核壳型SiO_(2)@松香基高分子高效液相制备柱的制备及其对喜树碱的分离纯化

Preparation of Core-shell SiO_(2)@Rosin-based Polymer High Performance Liquid Preparation Column and Purification of Camptothecin

首先将球形硅胶活化后进行烷基化改性。然后,以甲基丙烯酸为功能单体、马来海松酸丙烯酸乙二醇酯为交联剂、偶氮二异丁腈为引发剂,按一定比例混合后均匀涂覆到改性硅球表面,通过涂覆-悬浮自由基聚合法合成核壳型SiO_(2)@松香基高分子(SiO_(2)@R)色谱固定相。采用红外光谱、扫描电镜、透射电镜、热重分析、比表面积及微孔吸附分析等方法对色谱固定相进行了表征,证明了松香基高分子在硅胶表面键合成功,核壳型SiO_(2)@R色谱固定相已制备成功,且其细胞毒性低于商业树脂。采用湿法装柱制得了高效液相制备型色谱柱(30 mm×250 mm,10μm),对色谱柱性能进行了测试,结果表明:制备柱的理论塔板数为59992,具有较好的流通性和重复使用性。以甲醇-水(体积比95∶5)为流动相,检测波长254 nm,流速7 mL/min,进样体积1.2 mL(上样量约1.728 mg),在室温下对喜树碱进行了分离纯化,所得喜树碱纯度经HPLC检测,为88.08%,相比粗提物纯度提高了45.78个百分点。

The silanated silica gels was synthesised by modifing the activated spherical silica gels.Methacrylic acid was uesd as functional monomer,glycol maleated rosinate modified acrylic acid and azobisisobutyronitrile were used as crosslinking agent and initiator.They were mixed and evenly coated on the surface of the silanated silica gels,and then the core-shell SiO_(2)@rosin-based polymer(SiO_(2)@R)chromatography stationary phase was synthesised by coating-suspension free radical polymerization.The stationary phase was characterized by Fourier transform infrared spectroscopy,scanning electron microscopy,transmission electron microscopy,thermogravimetric analysis,specific surface area and microporous adsorption analysis.The results showed that the rosin polymer was successfully bonded on the surface of silica gel,the core-shell SiO_(2)@R stationary phase was successfully prepared,and the cytotoxicity of stationary phase was lower than that of commercial resin.The high performance liquid preparative chromatographic column(30 mm×250 mm,10μm)was prepared by wet packing.And its performance was tested.The results showed that the particle size of the theoretical plate number of the preparative column was 59992,it had good liquidity and reusability.Methanol-water(volume ratio 95∶5)was used as mobile phase,the detection wavelength was set at 254 nm,the flow rate was 7 mL/min,the injection volume was 1.2 mL(the loading amount was about 1.728 mg),and the camptothecin(CPT)was separated and purified at room temperature.And the purification rate of CPT was 88.08%,compared with the crude extract,the purity increased by 45.78 percentage points.