瑞斯托菌素的高效液相色谱分析

HPLC restocetin assay

目的：建立快速、简便、准确的瑞斯托菌素的高效液相检测方法,在瑞斯托菌素菌种选育、有效组分的分离纯化等研究工作中及时提供可靠的,有指导意义的检测数据。方法：采用Kromasil C18（4.6mm×250mm,5μm）色谱柱,0.1%三氟乙酸-甲醇（30∶70）为流动相,流速0.5mL·min-1,柱温45℃,紫外检测波长254nm的色谱条件。结果：瑞斯托菌素在0.01~2.5mg.mL-1浓度范围呈良好的线性关系,r=0.9987;最低检测限5ng,最小定量限20ng;精密度RSD=0.04%。结论：该方法测定瑞斯托菌素样品的含量准确可靠,在分离纯化过程中也可以有效地检测和控制瑞斯托菌素的纯度。

Objective：To establish an effective HPLC assay for determination both the content and the purity of ristocetin A,which would be helpful in the determination of the ristocetin A in fermentation broth for the improvement of producing strain,isolation and purification for ristocetin A.Methods：HPLC analysis condition：column：Kromasil C18（4.6 mm×250 mm,5 μm）;mobile phase：0.1% trifluoroacetic acid-methanol（30∶70）;flow rate：0.5 mL·min-1;temperature：45 ℃;detection wavelength：254 nm.Results：The linear range of ristocetin A was 0.01-2.5 mg·mL-1,r=0.9987;the LOD was 5 ng and the LOQ was 20 ng;RSD=0.04%.Conclusion：The method is accurate and reliable in determination the content and the purity for the ristocetin A.