建立UPLC法测定复方氨基酸胶囊(9-5)中氨基酸的含量

目的通过优化色谱条件建立UPLC法测定复方氨基酸胶囊(9-5)中氨基酸的含量。方法以9种氨基酸对照为外标物,异硫氰酸苯酯为柱前衍生剂,采用ACQUITY UPLCBEH C_(18)(2.1 mm×100 mm,1.7μm)柱,流动相A为0.1 mol/L的醋酸钠溶液(用冰醋酸调整pH至6.50)-乙腈(93∶7),流动相B为乙腈-水(4∶1),检测波长254 nm,流速为0.45 mL/min,柱温36℃。结果复方氨基酸胶囊(9-5)中9种氨基酸出峰时间与对照品出峰时间一致,其他物质无干扰;仪器精密度RSD为0.5%~1.2%,中间精密度RSD为1.3%~1.9%;各氨基酸的溶液浓度与其峰面积线性关系良好(r≥0.999),溶液稳定性RSD为0.6%~1.8%,各氨基酸的平均回收率在95.0%~105.0%之间。含量测定方法比对实验中UPLC法与HPLC法分析结果无明显差异,分析速度明显提高。结论建立的UPLC法高效,快速,灵敏,准确,可用于测定复方氨基酸胶囊(9-5)中氨基酸的含量。

Measuring the content of amino acid in compound amino acid capsules ( 9-5 ) by es- tablishing the method of UPLC

Objective Optimal method of UPLC was established to measure the content of amino acid in com-pound amino acid capsules by detailed investigation of Chromatographic parameters. Methods Nine kinds of amino acids were external standard,with phenyl isothiocyanate as the derivatization agent. Use ACQUITY UPLC ? BEH C18 (2. 1 mm × 100 mm,1. 7 μm) column. The mobile phase A was 0. 1 mol/L sodium acetate solution ( adjusted with acetic acid to pH 6. 50 )-acetonitrile (93:7);and acetonitrile-solution(4:1) was the mobile phase B. The detection wavelength was 254 nm;the flow rate was 0. 45 mL/min;the column temperature was 36 ℃. Results The peak time of nine kinds of amino acids in Compound amino acid capsule (9-5) was the same as that of reference substance. There was no interference for other ingredients. RSD of instrument precision was 0. 5% ~1. 2%,and intermediate precision RSD was 1. 3% ~1. 9%;there was a good linear relationship of nine kinds of amino acids in different concentrations ( r≥0. 999);the stability of solution RSD was 0. 6% ~1. 8%;the average recovery of each kind of amino acid rate was 95. 0% ~105. 0%. There was no obvious difference between the method of UPLC and HPLC based on the content de-termination. UPLC improved the speed of analysis significantly. Conclusion The method of UPLC can be used for measuring the content of amino acid in compound amino acid capsules(9-5),which is efficient and accurate.