p-Cymene, a natural antioxidant, in Canadian total diet foods: occurrence and dietary exposures.

BACKGROUND: There is a lack of information on the presence of volatile organic compounds including p-cymene in foods for dietary exposure assessment. p-Cymene is a monoterpene found in many plant species, which has been used as a flavouring agent in foods and also exhibits antioxidant and antimicrobial properties. While the Joint FAO/WHO Expert Committee on Food Additives (JECFA) has no safety concerns at current levels of intake when used as a flavouring, the current study examines p-cymene levels in foods in general, not just from possible uses as a flavouring, as there could still be a potential health concern if there were high levels of exposure. RESULTS: 159 food composite samples from the 2014 Canadian Total Diet Study were analysed for p-cymene using a gas chromatographic-mass spectrometric method after headspace solid-phase microextraction. Concentrations of p-cymene in the composite samples of most food types were generally low, with a maximum level of 73.5 µg g-1 , detected in the composite sample of herbs and spices. Dietary exposures to p-cymene were estimated for different age groups of Canadian populations. CONCLUSIONS: Although p-cymene was detected in various foods, estimated dietary exposures to p-cymene for different age groups are well below the human intake threshold of toxicological concern established by JECFA of 1800 µg per person per day for structural class I flavours, which includes p-cymene. © 2019 Society of Chemical Industry.

Bisphenol A induces DSB-ATM-p53 signaling leading to cell cycle arrest, senescence, autophagy, stress response, and estrogen release in human fetal lung fibroblasts.

Experimental and/or epidemiological studies suggest that prenatal exposure to bisphenol A (BPA) may delay fetal lung development and maturation and increase the susceptibility to childhood respiratory disease. However, the underlying mechanisms remain to be elucidated. In our previous study with cultured human fetal lung fibroblasts (HFLF), we demonstrated that 24-h exposure to 1 and 100 µM BPA increased GPR30 protein in the nuclear fraction. Exposure to 100 µM BPA had no effects on cell viability, but increased cytoplasmic expression of ERß and release of GDF-15, as well as decreased release of IL-6, ET-1, and IP-10 through suppression of NFκB phosphorylation. By performing global gene expression and pathway analysis in this study, we identified molecular pathways, gene networks, and key molecules that were affected by 100, but not 0.01 and 1 µM BPA in HFLF. Using multiple genomic and proteomic tools, we confirmed these changes at both gene and protein levels. Our data suggest that 100 µM BPA increased CYP1B1 and HSD17B14 gene and protein expression and release of endogenous estradiol, which was associated with increased ROS production and DNA double-strand breaks, upregulation of genes and/or proteins in steroid synthesis and metabolism, and activation of Nrf2-regulated stress response pathways. In addition, BPA activated ATM-p53 signaling pathway, resulting in increased cell cycle arrest at G1 phase, senescence and autophagy, and decreased cell proliferation in HFLF. The results suggest that prenatal exposure to BPA at certain concentrations may affect fetal lung development and maturation, and thereby affecting susceptibility to childhood respiratory diseases.

Bisphenol A exposure alters release of immune and developmental modulators and expression of estrogen receptors in human fetal lung fibroblasts.

Bisphenol A (BPA) has been shown to exert biological effects through estrogen receptor (ER)-dependent and ER-independent mechanisms. Recent studies suggest that prenatal exposure to BPA may increase the risk of childhood asthma. To investigate the underlying mechanisms in the actions of BPA, human fetal lung fibroblasts (hFLFs) were exposed to varying doses of BPA in culture for 24hr. Effects of BPA on localization and uptake of BPA, cell viability, release of immune and developmental modulators, cellular localization and expression of ERα, ERß and G-protein coupled estrogen receptor 30 (GPR30), and effects of ERs antagonists on BPA-induced changes in endothelin-1 (ET-1) release were examined. BPA at 0.01-100µmol/L caused no changes in cell viability after 24hr of exposure. hFLFs expresses all three ERs. BPA had no effects on either cellular distribution or protein expression of ERα, however, at 100µmol/L (or 23µmol/L intracellular BPA) increased ERß protein levels in the cytoplasmic fractions and GPR30 protein levels in the nuclear fractions. These paralleled with increased release of growth differentiation factor-15, decreased phosphorylation of nuclear factor kappa B p65 at serine 536, and decreased release of ET-1, interleukin-6, and interferon gamma-induced protein 10. ERs antagonists had no effects on BPA-induced decrease in ET-1 release. These data suggest that BPA at 100µmol/L altered the release of immune and developmental modulators in hFLFs, which may negatively influence fetal lung development, maturation, and susceptibility to environmental stressors, although the role of BPA in childhood asthma remains to be confirmed in in vivo studies.

Chloroform in food samples from 2014 Canadian total diet study: occurrence and dietary exposure.

Chloroform or trichloromethane is one of the trihalomethanes formed during disinfection of water with chlorine, and residues of chloroform can be detected in foods and food products due to the use of chlorinated drinking water and disinfecting food processing equipment with chlorine-based disinfectants. In this study, chloroform was detected in 37 (or 23%) of the 159 composite food samples from the 2014 Canadian Total Diet Study, but was not detected in cereals, fruits, fast foods, and most of the meat samples. Chloroform was detected in almost all 14 composite samples of dairy products, with the highest level (58 ng/g) observed in butter, followed by cream (26 ng/g), and cheese (12-21 ng/g). Chloroform was detected in tap water (23 and 29 ng/g) and most of the beverage samples, but concentrations were lower than those reported in other studies possibly due to evaporation during the preparation of the composite samples. Dietary exposures to chloroform are higher for younger age groups (0.51-1.41 µg/kg body weight/day) than for adults (0.25-0.42 µg/kg body weight/day). Drinking water contributed most to daily intakes for all age groups, accounting for between 62% and 86% of the total chloroform dietary intakes.

Development and Validation of A GC-MS Method Based on Solid Phase Extraction and Derivatization for Analysis of Free and Glucuronide-Conjugated Bisphenol F in Biological Samples.

BACKGROUND: As one of the speculated bisphenols to replace bisphenol A (BPA), bisphenol F (BPF), naturally present in mustard, is structurally similar to BPA and may have similar estrogenic activity, but information on its toxicity is very limited compared to BPA. OBJECTIVE: In order to support the toxicology study of BPF at Heath Canada, a gas chromatography (GC) with mass spectrometry (MS) method based on solid phase extraction and derivatization was developed for analysis of BPF in liver samples. METHODS: Samples were treated with ß-glucuronidase to convert BPF glucuronide to free BPF for analysis of total BPF. RESULTS: The method was validated for free BPF at different spiking levels, and recoveries ranged from 90.0-97.5% with relative standard deviations from 0.11-5.54%. The method was also validated for glucuronide-conjugated BPF at different spiking levels of BPF mono-ß-D-glucuronide, recoveries ranged from 72.3-93.3% with relative standard deviations from 1.7-8.94%. The method was used to analyse 60 liver tissue samples from the rats dosed with BPF at different levels in a toxicology study. Free and glucuronide-conjugated BPF were not detected in any of the control samples which were not dosed with BPF (average method detection limit: 0.31 ng/g), but detected in all the other liver tissue samples with levels increasing at higher doses. The percent of glucuronide-conjugated BPF in total BPF varied among the liver samples, from as low as 9.8% to as high as 77.9%, indicating the importance of analysing biological samples for BPF in both free and conjugated forms for total exposure. CONCLUSION: A GC-MS method based on solid phase extraction and derivatization was developed for analysis of both free and glucuronide-conjugated BPF in liver samples. This method was validated not only for free BPF, but also for mono-ß-D-glucuronide conjugated BPF for the first time to confirm the efficiency of the deconjugation procedure with enzyme. HIGHLIGHTS: This method can be adapted and applied for analysis of free and glucuronide-conjugated BPF in other biological samples with appropriate validation in target sample matrices.

Bisphenol A activates the Nrf1/2-antioxidant response element pathway in HEK 293 cells.

Bisphenol A (BPA) is used in the production of polycarbonate plastics and epoxy resins for baby bottles, liners of canned food, and many other consumer products. Previously, BPA has been shown to reduce the activity of several antioxidant enzymes, which may contribute to oxidative stress. However, the underlying mechanism of the BPA-mediated effect upon antioxidant enzyme activity is unknown. Antioxidant and phase II metabolizing enzymes protect cells from oxidative stress and are transcriptionally activated by Nrf1 and Nrf2 factors through their cis-regulatory antioxidant response elements (AREs). In this work, we have assessed the effect of BPA on the Nrf1/2-ARE pathway in cultured human embryonic kidney (HEK) 293 cells. Surprisingly, glutathione and reactive oxygen species (ROS) assays revealed that BPA application created a more reduced intracellular environment in cultured HEK 293 cells. Furthermore, BPA increased the transactivation activity of ectopic Nrf1 and Nrf2 and increased the expression of ARE-target genes ho-1 and nqo1 at high (100-200 µM) BPA concentrations only. Our study suggests that BPA activates the Nrf1/2-ARE pathway at high (>10 µM) micromolar concentrations.

Cohort profile: the maternal-infant research on environmental chemicals research platform.

BACKGROUND: The Maternal-Infant Research on Environmental Chemicals (MIREC) Study was established to obtain Canadian biomonitoring data for pregnant women and their infants, and to examine potential adverse health effects of prenatal exposure to priority environmental chemicals on pregnancy and infant health. METHODS: Women were recruited during the first trimester from 10 sites across Canada and were followed through delivery. Questionnaires were administered during pregnancy and post-delivery to collect information on demographics, occupation, life style, medical history, environmental exposures and diet. Information on the pregnancy and the infant was abstracted from medical charts. Maternal blood, urine, hair and breast milk, as well as cord blood and infant meconium, were collected and analysed for an extensive list of environmental biomarkers and nutrients. Additional biospecimens were stored in the study's Biobank. The MIREC Research Platform encompasses the main cohort study, the Biobank and follow-up studies. RESULTS: Of the 8716 women approached at early prenatal clinics, 5108 were eligible and 2001 agreed to participate (39%). MIREC participants tended to smoke less (5.9% vs. 10.5%), be older (mean 32.2 vs. 29.4 years) and have a higher education (62.3% vs. 35.1% with a university degree) than women giving birth in Canada. CONCLUSIONS: The MIREC Study, while smaller in number of participants than several of the international cohort studies, has one of the most comprehensive datasets on prenatal exposure to multiple environmental chemicals. The biomonitoring data and biological specimen bank will make this research platform a significant resource for examining potential adverse health effects of prenatal exposure to environmental chemicals.

Intermediate volatile organic compounds in Canadian residential air in winter: Implication to indoor air quality.

Intermediate volatile organic compounds (IVOCs) have recently been characterized for their contributions to the formation of secondary organic aerosol in atmospheric air. However, IVOCs in air in various indoor environments have not been characterized yet. In this study, we characterized and measured IVOCs, volatile organic compounds (VOCs) and semi-volatile organic compounds (SVOCs), in residential indoor air in Ottawa, Canada. IVOCs, including n-alkanes, branched-chain alkanes (b-alkanes), unspecified complex mixtures (UCM) IVOCs, and oxygenated IVOCs (such as fatty acids), were found to have a large impact on indoor air quality. The results indicate that the indoor IVOCs behave differently from those in the outdoor environment. IVOCs in the studied residential air ranged from 14.4 to 69.0 µg/m3, with a geometric mean of 31.3 µg/m3, accounting for approximately 20% of the total organic compounds (IVOCs, VOCs and SVOCs) in indoor air. The total b-alkanes and UCM-IVOCs were found to have statistically significant positive correlations with indoor temperature but have no correlations with airborne particulate matter less than 2.5 µm (PM2.5) as well as ozone (O3) concentration. However, indoor oxygenated IVOCs behaved differently from b-alkanes and UCM-IVOCs, with a statistically significant positive correlation with indoor relative humidity but no correlation with other indoor environmental conditions.

Trends of bisphenol A occurrence in canned food products from 2008-2020.

It is known that the industry already abandoned the use of bisphenol A (BPA)-based epoxy resins as internal coatings for some canned food products (e.g. infant formula, soups). The presence of bisphenol A (BPA) in foods has also been extensively investigated, especially since late 2000. However, information on temporal trends of BPA occurrence in foods is very limited. It is not clear if BPA-based epoxy resins are still being used in internal coating for many other canned foods and if the overall exposure to BPA through consumption of canned foods has significantly decreased. As part of the Canadian total diet study (TDS) program, we have been analysing food samples for BPA since 2008. In this study, results of BPA in samples of different composite canned foods from 2008-2020 TDS were reported. Clear temporal trends were observed for canned fish and soups, with BPA levels being significantly reduced since 2014 for canned fish products and 2017 for canned soups. Temporal trends were not observed for canned evaporated milk, luncheon meats, and vegetables, with even the highest levels of BPA being detected in recent samples for evaporated milk (57 ng/g), luncheon meats (56 ng/g), and baked beans (103 ng/g). This seems to indicate that BPA-based epoxy resins are still being used in the internal coatings for these canned foods products. Thus, analysis of canned food samples for BPA should be continued for exposure assessment.

Challenges and trends in the determination of selected chemical contaminants and allergens in food.

This article covers challenges and trends in the determination of some major food chemical contaminants and allergens, which-among others-are being monitored by Health Canada's Food Directorate and for which background levels in food and human exposure are being analyzed and calculated. Eleven different contaminants/contaminant groups and allergens have been selected for detailed discussion in this paper. They occur in foods as a result of: use as a food additive or ingredient; processing-induced reactions; food packaging migration; deliberate adulteration; and/or presence as a chemical contaminant or natural toxin in the environment. Examples include acrylamide as a food-processing-induced contaminant, bisphenol A as a food packaging-derived chemical, melamine and related compounds as food adulterants and persistent organic pollutants, and perchlorate as an environmental contaminant. Ochratoxin A, fumonisins, and paralytic shellfish poisoning toxins are examples of naturally occurring toxins whereas sulfites, peanuts, and milk exemplify common allergenic food additives/ingredients. To deal with the increasing number of sample matrices and analytes of interest, two analytical approaches have become increasingly prevalent. The first has been the development of rapid screening methods for a variety of analytes based on immunochemical techniques, utilizing ELISA or surface plasmon resonance technology. The second is the development of highly sophisticated multi-analyte methods based on liquid chromatography coupled with multiple-stage mass spectrometry for identification and simultaneous quantification of a wide range of contaminants, often with much less requirement for tedious cleanup procedures. Whereas rapid screening methods enable testing of large numbers of samples, the multi analyte mass spectrometric methods enable full quantification with confirmation of the analytes of interest. Both approaches are useful when gathering surveillance data to determine occurrence and background levels of both recognized and newly identified contaminants in foods in order to estimate human daily intake for health risk assessment.

Temporal trend of bisphenol S occurrence in meat samples from 2008-2020 Canadian total diet study.

In our previous study, bisphenol S (BPS) was detected unexpectedly and at high levels in meat samples from 2016 and 2020 Canadian total diet study (TDS). In this study, samples of meat and meat products from 2008-2015 and 2017-2019 TDS were also analysed to investigate the consistency of BPS occurrence in meat and identify possible trends and provide some information on the potential sources for BPS in meat. BPS was detected again with the highest levels observed in samples of fresh pork (105 ng/g) and veal cutlets (140 ng/g) from the 2008 TDS. This indicates that the occurrence of BPS in meat is not a recent phenomenon but rather an existing one since 2008 or even earlier. BPS concentrations in meat samples from the 2008 to 2020 TDS varied significantly, e.g. 1.2-118 ng/g in roast beef, 1.8-140 ng/g in veal cutlets, but no trend was observed. The lack of trend for BPS over the period of 13 years (2008-2020) does not support the speculation that BPS is being used to replace BPA in food packaging, and sources other than food packaging may be possible and should be investigated for BPS.

Bisphenol S in individual and composite meat and meat products and implication for its sources.

In this study, the occurrence of bisphenol S (BPS) in the meat and meat products from a recent Canadian Total Diet Study (TDS) was investigated in more detail. In addition to their composite samples, the individual raw meat and meat products were also analysed for BPS to investigate the variations of BPS levels and provide some information on the potential sources for BPS in meat. BPS was detected in all the 11 composite samples of different meat and meat products, with the highest level in roast beef (118.23 ng/g) and lowest in cured pork (0.14 ng/g) and cold cuts luncheon meats (0.18 ng/g). BPS was also detected in all the 84 individual raw meat and meat products, with the highest level of 257.61 ng/g in roast beef, followed by 190.41 ng/g in organ meats, 110.15 ng/g in beef steak, 27.91 ng/g in veal cutlets, 17.63 ng/g in wieners & sausages, and 15.27 ng/g in ground beef. However, significant variations of BPS levels were observed in the individual meat and meat product samples under the same category collected from different stores. This may indicate that packaging is unlikely the sources for BPS in meat otherwise BPS levels would have been more or less the same with the same type of packaging (Styrofoam and cling film) regardless where they were collected. Thus, sources other than food packaging, such as the contaminated feed and farming environment (e.g. grass) for animals (e.g. cow), may be possible and should be investigated in future studies.

GC-MS Analysis of Phthalates and Di-(2-thylhexyl) Adipate in Canadian Human Milk for Exposure Assessment of Infant Population.

BACKGROUND: Although more information has become available on the occurrence of phthalates and di(2-ethylhexyl) adipate (DEHA) in foods including cow's milk, information on their presence in human milk, the important and recommended sole diet for infants up to six months of age, is very limited, especially for DEHA. OBJECTIVE: To develop a GC-MS method for simultaneous analysis of DEHA and phthalates in human milk samples and generate occurrence data for exposure assessment. METHOD: Human milk samples were extracted with acetonitrile followed by dispersive solid-phase extraction and GC-MS analysis. RESULTS: Among the 305 human milk samples collected from the Canadian Maternal-Infant Research on Environmental Chemicals Study, some phthalates (DHxP, BBzP, and DOP) were not detected in any of the samples, while DEHA and the other phthalates (DMP, DEP, DBP, DiBP, and DEHP) were detected at low frequencies with levels from 30.4-237 ng/g in up to 31 of the 305 human milk samples. CONCLUSIONS: In general, DEHA and phthalates were detected at low frequencies and low levels in the 305 human milk samples. HIGHLIGHTS: A GC-MS method based on dispersive solid phase extraction was developed for analysis of DEHA and eight phthalates in 305 human milk samples for exposure assessment.

Sources of low concentrations of bisphenol A in canned beverage products.

Although migration from can coatings is likely the source of bisphenol A (BPA) for the canned soft drink products with relatively high BPA concentrations, questions have been raised concerning the exact sources of BPA for those canned soft drink products with low BPA concentrations. Information is also needed for BPA concentrations in canned beer products to conduct proper exposure assessment for BPA under the Government of Canada's Chemicals Management Plan. In this work, 22 soft drink samples and 16 beer samples in both cans and plastic and/or glass bottles were analyzed for BPA. BPA was not detected in any of the soft drink samples in either plastic or glass bottles except for one product with a BPA concentration (0.018 microg/liter) close to the limit of quantification (0.015 microg/liter). BPA was detected in all of the corresponding soft drink products in cans, indicating that migration from can coatings is the likely source for BPA in canned products. Because considerable interference with ions m/z 213 and m/z 228 from sample matrices was observed for all beer samples, BPA concentrations in beer samples were measured using the ion m/z 270 instead. BPA was detected in only one of the seven beer products in glass bottles (0.054 microg/liter) but was detected in all corresponding beer samples in cans at low concentrations ranging from 0.081 to 0.54 microg/liter, indicating that migration from can coatings is likely the source of BPA in canned beer products.

Bisphenol a in canned food products from canadian markets.

A method based on solid phase extraction followed by derivatization and gas chromatography-mass spectrometry analysis was validated for the determination of bisphenol A (BPA) in canned food products. This method was used to analyze 78 canned food products for BPA. Concentrations of BPA in canned food products differed considerably among food types, but all were below the specific migration limit of 0.6 mg/kg set by the European Commission Directive for BPA in food or food simulants. Canned tuna products had the highest BPA concentrations in general, with mean and maximum values of 137 and 534 ng/g, respectively. BPA concentrations in the condensed soup products were considerably higher than those in the ready-to-serve soup products, with mean and maximum values of 105 and 189 ng/g, respectively, for the condensed soups and 15 and 34 ng/g, respectively, for the ready-to-serve soups. BPA concentrations in canned vegetable products were relatively low; about 60% of the products had BPA concentrations of less than 10 ng/g. Canned tomato paste products had lower BPA concentrations than did canned pure tomato products. The mean and maximum BPA concentrations were 1.1 and 2.1 ng/g, respectively, for tomato paste products and 9.3 and 23 ng/g, respectively, for the pure tomato products.

Phthalate Esters in Foods: Sources, Occurrence, and Analytical Methods.

Phthalates are a group of diesters of ortho-phthalic acid (dialkyl or alkyl aryl esters of 1,2-benzenedicarboxylic acid). Higher-molecular-weight phthalates, such as di-2-ethylhexyl phthalate (DEHP), are primarily used as plasticizers to soften polyvinyl chloride (PVC) products, while the lower-molecular-weight phthalates, such as diethyl phthalate (DEP), di-n-butyl phthalate (DBP), and butyl benzyl phthalate (BBzP), are widely used as solvents to hold color and scent in various consumer and personal care products. Phthalates have become ubiquitous environmental contaminants due to volatilization and leaching from their widespread applications, and thus contamination of the environment has become another important source for phthalates in foods in addition to migration from packaging materials. Human exposure to phthalates has been an increased concern due to the findings from toxicology studies in animals. DEHP, one of the important and widely used phthalates, is a rodent liver carcinogen. DEHP, DBP, BBzP, and several phthalate metabolites, such as monobutyl phthalate, monobenzyl phthalate, and mono-(2-ethylhexyl) phthalate, are teratogenic in animals. Since foods are the major source of exposure to phthalates, information on levels of phthalates in foods is important for human exposure assessment. The objective of this review is to identify the knowledge gaps for future investigations by reviewing levels of a wide range of phthalates in a variety of foods, such as bottled water, soft drinks, infant formula, human milk, total diet foods, and others, migration of phthalates from various food-packaging materials, and traditional and new methodologies for the determination of phthalates in foods.

Migration of bisphenol A from can coatings to liquid infant formula during storage at room temperature.

Information on migration of bisphenol A (BPA) from can coatings to foods during storage at room temperature is very limited, and the conclusions from the available studies are not always consistent. To investigate the effect of storage time on BPA migration from can coatings to liquid infant formula at room temperature, samples of 21 canned liquid infant formula products from different cans but the same lot as those analyzed for BPA previously were analyzed for BPA again after storage at room temperature for 10 months. Additional migration of BPA from can coatings to liquid formula during the 10-month storage period at room temperature was observed for 9 of the 21 products, with increases in BPA levels ranging from 29.8 to 110%. Significant differences between the 2007 and 2008 results (P = 0.026) were observed for only one brand of product, which had the lowest BPA levels in the 2007 survey. The BPA levels in the milk-based formula products analyzed in 2008 (mean, 6.8 ng/g) were significantly higher (P = 0.00023) than those in the milk-based formula products analyzed in 2007 (mean, 5.0 ng/g), whereas the differences in BPA levels between the soya-based formula products analyzed in 2008 (mean, 5.3 ng/g) and those analyzed in 2007 (mean, 5.8 ng/g) were not significant (P = 0.097). No obvious correlation between the product expiration date and the level of BPA migration from can coatings was observed.

Levels of bisphenol A diglycidyl ether (BADGE) and bisphenol F diglycidyl ether (BFDGE) in canned liquid infant formula products in Canada and dietary intake estimates.

A method based on solid-phase extraction followed by HPLC analysis with fluorescence detection was developed for the determination of bisphenol A diglycidyl ether (BADGE) and bisphenol F diglycidyl ether (BFDGE) in liquid infant formula. In this method, instead of trying to isolate and measure each individual form of the molecules, hydrolysis of BADGE, BFDGE, BADGE x H2O, and BFDGE x H2O was forced to completion to their stable forms, BADGE x 2H2O and BFDGE x 2H2O, before extraction. The method LODs were 2.0 ng/g for BADGE and 3.0 ng/g for BFDGE. Extraction recoveries were 61-91% for BADGE, and 55-82% for BFDGE over the concentration range of 10 to 50 ng/g. The method was used to analyze samples of 21 canned liquid infant formula products for BADGE and BFDGE. BADGE was detected in samples of all products at levels ranging from as low as 2.4 ng/g to as high as 262 ng/g. BFDGE was detected in only one product (40 ng/g), and this product also had the highest BADGE level (262 ng/g). HPLC/MS/MS with a similar LOD was also used to confirm the results. The probable daily intakes (PDI) of BADGE and BFDGE due to consumption of canned liquid infant formula were estimated for infants from premature to 12-18 months of age. The maximum PDI of BADGE was 22 microg/kg body weight/day for the 12-18 months old with the maximum formula intake. The maximum PDI of BFDGE was < 3.4 microg/kg body weight/day.