Studies on Standardization and Automation of Chemical Analysis of Paralytic Shellfish Toxins.
| Project/Area Number |
07556049
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Fisheries chemistry
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| Research Institution | Tohoku Unicersity |
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Principal Investigator |
OSHIMA Yasukatsu Tohoku University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (60111267)
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| Co-Investigator(Kenkyū-buntansha) |
TSUKADA Katsuo Hitachi, Ltd.Instrument Division, Department General Manager, 計測事業部, 研究員
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1995: ¥3,200,000 (Direct Cost: ¥3,200,000)
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| Keywords | HPLC analysis / Paralytic shellfish toxins / Reference standards / Automation |
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| Research Abstract |
Paralytic shellfish poisoning is a serious toxicological problem affecting the shellfish industries in many countries. Currently, determination of the toxins has been performed using the mouse bioassay, but alternative methods have been strongly required due to the drawbacks of the bioassay. We have developed an post column derivatization HPLC method which enable to analyze almost all saxitoxin analogs at high sensitivity. However, utilition of the methods by public health institutions has been hampered due to the lack of standard toxins. In this study, the standardization of the HPLC analysis, including extraction and cleanup procedures, was inverstigated. Special emphasis was laid on the new preparative methods of the reference standards. Development of automated system was also explored. Results of the studies are summarized as follows.
1.Problem of precipitation in the oxidizing reagent which causes erratic operation of HPLC was solved by replacing sodium phosphate to potassium phosphate in the oxidizing solution.
2.A protocol for the preparation of test solution from bivalves was achieved, including acid extraction and cleanup with an ODS cartridge column and ultrafiltration. Extraction methods from phytoplankton and seawater samples were also developed.
3.Effective toxins purification method from toxic bivalves was developed using several steps of column chromatography including new ones. After purity was determined by HPLC and NMR,concentration of the toxin was determined by measurement of nitrogen content with a combustion analysis.
4.Specific toxicity of each components, which is essential to evaluate seafood toxicity from the chemical data, were determined.
Complete automated system with column and mobile phase switching mechanism was developed. The system showed good reliability and could perform the analysies of more than 20 samples perday.
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Report
(3 results)
Research Products
(10 results)
