Methylmercury Determination in Fish by Direct Mercury Analyzer.

BACKGROUND: There are two methods for quantifying methylmercury (MeHg) in fish using GC-electron capture detection (ECD): AOAC INTERNATIONAL Official Method 988.11 and the Korean Food Code (KFC) method. Both of these methods consume a large amount of chemicals and require long pretreatment times because of several complicated MeHg extraction steps. OBJECTIVE: In this study, a new method for the simple and rapid determination of MeHg in fish has been developed. The method is based on the investigation of oxygen combustion-gold amalgamation using a direct mercury analyzer (DMA) after the complete removal of MeHg by organic extraction and back-extraction to an aqueous medium. METHODS: The DMA is suitable for the analysis of both solid and liquid materials and has a good detection limit. The developed method was validated by comparing the MeHg recoveries (%) of both certified reference materials and the market-purchased fish samples with the MeHg concentration obtained using the KFC method. RESULTS: The following parameters pertaining to the developed method were established: detection limit, 1.02 µg/kg; LOQ, 3.09 µg/kg; linearity (r), 0.9998; range, 0.1-300 µg/kg; and recovery, 95-97%. CONCLUSIONS: Our method is a promising alternative by virtue of its much simpler and faster sample pretreatment procedure, with a MeHg recovery as high as that of the KFC method. HIGHLIGHTS: The developed method enables the simultaneous analysis of total Hg and MeHg with only DMA equipment.

Acute toxicity and pharmacokinetics of 13 nm-sized PEG-coated gold nanoparticles.

In general, gold nanoparticles are recognized as being as nontoxic. Still, there have been some reports on their toxicity, which has been shown to depend on the physical dimension, surface chemistry, and shape of the nanoparticles. In this study, we carry out an in vivo toxicity study using 13 nm-sized gold nanoparticles coated with PEG (MW 5000). In our findings the 13 nm sized PEG-coated gold nanoparticles were seen to induce acute inflammation and apoptosis in the liver. These nanoparticles were found to accumulate in the liver and spleen for up to 7 days after injection and to have long blood circulation times. In addition, transmission electron microscopy showed that numerous cytoplasmic vesicles and lysosomes of liver Kupffer cells and spleen macrophages contained the PEG-coated gold nanoparticles. These findings of toxicity and kinetics of PEG-coated gold nanoparticles may have important clinical implications regarding the safety issue as PEG-coated gold nanoparticles are widely used in biomedical applications.

Simple validated method for simultaneous determination of deoxynivalenol, nivalenol, and their 3-ß-D-glucosides in baby formula and Korean rice wine via HPLC-UV with immunoaffinity cleanup.

A simple and reliable method for the simultaneous determination of major type B trichothecene mycotoxins, deoxynivalenol (DON) and nivalenol (NIV), along with their 3-ß-d-glucosides (DON-3-glucoside (DON3G) and NIV-3-glucoside (NIV3G)) in baby formula and Korean rice wine was validated in the present study. The method was based on immunoaffinity cleanup followed by analysis using an HPLC-UV technique. The method was validated in-house for two matrices as follows: linearity (R2 > 0.99) was established in the range of 20-1000 µg kg-1; accuracy (expressed as recovery) ranged from 78.7 to 106.5% for all the analytes; good intermediate precision (relative standard deviation < 12%), and adequate detection and quantitation limits (< 4.4 and < 13.3 µg kg-1, respectively) were achieved. Furthermore, the estimated measurement expanded uncertainty was determined to be 4-24%. The validated method was successfully applied to the analysis of 31 baby formulas and Korean rice wines marketed in Korea.

Exposure assessment to paralytic shellfish toxins through the shellfish consumption in Korea.

Paralytic shellfish poisoning is caused by saxitoxin and its analogues. The paralytic shellfish toxins (PSTs) are produced by marine dinoflagellates and can be accumulated in filter feeding shellfish, such as mussel, clam, oyster and ark shell. The worldwide regulatory limits for PSTs in shellfish are set at 80 µg STX eq./100 g meat and this is widely accepted as providing adequate public health protection. In this study, we have determined five individual PSTs (STX, GTX1, GTX2, GTX3 and GTX4) in shellfish using LC-MS/MS and assessed the human acute and chronic exposures to PSTs through shellfish consumption. Food consumption data was obtained from the Korea National Health and Nutrition Examination Survey (KNHANES 2010-2015). The acute exposure using a large portion size of 88 g/day (95th percentile for consumers only) with maximum toxin level of 198.7 µg/kg was 0.30 µg/kg bw. Even though we estimated the acute exposure with a conservative manner, it was below the ARfDs (0.5 or 0.7 µg STX eq./kg bw) proposed by the international organizations, representing 43-60% of the ARfDs. The chronic exposures using mean consumption data for whole population with mean concentration of PSTs were ranged from 0.002 to 0.026 µg STX eq./kg bw/day. For consumers only, the chronic exposures were in the range of 0.012-0.128 µg STX eq./kg bw/day.

Determination and risk characterization of polycyclic aromatic hydrocarbons of tea by using the Margin of Exposure (MOE) approach.

Tea is one of the most frequently consumed drinks due to its favourite taste and the health benefit. Tea is produced by several processes and drying is very important step to develop the flavour and destroys the enzymes in tea. However, during drying tea, polycyclic aromatic hydrocarbons some of which are carcinogen and genotoxin are naturally produced. The risk of PAHs by drinking tea was characterized by determining contents of 4 PAHs in tea. 4 PAHs including Benz(a)anthracene (BaA), Chrysene (CHR), Benzo(b)fluoranthene (BbF) and Benzo(a)pyrene (BaP) were investigated by GC-MS in total 468 tea products, which were contaminated up to 4.63 ng g-1. Mate tea was the most highly contaminated by BaA, CHR, BbF and BaP and followed by Solomon's seal and Chrysanthemum. The Margin of Exposures calculated by the concentration of BaA, CHR, BbF and BaP and consumption amount of tea were higher than 10,000, and the risk of PAHs in tea were low concern to public health.

Multiclass mycotoxin analysis in edible oils using a simple solvent extraction method and liquid chromatography with tandem mass spectrometry.

A simple and rapid method for the simultaneous determination of 11 mycotoxins - aflatoxins B1, B2, G1 and G2; fumonisins B1, B2 and B3; ochratoxin A; zearalenone; deoxynivalenol; and T-2 toxin - in edible oils was established using liquid chromatography tandem mass spectrometry (LC-MS/MS). In this study, QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe), QuEChERS with dispersive liquid-liquid microextraction, and solvent extraction were examined for sample preparation. Among these methods, solvent extraction with a mixture of formic acid/acetonitrile (5/95, v/v) successfully extracted all target mycotoxins. Subsequently, a defatting process using n-hexane was employed to remove the fats present in the edible oil samples. Mass spectrometry was carried out using electrospray ionisation in polarity switching mode with multiple reaction monitoring. The developed LC-MS/MS method was validated by assessing the specificity, linearity, recovery, limit of quantification (LOQ), accuracy and precision with reference to Commission Regulation (EC) 401/2006. Mycotoxin recoveries of 51.6-82.8% were achieved in addition to LOQs ranging from 0.025 ng/g to 1 ng/g. The edible oils proved to be relatively uncomplicated matrices and the developed method was applied to 9 edible oil samples, including soybean oil, corn oil and rice bran oil, to evaluate potential mycotoxin contamination. The levels of detection were significantly lower than the international regulatory standards. Therefore, we expect that our developed method, based on simple, two-step sample preparation process, will be suitable for the large-scale screening of mycotoxin contamination in edible oils.

Antifungal and Antiaflatoxigenic Methylenedioxy-Containing Compounds and Piperine-Like Synthetic Compounds.

Twelve methylenedioxy-containing compounds including piperine and 10 piperine-like synthetic compounds were assessed to determine their antifungal and antiaflatoxigenic activities against Aspergillus flavus ATCC 22546 in terms of their structure-activity relationships. Piperonal and 1,3-benzodioxole had inhibitory effects against A. flavus mycelial growth and aflatoxin B1 production up to a concentration of 1000 µg/mL. Ten piperine-like synthetic compounds were synthesized that differed in terms of the carbon length in the hydrocarbon backbone and the presence of the methylenedioxy moiety. In particular, 1-(2-methylpiperidin-1-yl)-3-phenylprop-2-en-1-one had potent antifungal and antiaflatoxigenic effects against A. flavus up to a concentration of 1 µg/mL. This synthetic compound was remarkable because the positive control thiabendazole had no inhibitory effect at this concentration. Reverse transcription-PCR analysis showed that five genes involved in aflatoxin biosynthesis pathways were down-regulated in A. flavus, i.e., aflD, aflK, aflQ, aflR, and aflS; therefore, the synthetic compound inhibited aflatoxin production by down-regulating these genes.

Harmonized Collaborative Validation of Aflatoxins and Sterigmatocystin in White Rice and Sorghum by Liquid Chromatography Coupled to Tandem Mass Spectrometry.

An interlaboratory study was performed in eight laboratories to validate a liquid chromatography-tandem mass spectrometry (LC/MS/MS) method for the simultaneous determination of aflatoxins and sterigmatocystin (STC) in white rice and sorghum (Sorghum bicolor). Fortified samples (at three different levels) of white rice and sorghum were extracted, purified through a solid-phase extraction (SPE) column, and then analyzed by LC/MS/MS. The apparent recoveries (ARs) ranged from 78.8% to 95.0% for aflatoxins and from 85.3% to 96.7% for STC. The relative standard deviations for repeatability (RSDr) and reproducibility (RSDR) of aflatoxins were in the ranges 7.9%-33.8% and 24.4%-81.0%, respectively. For STC, the RSDr ranged from 7.1% to 40.2% and the RSDR ranged from 28.1% to 99.2%. The Horwitz ratio values for the aflatoxins and STC ranged from 0.4 to 1.2 in white rice and from 0.3 to 1.0 in sorghum, respectively. These results validated this method for the simultaneous determination of aflatoxins and STC by LC/MS/MS after SPE column cleanup. The percentages of satisfactory Z-score values (|Z| ≤ 2) were the following: for white rice, 100% for aflatoxins and STC; for sorghum, 100%, except in data from two laboratories for STC (0.3 µg/kg). This validated that the LC/MS/MS method was successfully applied for the determination of aflatoxins and STC in 20 white rice and 20 sorghum samples sourced from Korean markets.

Effects of grilling procedures on levels of polycyclic aromatic hydrocarbons in grilled meats.

Polycyclic aromatic hydrocarbons (PAHs) are chemicals formed when muscle meat is cooked using high-temperature methods, such as grilling directly over an open flame. PAHs have been found to be mutagenic-that is, they cause changes in DNA that may increase the risk of cancer. We investigated the effects of grilling procedures on the level of 4 PAHs; benzo[a]anthracene (B[a]A), chrysene (Chr), benzo[b]fluoranthene (B[b]F), and benzo[a]pyrene (B[a]P). PAHs were extracted and determined by gas chromatography with mass detection (GC-MS). With regard to barbecuing successive meat samples with the same batch of burning charcoal, it was observed that stable combustion contribute to reduction of PAHs. Significant reductions in the sum of the four PAHs were observed through treatments which removed meat drippings and smoke with alternative grilling apparatus. The sums of 4 PAHs were reduced 48-89% with dripping removed and 41-74% with the smoke removal treatment in grilled pork and beef meats than conventional grilling. We investigated the components of meats drippings. The major constituent of meat dripping was fat. The most important factor contributing to the production of PAHs in grilling was smoke resulting from incomplete combustion of fat dripped onto the fire.

Total Diet Studies as a Tool for Ensuring Food Safety.

With the diversification and internationalization of the food industry and the increased focus on health from a majority of consumers, food safety policies are being implemented based on scientific evidence. Risk analysis represents the most useful scientific approach for making food safety decisions. Total diet study (TDS) is often used as a risk assessment tool to evaluate exposure to hazardous elements. Many countries perform TDSs to screen for chemicals in foods and analyze exposure trends to hazardous elements. TDSs differ from traditional food monitoring in two major aspects: chemicals are analyzed in food in the form in which it will be consumed and it is cost-effective in analyzing composite samples after processing multiple ingredients together. In Korea, TDSs have been conducted to estimate dietary intakes of heavy metals, pesticides, mycotoxins, persistent organic pollutants, and processing contaminants. TDSs need to be carried out periodically to ensure food safety.

The impact of size on tissue distribution and elimination by single intravenous injection of silica nanoparticles.

Many approaches for the application of nano-sized particles to the human body as nanotechnology have been recently developed. The size of nanoparticles is related to their useful character and also plays a key role in toxicity. Since this surface area can interact with biological components of cells, nanoparticles can be more reactive in than larger particles. In the present study, a fluorescence dye-labeled 50, 100 and 200 nm-sized silica particle suspension was intravenously injected into mice to identify the toxicity, tissue distribution and excretion of silica nanoparticles in vivo. Incidence and severity of inflammatory response was transiently increased with injection of 200 and 100 nm silica nanoparticles within 12h. But there was no significant response related to injection of 50 nm particles. The silica particles of 50, 100 and 200 nm were cleared via urine and bile. The 50 nm silica nanoparticles cleared to urine and bile than 100 nm and particles of 200 nm existed at lower concentration than other two smaller particles in urine and feces. Silica nanoparticles were trapped by macrophages in the spleen and liver and remained there until 4 weeks after the single injection.

Investigation of the bovine leukemia virus proviral DNA in human leukemias and lung cancers in Korea.

The bovine leukemia virus (BLV) is the causative agent of enzootic bovine leucosis. This study investigated the presence of the BLV in leukemia (179 acute lymphoblastic leukemia, 292 acute myeloid leukemia and 46 chronic myelogenous leukemia cases) and 162 lung cancer patients (139 adenocarcinoma, 23 squamous cell carcinoma) to determine if the BLV is a causative organism of leukemia and lung cancer in Koreans. A BLV infection was confirmed in human cells by PCR using a BLV-8 primer combination. All 517 cases of human leukemia and 162 lung cancer were negative for a PCR of the BLV proviral DNA. In conclusion, although meat has been imported from BLV endemic areas, the BLV infection does not appear to be the cause of human leukemia or lung cancer in Koreans. These results can be used as a control for further studies on the BLV in Koreans.