The tissues and biological fluid of bovine that had apparently been poisoned by the corm of Colchicum autumnale L. were examined. The test solutions were prepared by the following steps: (1) colchicine-alkaloid extraction with acetone, (2) defatting of the extract with petroleum ether, (3) removal of protein from the extract with 5% zinc sulfate solution and 0.3N barium hydroxide, (4) extract of colchicine-alkaloid with chloroform. High performance liquid chromatography was performed with a LiChrosorb RP-18 column using acetonitrile-methanol-phosphate buffer (pH 6.0) (32:5:63) as the mobile phase with detection by absorbance measurement at 350nm.
Colchicine (COL) and demecolcine (DEM) contents in the muscle were in the range of 0.05-0.12ppm and 0.03-0.05ppm, respectively. In the spleen, COL and DEM levels were 0.76-1.17ppm and 0.71-1.19ppm, respectively. The contents of COL and DEM in the spleen were higher than those in the liver and kidney.
The determination limits of COL and DEM were 0.01 and 0.02ppm, respectively, in muscles. At the level of 5ppm, the recoveries of COL and DEM added to beef were 77.1% and 72.7%, and those of COL and DEM added to milk were 77.0% and 75.0%, respectively.

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The determination of multicomponent residues of organophosphorus pesticides was performed by using a combination of gradient silica gel dry column chromatography (SDC) with gas chromatography (GC). Two kinds of solvent systems were used for the SDC: system I, n-hexane, benzene and acetone; system II, benzene and acetone.
Elution patterns in SDC and relative retention times in GC were examined for 40 kinds of organophosphorus pesticides. Several organophosphorus pesticides which had the same or similar relative retention times in GC could be separated by SDC.
SDC is convenient because the activity of the silica gel is stable and a small amount of silica gel can be used repeatedly. Recoveries in the SDC step were in the ranges of 23-95% (system I) and 35-101% (system II).

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The toxic effects of corrosion tin from canned juice on the bone and blood of rats were examined. Wistar male rats were divided into three groups and treated with 0, 230 or 454ppm of corrosion tin in the drinking water for four weeks. After the treatment all the rats were killed by bleeding and the femurs, humeri and tibiae were dissected out. The bones were cleaned of soft tissues and subjected to compression and bending tests.
The following results were obtained.
1) In the 454ppm group, the compressive strength of the distal end of the femur and the proximal end of the humerus were significantly decreased. There were no significant changes in the bending strength of the diaphysis of the femur, humerus and tibia.
2) No significant difference was observed in the mechanical strength of bones of the 230ppm group compared with the control group.
3) Anemia was observed in the 454ppm group, but not in the 230ppm group.

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The effectiveness of CIN medium as a selective differential medium for Yersinia enterocolitica and that of alkali treatment for recovery of Yersinia species after PBS cold enrichment were evaluated using a total of 94 strains; 45 strains of Yersinia species and 49 strains of other species.
All 45 strains of Yersinia grew on CIN medium forming colonies with a deep red center and translucent in the periphery. Of the 49 non-Yersinia strains, 21 strains of Citrobacter, Morganella, Serratia or Pseudomonas grew on the medium, but the colony forms of these strains were clearly different from that of Yersinia.
Yersinia showed comparatively high resistance to alkali and the viable cell count was reduced to only 1/10-1/100 after treatment with 0.4% potassium hydroxide solution for 15-30 seconds. On the other hand, many of the non-Yersinia strains did not survive treatment with 0.4% or less potassium hydroxide solution for a short time. Under our experimental conditions with Yersinia and non-Yersinia strains, treatment with 0.4% potassium hydroxide solution for 30 seconds is appropriate for the recovery of Yersinia.
It is suggested that combined use of alkali treatment after PBS cold enrichment and isolation with CIN medium is the most effective isolation procedure for Y. enterocolitica from meat.

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A simple and rapid method for determination of tetracycline (TC), oxytetracycline (OTC), chlortetracycline (CTC) and tylosin (TS) in meats by high performance liquid chromatography (HPLC) was developed. After extraction with 0.5% metaphosphoric acid-methanol (8:2) from homogenized samples, the extract was concentrated. The concentrated solution was passed through a Sep-pak C₁₈ cartridge and then eluted with methanol. The eluate was evaporated to dryness. The residue was dissolved in distilled water-acetonitrile (75:25), and the solution was injected into an HPLC column packed with Nucleosil CN. The column was eluted with a mixture of 0.01M monobasic sodium phosphate-acetonitrile (75:25) (pH 2.5). The antibiotics were detected with a UV-photometer set at 268nm. The calibration graphs were rectilinear from 0.5μg/ml to 5.0μg/ml of TC and OTC, and 1.0μg/ml to 10.0μg/ml of CTC and TS. The average recoveries of TC, OTC, CTC and TS from meat samples fortified at the level of 1.0ppm or 2.0ppm were 75.1-92.1%

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The toxicity levels of the livers and ovaries of 66 specimens (40 males, 26 females) of pufferfish, Fugu rubripes chinensis (“karasu”), were assayed and compared with those of F. rubripes rubripes (“torafugu”) specimens. The karasu males had a toxic liver frequency of 15.0% with the highest toxicity score of 100MU/g, whereas the females showed a toxic liver frequency of 23.1% and the highest toxicity of 1, 500MU/g. In the case of torafugu, the toxic liver frequency and the highest toxicity were 9.5% and 100MU/g, respectively, for males, and 21.7% and 510MU/g, respectively, for females. On the other hand, the toxic ovary frequency was 57.7% and the highest toxicity was 2, 000MU/g in karasu, and the corresponding values were 65.2% and 820MU/g in torafugu.
It follows therefore that females are more toxic than males in both species, that the average toxicity of karasu is roughly comparable to that of torafugu, and that the livers are sometimes toxic enough to kill a man even when ingested in a small amount.

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Attempts were made to identify sixteen species of puffers electrophoretically. The myogen fractions were prepared from the ordinary muscle of the puffers, and subjected to polyacrylamide gel electrophoresis. After the runs, the gels were stained for protein with Coomassie brilliant blue, and for some enzymes such as malate dehydrogenase (MDH) and phosphoglucomutase (PGM) with specific reagents. The results obtained showed that the protein patterns are species-specific enough to permit identification of most of the puffers. The MDH and PGM isozyme patterns were less species-specific. The puffers were hardly distinguishable from each other when the myogen fractions were subjected to cellulose acetate membrane electrophoresis. On the other hand, the myogen fractions of most puffers exhibited species-specific protein patterns when subjected to isoelectric focusing, followed by Coomassie brilliant blue staining.
It was concluded from the above results that puffers can be practically identified by the above electrophoretic techniques except cellulose acetate membrane electrophoresis, and that a more reliable identification can be attained by the combined use of these techniques.

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