柱前衍生化HPLC测定复方谷氨酰胺肠溶胶囊中的L-谷氨酰胺

Determination of L-Glutamine in Compound Glutamine Entresoluble Capsules by Pre-column Derivatization HPLC

关键词:  复方谷氨酰胺肠溶胶囊  L-谷氨酰胺  柱前衍生  高效液相色谱法

Key words:  Compound Glutamine Entresoluble capsules  L-glutamine  pre-column derivatization  HPLC

摘要:

目的 建立柱前衍生化HPLC测定复方谷氨酰胺肠溶胶囊中L-谷氨酰胺的含量。方法 采用CAPCELL PAK C₁₈色谱柱(150 mm×4.6 mm，5 μm)，以乙酸钠缓冲液(pH 6.4)-乙腈为流动相，梯度洗脱，检测波长为254 nm。以牛磺酸为内标物质，异硫氰酸苯酯为衍生剂，采用单因素和正交试验确定衍生反应最佳条件。结果 L-谷氨酰胺在0.015 9~0.143 mg·mL^-1内线性关系良好(r²=0.999 9)，加样回收率为100.3%，RSD为0.8%(n=9)。结论 本方法准确、重复性好，可用于复方谷氨酰胺肠溶胶囊中L-谷氨酰胺的含量测定。

Abstract:

OBJECTIVE To establish a pre-column derivatization HPLC for determination of L-glutamine in Compound Glutamine Entresoluble capsules.METHODS HPLC experiment was performed on a CAPCELL PAK C₁₈ column (150 mm×4.6 mm, 5 μm). The mobile phase were sodium acetate buffer solution(pH 6.4) and acetonitrile with gradient elution. The detection wavelength was set at 254 nm. Taurine was used as an internal standard substance, the sample and taurine were derivatized with phenyl isothiocyanate. The combination of single factor and orthogonal experiments were used to optimize the derivatization experiment conditions.RESULTS The L-glutamine had good linearity in the range of 0.015 9-0.143 mg·mL^-1(r²=0.999 9). The average recoveries was 100.3% with RSD of 0.8%(n=9).CONCLUSION The HPLC method is accruate with high repeatability, which is helpful for the determination of L-glutamine in Compound Glutamine Entresoluble capsules.