[Survey of Total Mercury Content in Fishery Products].

The total mercury content in 112 fishery products, purchased from Osaka city during 2013-2018, was measured using a heating vaporization mercury analyzer. The average total mercury content in all tuna processed products was found out to be 0.115 µg/g (median=0.070 µg/g). Notably, albacore tuna exhibited the highest mercury content with an average concentration of 0.301 µg/g (median=0.296 µg/g). The total mercury concentration of the fishery products, except processed tuna and Kezuribushi (comprising shavings of dried and smoked mackerel, sardine, as well as horse mackerel), was generally low (<0.1 µg/g).The provisional tolerable weekly intake (PTWI) for mercury as set by the Joint FAO/WHO Expert Committee on Food Additives is 4.0 µg/kg body weight/week. On an average, for Japanese diet, the total weekly mercury intake from fishery products for a person weighing 50 kg was estimated to be 0.13 µg/kg body weight/week, which was 3.3% of the PTWI. These values indicate that it was not a problem to consume fishery products normally. However, albacore tuna cans have a relatively high total mercury concentration; thus, pregnant women consuming them daily may exceed the tolerable weekly intake of methylmercury (2 µg/kg body weight/week) as set by the Food Safety Commission of Japan.

Comparison between mouse bioassay and HILIC-MS/MS for quantification of paralytic shellfish toxin in Japanese basket clams and mussels caught off coastal Osaka Bay in Japan.

The content and composition of paralytic shellfish toxins (PSTs) in Japanese basket clam (Corbicula japonica) and mussels (Mytilus galloprovincialis) from Osaka Bay, Japan, were investigated using a mouse bioassay (MBA) and hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS), and the association between toxicity values of MBA and HILIC-MS/MS was verified based on research data. The overall toxicity in Japanese basket clam was lower than that in the mussel. The PSTs of Japanese basket clam and mussel consisted mainly of C1, C2, and gonyautoxins 1-4 (GTX1-4) taking toxins compositional differences as mol%. When multiplying the content of different toxins by the toxic equivalent factor (TEF), C2 and GTX1-4 accounted for more than 90% of total toxicity (MU TEF/g) based on the MU TEF score converted by TEF for the two species. The total content of C2 and GTX1-4 converted to toxicity was significantly correlated with the toxicity determined by MBA for the two species (r2 > 0.983). This study provides a suitable and ethical monitoring method to investigate toxicity in bivalves contaminated with A. tamarense by analysis of only predominant toxins, along with reducing use of MBA.

Polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and dioxin-like coplanar polychlorinated biphenyls in mackerel obtained from the Japanese market, 1999-2003.

This study analysed the concentrations and toxic-equivalent (TEQ) levels of dioxin congeners in mackerel commercially available in Japan in early 2000s. The content of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin-like coplanar polychlorinated biphenyls (DL-PCBs) was determined in 17 mackerel samples from different areas. TEQ levels in mackerel muscle were 0.78 pg TEQ g-1 wet weight (wt) on average (range = 0.21-2.26 pg TEQ g-1 wet wt) for PCDD/Fs (PCDDs plus PCDFs), 2.81 pg TEQ g-1 wet wt on average (range = 1.02-8.5 pg TEQ g-1 wet wt) for DL-PCBs, and 3.59 pg TEQ g-1 wet wt on average (range = 1.24-10.8 pg g-1 wet wt) for dioxins (PCDD/Fs plus DL-PCBs). The results revealed somewhat higher TEQ levels for dioxins compared with the other data for fish and shellfish in the Japanese market. TEQ levels were well correlated with mackerel body weight; the main contributors were tetra- and penta-CDD/Fs, some hexa-CDD/Fs, and all 12 DL-PCBs, which are known to have high bioaccumulation potential. In particular, PCB 126 was mostly responsible for the observed correlation of DL-PCB and dioxins-TEQ levels with mackerel body weight. The average daily intake of dioxins in terms of TEQ through mackerel consumption was estimated to be 4.81 pg TEQ/person/day, accounting for 7% of the total intake of dioxins-TEQ through fish and shellfish in Japan.

Validation Study on a Rapid Method for Simultaneous Determination of Pesticide Residues in Vegetables and Fruits by LC-MS/MS.

A validation study was carried out on a rapid method for the simultaneous determination of pesticide residues in vegetables and fruits by LC-MS/MS. Preparation of the test solution was performed by a solid-phase extraction technique with QuEChERS (STQ method). Pesticide residues were extracted with acetonitrile using a homogenizer, followed by salting-out and dehydration at the same time. The acetonitrile layer was purified with C18 and PSA mini-columns. The method was assessed for 130 pesticide residues in 14 kinds of vegetables and fruits at the concentration level of 0.01 µg/g according to the method validation guideline of the Ministry of Health, Labour and Welfare of Japan. As a result 75 to 120 pesticide residues were determined satisfactorily in the tested samples. Thus, this method could be useful for a rapid and simultaneous determination of multi-class pesticide residues in various vegetables and fruits.

The Contents and Composition of Tetrodotoxin and Paralytic Shellfish Poisoning Toxins in Marine Pufferfish Canthigaster rivulata.

The contents and composition of tetrodotoxin (TTX) and paralytic shellfish toxins (PSTs) in skin, muscle, and internal organs of two samples of marine puffer fish Canthigaster rivulata from Wakayama prefecture, Japan, were analyzed. Liquid chromatography-mass spectrometry (LC-MS) and high-performance liquid chromatography with post-column derivatization and fluorescence detection (LC-FLD) were used for the analysis of TTX and PSTs, respectively. For both samples, TTX and two analogues of PSTs, saxitoxin (STX) and decarbamoyl STX (dcSTX), were detected at levels over the limit of quantization (LOQ) only in the skin. These toxins in the muscle and internal organs were at trace levels, or not detected (ND). TTX contents were 11,000 and 13,000 ng/g (or 35 and 41 nmol/g), while PSTs contents were 168 and 460 ng/g (or 0.63 and 1.72 nmol/g) in the two skin specimens. The compositions of total toxin content were 98.2 and 96.0 mol% TTX and 1.8 and 4.0 mol% PSTs, respectively. Thus, the main contributor to toxin content in C. rivulata skin was TTX and the levels of PSTs toxicity in C. rivulata were very low. When the PSTs contents were converted into mouse unit score from the LC-FLD results, the resulting values of 1.0 and 2.8 MU/g of PSTs in C. rivulata skin were similar to those in Takifugu poecilonotus and Takifugu vermicularis in Japan, as determined in previous studies.

A survey of dietary intake of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and dioxin-like coplanar polychlorinated biphenyls from food during 2000-2002 in Osaka City, Japan.

Studies of the dietary intake of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like coplanar polychlorinated biphenyls (dioxin-like PCBs), collectively referred as dioxins, were conducted using the total diet study (TDS) method with food purchased in Osaka City, Japan during 2000-2002. The daily intake of dioxin-TEQ (toxic equivalent) from food per adult person was estimated respectively as 104.24 pg TEQ/person/day in 2000, 72.73 pg TEQ/person/day in 2001, and 87.28 pg TEQ/person/day in 2002, corresponding to 2.08, 1.45, and 1.74 pg TEQ/kg body weight (bw)/day for an adult weighing 50 kg. The highest contribution ratio to the total intake of dioxin-TEQ was from fish and shellfish (group 10) in each year, accounting for 77-92%. The next highest contributor was meat and eggs (groups 11-A and 11-B). An annual decrease of the intake of dioxin-TEQ was not observed clearly. Otherwise, the dietary intake of non-2,3,7,8-substituted PCDDs/PCDFs were estimated together. The intake of 1,3,6,8-TeCDD came mostly from intake of fish and shellfish (group 10), green vegetable (group 7), and the other vegetables, including mushrooms and seaweed (group 8). In addition, the intake of 1,3,6,8-TeCDF, which reportedly antagonizes 2,3,7,8-TeCDD-mediated aryl hydrocarbon hydroxylase (AHH) and ethoxyresorufin O-deethylase (EROD) induction activities, was an insufficient amount to suppress 2,3,7,8-TeCDD-mediated activities. Furthermore, we discussed different TEQ compositions of PCDDs/PCDFs to dioxin-like PCBs from food intake and in human samples and inferred that the difference was caused by low bioaccumulation properties of 3,3',4,4',5-PeCB (PCB 126).

Contamination levels of PCDDs, PCDFs and CoPCbs in fish oils used as raw materials for margarine and shortening and retailed fish.

Toxic equivalent (TEQ) levels of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs/DFs) and coplanar polychlorinated biphenyls (CoPCBs) were analyzed in both crude (n = 5) and refined-hardening fish oils (n = 2), which are used in part as the raw materials of margarine and shortening, and in some retailed fish (n = 6), which may be similar to those used as sources of fish oil. PCDDs/DFs-TEQ levels ranged from 0.1 to 13.5 pg TEQ/g fat weight in crude fish oils, from 0.5 to 1.6 pg TEQ/g fat weight in refined-hardening fish oils, and ranged from 1.4 to 31.2 pg TEQ/g fat weight in retailed fish. The ranges of PCDDs/DFs-TEQ in crude fish oils, in refined-hardening fish oils and in retailed fish overlapped each other. In addition, CoPCBs-TEQ levels ranged from 4.9 to 19.6 pg TEQ/g fat weight in crude fish oils, from 0.2 to 1.4 pg TEQ/g fat weight in refined-hardening fish oils, and from 2.7 to 165.8 pg TEQ/g fat weight in retailed fish. TEQ levels of CoPCBs in refined-hardening fish oils were much lower than those in crude fish oils and in retailed fish. This was due to the lower concentrations of less-chlorinated congeners in refined-hardening fish oils, as compared to those in crude fish oils and retailed fish. It appears that less-chlorinated PCBs congeners in refined-hardening fish oils are partly removed during the refining process.

New Dihydroquinolinone Toxic to Artemia salina Produced by Penicillium sp. NTC-47.

Penicillium sp. NTC-47, which had been isolated from a soil sample, produced novel dihydroquinolinones when cultured with okara (the insoluble residue of whole soybean). These metabolites, 1 and 2, were crystalline products with molecular formulas of C17H17NO5 and C17H17NO4, respectively. The structure of 1 was established to be 3-methoxy-4,5-dihydroxy-4-(4'-methoxyphenyl)-quinolinone by spectroscopic evidence and by an X-ray crystal-lographic analysis. Spectral data indicated the structure of 2 to be 5-deoxy-l. Compound 1 demonstrated toxicity against brine shrimp with an LC50 value of 20µg/ml, but 2 exhibited no activity at a dose of 100 µg/ml.