HPLC法同时测定不同厂家珍珠明目滴眼液中17种氨基酸的含量

Simultaneous Determination of Seventeen Kinds of Amino Acids in Zhenzhu Mingmu Eye Drops from Different Manufacturers by HPLC

目的建立HPLC法同时测定珍珠明目滴眼液中17种氨基酸的含量,比较9个不同生产厂家的19批珍珠明目滴眼液中17种氨基酸的含量差异。方法以异硫氰酸苯酯为衍生化试剂,对样品进行衍生处理后,采用HPLC法测定。色谱柱为Venusil-AA氨基酸分析柱(250 mm×4.6 mm,5μm);流动相A为0.1 mo L·L^(-1)醋酸钠缓冲液(pH 6.5)-乙腈(93∶7),流动相B为乙腈-水(4∶1),梯度洗脱;流速:1.0 mL·min^(-1);检测波长:254 nm;柱温:38℃。结果 17种氨基酸质量浓度在相应的范围内呈良好的线性关系(>0.993 0);17种氨基酸平均回收率在90.3%~108.4%之间,RSD在0.30%~3.31%之间。结论不同厂家的珍珠明目滴眼液中游离氨基酸含量存在较大差异。

Objective To establish a method for simultaneous determination of 17 kinds of amino acids in Zhenzhu Mingmu Eye Drops by HPLC, and to investigate the difference of the contents of 17 kinds of amino acids in 19 batches of the samples from 9 manufacturers. Methods Phenyl isothiocyanate （PITC） was used as the reagent for sample derivatization, and then the samples were analyzed by high performance liquid chromatography（ HPLC ）. The samples were analyzed on a Venusil-AA amino acid analysis column（250 mm×4.6 mm,5μm） with gradient elution of 0.1moL·L^-1 sodiumacetate solution（pH 6.5）- acetonitrile（93 ： 7）（A） and acetonitrile-water（4 ： 1）（B ） at the flow rale of 1.0 mL·min^-1. The detection wavelength was 254 nm and the column temperature was 38℃. Results Linear relationship was present in 17 kinds of amino acids （r〉 0.993 0）. Their average recoveries were between 90.3 % and 108.4 %, and RSDs were between 0.30 %-3.31%. Conclusion The contents of 17 kinds of amino acids in Zhenzhu Mingmu Eye Drops from different manufacturers are various.