柱前衍生化RP-HPLC测定鸡内金中16种氨基酸的含量

Determination of amino acids in Galli Gigerii Endothelium Corneum by HPLC with pre-column derivatization

采用酸水解法制备水解氨基酸样品溶液，以异硫氰酸苯酯（PITC）柱前衍生化后进行HPLC分析。采用Ultimate Prime C18色谱柱（4．6mm×250mm，5tzm）；流动相A0．1mol·L-1醋酸钠溶液（调pH至6．5）-乙腈（93：7），流动相B乙腈-水（8：2）；流速1．0mL·min-1；柱温40oC；检测波长254nm；梯度洗脱。氨基酸衍生溶液在36h内保持稳定，16种氨基酸线性相关系数均大于0．9995，加样回收率在98．01％～101．8％。该方法操作简便，精密度和重复性良好，可用于鸡内金中16种水解氨基酸含量的测定。

Hydrolytic amino acids were extracted by acid hydrolysis method, then derivatized with phenyl isothiocyanate （PITC）. And the samples were analysed by HPLC on an Ultimate Prime C18（4. 6 mm × 250 mm ,5 μm）column with gradient elution of 0. 1 mol L- 1 sodium acetate buffer solution （ adjusted to pH 6. 5 ） -acetonitrile （93： 7 ） （A） and acetonitrile-water （ 8： 2） （B） at a flow rate of 1.0 mL min-1. Column temperature was 40 ℃ and the detected wavelength was 254 nm. Amino acids derivative solution remained stable in 36 hours. The response was linear for 16 amino acids with a correlation coefficient r 〉0. 999 5. The average recover- ies were 98.01% -101.8%. The method is reliable with good accuracy and repeatability, which is useful for the determination of amino acids in Galli Gigerii Endothelium Corneum.