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

本文对鹅藤壶（Mitella Mitella）软组织中的甜菜碱进行了研究，建立了快速高效液相色谱分离甜菜碱的方法，并对藤壶组织提取物中未知的Dragendorff阳性物质的化学结构进行了研究。采用Dowex 50-X4柱对80%甲醇的藤壶组织提取液进行分馏，逐步增加HCl浓度。洗脱液的薄层色谱显示至少有10种阳性物质存在。为快速分离甜菜碱（ⅰ）、< β >-丙氨酸-甜菜碱（ⅱ）、< β >-丁基甜菜碱（ⅲ）、甘氨酸（ⅳ）和葫芦巴碱（ⅴ），建立了以Magnum 9-SCX和Custom 2616为色谱柱的高效液相色谱方法，并应用于藤壶提取物中。通过与正品甜菜的光谱数据、HPLC保留时间和TLC Rf值的比较，分离得到的5种Dragendorff阳性物质鉴定为I-V。在藤壶提取物的Dowex 50-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，并通过HPLC分离得到。质谱、1H NMR、13bbbc NMR、IR、UV等光谱数据证实其与Ackermann于1953年从海葵Anemonia sulcata中分离得到的zooanemonin一致。这可能是首次报道在节肢动物中出现动物贫血素。在另一种藤壶（Tetraclita Squamosa japonica）和两种海葵（Actinia equina）和日本胸膜（Anthopleura japonica）中也检测到动物厌氧酶。

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.