青葙子中氨基酸的柱前衍生化HPLC-DAD法测定

Determination of amino acids in Celosiae Semen by pre- column derivation and HPLC-DAD method

目的建立柱前衍生HPLC-DAD法测定中药青葙子中游离氨基酸和水解氨基酸的含量方法。方法采用超声提取法和盐酸水解法制备青葙子的2类氨基酸样品溶液,以异硫氰酸苯酯柱前衍生HPLC-DAD法分析青葙子中的氨基酸。结果 18种氨基酸的分离良好,空白无干扰,线性关系良好,精密度RSD在0.30%-1.58%之间,重复性RSD在0.47%-1.92%之间,稳定性RSD在0.42%-1.71%之间,加样回收率96%-103%之间且RSD均小于5%,游离和水解氨基酸的含量分别为0.4022%、6.9082%,药用氨基酸和必需氨基酸含量较高。结论该方法简便、快捷,能准确地测定青葙子中游离和水解氨基酸的含量,为青葙子及其制剂的质量控制提供了依据。

Objective To establish a phenyl isothiocyanate( PITC) pre- column derivation and HPLC- DAD method for the determination of free and hydrolysis amino acids in Celosiae Semen. Methods The free amino acids were extracted by ultrasonic and the hydrolysis amino acids by 6mol·L- 1hydrochloric acid,PITC as the pre- column derivating reagent,the content of the amino acids were determined by HPLC- DAD method. Results The linear relationship was good,the precision RSD was between 0. 30% and 1.58%,the repeatability RSD was between 0. 47% and 1. 92%,the stability RSD was between 0. 42% and 1. 71%,the sample recovery was between 95% and 102%,and the RSD was less than 5%. The contents of free and hydrolysis amino acids were 0. 4022% and6. 9082%. Medicinal and essentialamino acids have higher levels of quantitation. Conclusion The method can be very accurate,sensitive and reliable,determinate of contents of the free and hydrolysis amino acids in Celosiae Semen and its preparation.