HPLC法测定复方阿胶膏中4种氨基酸的含量

Determination of four amino acids in compound ejiao paste by HPLC

目的建立复方阿胶膏中4种的含量测定方法。方法采用HPLC法,Shimpack ODS C18柱(4.6mm×250mm,5μm),以乙腈-0.1mol/L乙酸钠溶液(用冰乙酸调节pH值至6.5,7∶93)为流动相A,以乙腈-水(4∶1)为流动相B,进行梯度洗脱。结果 L-羟脯氨酸、甘氨酸、丙氨酸、L-脯氨酸分别在12.46~124.6μg/mL、23.62~236.2μg/mL、8.47~84.7μg/mL、19.14~191.4μg/mL范围内浓度与峰面积呈良好的线性关系(r=0.9999,n=6),平均加样回收率分别为97.91%、97.77%、97.67%、97.65%,RSD分别为1.70%、1.18%、1.31%、1.29%(n=9)。结论本法准确可靠,可用于复方阿胶膏的质量控制。

Objective To develop a method for determining the content of four amino acids in Compound Ejiao Paste. Methods HPLC method was used to analyze quantitatively the content of four acids with a Shimpack C518（250mm ×4.6mm,5 μ m）column.The mobile phase A was acetonitrile-0.1mol/L sodium acetate solution（adjust pH=6.5 by acetic acid, 7 ： 93）.The mobile phase B was acetonitrile-water（4 ： 1）,gradient elution. Results The calibration curves of L-hydroxyproline,glycine,alanine and L-proline were linear in the range 12.46- 124.6μ g/mL,23.62-236.2 μg/mL,8.47-84.7 μg/mL and 19.14 - 191.4μ g/mL （r=0.9999,n=6）respectively. The average recoveries were 97.91%（RSD=l.70%,n=9）for L-hydroxyproline,97.77%（RSD=1.18%,n=9）for glycine,97.67%（RSD=1.31%,n=9）for alanine and 97.65% （RSD=1.29%,n=9）for L-proline. Conclusion This method was accurate and reliable.It can be used for the quality control of compound ejiao paste.