Studies on Chemical Structure and Biosynthesis of Unknown Dragendorff Positive Substances in A Goose Barnacle, Mitella mitella.

本研究对鹅藤壶软组织中的甜菜碱进行了调查，建立了一种快速分离甜菜碱的高效液相色谱方法，并阐明了藤壶组织提取物中发现的未知Dragendorff阳性物质的化学结构。藤壶组织的80%甲醇提取物在Dowex50-X4色谱柱上进行分级，逐步增加盐酸浓度。洗脱物的TLC显示至少存在10个Dragendorff阳性物质。为了快速分离甜菜碱，如甜菜碱(I)，-lt；β&gt；-丙氨酸-甜菜碱(II)，&lt；γ&gt；-丁基甜菜碱(III)，高马豆碱(IV)和胡芦巴碱(V)，建立了采用Magnum 9-SCX和Custom 2616色谱柱的高效液相色谱法，并应用于藤壶提取物中。通过对分离得到的5种Dragendorff阳性物质的光谱数据、高效液相色谱保留时间和薄层Rf值与贝塔正品进行比较，鉴定其为I-V。在藤壶提取物的Dowex50-X4柱层析中，用1N-HCl洗脱的部分中检测到相当数量的未知Dragendorff阳性物质，并用高效液相色谱分离得到。经MS、~1H、~(13)C、~(13)H、~(13)C、~(13)C、~这可能是首次在节肢动物中发现动物海葵素的报道。在另一种藤壶Tetraclita Squamosa Japan和两种海葵Actinia equina和Anthopleura Japan onica中也检测到了动物海葵素。

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.