Lactoferrin is a protein found in many mammalian milks, especially in the first breast milks. The Japanese Analytical Methods of Food Additives in Foods lists an analysis method for lactoferrin and specifies to use a butylated polyvinyl alcohol polymer gel column. In this application, a polymer-base reversed phase column, Asahipak C4P-50 4D, was used to analyze lactoferrin according to the Analytical Methods of Food Additives in Foods. A lactoferrin concentrate was analyzed as an actual sample. The Analytical Methods of Food Additives in Foods states that lactoferrin concentrates to contain 85% or more of lactoferrin. It was confirmed that the sample analyzed met this requirement.

(Sample Preparation)
1) Measure out 0.1 g of a commercial lactoferrin concentrate.
2) Add 50 mL of 0.3 % NaCl solution to dissolve the concentrate.
3) Use the dissolved-solution as the injection sample.


Sample : 25 µL

1. Lactoferrin
(Standard solution)
4000 mg/L

Column       : Shodex Asahipak C4P-50 4D (4.6 mm I.D. x 150 mm)
Eluent       :(A); 0.3 % NaCl/CH3CN/TFA=9000/1000/3
              (B); 0.3 % NaCl/CH3CN/TFA=5000/5000/3
               Linear gradient: 50 to 100 % B (0 to 25 min)
Flow rate    : 0.8 mL/min 
Detector     : UV (280 nm) 
Column temp. : 40 °C