牛乳铁蛋白的反相高效液相色谱法含量测定

RP - HPLC Determination of Bovine Lactoferrin

目的:运用反相高效液相色谱法分离测定牛乳铁蛋白(BLF)的含量。方法:采用Vydac C4色谱柱(250 mm ×4.6 mm,5μm),柱温为25℃,检测波长为220 nm,流动相A为0.1%三氟乙酸(TFA)水溶液;B为乙腈-水-TFA(95:5:0.1),梯度洗脱,流速为1.0 mL·min-1。结果:牛乳铁蛋白的浓度在25～1000μg·mL-1,r=0.9999。结论:本法操作简便,精密度好,结果准确可靠。

Objective: To establish a RP - HPLC method for determining and separating bovine lactoferrin. Method: Bovine lactoferrin was chromatographed on a Vydac C4 column(250 mm ×4. 6 mm,5μm)with the column temperature of 25℃ and detected at 220 nm. The mobile phase used were A:0. 1% trifluoroacetic acid(TFA)in water , B: acetonitrile -water - TFA(95: 5:0. 1) , using gradient elution with flow rate 1. 0 mL·min-1. Results: The standard curve was linear in the range of 25 - 1000 μg· mL-1 and the correlation coefficient was 0. 9999. Conclusion: The proposed method is simple, highly accurate and reliable.