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目的采用HPLC法与超高压液相色谱(UPLC)法测定低分子量鱼精蛋白(LMWP)的含量。方法分别对两种测定方法的条件进行了优化,建立了测定的方法学。结果 HPLC采用AgelaVenusil ASB C18色谱柱(250 mm×4.6 mm,5μm),流动相为含0.1%三氟乙酸的水-乙腈(14∶86),流速1 mL.min-1,检测波长214 nm,柱温40℃。UPLC采用Acquity UPLC BEHC18色谱柱(50 mm×2.1 mm,1.7μm),流动相为含0.1%三氟乙酸的水-乙腈(1∶9),流速0.4 mL.min-1,检测波长214 nm,柱温40℃。结论 HPLC和UPLC都能用于LMWP的检测,但UPLC的检测时间更短、检测限更低。
Objective To determine the content of low molecular weight protamine (LMWP) by HPLC and ultra high pressure liquid chromatography (UPLC). Methods The conditions of the two methods were optimized respectively, and the methodology of determination was established. Results The HPLC was carried out on AgelaVenusil ASB C18 column (250 mm × 4.6 mm, 5 μm) with a mobile phase of water-acetonitrile (14:86) containing 0.1% trifluoroacetic acid at a flow rate of 1 mL · min- Column temperature 40 ℃. UPLC was performed on an Acquity UPLC BEHC18 column (50 mm × 2.1 mm, 1.7 μm) with a mobile phase of 0.1% trifluoroacetic acid in water-acetonitrile (1: 9) at a flow rate of 0.4 mL · min- Column temperature 40 ℃. Conclusion Both HPLC and UPLC can be used for the detection of LMWP, but the detection time of UPLC is shorter and the detection limit is lower.