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Toxicol Lett. 2005 Apr 10;156(2):241-51.

The effect on human eye blink frequency of exposure to limonene oxidation products and methacrolein.

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  • 1National Institute of Occupational Health, Lersø Parkallé 105, DK-2100 Copenhagen Ø, Denmark.


Oxidation products of terpenes (e.g. limonene) contain unidentified irritants, which may be responsible for a fraction of the reported eye and airway complaints in indoor environments. Here we report exposure to parts per billion (ppb) levels of limonene oxidation products (LOPs) and the terpene oxidation product methacrolein using blink frequency (BF) as a measure of trigeminal stimulation of the human eye. Ten male subjects averaging 43 (standard deviation 10.5) years were exposed for 20 min to LOPs, methacrolein, and clean air, respectively. A baseline BF was measured prior to and following each exposure (8 min and 4 min, respectively). The subjects were exposed locally in the non-dominant eye and single blind at 20% relative humidity (RH), while viewing an educational film. Blinking was video recorded and evaluated for full sessions of 36 min. Mean BF increased significantly during exposure to LOPs and methacrolein compared to the baseline of clean air, and the findings coincided with weak eye irritation symptoms. Lowest observed effect levels were 286 ppb methacrolein and a 10-min-old LOPs mixture of initially 92 ppb limonene and 101 ppb ozone (O3), which increased the BF comparably by 18% (p=0.001) and 17% (p=0.003), respectively. The increase in BF was smaller, although not significantly different, during exposure to LOPs at 50% RH to 20% RH in mixtures prepared from ca. 350 ppb limonene and 300 ppb O3. LOPs may cause trigeminal stimulation and possibly eye irritation at O3 and limonene concentrations, which are close to high-end values measured in indoor settings. The effects may be exacerbated by low RH.

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