HPLC测定转移因子注射液中游离氨基酸含量探讨

Content determination of free amino acid in transfer factor injection by HPLC

目的探讨转移因子注射液中游离氨基酸含量的测定方法。方法采用高效液相色谱(HPLC)法进行含量测定,色谱柱为Diamonsil C18(250.0 mm×4.6 mm,5μm);检测波长:360 nm;进样量:20μL;柱温:(40±1)℃,流速:1.0mL/min;梯度洗脱;流动相:A液为0.1%三氟乙酸,B液为0.1%三氟乙酸乙腈,用0.45μm滤膜过滤并脱气,2,4-二硝基氟苯柱前衍生,梯度洗脱。结果游离氨基酸面积线性关系良好,r2=0.997 7,相对标准偏差为0.6%。结论 HPLC法测定转移因子注射液中游离氨基酸含量简便、快捷、准确,值得推广应用。

Objective To explore the method for determining the content of free amino acid in transfer factor injection.Methods High performance liquid chromatography（HPLC） was adopted for the content determination. The method was as follows：chromatographic column：C18Diamonsil（250.0 mm×4.6 mm,5 μm）;detection wavelength：360 nm;sample size：20 μL;column temperature：（40 ±1）℃;flow velocity：1.0 mL/min;gradient elute;A mobile phase was 0.1% trifuoroacetic acid,and B mobile phase was 0.1% trifluoroacetic acid acetonitrile with 0.45 μm filter membrane for filtration and degrassing;2,4-dinitrofluorobenzene was undergone precolumn derivatization,gradient elution. Results Free amino acid area showed a good linear relationship（r^2=0.9977,RSD=0.6%）. Conclusion The HPLC for the content determination of free amino acid in transfer factor injection is simple,fast and accurate,and it is worthy of popularization and application.