HPLC柱前程序衍生法测定复方α-酮酸片中5种氨基酸的含量

Content Determination of 5 Kinds of Amino Acids in Compound α-Ketoacid Tablets by HPLC after Pre-column Programming Derivatization

目的:采用HPLC柱前程序衍生法测定复方α-酮酸片中5种氨基酸的含量。方法:以辛烷基硅烷键合硅胶为填充剂,梯度洗脱方式,柱温为40℃,以邻苯二甲醛为衍生试剂,检测波长为338 nm。检测复方α-酮酸片中L-醋酸赖氨酸、L-苏氨酸、L-色氨酸、L-组氨酸、L-酪氨酸的含量。结果:L-醋酸赖氨酸、L-苏氨酸、L-色氨酸、L-组氨酸和L-酪氨酸检测浓度的线性范围分别为0.10～5.25(r=0.999 9)、0.052～2.65(r=0.999 8)、0.022～1.15(r=0.999 9)、0.038～1.91(r=0.999 7)、0.030～1.51(r=1.000 0)mg·mL-1;平均回收率为99.8%～100.1%(RSD均为0.1%)。结论:该方法灵敏度高,专属性强,结果准确可靠,可用于测定复方α-酮酸片中氨基酸的含量。

OBJECTIVE： To establish an HPLC method with pre-column programming derivatization for the content determination of 5 kinds of amino acids in Compound α-ketoacid tablets.METHODS： The separation was performed on C8 column（gradient elution） with column temperature at 40 ℃.OPA was used as the derivation reagent.The detection wavelength was set at 338 nm.The contents of L-lysine acetate,L-threoine,L-tryptophan,L-histidine,L-tyrosine in Compound α-ketoacid tablets were determined.RESULTS： The linear ranges of L-lysine acetate,L-threoine,L-tryptophan,L-histidine,L-tyrosine were 0.10～5.25 mg·mL^-1（r=0.999 9）,0.052～2.65 mg·mL^-1（r=0.999 8）,0.022～1.15 mg·mL^-1（r=0.999 9）,0.038～1.91 mg·mL^-1（r=0.999 7）,0.030～1.51 mg·mL^-1（r=1.000 0）,respectively.The average recovery was 99.8%～100.1%（RSD=0.1%）.CONCLUSION： This method is exclusive,sensitive and reliable for the content determination of amino acids in Compound α-ketoacid tablets.