녹용의 지용성 성분에 관한 생화학적 연구의 일환으로 대만산 꽃사슴(Cervus nippon taiouans)의 녹용을 직접 절단하여 동결시킨 후, 벨?殼?(velvet layer)과 해면상 골조직층(spongybone layer)을 분리하였다. 조직을 Virtis homogenizer로 분쇄한 후 $CHCl_3$ : $CH_3OH$ (1 : 2) 및 (2 : 1) 비율의 혼합 용매로 지용성 성분을 추출하고, Rouser 법에 따라서 Sephadex G-25 column chromatography를 행하여 총지질로부터 ganglioside 및 nonlipid를 분리하고 이어서 Rouser의 방법에 준하여 silicic acid column chromatography를 행하여 총지질을 중성지질, 당지질, 인지질로 분리하였다. 대조시험으로, 녹용의 수성에탄올 제제인 Pantocrin을 질소 기류하에서 감압 건조한후, 상기와 동일한 방법으로 지질 성분을 분획하였다. 지용성 성분의 건조 중량에 대한 각 분획의 백분율은 녹용의 경우 neutrallipid 65.30%, glycolipid 5.22%, phospholipid 12.86%, ganglioside 6.12%, nonlipid(proteolipid & peptide) 10.51%의 비율로 합류되어 있었으며, Pantocrin은 neutral lipid 41.7%, glycolipid 0.62%, phospholipid 5.39%, ganglioside 49.32%, nonlipid(proteolipid & peptide) 2.93%의 함량이었다. 녹용의 glycolipid는 DEAE Sephadex A-25 column chromatography를 행하여 glycolipid의 전체양에 대하여 cerebroside 63.5%, sulfatide 36.5%를 얻었으며 각각을 표준품과 비교하여 T.L.C.를 행하여 7종의 spots를 얻었으며 그중 2종은 cerebroside로 3종은 sulfatide로 확인되었으나 2종은 미지물질로 확인되지 않았다. Pantocrin에서는 1종의 cerebroside를 확인하였다. 녹용과 Pantocrin에 존재하는 ganglioside는 Sephadex G-25 column chromatography법보다는 용매 분획법에 의해 얻은 시료에서 T.L.C.법으로 확인하였다, 또 녹용에서 얻은 phospholipid로는 phosphatidyl ethanolamine, phosphatidyl choline, sphingomyelin, phosphatidic acid, lysophosphatidyl ethanolamine 및 lysophosphatidyl choline등으로 동정되었으며, Pantocrin에서는 phosphatidyl choline, sphingomyelin을 검출하였다.
The lipid soluble fraction of Antler velvet layer (cervus nippon taiouanous) was extracted and compared to that of pantocrin (ethanol preparation of Antler, commercially available). Lipid soluble components (801mg/24.5g from Antler velvet ayer and 979.1mg/143ml from pantocrin) were fractionated by gel filteration using a sephadex CT-25 into neutral lipids, sulfatides, cerebrosides, phospholipids and non-lipid components (lipoprotein and peptide). A silicic acid and a DEAE-sephadex A-25 column chromatography were followed from refractionation and purification. Each fraction was identified by thin layer chromatography using standard materials. Typical lipid soluble fraction of Antler contained 65.3% neutral lipid, 5.2% glycolipid, 12.9% phospholipid, 6.1% ganglioside and 2.9% non-lipid components. The glycolipid was composed of 63.5% cerebroside and 36.5% sulfatide. The cerebroside fraction of the Antler velvet layer showed four spots while pantocrin gave only one spot on a thin layer chromatogram. Among the four spots of Antler cerebroside two had almost same migration with standard but two had quite different mobility compared to the standard. The latter fractions were hydrolyzed by 0.025M methanolic HCl in order to desulfate. The two unknown materials were identified as sulfatides by the method of IR spectroscopy and thin layer chromatography. By two dimensional thin layer chroma tography, it was found that phospholipid of the Antler velvet layer were composed of phosphatidylethanolamine, sphingomyelin, phosphatidylcholine, lysophospahtidylethanolamine, and lysophosphatidylcholine and phosphatidic acid. On the other hand pantocrin Ccontained phosphatidylcholine and sphingomyelin as phospholipids.
