AQC柱前衍生化高效液相色谱法测定宝乐果中的游离氨基酸

摘要

To determine and analyze free amino acids in Borojo.The sample was derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) and then was analyzed with high-performance liquid chromatography (HPLC).HPLC was performed on a Phenomenex C18 (250mm × 4.6 mm,5 μm) column with gradient elution of deionized water (A),acetonitrile (B) and 10 mmol/L ammonium acetate buffer solution (adjusted to pH 5.10 with glacial acetic acid) (C) at the flow rate of 0.8 ～1.0 mL/min,detected with Ultra Voilet (UV) at 248 nm.Linear relationship equations were obtained with 18kinds of amino acids in the range of 0.02 ～ 0.5 mmoL/L,and their correlation coefficients were in the range of 0.998 3 ～ 0.999 2.Fourteen amino acids were detected and average recoveries of those added were in the range of 96.9 ％ ～ 106.9 ％ and RSD values were in the range of 0.67 ％ ～ 3.07 ％.The method could be used for determination of content of free amino acids in Borojo.%为了测定宝乐果中游离氨基酸的种类以及含量.该文的样品用6-氨基喹啉-N-羟基琥珀酰亚胺基-氨基甲酸酯(AQC)柱前衍生后进行高效液相色谱(HPLC)分析.采用Phenomenex Luna C18 (250 mm×4.6 mm,5 μm)色谱柱,柱温为35℃；流动相A为去离子水,流动相B为乙腈,流动相C为10 mmol/L醋酸铵缓冲液(以冰醋酸调至pH 5.10),梯度洗脱,流速为0.8～1.0 mL/min;检测波长为248 nm,同时测定宝乐果中的游离氨基酸.结果得:18种氨基酸在0.02～0.5 mmol/L浓度范围内有良好的线性关系,r均在0.998 3 ～0.999 2之间；宝乐果中测得14种游离氨基酸,其含量范围为0.087 ～2.680 mg/g,平均加样回收率为96.9％～106.9％,RSD为0.67％～3.07％.该方法可用于检测宝乐果中游离氨基酸含量,测定的结果为宝乐果质量研究和评价提供了依据.