Abstract: A new method to analyse and test vitamin K₁ in milk powder was reported,it not only saves testing time(the remain time is about 11 min,the old method is about 55 min),but also reduces the limits of determination(0.02 μg/100 g),and neighborhood peaks can be(sepa)rated completely.The mobile phase is composed of acetonitrile+methylalcohol+iso-propyl-alcohol=20+65+15.The method is fit for the determination of vitamin K₁ in the milk powder.