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.
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
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