| Budget Amount *help |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1987: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1986: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Research Abstract |
The present studies were undertaken to survey betaines in the soft tissues of a goose barnacle, Mitella mitella, to develop a rapid HPLC method for separation of betaines, and to elucidate the chemical structure of unknown Dragendorff positive substances found in the extract of the barnacle tissues.
1. The 80% methanol extract of the barnacle tissues was fractionated on a Dowex 50-X4 column with stepwise increase of HCl concentration. TLC of the eluates revealed the presence of at least ten Dragendorff positive substances.
2. For rapid separation of betaines, such as glycinebetaine(I), <beta>-alanine- betaine(II), <gamma>-butyrobetaine(III), homarine(IV) and trigonelline(V), HPLC methods with Magnum 9-SCX and Custom 2616 column were developed and applied to the barnacle extract.
3. Five Dragendorff positive substances isolated were identified as I-V on the basis of comparison of their spectral data, retention times on HPLC and Rf values on TLC with the authentic betaines.
4. In the Dowex 50-X4 column chromatography of the barnacle extract, a fairly large amount of an unknown Dragendorff positive substance was detected in the fraction eluted with 1N HCl and it was isolated by HPLC. The spectral data including MS, ^1H NMR, ^<13>C NMR, IR and UV, demonstrated its identity with zooanemonin isolated by Ackermann from the sea anemone Anemonia sulcata in 1953. This may be the first to report the occurrence of zooanemonin in the Arthropoda.
5. Zooanemonin was also detected in another species of barnacle Tetraclita Squamosa japonica as wall as two species of sea anemones Actinia equina and Anthopleura japonica.
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