with observations for deaths between 4 hours and 6 days. [1]

Dermal acute toxicity: Neat Benzyl cinnamate was applied to healthy albino rabbits (4/dose) weighing 2-3 kg for 24 hours under occlusion at dose levels of 1.0, 2.0, and 3.0 g/kg body weight. Applications were made to clipped intact and abraded skin areas, covered with a rubber sleeve or dam. Animals were observed for 14 days. Initial and final hematogram values were compared. [1]

Human repeated insult patch test (HRIPT) for irritation: During the induction phase, 0.3 mL of 4% Benzyl cinnamate in 3:1 diethyl phthalate:ethanol was applied for 24 hours under occlusion using a 25 mm patch on the backs of 101 volunteers. A total of nine induction applications were made on a Monday/Wednesday/Friday schedule. [1]

Maximization pre-test in humans: 8% Benzyl cinnamate in petrolatum was applied for 48 hours under occlusion on the backs of five healthy male volunteers, and separately on five male and female volunteers. [1]

Animal skin irritation pre-test (open epicutaneous): 0.025 mL of Benzyl cinnamate (vehicle not specified) was applied to a 2 cm² area on clipped flank of Himalayan white-spotted guinea pigs (6-8/sex/group). Single application, uncovered, reactions read after 24 hours. Concentrations ranged; 3% was the minimal irritating concentration (lowest concentration producing mild erythema in at least 25% of animals). [1]

Animal skin irritation during induction (open epicutaneous): 0.1 mL of Benzyl cinnamate at a range of concentrations (vehicle not specified) was applied to an 8 cm² area on clipped flank of 6-8 male and female guinea pigs for 21 daily applications. Uncovered, reactions read after 24 hours. Minimal irritating concentration after 21 applications was 3%. [1]

Guinea pig maximization test (GPMT) for sensitization: Outbred white-spotted Himalayan guinea pigs (400-500 g) were used. Induction: intradermal injections on day 0 (each injection given twice) of 0.1 mL of 5% Benzyl cinnamate; 0.1 mL of 5% emulsion of Benzyl cinnamate in Freund’s Complete Adjuvant (FCA); and 0.1 mL of FCA alone. On day 8, topical induction with 25% Benzyl cinnamate in petrolatum for 48 hours under occlusion. On day 21, challenge with a 24-hour closed patch of Benzyl cinnamate at a subirritant concentration in petrolatum applied to clipped flank. Reactions read 24 and 48 hours after patch removal. [1]

Open epicutaneous test in guinea pigs: Benzyl cinnamate at 3% and 8% (vehicle not specified) was evaluated for sensitization. No reactions were observed. Another study reported reactions at 0.3% and 3%; 0.3% was the minimum eliciting concentration, 3% the minimum sensitizing concentration. [1]

Freund's Complete Adjuvant Test (FCAT): Male and female outbred Himalayan guinea pigs (400-500 g) received five intradermal injections of 0.1 mL neat Benzyl cinnamate and FCA as a 50:50 mixture on days 0,2,4,7,9. On days 21 and 35, a 24-hour closed challenge patch at subirritant concentration in petrolatum was applied to the flanks. Sensitization was observed. [1]

Modified FCAT: Female Pirbright white guinea pigs (10/dose) received six intradermal injections of Benzyl cinnamate in physiologic saline containing FCA into the shoulder area on days 1,5,9 (total 4.5 mg test material). Challenge 11 days after induction with open applications of 0.05 mL Benzyl cinnamate at subirritant dose in acetone. Reactions read at 24,48,72 hours. 3% in acetone gave 2/10 reactions; 10% in acetone gave 4/10 reactions. [1]

Modified Draize test: Male and female outbred Himalayan guinea pigs (400-500 g) received 10 intradermal injections on alternate days (starting day 0) with 0.05 mL of 0.1% solution of Benzyl cinnamate in isotonic saline. Challenge on days 35 and 49 with intradermal injection of 0.05 mL of 0.1% solution in saline. Sensitization was observed. [1]

Local Lymph Node Assay (LLNA): Groups of four female CBA mice were tested with Benzyl cinnamate at concentrations of 2.5, 5, 10, 25, or 50% w/v in (1:3) ethanol:diethylphthalate. 25 μL of test material was applied to the dorsum of each ear for three consecutive days. Control group received vehicle. After third application, mice were injected in the tail vein with 250 μL phosphate buffered saline containing approximately 20 μCi of ³H-methyl thymidine (specific activity 2.0 Ci/mmol). After 5 hours, animals were sacrificed; draining auricular lymph nodes were removed, single cell suspensions prepared, washed, suspended in trichloroacetic acid (TCA), precipitated overnight at 4°C, pelleted, resuspended in 1 mL TCA, transferred to scintillation vials with 10 mL scintillant for β-counting. A 3-fold or greater increase in isotope incorporation relative to vehicle control indicated sensitization. The EC3 (concentration giving 3-fold increase) was calculated as 18.4% w/v (4600 μg/cm²). [1]

In vivo percutaneous absorption: Skin absorption of Benzyl cinnamate was studied in five male mice. A 2.2 cm² area on shaved abdominal skin was used. Eserine (0.23%) was used as an indicator, and Benzyl cinnamate as a carrier. The latency between application and appearance of eserine effect in periodically stimulated masticatory muscles was used to measure absorption rate. No evidence of absorption was found. [1]

Subchronic oral toxicity (19-week study): Weanling Osborne-Mendel rats (5/sex/dose) received Benzyl cinnamate by dietary admixture at doses of 0, 1000, and 10,000 ppm (approximately 0, 50, and 500 mg/kg body weight/day) for 19 weeks. Weight, food intake, general condition recorded weekly. Hematological examinations (white cell counts, red cell counts, hemoglobin, hematocrits) at termination. All surviving animals sacrificed and examined macroscopically; organ weights recorded; tissues preserved for histopathology. Detailed microscopic examinations on six or eight animals (evenly divided by sex) in high dose group. Another group of 10 rats (5/sex) were fed same doses for 19 weeks with 20 controls; gross and microscopic examinations performed. [1]

In vitro metabolism: Using pancreatin in simulated intestinal fluid, 80% hydrolysis of 18 mg/L Benzyl cinnamate was measured after 2 hours at pH 7.5 and 37°C. [1]

In vitro skin absorption (animal): Benzyl cinnamate was evaluated as a skin-penetrating agent in excised guinea pig skin by assessing the depth to which Rhodamine B penetrated in the presence of 50% Benzyl cinnamate (vehicle possibly ethylene glycol). Histological findings evaluated in epithelium, hair follicles, corium, and subcutis. After 2 hours, Rhodamine B was slightly detectable in the epithelium. Benzyl cinnamate did not enhance skin penetration of Rhodamine B. [1]

Physical properties relevant to ADME: Calculated Log KOW = 4.06; calculated water solubility = 9.269 mg/L at 25°C; calculated Henry’s Law constant = 0.000000334 atm·m³/mol at 25°C; calculated vapor pressure < 0.001 mmHg at 20°C. [1]

Non-Human Toxicity Values
Oral LD50 in rats: 5.38 g/kg

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Acute oral toxicity (rat, Charles River): LD50 = 3.28 g/kg (19/20 CI 2.62-4.10 g/kg). At 2.0 and 2.25 g/kg, 2/10 deaths; at 3.0 g/kg, 4/10 deaths; at 5.0 g/kg, 8/10 deaths within 24 hours post-dosing with CNS effects observed. [1]

Acute oral toxicity (rat, Osborne-Mendel): LD50 = 5.53 g/kg (95% CI 3.10-7.74 g/kg). Deaths occurred between 4 hours and 5 days; clinical signs included depression and coma persisting 24 hours in some animals. [1]

Acute oral toxicity (guinea pig): LD50 = 3.76 g/kg (95% CI 2.34-6.05 g/kg). Deaths between 4 hours and 6 days; clinical signs included depression, gastrointestinal tract irritation, and rectal bleeding. [1]

Acute dermal toxicity (rabbit): LD50 > 3.0 g/kg. No toxic effects or deaths at doses up to 3.0 g/kg. [1]

Skin irritation – human studies: In HRIPT induction phase (4% in 3:1 DEP:ethanol), 0/101 volunteers showed irritation. In maximization pre-tests (8% in petrolatum), 0/5 and 0/5 volunteers showed irritation. [1]

Skin irritation – animal studies: In rabbits (100% neat), moderate erythema observed on day 1, decreasing after 24 hours, returning to normal after 48 hours. In guinea pigs, 3% was minimal irritating concentration (mild erythema in ≥25% animals) in both single 24-hour and 21-day repeated applications. [1]

Skin sensitization – human predictive studies: HRIPT with 4% (4720 μg/cm²) on 101 volunteers: 0/101 sensitization reactions. Maximization tests with 8% (5520 μg/cm²) on 25 volunteers (RIFM 1975) and 25 male volunteers (RIFM 1972b): 0/25 sensitization in each. [1]

Skin sensitization – human diagnostic studies: Patch test with 5% in petrolatum on 110 eczema patients previously reactive to balsam of Peru: 21/110 reactions. Another study on 103 subjects: 19/103 reactions. Patch test with 8% in petrolatum on 182 patients suspected of cosmetic contact sensitization: 3.2% (6/182) reactions. [1]

Skin sensitization – guinea pig studies: GPMT gave sensitization. Open epicutaneous test: 3% and 8% gave no reactions; however, 0.3% was minimum eliciting concentration and 3% minimum sensitizing concentration in another study. FCAT gave sensitization. Modified FCAT: weak allergen; 3% in acetone gave 2/10 reactions, 10% gave 4/10 reactions. Modified Draize test gave sensitization. [1]

Local Lymph Node Assay (LLNA): EC3 = 18.4% (4600 μg/cm²); Benzyl cinnamate was considered a sensitizer. [1]

Phototoxicity/photoallergy: UV spectra showed peak absorption within 245-278 nm range and minor absorption in 290-320 nm region. No data on phototoxicity or photoallergy. [1]

Subchronic oral toxicity (rat): NOEL = 10,000 ppm (approximately 500 mg/kg body weight/day) in two 19-week studies. No mortalities, adverse clinical signs, effects on growth or hematology, or macroscopic/microscopic changes in tissues. [1]

Mutagenicity/genotoxicity: In modified Ames test (Salmonella typhimurium TA98, TA100, TA1535, TA1537) with and without S9, Benzyl cinnamate at 715 μg/plate precipitated and showed no effects. In rec assay (Bacillus subtilis H17 and M45), 1000 μg/disk produced no effects. [1]

Reproductive and developmental toxicity: No data available. [1]

Carcinogenicity: No data available. [1]

[1]. Fragrance material review on benzyl cinnamate. Food Chem Toxicol. 2007;45 Suppl 1:S40-8. Epub 2007 Sep 14.

Benzyl cinnamate is a cinnamic ester derived from cinnamic acid and benzyl alcohol. It is found in Peruvian balsam and Tolu balsam, as well as Sumatran and Penang benzoin, and is a major component of Cuban balsam. Benzyl cinnamate is commonly used in rich oriental perfumes as a fixative and flavoring agent. It has multiple functions, including flavoring, fragrance, fixative, antigen, and epitope. Benzyl cinnamate has also been reported in Friesodielsia velutina and Isotachis japonica, but relevant data are still unclear.

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Regulatory status: Council of Europe (COE No. 331) – may be used in foodstuffs; FDA approved as flavor (21 CFR 172.515); FEMA GRAS 3 (No. 2142); JECFA No. 670 – no safety concern at current intake as flavoring agent. [1]

Physical form: White to pale yellow fused solid or crystal melting to yellow liquid at very warm room temperature; sweet balsamic odor. Acid value max 1.0; boiling point 350°C; congealing point 33.0-34.5°C; flash point >212°F. [1]

Usage: Maximum skin level in fine fragrances reported as 0.89% (assuming fragrance oil up to 20% in final product). 97.5 percentile use level in cosmetics 0.0854%, resulting in calculated maximum daily skin exposure of 0.0022 mg/kg for high-end users. [1]

IFRA Standard based on QRA: Limits in finished product: Category 1: 0.1%; Category 2: 0.2%; Category 3: 0.7%; Category 4: 2.1%; Category 5: 1.1%; Category 6: 3.4%; Category 7: 0.4%; Category 8: 2.0%; Category 9: 5.0%; Category 10: 2.5%; Category 11: usual concentration of fragrance compound (e.g., if fragrance compound at 5% in candle, then Benzyl cinnamate ≤5%). [1]

Sensitization potency classification: Weight of Evidence NESIL (No Expected Sensitization Induction Level) = 4700 μg/cm²; potency classification = weak. [1]

Conflict of interest: Authors are employees of the Research Institute for Fragrance Materials (RIFM), an independent research institute supported by manufacturers of fragrances and consumer products containing fragrances. This research was supported by RIFM. [1]

C16H14O2

238.2812

238.099

103-41-3

Benzyl cinnamate-d5;347840-02-2

5273469

Off-white to light yellow <34°C powder,>37°C liquid

1.1±0.1 g/cm3

195-200 ºC (5 mmHg)

34-37 °C(lit.)

225.7±10.4 °C

0.0±0.8 mmHg at 25°C

1.4025-1.4045

3.65

0

2

5

18

271

0

O(C(C([H])=C([H])C1C([H])=C([H])C([H])=C([H])C=1[H])=O)C([H])([H])C1C([H])=C([H])C([H])=C([H])C=1[H]

NGHOLYJTSCBCGC-VAWYXSNFSA-N

InChI=1S/C16H14O2/c17-16(12-11-14-7-3-1-4-8-14)18-13-15-9-5-2-6-10-15/h1-12H,13H2/b12-11+

benzyl (E)-3-phenylprop-2-enoate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~419.67 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (10.49 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (10.49 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (10.49 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)