水蛭活性肽的纯化工艺

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·mL^-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·mL^-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。

Purification process of leech bioactive peptides

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·mL^-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·mL^-1, at a flow velocity of 3.0 BV·h^-1, eluted with 2.5% NaCl 25% ethanol, 2.5% NaCl 50% ethanol, 2.5% NaCl 75% ethanol. The 2.5% NaCl 25% ethanol and 2.5% NaCl 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% NaCl 25% ethanol and 2.5% NaCl 50% elution sites show high activities, good degrees of separation.