Duck Soo Lim

Safety Evaluation And Risk Assessment Of d-Limonene

d-Limonene, a major constituent of citrus oils, is a monoterpene widely used as a flavor/fragrance additive in cosmetics, foods, and industrial solvents as it possesses a pleasant lemon-like odor. d-Limonene has been designated as a chemical with low toxicity based upon lethal dose (LD50) and repeated-dose toxicity studies when administered orally to animals. However, skin irritation or sensitizing potential was reported following widespread use of this agent in various consumer products. In experimental animals and humans, oxidation products or metabolites of d-limonene were shown to act as skin irritants. Carcinogenic effects have also been observed in male rats, but the mode of action (MOA) is considered irrelevant for humans as the protein α2u-globulin responsible for this effect in rodents is absent in humans. Thus, the liver was identified as a critical target organ following oral administration of d-limonene. Other than the adverse dermal effects noted in humans, other notable toxic effects of d-limonene have not been reported. The reference dose (RfD), the no-observed-adverse-effect level (NOAEL), and the systemic exposure dose (SED) were determined and found to be 2.5 mg/kg/d, 250 mg/kg//d, and 1.48 mg/kg/d, respectively. Consequently, the margin of exposure (MOE = NOAEL/SED) of 169 was derived based upon the data, and the hazard index (HI = SED/RfD) for d-limonene is 0.592. Taking into consideration conservative estimation, d-limonene appears to exert no serious risk for human exposure. Based on adverse effects and risk assessments, d-limonene may be regarded as a safe ingredient. However, the potential occurrence of skin irritation necessitates regulation of this chemical as an ingredient in cosmetics. In conclusion, the use of d-limonene in cosmetics is safe under the current regulatory guidelines for cosmetics.

Potential Risk of Bisphenol a Migration From Polycarbonate Containers After Heating, Boiling, and Microwaving

The migration levels of bisphenol A (BPA) were analyzed in food samples by high-performance liquid chromatography (HPLC) from polycarbonate (PC) bottles subjected to simulated use by heating with microwave, heating in a boiling water bath, or filling them with boiling hot water (100°C). Migration testing performed in PC bottles filled with steamed rice or hot cooked pork, standing at room temperature, or heated in a boiling water bath (100°C) showed that BPA was not detected at the limit of detection (LOD) of 1 μg/L (ppb). In contrast, heating by microwaving to 100°C for 9 min increased BPA migration levels from 6 to 18 ppb and from 5 to 15 ppb for steamed rice or for cooked pork, respectively. In addition, 3 different PC bottles were tested by filling them with boiling hot water (100°C) and leaving them to stand at room temperature for up to 3 h. The mean BPA levels from the bottles increased in a time-dependent manner, with the range of not detected (ND) to 2.5 ppb after 60 min. However, none of the PC bottles released BPA at levels that exceed the recently established specific migration limits (SML) of 600 ppb established by European Union and Korea Food and Drug Administration (KFDA). Data suggest that the use of PC plastic bottles in our daily life is considered safe in Korea.

Risk Assessment of Bisphenol a Migrated from Canned Foods in Korea

Exposure and risk assessment of bisphenol A (BPA) was conducted on consumption of canned foods in Korean adults. Sixty-one canned food items with different brands purchased from retail outlets in markets were analyzed for BPA concentration by high-performance liquid chromatography (HPLC) coupled with fluorescence detection. Limits of detection (LOD) were 3 μg/kg for solid and 2 μg/kg for liquid foods. BPA was detected from 7 groups of food items, such as tuna (n = 8), fish (n = 11), fruits (n = 9), vegetables (n = 12), meats (n = 13), coffee (n = 5), and tea (n = 3) in the range from not detected (ND) to 136.14 μg/kg. Mean concentrations of BPA were 3.1 μg/kg (ND−21.5 μg/kg) for vegetables, 8.3 μg/kg (ND−14.26) for tea, 8.6 μg/kg (ND−54.56 μg/kg) for fruits, 24.49 μg/kg (ND−98.30 μg/kg) for meats, 39.78 μg/kg (ND−125.25 μg/kg) for fish, 43.7 μg/kg (ND−116.88 μg/kg) for tuna, and 45.51 μg/kg (ND−136.14 μg/kg) for coffee, in the order of magnitude. Based on daily dietary intake of canned food items and concentrations of BPA, human exposure level to BPA was estimated to be 1.509 μg/kg body weight (bw)/d, well below the tolerable daily intake (TDI) or reference dose (RfD) of 50 μg/kg, bw/d set by the European Commission, U.S.EPA, and South Korea, . Therefore, the potential risk for BPA contamination due to consumption of each canned food items was calculated to be (1.509 μg/kg bw/d)/(50 μg/kg bw/d) = 0.03, which is the hazard index [HI = exposure level/(RfD or TDI)]. Evidence indicates that the levels of BPA levels in canned foods are not likely to constitute a safety concern for consumers in Korea.

Risk Assessment of Volatile Organic Compounds Benzene, Toluene, Ethylbenzene, and Xylene (BTEX) in Consumer Products

Exposure and risk assessment was performed by evaluating levels of volatile organic compounds (VOC) benzene, toluene, ethylbenzene, and xylene (BTEX) in 207 consumer products. The products were categorized into 30 different items, consisting of products of different brands. Samples were analyzed for BTEX by headspace-gas chromatography/mass spectrometry (headspace-GC/MS) with limit of detection (LOD) of 1 ppm. BTEX were detected in 59 consumer products from 18 item types. Benzene was detected in whiteout (ranging from not detected [ND] to 3170 ppm), glue (1486 ppm), oil-based ballpoint pens (47 ppm), and permanent (marking) pens (2 ppm). Toluene was detected in a leather cleaning product (6071 ppm), glue (5078 ppm), whiteout (1130 ppm), self-adhesive wallpaper (15–1012 ppm), shoe polish (806 ppm), permanent pen (609 ppm), wig adhesive (372 ppm), tapes (2–360 ppm), oil-based ballpoint pen (201 ppm), duplex wallpaper (12–52 ppm), shoes (27 ppm), and air freshener (13 ppm). High levels of ethylbenzene were detected in permanent pen (ND–345,065 ppm), shoe polish (ND–277,928 ppm), leather cleaner (42,223 ppm), whiteout (ND–2,770 ppm), and glue (ND–792 ppm). Xylene was detected in permanent pen (ND–285,132 ppm), shoe polish (ND–87,298 ppm), leather cleaner (12,266 ppm), glue (ND–3,124 ppm), and whiteout (ND–1,400 ppm). Exposure assessment showed that the exposure to ethylbenzene from permanent pens ranged from 0 to 3.11 mg/kg/d (men) and 0 to 3.75 mg/kg/d (women), while for xylene, the exposure ranges were 0–2.57 mg/kg/d and 0–3.1 mg/kg/d in men and women, respectively. The exposure of women to benzene from whiteout ranged from 0 to 0.00059 mg/kg/d. Hazard index (HI), defined as a ratio of exposure to reference dose (RfD), for ethylbenzene was 31.1 (3.11 mg/kg/d/0.1 mg/kg/d) and for xylene (2.57 mg/kg/d/0.2 mg/kg/d) was 12.85, exceeding 1 for both compounds. Cancer risk for benzene was calculated to be 3.2 × 10−5 based on (0.00059 mg/kg/d × 0.055 mg/kg-d−1, cancer potency factor), assuming that 100% of detected levels in some products such as permanent pens and whiteouts were exposed in a worst-case scenario. These data suggest that exposure to VOC via some consumer products exceeded the safe limits and needs to be reduced.

An Exposure Assessment of DI-(2-Ethylhexyl) Phthalate (DEHP) and DI-n-Butyl Phthalate (DBP) in Human Semen

Levels of the phthalates such as di(2-ethylhexyl) phthalate (DEHP), mono(2-ethylhexyl) phthalate (MEHP, a major metabolite of DEHP), di-n-butyl phthalate (DBP), mono-n-butyl phthalate (MBP, a major metabolite of DBP), and phthalic acid (P, (a common metabolite of phthalates, including DEHP and DBP) were determined in the semen samples of 99 healthy volunteers without known prior medicosurgical history. Samples were obtained from young men (age 20–25 yr) who visited a clinic, and the semen concentrations of phthalates were measured using ultra-performance liquid chromatography (UPLC) and tandem mass spectrometry (MS/MS). UPLC/MS/MS showed that mean concentrations in semen samples were 1.07 μg/ml for MEHP, 0.61 μg/ml for DEHP, 0.39 μg/ml for PA, 0.06 μg/ml for MBP, and 0.003 μg/ml for DBP. The concentration of MEHP (the metabolite of DEHP) was highest, and the concentrations of the metabolites including MEHP, MBP, and PA were higher than actual concentrations of parent DEHP and DBP. These findings suggest the detection of phthalates in healthy human semen might require further investigation for effects on human fertility.

Chemopreventive mechanisms of methionine on inhibition of benzo(a)pyrene-DNA adducts formation in human hepatocellular carcinoma HepG2 cells.

This study was designed to investigate the molecular mechanism underlying the chemopreventive effects of methionine on benzo[a]pyrene (B[a]P)-DNA adducts formation in HepG2 cells. Methionine significantly inhibited B[a]P-DNA adduct formation in HepG2 cells. Methionine significantly decreased the cellular uptake of [(3)H] B[a]P, but increased the cellular discharge of [(3)H] B[a]P from HepG2 cells into the media. B[a]P significantly lowered total cellular glutathione (GSH) level, but co-cultured with B[a]P and methionine, gradually attenuated intracellular GSH levels in a concentration-dependent manner, which was markedly higher at 20-500μM methionine. The cellular proteins of treated cells were resolved by 2D-polyacrylamide gel electrophoresis. Proteomic profiles showed that phase II enzymes such as glutathione S-transferase (GST) omega-1, GSTM3, glyoxalase I (GLO1) and superoxide dismutase (SOD) were down-regulated by B[a]P treatment, whereas cathepsin B (CTSB), Rho GDP-dissociation inhibitor alpha (Rho-GDP-DIA), histamine N-methyltransferase (HNMT), spermidine synthase (SRM) and arginase-1 (ARG1) were up-regulated by B[a]P. B[a]P and methionine treatments, GST omega-1, GSTM3, GLO1 and SOD were significantly enhanced compared to B[a]P alone. Similarly, methionine was effective in diminishing the B[a]P-induced up-regulation of CTSB, Rho-GDP-DIA, HNMT, SRM and ARG1. Our data suggests that methionine might exert a chemoprotective effect on B[a]P-DNA adduct formation by attenuating intracellular GSH levels, blocking the uptake of B[a]P into cells, or by altering expression of proteins involved in DNA adduct formation.

Risk assessment of zinc oxide, a cosmetic ingredient used as a UV filter of sunscreens

ABSTRACT Zinc oxide (ZnO), an inorganic compound that appears as a white powder, is used frequently as an ingredient in sunscreens. The aim of this review was to examine the toxicology and risk assessment of ZnO based upon available published data. Recent studies on acute, sub-acute, and chronic toxicities of ZnO indicated that this compound is virtually non-toxic in animal models. However, it was reported that ZnO nanoparticles (NP) (particle size, 40 nm) induced significant changes in anemia-related hematologic parameters and mild to moderate pancreatitis in male and female Sprague-Dawley rats at 536.8 mg/kg/day in a 13-week oral toxicity study. ZnO displayed no carcinogenic potential, and skin penetration is low. No-observed-adverse-effect level (NOAEL) ZnO was determined to be 268.4 mg/kg/day in a 13-week oral toxicity study, and a maximum systemic exposure dose (SED) of ZnO was estimated to be 0.6 mg/kg/day based on topical application of sunscreen containing ZnO. Subsequently, the lowest margin of safety (MOS) was estimated to be 448.2, which indicates that the use of ZnO in sunscreen is safe. A risk assessment was undertaken considering other routes of exposure (inhalation or oral) and major product types (cream, lotion, spray, and propellant). Human data revealed that MOS values (7.37 for skin exposure from cream and lotion type; 8.64 for skin exposure of spray type; 12.87 for inhalation exposure of propellant type; 3.32 for oral exposure of sunscreen) are all within the safe range (MOS > 1). Risk assessment of ZnO indicates that this compound may be used safely in cosmetic products within the current regulatory limits of 25% in Korea.

Potential metabolomic biomarkers for evaluation of adriamycin efficacy using a urinary 1H-NMR spectroscopy.

A metabolomics approach using proton nuclear magnetic resonance (NMR) was applied to investigate metabolic alterations following adriamycin (ADR) treatment for gastric adenocarcinoma. After BALB/c‐nu/nu mice were implanted with human gastric adenocarcinoma, ADR (1 or 3 mg kg−1 per day) was intraperitoneally administered for 5 days. Urine was collected on days 2 and 5 and analyzed by NMR. The levels of trimethylamine oxide (TMAO, ×0.3), hippurate (×0.3) and taurine (×0.6) decreased significantly (P < 0.05), whereas the levels of 3‐indoxylsulfate (×12.6), trigonelline (×1.5), citrate (×2.5), trimethylamine (TMA, ×2.0) and 2‐oxoglutarate (×2.3) increased significantly (P < 0.05) in the tumor model. After ADR treatment, TMAO, hippuarte and taurine were increased significantly on day 5 compared with those of the tumor model. The levels of 2‐oxoglutarate, 3‐indoxylsulfate, trigonelline, TMA and citrate, which increased in the tumor model, significantly decreased to those of normal control by ADR treatment. Furthermore, the ratio between TMA and TMAO was dramatically altered in both tumor and ADR‐treated groups. Overall, metabolites such as TMAO, TMA, 3‐indoxylsulfate, hippurate, trigonelline, citrate and 2‐oxoglutarate related to the tricarboxylic acid (TCA) cycle might be considered as therapeutic targets to potentiate the efficacy of ADR. Thus, these results suggest that the metabolomics analysis of tumor response to ADR treatment may be applicable for demonstrating the efficacy of anticancer agent, ADR and treatment adaptation. Copyright

Potential Application of Benzo(a)Pyrene-Associated Adducts (Globin or Lipid) as Blood Biomarkers for Target Organ Exposure and Human Risk Assessment

In order to investigate the potential application of blood biomarkers as surrogate indicators of carcinogen–adduct formation in target-specific tissues, temporal formation of benzo[a]pyrene (BaP)-associated DNA adducts, protein adducts, or lipid damage in target tissues such as lung, liver, and kidney was compared with globin adduct formation or plasma lipid damage in blood after continuous intraperitoneal (ip) injection of [3H]BaP into female ICR mice for 7 d. Following treatment with [3H]BaP, formation of [3H]BaP–DNA or –protein adducts in lung, liver, and kidney increased linearly, and persisted thereafter. This finding was similar to the observed effects on globin adduct formation and plasma lipid damage in blood. The lungs contained a higher level of DNA adducts than liver or kidneys during the treatment period. Further, the rate of cumulative adduct formation in lung was markedly greater than that in liver. Treatment with a single dose of [3H]BaP indicated that BaP–globin adduct formation and BaP–lipid damage in blood reached a peak 48 h after treatment. Overall, globin adduct formation and lipid damage in blood were significantly correlated with DNA adduct formation in the target tissues. These data suggest that peripheral blood biomarkers, such as BaP–globin adduct formation or BaP–lipid damage, may be useful for prediction of target tissue-specific DNA adduct formation, and for risk assessment after exposure.

Uterotrophic and Hershberger Assays for Endocrine Disruption Properties of Plastic Food Contact Materials Polypropylene (PP) and Polyethylene Terephthalate (PET)

Plasticizers or plastic materials such as phthalates, bisphenol-A (BPA), and styrene are widely used in the plastic industry and are suspected endocrine-disrupting chemicals (EDC). Although plastic materials such as polypropylene (PP) and polyethylene terephthalate (PET) are not EDC and are considered to be safe, their potential properties as EDC have not been fully investigated. In this study, plastic samples eluted from plastic food containers (PP or PET) were investigated in Sprague-Dawley rats using Hershberger and uterotrophic assays. In the Hershberger assay, 6-wk-old castrated male rats were orally treated for 10 consecutive days with plastic effluent at 3 different doses (5 ml/kg) or vehicle control (corn oil, 1 ml/100 g) to determine the presence of both anti-androgenic and androgenic effects. Testosterone (0.4 mg/ml/kg) was subcutaneously administered for androgenic evaluation as a positive control, whereas testosterone (0.4 mg/ml/kg) and flutamide (3 mg/kg/day) were administered to a positive control group for anti-androgenic evaluation. The presence of any anti-androgenic or androgenic activities of plastic effluent was not detected. Sex accessory tissues such as ventral prostate or seminal vesicle showed no significant differences in weight between treated and control groups. For the uterotrophic assay, immature female rats were treated with plastic effluent at three different doses (5 ml/kg), with vehicle control (corn oil, 1 ml/100 g), or with ethinyl estradiol (3 μg/kg/d) for 3 d. There were no significant differences between test and control groups in vagina or uterine weight. Data suggest that effluents from plastic food containers do not appear to produce significant adverse effects according to Hershberger and uterotrophic assays.