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目的:建立鉴别白眉蝮蛇降纤酶和尖吻蝮蛇降纤酶的反相高效液相色谱方法。方法:采用汉邦Lichrospher C_4(4.6 mm×250 mm,5μm)色谱柱,以A:0.1%三氟乙酸、B:含0.1%三氟乙酸乙腈-水(90:10)为流动相,梯度洗脱,检测波长280 nm,流速1.0 ml·min~(-1),柱温:40℃,进样量:100μl。结果:白眉蝮蛇降纤酶与尖吻蝮蛇降纤酶在色谱图上存在明显差异。结论:该方法简便准确,专属性强,可用于白眉蝮蛇与尖吻蝮蛇两种不同来源降纤酶的鉴别。
OBJECTIVE: To establish a reversed-phase high-performance liquid chromatography (HPLC) method for the identification of Agkistrodon halysii defibrase and Agkistrodon halysii defibrase. Methods: The mobile phase consisted of Lichrospher C_4 (4.6 mm × 250 mm, 5 μm) column with mobile phase of A: 0.1% trifluoroacetic acid and B: acetonitrile containing 0.1% trifluoroacetic acid and water (90:10) Off, detection wavelength 280 nm, flow rate 1.0 ml · min -1, column temperature: 40 ℃, injection volume: 100μl. Results: There was a significant difference in the chromatograms between the Agkistrodon acutus decapitation enzyme and the Agkistrodon acutus decapitation enzyme. Conclusion: The method is simple, accurate and specific. It can be used for the identification of defibrase from two different origins of Agkistrodon acutus and Agkistrodon acutus.