柱前衍生-高效液相色谱快速测定皮革和头发水解液中的氨基酸

目的: 建立皮革和头发中17种蛋白水解氨基酸的快速高效液相色谱分析方法。 方法: 氨基酸用AccQ Tag衍生试剂衍生后,用Acclaim C₁₈(4.6 mm×150 mm,5 μm)柱分离,三元梯度洗脱。三元流动相为pH为4.45的140 mmol·L^-1醋酸钠溶液、乙腈-水(60∶ 40,v/v)、pH为4.95的140 mmol·L^-1醋酸钠溶液,以紫外检测器检测,在32 min内可简单快速地完成17种氨基酸的分析。 结果: 17种氨基酸衍生物保留时间的RSD≤1.1%(n=5),峰面积的RSD≤5.0%(n=5)。方法检测限(S/N=3)为2.92~4.51 pmol,线性范围2.5~500 μmol·L^-1(其中胱胺酸为1.25~250 μmol·L^-1),相关系数R²≥0.998,皮革样品加标回收率在80.1%~102.3%,头发样品加标回收率在85.3%~103.2%。 结论: 该方法不仅简化了流动相的配制过程,降低了实验成本,而且专属性高,样品基体干扰小,可完全满足皮革和头发水解液中氨基酸的测定。

Objective: To develop a fast HPLC method for determining 17 common amino acids in hydrolysate of leather and hair. Methods: Amino acids were separated on Dionex Acclaim C₁₈(4.6 mm×150 mm,5 μm) column within 32 min using 140 mmol·L^-1 sodium acetate buffer solution at pH 4.45 and 4.95 for gradient elution with acetonitrile-water(60∶ 40,v/v) and UV for detection after derivatization with AccQ Tag derivatization reagent. Results: The RSDs(n=5) of retention time and peak area of all amino acids were less than 1.1% and 5.0%,respectively.The limit of detection(S/N=3) was in the range of 2.92-4.51 pmol,the linear range was in the range of 2.5-500 μmol·L^-1(1.25-250 μmol·L^-1 for cystine) with R²≥0.998.The spiked recovery of leather and hair were in the range of 80.1%-102.3% and 85.3%-103.2%,respectively. Conclusion: The main advantages of the method are simplified for the preparation of mobile phase,cost saving,high speed and less matrix interference.It can be used for amino acids determination in hydrolysate of leather and hair effectively.