Di-(2-ethylhexyl) adipate (DEHA) and phthalates are commonly used as plasticizers to soften polyvinyl chloride products. Because both DEHA and certain phthalates have been identified as priority chemicals for assessment of human health risk under the Government of Canada's Chemicals Management Plan, a comprehensive targeted survey was conducted to investigate the occurrence of DEHA and eight phthalates (di-methyl phthalate, di-ethyl phthalate, di-n-butyl phthalate, di-iso-butyl phthalate, butyl benzyl phthalate, di-n-hexyl phthalate, d-(2-ethylhexyl) phthalate, and di-n-octyl phthalate) in a total of 118 samples of meat (beef, pork, and chicken), fish, and cheese packaged mostly in cling films. The eight phthalates were not detected in any of the food packaging, but DEHA was detected in most of the cling films, indicating that although DEHA-plasticized films (e.g., polyvinyl chloride film) are currently being used by most grocery stores, nonplasticized cling films such as polyethylene film, are also being used by some stores. DEHA was not detected in any of the 10 cheese samples packaged in nonplasticized rigid plastics but was detected in all 30 cheese samples packaged in DEHA-plasticized cling films at levels from 0.71 to 879 μg/g, with an average of 203 μg/g. Only DEHA was detected in the beef, pork, chicken, and fish samples packaged in DEHA-plasticized cling films but at considerably lower levels than those found in cheese, with averages of 6.3, 9.1, 2.5, and 5.9 μg/g, respectively. Among the eight phthalates, only di-(2-ethylhexyl) phthalate (DEHP) was detected in a few cheese samples at levels from 0.29 to 15 μg/g, with an average of 2.8 μg/g; these levels were very likely due to environmental contamination. Levels of DEHA found in most of the cheese samples from this study are above the European specific migration limit of 18 mg/kg for DEHA in food or food simulants, and levels of phthalates (i.e., DEHP) were low.
Listeria monocytogenes is an emerging foodborne pathogen that is responsible for about 28% of the food-related deaths in the United States. It causes meningitis, septicaemia and in pregnant women, abortions and stillbirths. It secretes the toxin listeriolysin O (LLO) that allows the bacteria to enter the cytoplasm of host cells, where they can replicate and cause further infection. The rapid and sensitive detection of LLO in food samples is a key to monitoring and prevention of listeriosis. To facilitate the development of an assay for the specific detection of LLO, a source of LLO is essential. We outline a method of producing a large amount of functional LLO by expressing the hlyA gene (encoding LLO) in Escherichia coli and purifying the recombinant LLO using a one-step purification method. Purification of the protein takes only about 4 h. We compared three different expression constructs for the production of the toxin, which tends to interact strongly with a number of column surfaces. The first construct, using an intein fusion system, could not be purified from the column. The second LLO construct contained an N-terminus His tag; it gave a yield of 3.5-8 mg l(-1). The third contained a C-terminus His tag; it gave a yield of 2.5 mg l(-1) LLO. The purified LLO from the latter two constructs retained its activity at 4 degrees C for over a year as determined by bovine red blood cell hemolysis assay. This paper provides a much-needed, high-yield, one-step purification method of recombinant LLO, and is the first to provide evidence of long-term stability of the toxin for further applications.
Polyvinyl chloride (PVC) food-wrapping films plasticized with di-(2-ethylhexyl) adipate (DEHA) are commonly used by grocery stores in Canada to rewrap meat, poultry, fish, cheese, and other foods. DEHA was assessed as part of the Government of Canada's Chemicals Management Plan. The main source of exposure for most age groups was expected to be food. Although the margin of exposure from food and beverages is considered to be adequately protective, the Government of Canada committed to performing targeted surveys of DEHA in foods and food packaging materials to better define Canadian exposure to DEHA through dietary intake. In order to determine whether more-comprehensive targeted surveys on DEHA in foods should be conducted, 26 food composite samples from the 2011 Canadian total diet study were selected and analyzed for DEHA using a method based on solvent and dispersive solid-phase extraction and gas chromatography-mass spectrometry. These 26 food composites include cheese, meat, poultry, fish, and fast foods, and PVC films were likely used in packaging the individual foods used to make the composites. DEHA was detected in most of the meat, poultry, and fish composite samples, with the highest concentration found in ground beef (11 μg/g), followed by beef steak (9.9 μg/g), freshwater fish (7.8 μg/g), poultry liver pâté (7.4 μg/g), fresh pork (6.9 μg/g), cold cuts and luncheon meats (2.8 μg/g), veal cutlets (2.1 μg/g), roast beef (1.3 μg/g), lamb (1.2 μg/g), and organ meats (0.20 μg/g). Targeted surveys should be conducted to investigate the presence of DEHA in various foods packaged with PVC films in more detail and provide updated occurrence data for accurate human exposure assessment.
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