摘要
目的:改进测定门冬氨酸鸟氨酸原料药中有关物质的方法。方法:采用高效液相色谱法。色谱柱为Phenomenex Luna NH2(两根串联),流动相为0.02 mol/L磷酸二氢钾缓冲液(p H 5.6)-乙腈(40∶60,V/V),流速为1.0 m L/min,检测波长为205 nm,柱温为35℃,进样量为20μL。结果:马来酸、3-氨基-2-哌啶酮、琥珀酸、苹果酸、富马酸、β-门冬氨酰门冬氨酸检测质量浓度线性范围分别为5.398~26.992μg/m L(r=0.999 1)、5.204~26.021μg/m L(r=0.999 3)、5.149~25.749μg/m L(r=0.999 6)、5.174~25.874μg/m L(r=0.999 3)、5.164~25.823μg/m L(r=0.999 3)、5.194~25.973μg/m L(r=0.999 5);定量限分别为0.22、0.53、51.50、103.50、0.26、8.31 ng,检出限分别为0.13、0.26、10.30、20.70、0.13、2.08 ng;精密度、稳定性、重复性试验的RSD均不超过2%;加样回收率分别为99.30%~102.31%(RSD=1.02%,n=9)、99.88%~100.89%(RSD=0.37%,n=9)、99.36%~101.53%(RSD=0.70%,n=9)、99.13%~102.65%(RSD=1.15%,n=9)、100.18%~101.45%(RSD=0.38%,n=9)、99.39%~100.81%(RSD=0.58%,n=9)。6批样品均未检出马来酸、琥珀酸和β-门冬氨酰门冬氨酸;3-氨基-2-哌啶酮含量为0.006%~0.056%,苹果酸含量为0.014%~0.071%,富马酸含量为0.000 5%~0.003 4%,最大未知杂质含量为0.013%~0.110%,未知杂质总含量为0.013%~0.120%。结论:该方法操作简便,精密度、稳定性、重复性均较好,可用于门冬氨酸鸟氨酸原料药中有关物质的测定。
OBJECTIVE:To improve the determination method of related substances in ornithine aspartate raw material.METHODS:HPLC method was adopted.The determination was performed on Phenomenex Luna NH2 column(two seriesconnected)with mobile phase consisted of 0.02 mol/L potassium dihydrogen phosphate buffer solution(pH 5.6)-acetonitrile(40∶60,V/V)at the flow rate of 1.0 mL/min.The detection wavelength was set at 205 nm,and column temperature was 35℃.The sample size was 20μL.RESULTS:The linear range of maleic acid,3-amino-2-piperidinone,succinic acid,malic acid,fumaric acid andβ-aspartyl-aspartic acid were 5.398-26.992μg/mL(r=0.999 1),5.204-26.021μg/mL(r=0.999 3),5.149-25.749μg/mL(r=0.999 6),5.174-25.874μg/mL(r=0.999 3),5.164-25.823μg/mL(r=0.999 3),5.194-25.973μg/mL(r=0.999 5).The limits of quantitation were 0.22,0.53,51.50,103.50,0.26 and 8.31 ng;the limits of detection were 0.13,0.26,10.30,20.70,0.13 and 2.08 ng.RSDs of precision,stability,and reproducibility tests were no more than 2%.The average recoveries were 99.30%-102.31%(RSD=1.02%,n=9),99.88%-100.89%(RSD=0.37%,n=9),99.36%-101.53%(RSD=0.70%,n=9),99.13%-102.65%(RSD=1.15%,n=9),100.18%-101.45%(RSD=0.38%,n=9),99.39%-100.81%(RSD=0.58%,n=9).Neither maleic acid,succinic acid andβ-aspartyl-aspartic acid were found in 6 batches of samples.The contents of 3-amino-2-piperidinone were 0.006%-0.056%;the contents of malic acid were 0.014%-0.071%;the contents of fumaric acid were 0.000 5%-0.003 4%;the contents of maximal unknown impurity were 0.013%-0.110%;total contents of unknown impurity were 0.013%-0.120%.CONCLUSIONS:The method is simple,precise,stable and reproducible,and can be used for the determination of the related substances in ornithine aspartate raw material.
作者
陈宇堃
陈华
梁蔚阳
CHEN Yukun;CHEN Hua;LIANG Weiyang(Guangdong Institute for Drug Control,Guangzhou 510180,China)
出处
《中国药房》
CAS
北大核心
2018年第18期2509-2515,共7页
China Pharmacy
基金
广东省科技计划项目(No.粤科规财字[2016]48号-2016A010121004
粤科规财字[2015]150号-2015A030402004)
广东省医学科学技术研究基金项目(No.粤卫函[2016]568号-B2016121
B2016106)
