论文部分内容阅读
建立了一种用于测定中药制剂中氨基酸成分的毛细管电泳-荧光检测方法.用含有α-环糊精(α-CD)的硼砂缓冲溶液为背景电解质,经异硫氰酸荧光素(FITC)衍生的5种氨基酸在50min内可以得到很好的分离和测定.考查了各个分离参数对分离的影响,得到的优化条件为:含45mmol/L的α-环糊精的80mmol/L硼砂缓冲溶液(pH值9.2)作为背景电解质,分离电压20kV;柱温22℃.衍生试剂FITC与单个氨基酸的化学计量比为4∶1时,能够获得稳定荧光强度的氨基酸衍生物.在优化条件下,各氨基酸成分在73.5～2900nmol/L的浓度范围内呈良好的线性关系(相关系数r2为0.9906～0.9998).保留时间和峰面积的相对标准偏差分别为0.8%～3.0%和0.7%～5.7%,检测限(3倍信噪比)为3.5～35nmol/L.该方法准确可靠,可用于质量控制为目的的中药制剂中氨基酸成分的定量测定.
A capillary electrophoresis and fluorescence detection method was developed for the determination of amino acids in traditional Chinese medicine preparations.The borax buffer solution containing α-cyclodextrin (α-CD) was used as the background electrolyte and fluorescein isothiocyanate (FITC) The five kinds of amino acid derivatives were well separated and determined within 50 min.The influence of each separation parameter on the separation was investigated.The optimal conditions were as follows: 80 mmol / L borax buffer containing 45 mmol / L α-cyclodextrin (pH 9.2) as the background electrolyte, the separation voltage was 20kV and the column temperature was 22 ℃ .Amino acid derivatives with stable fluorescence intensity were obtained when the stoichiometric ratio of FITC to single amino acid was 4:1. Under optimized conditions, The amino acid composition showed a good linearity in the range of 73.5 ~ 2900 nmol / L (correlation coefficient r2 was 0.9906 ~ 0.9998) .The relative standard deviations of retention time and peak area were 0.8% ~ 3.0% and 0.7% ~ 5.7%, respectively, The detection limit (3 times signal to noise ratio) is 3.5 ~ 35nmol / L. This method is accurate and reliable, and can be used for quantitative determination of amino acid composition in traditional Chinese medicine preparation for quality control.