柱前衍生HPLC同时测定驴胶补血颗粒中6种水解氨基酸

目的:建立柱前衍生反相高效液相色谱法同时测定驴胶补血颗粒中6种水解氨基酸含量的方法.方法:采用Hypersil BDS C18色谱柱(4.6 mm×200 mm,5μm),异硫氰酸苯酯为柱前衍生剂,梯度洗脱,流速1.0 mL· min-1,检测波长254nm,柱温30℃.结果:L-谷氨酸的线性范围0.169 ～1.014 μg(r =0.9995),平均回收率101.43％,RSD 3.20％；L-羟脯氨酸的线性范围0.178～ 1.068 μg(r =0.999 9),平均回收率101.23％,RSD 3.25％；甘氨酸的线性范围0.381～2.286 μg(r=0.9995),平均回收率102.87％,RSD 2.32％;L-丙氨酸的线性范围为0.162 ～0.972 μg(r=0.999 5),平均回收率102.37％,RSD 3.36％；L-精氨酸的线性范围0.118 ～0.708 μg(r =0.999 6),平均回收率102.73％,RSD 2.41％；L-脯氨酸的线性范围0.253 ～ 1.518 μg(r =0.9999),平均回收率98.69％,RSD 2.34％.结论:建立的方法可靠,可用于驴胶补血颗粒中氨基酸的含量检测.

Determination of 6 Amino Acids in Lujiao Buxue Keli by Pre-Column Derivation HPLC

Objective:To develop a HPLC method for determining 6 amino acids in Lujiao Buxue keli.Method: The sample was derived with phenyl isothiocyanate(PITC).Amino acids were separated on Hypersil BDS C18(4.6 mm×200 mm,5 μm) column at the flow rate of 1.0 mL · min-1,detected at 254 nm.Result: L-Glu had good linearity in the range of 0.169-1.014 μg(r=0.999 5).The average recovery was 101.43%,RSD was 3.20%;L-Hyp had good linearity in the range of 0.178-1.068 μg(r=0.999 9).The average recovery was 101.23%,RSD was 3.25%;Gly had good linearity in the range of 0.381-2.286 μg(r=0.999 5).The average recovery was 102.87%,RSD was 2.32%;L-Ala had good linearity in the range of 0.162-0.972 μg(r=0.999 5).The average recovery was 102.37%,RSD was 3.36%;L-Arg had good linearity in the ranges of 0.118-0.708 μg(r=0.999 6).The average recovery was 102.73%,RSD was 2.41%;L-Pro had good linearity in the range of 0.253-1.518 μg(r=0.999 9).The average recovery was 98.69%,RSD was 2.34%. Conclusion: The method is reliable,which is helpful for the determination of amino acids in Lujiao Buxue keli.