不同方法测定赖氨葡锌颗粒中盐酸赖氨酸含量的比较

Comparison of Three Methods for the Determination of Lysine Hydrochloride in Lysine Glucozinc Granules

目的建立HPLC直接测定法测定赖氨葡锌颗粒中盐酸赖氨酸的含量,并将测定结果与比色法(茚三酮显色)和柱后衍生法测定结果进行比较,确定3种方法的优劣。方法采用APS-HYPERSIL NH 2色谱柱(25 cm×4.6 mm,5μm);流动相:0.05 mol·L^(-1)磷酸二氢钾溶液(用磷酸调节pH至5.0)-乙腈(40∶60);柱温:30℃;流速:1.0 mL·min^(-1);检测波长:200 nm;进样量:20μL;用3种方法测定108批样品中盐酸赖氨酸的含量并对测定结果进行统计分析。结果新建HPLC直接测定法线性关系良好,准确度高,重复性、精密度良好。比色法与HPLC直接测定法的测定结果存在显著性差异(P=0.002<0.05),柱后衍生法和新建HPLC法的测定结果不存在显著性差异(P=0.472>0.05),结论新建HPLC法和柱后衍生法测定盐酸赖氨酸的含量,含量结果一致,与比色法比较,结果不一致。HPLC直接测定法重复性好,操作简便,较柱后衍生法成本更低,更为适用。

OBJECTIVE To establish a HPLC method for the determination of lysine hydrochloride in lysozyme granules,and compare the results with those of Colorimetry(ninhydrin color)and post column derivatization to determine the advantages and disadvantages of the three methods.METHODS The aps-hypersil NH 2 column(25cm×4.6 mm,5μm)was used.Mobile phase:0.05 mol·L^(-1) potassium dihydrogen phosphate solution(adjusted pH to 5.0 with phosphoric acid)-acetonitrile(40∶60);column temperature:30℃;flow rate:1.0 mL·min^(-1);detection wavelength:200 nm;injection volume:20μL;Lysine hydrochloride content in 108 batches of samples was determined by three methods and the results were analyzed statistically.RESULTS The new HPLC method had good linearity,high accuracy,repeatability and precision.There was significant difference between colorimetric method and HPLC direct method(P=0.002<0.05).There was no significant difference between post column derivatization method and new HPLC method(P=0.472>0.05).CONCLUSION The content of lysine hydrochloride determined by HPLC and post column derivatization is the same as that determined by colorimetry.HPLC is more suitable than post column derivatization because of its good repeatability,simple operation and lower cost.