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目的通过对竹节参总皂苷进行分段研究,确定其发挥佐剂效应的最佳段位。方法正丁醇萃取的总皂苷过D101大孔吸附树脂,通过用不同醇浓度洗脱,得到不同洗脱段,将各个段所得皂苷与蛋白共同注入小鼠体内进行免疫,每周免疫1次,共3次,最后1次免疫后5天处死小鼠取血,测定抗体效价;然后根据HPLC色谱分析及含量测定确定。结果 Elisa实验表明90%浓度的醇过大孔树脂所得皂苷的免疫佐剂活性最强,10%次之,30%和60%相当;HPLC及含量测定证明了皂苷的佐剂活性。结论 90%醇浓度洗脱的皂苷具有显著的佐剂效应。
OBJECTIVE: To determine the optimum stage of adjuvant effect by studying the total saponin of S. japonicus. Methods Total saponin extracted with n-butanol was adsorbed on D101 macroporous resin. Different fractions were eluted with different alcohol concentrations. Saponin and protein from each segment were injected into mice to immunize them once a week, A total of 3 times, 5 days after the last immunization mice were sacrificed to determine the antibody titer; and then determined by HPLC chromatographic analysis and content determination. Results Elisa experiments showed that 90% of the macroporous resin obtained saponin immune adjuvant activity strongest, 10%, 30% and 60% of the corresponding; HPLC and content determination proved adjuvant activity of saponins. Conclusion Saponins eluted at 90% alcohol concentration have a significant adjuvant effect.