反相液相制备色谱法结合超临界流体制备色谱法分离纯化海风藤中的化合物

摘要

建立了基于反相液相制备色谱和超临界流体制备色谱的组合方法,用于分离纯化醇提水沉后石油醚层中的海风藤.首先以甲醇作为改性剂,采用醇提水沉法去除海风藤甲醇提取物中的叶绿素,加入硅藻土后用石油醚回流富集目标成分.选用反相C18制备色谱柱将其分为18个组分,然后将组分在SFC模式下进行制备.选用酰胺色谱柱,以甲醇为改性剂,在柱温30℃、背压15.0MPa的条件下进行分离.基于反相色谱和超临界流体色谱不同的分离选择性,最后分离得到6个高纯度化合物.该法展示了反相制备色谱和超临界流体制备色谱在海风藤分离纯化方面的优势,特别是超临界流体色谱在天然产物的分析和制备方面的巨大潜力.%A method based on preparative reversed-phase liquid chromatography(prep-RPLC) and preparative supercritical fluid chromatography(prep-SFC)was developed for the separa-tion and purification of compounds from piper kadsura. A pretreatment method was first devel-oped,including methanol extraction,water precipitation,petroleum ether extraction,etc. Chlorophyll and other strong polar impurities were removed from the piper kadsura samples, and the target components were enriched in petroleum ether extracts. The piper kadsura sam-ples were separated into 18 fractions on a Unitary C18 column(250 mm×20 mm,5 μm)with water and methanol as the mobile phases. Then,the SFC parameters,including the column, modifier,temperature,and backpressure were optimized. The optimized conditions for prep-SFC were as follows:XAmide column(250 mm×20 mm,5 μm),methanol as the modifier,30℃ column temperature,and 15.0 MPa backpressure. Because of the good orthogonality of RPLC and SFC,six highly pure compounds were isolated,including kadsurenone,wallichinine, denudatin B,pellitorine,2E-decenoic acid N-isobutylamide,and futoxide.