RP-HPLC法同时测定石韦中4种氨基酸的含量

Contents Determination of 4 Amino Acids in Pyrrosia lingua by RP-HPLC

目的：建立同时测定石韦中4种氨基酸含量的方法。方法：采用反相高效液相色谱法。色谱柱为Welchrom C18,流动相A为0.1 mol/L的醋酸钠缓冲液（以醋酸钠调节p H至6.5）-乙腈（93∶7,V/V）,流动相B为乙腈-水（4∶1,V/V）,梯度洗脱,流速为1.0ml/min,检测波长为254 nm,柱温为30℃,进样量为5μl。结果：谷氨酸、甘氨酸、丙氨酸、亮氨酸的质量浓度分别在4.92～49.2、3.168～31.68、2.6～26、2.88～28.8μg/ml范围内与各自峰面积呈良好的线性关系（r=0.999 1、0.999 9、0.999 4、0.999 9）;精密度、稳定性、重复性试验的RSD均≤2.2%;平均加样回收率分别为98.4%、99.6%、98.3%、100.1%,RSD分别为2.2%、2.5%、2.2%、2.3%（n=9）。结论：该方法简便、准确、重复性好,可用于石韦中4种氨基酸的含量测定。

OBJECTIVE ： To establish a method for the contents determination of 4 amino acids in Pyrrosia lingua. METHODS ： The samples were analyzed by RP-HPLC. It was performed on a column of Welchrom C18with the mobile phase A of 0.1 mol/L So- dium acetate buffer （pH6.5）- acetonitrile（93 ： 7, V/V） and mobile phase B of acetonitrile-water（4 ： 1, V/V） （gradient elution） at the flow rate of 1.0 ml/min, detection wavelength was 254 nm, column temperature was 30 ℃ and the sample size was 5 μl. RE- SULTS：The linear range was 4.92-49.2 μg/ml for glutamate（r=0.999 1）, 3.168-31.68 pg/ml for glycine（r=0.999 9）,2.6-26μg/ml for alanine（r=0.999 4） and 2.88-28.8 μg/ml for leucine （r=0.999 9）; RSDs of precision, stability and reproducibility tests were no more than 2.2%; average recoveries were 98.4% （RSD=2.2% ,n=9）, 99.6% （RSD=2.5% ,n=9）, 98.3% （RSD=2.2% ,n= 9） and 100.1% （RSD=2.3% ,n=9）, respectively. CONCLUSIONS： The method is simple, accurate and reproducible, and can be used for the contents determination of 4 amino acids in P. lingua.