柱前衍生化RP-HPLC测定铁皮石斛中的氨基酸的含量  

Pre-column Derivatization RP-HPLC Determination of Amino Acids in Dendrobii Officinalis Caulis

作　　者：张亮[1] 鲁芹飞[1] 张扬[1] 徐香琴[1] 徐兰芳[1] 曾毅[1] 黄松[1,2] 

机构地区：[1]广州中医药大学,广州510006 [2]东莞广州中医药大学中医药数理工程研究院,广东东莞523808

出　　处：《中国实验方剂学杂志》2014年第22期61-65,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：广东省科技计划项目(2008A030101009);广东省科技计划项目(2011A030100011);广东省教育部产学研结合项目(2010B090400474)

摘　　要：目的：测定不同产地的铁皮石斛样品中氨基酸的含量。方法：样品以异硫氰酸苯酯（PITC）为衍生化试剂,与铁皮石斛中氨基酸柱前衍生,采用高效液相色谱法测定其含量,采用CAPCELL PAK C18SG300色谱柱（4.6 mm×250 mm,5μm）,流动相[0.05 mol·L^-1乙酸钠缓冲液（以乙酸调p H 6.5）-乙腈（1∶1）]（A）-[0.05 mol·L^-1乙酸钠缓冲液（以乙酸调p H6.5）]（B）梯度洗脱,柱温25℃,流速0.6 m L·min^-1,检测波长254 nm。结果：游离氨基酸和总的氨基酸的回收率范围分别为92.48%-97.30%和90.95%-95.63%,RSD都小于3.0%,9个不同产地的样品游离氨基酸的含量范围在0.210 5%-0.571 2%,总的氨基酸含量范围在3.670 8%-6.890 4%,必需氨基酸占总的游离氨基酸的比例为0.094 4%-0.186 0%;必需氨基酸占总的氨基酸的比例范围为0.338 5%-0.374 1%。结论：该方法灵敏、准确,具有良好的重复性和稳定性,可用于铁皮石斛中氨基酸的检测,不同产地的铁皮石斛样品其氨基酸的含量存在差异,必需氨基酸的比例也存在差异。Objective： To determine the contents of amino acids of different origin of Dendrobii Officinalis Caulis. Method： The amino acids in Dendrobii Officinalis Caulis pretreated with phenyl isothiocyanate（ PITC） and the content of derivatives was determined by HPLC. The determination was performed on CAPCELL PAK C18SG300（ 4. 6 mm × 250 mm,5 μm） column with mobile phase A consisted of sodium acetate-acetic acid buffer solution（ adjusted p H to 6. 5 with acetic acid）-1% acetonitrile and mobile phase B consisted of sodium acetate-acetic acid buffer solution（ adjusted p H to 6. 5 with acetic acid）; gradient elution. The column temperature was 25 ℃, flow rate was 0. 6 m L·min^-1,the detection wavelength was 254 nm. Result： The recovery of free amino acid and total amino acids were in the range of 92. 48%-97. 30% and 90. 95%-95. 63%,the RSD values are less than 3%. Different origin of free amino acid content of 10 samples in the range of 0. 2105%-0. 571 2%,the total amino acid content in the range of 3. 670 8%-6. 890 4%,the essential amino acids accounted for the proportion of total free amino acids in the range of 0. 094 4%-0. 186 0% and accounted for the proportion of total amino acids in the range of 0. 338 5% to 0. 374 1%. Conclusion： The method is sensitive,accurate,with good repeatability and stability,can be used for the detection of amino acids in Dendrobii Officinalis Caulis,there was significant difference between the different origin of Dendrobium candidum samples of amino acid,there are differences in the proportion of essential amino acids.

关 键 词：铁皮石斛 柱前衍生化 氨基酸 高效液相色谱法 

分 类 号：R284.1[医药卫生—中药学]