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目的:建立虎掌南星中各氨基酸含量测定的方法。方法:采用柱前衍生化后进行高效液相色谱法定量分析。色谱柱为Diamonsil C_(18)(250mm×4.6mm,5μm),柱温35℃;以异硫氰酸苯酯(PITC)为衍生化试剂;乙腈-醋酸铵缓冲液(pH=6.5)为流动相体系,梯度洗脱,流速1.0mL·min~(-1);检测波长254nm。结果:谷氨酸和色氨酸在2μg·mL~(-1)~100μg·mL~(-1)浓度范围内,精氨酸在6~300μg·mL~(-1)浓度范围内,其他氨基酸在0.8~40μg·mL~(-1)浓度范围内均与峰面积呈良好线性关系,相关系数均不低于0.999 5,平均回收率均在93.43%~111.01%之间,RSD均小于3%。结论:该方法灵敏、准确,重现性好,可用于虎掌南星中各氨基酸含量的测定。
Objective: To establish a method to determine the content of amino acids in Platycladus orientalis. Methods: Pre-column derivatization was used for quantitative analysis by high performance liquid chromatography. The column was Diamonsil C 18 (250 mm × 4.6 mm, 5 μm) and the column temperature was 35 ° C. Phenylisothiocyanate (PITC) was used as a derivatization reagent. Acetonitrile-ammonium acetate buffer (pH = 6.5) Phase system, gradient elution, flow rate 1.0mL · min ~ (-1); detection wavelength 254nm. Results: The concentration of glutamic acid and tryptophan in the range of 2μg · mL -1 -100μg · mL -1, the concentration of arginine in the range of 6-300μg · mL -1, The amino acid showed a good linear relationship with the peak area in the concentration range of 0.8 ~ 40μg · mL ~ (-1), the correlation coefficient was no less than 0.999 5, the average recovery was between 93.43% ~ 111.01%, RSD was less than 3 %. Conclusion: The method is sensitive, accurate and reproducible. It can be used to determine the content of amino acids in Platycodon grandiflorum.