HPLC柱前衍生法测定谷丙甘氨酸咀嚼片含量  

Determination of three kinds of amino acid content in the glu-ala-glycine chewable lablet with Pre-column derivatization by high performance liquid chromatography

作　　者：邱宇[1] 刘艳丽[1] 孙华[1] 曲福军[1] 

机构地区：[1]哈尔滨医科大学第二临床医学院药学部,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2006年第3期260-263,共4页Journal of Harbin Medical University

摘　　要：目的建立谷丙甘氨酸中谷氨酸、丙氨酸及甘氨酸的高效液相分析方法，并用该方法测定咀嚼片中3种氨基酸的含量，为该制剂的质量标准制定提供依据。方法谷丙甘氨酸咀嚼片中3种氨基酸含量测定采用异硫氰酸基苯酯柱前衍生化高效液相色谱法。色谱条件：色谱柱为Inertsil ODS-3V（4．6mm×250mm，5μm）；流动相A为0．1mol／L醋酸钠-醋酸缓冲液（pH6．5）／乙腈（93：7），流动相B为蒸馏水／乙腈（20：80）二元梯度洗脱分离；检测波长为254nm；流速为1ml／min；柱温为40℃。外标法计算3种氨基酸的含量。结果谷丙甘氨酸咀嚼片中谷氨酸、丙氨酸及甘氨酸分离度良好，在所考查的浓度范围内浓度对峰面积线性回归r〉0．99，平均回收率为98．7％，日内及日间RSD〈1．3％，三批样品中3种氨基酸的含量均在95％-105％之间。结论用HPLC柱前衍生方法可测定谷丙甘氨酸制剂中谷氨酸、丙氨酸及甘氨酸含量。Objective To establish the high performance liquid chromatography method that determines the content of glutanic acid, alanin and glycine in the glu-ala-glycine, and then determine three kinds of amino acid content in the chewable lablet with this method, provide the basis for this prepaxation quality specification formulation. Methods Isothioeyanic acid base phenyl ester pre-column derivatization was used for the determination of three kinds of amino acids contents in the glu-ala-glycine chewable lablet by high performance liquid daromatography, the chromatography condition was inertsil ODS-3V （4.6 × 250mm, 5μm） ; flows A was 0.1 mol/L sodium acetate-acetic acid buffer solution （pH6.5）/acetonitrile （93：7）, and flows B was the distilled water/acetonitrile （20： 80） dual gradient elution sepaxation; the examination wave length was 254nm; the speed of flow was 1ml/min;the column temperature was 40℃. The external standard law was used to calculate three kinds of amino acid contents. Results The dissociation degree of the glutanic acid, the alanin and the glycine was good in the glu-ala-glycine chewable lablet. The calibration curve was linear within the range of concentration, r 〉 0.99, and the average returns ratio was 98.7 % with the within-day and inter-day RSD 〈 1.3 %, and the contents of three kinds of amino acid was with in 95 % - 105 % among three batch of samples. Conclusion Three kinds of amino acid content in the glu-ala-glycine chewable lablet can be determined with pre-column derivatization by high performance liquid chromatography.

关 键 词：高效液相色谱法 谷丙甘氨酸 咀嚼片 

分 类 号：R927[医药卫生—药学]