Lactoferrin is a protein found in many mammalian milks, especially in the first breast milks. In this application, a supplement containing lactoferrin was analyzed using a polymer-base reversed phase column, Asahipak C4P-50 4D. C4P-50 4D was proved to effectively detect lactoferrin in a supplement. It also provided a good linear calibration curve in the concentration range 20 to 4,000 mg/L.

(Sample Preparation)
1) Grind three tablets of a commercial lactoferrin supplement in a mortar.
2) Measure out 0.2 g of the powder and add 50 mL of ultra-pure water.
   Sonicate for 10 minutes.
3) Centrifuge and filter it with a No.5B filtering paper.
4) Discard the first 10 mL and collect the rest.
5) Filter using a 0.45-µm membrane filter and use it as the injection sample.


Sample : 25 µL

1. Lactoferrin
(Standard solution)
1000 mg/L

Column       : Shodex Asahipak C4P-50 4D (4.6 mm I.D. x 150 mm)
Eluent       : (A); H2O/TFA=100/0.05(v/v)                    
               (B); CH3CN/TFA=100/0.05(v/v)
               Linear gradient; 0 to 50 % B (0 to 10min), 50 % B (10 to 15 min),
　　　　　                      50 to 0 % B (15 to 16 min), 0 % B (16 to 20 min) 
Flow rate    : 0.8 mL/min 
Detector     : UV (280 nm) 
Column temp. : 40 °C