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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0.169～1.014μg(r=0.999 5),平均回收率101.43%,RSD 3.20%;L-羟脯氨酸的线性范围0.178～1.068μg(r=0.999 9),平均回收率101.23%,RSD 3.25%;甘氨酸的线性范围0.381～2.286μg(r=0.999 5),平均回收率102.87%,RSD 2.32%;L-丙氨酸的线性范围为0.162～0.972μg(r=0.999 5),平均回收率102.37%,RSD 3.36%;L-精氨酸的线性范围0.118～0.708μg(r=0.999 6),平均回收率102.73%,RSD 2.41%;L-脯氨酸的线性范围0.253～1.518μg(r=0.999 9),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。
OBJECTIVE: To establish a method for the simultaneous determination of six kinds of hydrolyzed amino acids in Jiaojiao Buxue Granules by precolumn derivatization reversed-phase high performance liquid chromatography. METHODS: Hypersil BDS C18 column (4.6 mm × 200 mm, 5 μm) and phenylisothiocyanate as pre-column derivatization reagent were used. The flow rate was 1.0 mL · min-1. The detection wavelength was set at 254 nm and the column temperature was set at 30 ℃ . Results: The linear range of L-glutamic acid was 0.169-1.014μg (r = 0.999 5), the average recovery was 101.43% and the RSD was 3.20%. The linear range of L-hydroxyproline was 0.178-1.068μg (r = 0.999 9) , The average recovery was 101.23% and the RSD was 3.25%. The linear range of glycine was 0.381 ~ 2.286μg (r = 0.999 5) with the average recovery of 102.87% and RSD 2.32%. The linear range of L-alanine was 0.162 ~ 0.972μg The average recovery was 102.37% and RSD was 3.36% respectively. The linear range of L-arginine was 0.118-0.708μg (r = 0.999 6), the average recovery was 102.73% and the RSD was 2.41% The linear range of 0.253 ~ 1.518μg (r = 0.999 9), the average recovery rate of 98.69%, RSD 2.34%. Conclusion: The established method is reliable and can be used for the determination of amino acids in donkey jelly blood granules.