摘要
目的:对水蛭活性肽进行纯化,并对水蛭活性肽进行树脂纯化条件考察。方法:采用DA201-C大孔吸附树脂进行极性分离,后经D201阴离子交换树脂进行电荷分离,最终得到纯化的水蛭活性肽组分。结果:DA201-C大孔树脂,上样肽浓度20 mg·m L-1,流速1.0 BV·h-1,依次用25%乙醇、50%乙醇和75%乙醇溶液洗脱,各洗脱部位得率及活力较高,但总体活性成分被分散,以上各洗脱部位经D201离子交换树脂,pH 4.0,上样肽浓度30 mg·m L-1,流速3.0 BV·h-1,分别用2.5%氯化钠的25%乙醇、2.5%氯化钠的50%乙醇、2.5%氯化钠的75%乙醇溶液洗脱,2.5%氯化钠的25%乙醇和2.5%氯化钠的50%乙醇洗脱部位有明显的抗凝活性,经过分析性RP-HPLC验证后,基线平稳,各组分分离度良好。结论:经DA201-C大孔树脂和D201离子交换树脂纯化后的水蛭活性肽,2.5%氯化钠的25%乙醇和2.5%氯化钠的50%乙醇洗脱部位活力较高,分离度好。
OBJECTIVE To purify leech bioactive peptides and study on purification conditions of leech peptides. METHODS Separation with DA201-C resin and D201 resin were performed to obtain the purified leech bioactive peptides. RESULTS DA201-C sample was injected at 20 mg·m L-1,at a flow velocity of 1. 0 BV·h-1,eluted with 25% ethanol,50% ethanol and 75% ethanol,and the active ingredients were dispersed. D201 sample was injected at 30 mg·m L-1,at a flow velocity of 3. 0 BV·h-1,eluted with2. 5% Na Cl 25% ethanol,2. 5% Na Cl 50% ethanol,2. 5% Na Cl 75% ethanol. The 2. 5% Na Cl 25% ethanol and 2. 5% Na Cl 50%elution sites showed obvious anticoagulant activities. Analysis of RP-HPLC showed that the baseline was stable and the separation degree of each component was good. CONCLUSION DA201-C resin and D201 resin can purify leech peptide components. The 2. 5%Na Cl 25% ethanol and 2. 5% Na Cl 50% elution sites show high activities,good degrees of separation.
出处
《中国医院药学杂志》
CAS
北大核心
2017年第23期2322-2325,共4页
Chinese Journal of Hospital Pharmacy
基金
国家自然科学基金(编号:81073031)
