骆驼蓬中游离氨基酸的 HPLC 含量测定方法及超声提取工艺研究

摘要

Objective To establish the ultrasonic extraction process of free amino acids in the Peganum harmala herb,and determine the free amino acids in the Peganum harmala herb by HPLC.Methods The determination of amino acids in the Peganum harmala herb was performed on VP-ODS column (4.6× 250 mm,5 μm)and mobile phase consisted of acetonitrile- ammonium acetate buffer solution(PH 6.5). The column temperature was 40℃.The flow rate was 1.0 mL/min and the UV detective wavelength was set at 254 nm.The total content of free amino acids was used as index,the single-factor test and the or-thogonal test were adopted to optimize the extraction conditions of ultrasonic extraction methods of the Peganum harmala herb.Results The simultaneous determination of amino acids in the Peganum harmala herb could be used as quality control method.The results of amino acids of linearity,stability, reproducibility and precisions were good.The recoveries were 98%~102%.Ultrasonic extraction method was the suitable extraction of free amino acids in Peganum harmala herb:the 80% ethanol as the extrac-tion solvent and solvent multiple was 60,ultrasonic power under the condition of 400 Hz for 40 min.Con-clusion Established the method of qualitative and quantitative is precise and stable.The optimized ultra-sonic extraction method is stable,feasible,and reasonable,which can be used to extract the amino acids in the Peganum harmala herb in lab.%目的：研究新疆骆驼蓬药材中游离氨基酸的高效液相含量测定方法，建立新疆骆驼蓬游离氨基酸的超声提取工艺。方法采用日本岛津 VP-ODS 色谱柱（4．6 mm×250 mm，5μm），以0．10 mol／L 乙酸铵溶液（pH 6．5）-乙腈为流动相梯度洗脱，流速1．0 mL／min，检测波长254 nm，柱温40℃，测定新疆骆驼蓬药材地上部分游离氨基酸的含量，并进行方法学考察。以骆驼蓬游离氨基酸含量之和为考察指标，采用 L9（34）正交试验设计优选新疆骆驼蓬游离氨基酸超声提取工艺。结果测定的12种氨基酸分别在其线性范围内且线性关系良好，精密度、重复性、稳定性、衍生化重复性均良好，RSD 均＜2．0％，12种氨基酸的平均回收率为98％～102％。结合单因素和正交试验结果，超声提取法优选的工艺条件为：80％乙醇60倍量，超声功率400 Hz 条件下下超声40 min。结论建立的 HPLC 测定游离氨基酸含量的方法准确可靠，优选的最佳提取工艺合理、可行、稳定。