Quality Control And Stability Study For Bovine Lactoferrin

Bovine Lactoferrin ( BLF ) mainly exists in numerous exocrine secretions of mammals, and its content in milk is the higest. Lactoferrin not only has broad spectrum of antimicrobial properties, but also has the ability to improve antiviral activity, and anticancer activity, and immunoregulatory activity of the body to prevent and cure infectious disease. Occident, Japan and few developed country exploit lactoferrin to develop drug, immune accelerant and infant food which prevent human immunodeficiencyvirus (HIV), cytomegalovirus (CMV), herpessimplexvirus (HSV), human hepatitis C virus(HCV), helicobacter pylori eta.Bovine lactoferrin is the major glycosylated proteins from colostrum, milk, nonlactating, bovine mammary secretions, an iron-binding glycoprotein with a molecular mass of about 80 kDa and pI = 8, which has broad-spectrum antimicrobial, antiviral, inhibition of oxygen free radicel, regulation of iron absorption in gastrointestinal tract, immune-modulating and other biological functions.The experiment treated Bovine lactoferrin as research objective, made it into tablet, discussed the quality criterion and stability of tablet and raw drug, andprovided reliable quality criterion for the development of Bovine lactoferrin new drug.The contents of Bovine lactoferrin and related protein in raw drug and tablet were studied by HPLC and HPCE.HPLC method for determining bovine lactoferrin : Bovine lactoferrin was chromatographed on a vydac C4 column (250 mm X 4.6 mm, 5 u m) with the column temperature of 25 °C and detected at 220nm. 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'.The result is that the standard curve was linear in the range of 25 ~ 600 u g ? mL-1. The linearity regression equation was A = 6.693 X 102 C — 5.403 X 103,r = 0.999 5.LOD was 20 ug .mL"1, the percentage of BLF was about 93% and the percentage of related proteins was 1~ 3%.HPCE method for determining bovine lactoferrin: fused-silica capillary column used :57 cm (efficiency length 50 cm) X 50 pm, running buffer : 33 mmol'L"1 KH2PO4 — 10 mmol'L'1 H3PO4 solution (pH = 2.50);eletroinjection: 1.0 psi X 5 s;running voltage:25 kV;detection wavelength:214 nm;column temperature :25 °C. The result is that the standard curve was linear in the range of 25 ~ 1 000 u g ? mL"1. The linearity regression equation was A= 1.123 X 104C - 2.146 X105, r = 0.999 9.Bovine lactoferrin and related protein were determined by HPLC and HPCE;Stablity of Bovine lactoferrin was conducted by accelerated test and inspection for a long time, and amino acid constitution of Bovine lactoferrin and identity of impurity protein were studied primarily.