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目的:采用柱前衍生化法测定板蓝根颗粒中6种游离氨基酸的含量。方法:采用6-氨基喹啉基-N-羟基琥珀酰亚氨基甲酸酯(AQC)进行柱前衍生化,超高效液相色谱法测定板蓝根颗粒中精氨酸、脯氨酸、苏氨酸、丙氨酸、γ-氨基丁酸、缬氨酸的含量。Waters AccQ Tag TM Ultra C18色谱柱(2.1 mm×100 mm,5μm),流动相AccQ Tag Ultra Eluent A与AccQ Tag Ul-tra Eluent B进行梯度洗脱,流速0.7 mL.min-1,柱温55℃,检测波长260 nm。结果:精氨酸的线性范围为4.155~49.86μg,r=0.999 9;苏氨酸的线性范围为0.595~5.95μg,r=0.999 8;丙氨酸的线性范围为0.445~4.45μg,r=0.999 9;γ-氨基丁酸的线性范围为0.515~5.15μg,r=0.999 9;脯氨酸的线性范围为8.858~106.3μg,r=0.999 9;缬氨酸的线性范围为0.585~5.85μg,r=0.999 8。平均回收率分别为100.6%,98.34%,98.35%,100.2%,98.44%,98.18%;平均RSD分别为1.8%,1.5%,1.9%,2.0%,2.4%,2.0%(n=6)。5家企业的的板蓝根颗粒中氨基酸类成分的含量存在差异。结论:该方法操作简单,重复性好,灵敏度高,结果准确可靠。
OBJECTIVE: To determine the content of six free amino acids in Banlangen granules by precolumn derivatization. METHODS: Pre-column derivatization was performed using 6-aminoquinolyl-N-hydroxysuccinimide (AQC) and the contents of arginine, proline and threonine in Banlangen granules were determined by ultra performance liquid chromatography , Alanine, γ-aminobutyric acid, valine content. Waters AccQ Tag ™ Ultra C18 column (2.1 mm × 100 mm, 5 μm). The mobile phase was AccQ Tag Ultra Eluent A and AccQ Tag Ul-tra Eluent B. The flow rate was 0.7 mL.min-1 and the column temperature was 55 ℃ , Detection wavelength 260 nm. Results: The linear range of arginine was 4.155 ~ 49.86μg, r = 0.999 9; the linear range of threonine was 0.595 ~ 5.95μg, r = 0.999 8; the linear range of alanine was 0.445 ~ 4.45μg, r = 0.999 9, the linear range of γ-aminobutyric acid was 0.515-5.15μg, r = 0.999 9, the linear range of proline was 8.858-106.3μg, r = 0.999 9, and the linear range of valine was 0.585-5.85μg , r = 0.999 8. The average recovery rates were 100.6%, 98.34%, 98.35%, 100.2%, 98.44% and 98.18%, respectively. The average RSDs were 1.8%, 1.5%, 1.9%, 2.0%, 2.4% and 2.0%, respectively. There are differences in the contents of amino acids in Banlangen granules of five companies. Conclusion: The method is simple, reproducible, sensitive and the results are accurate and reliable.