摘要
目的:建立测定复方氨基酸(15)双肽(2)注射液中有关物质含量的高效液相色谱法。方法:采用Ultimate AQ-C_(18)(4.6 mm×250 mm,5μm)色谱柱,以0.01 mol·L^(-1)磷酸氢二铵溶液(pH 2.0)-甲醇(98∶2)为流动相,流速为0.7 mL·min^(-1),检测波长为205 nm,进样量为50μL。结果:在选定的色谱条件下,环-(甘氨酰谷氨酰胺)峰、焦谷氨酸峰和环-(甘氨酰酪氨酸)峰与各自相邻色谱峰可以有效分离;环-(甘氨酰谷氨酰胺)、焦谷氨酸、环-(甘氨酰酪氨酸)分别在7.863~52.42μg·mL^(-1)、5.256~35.04μg·mL^(-1)、0.939 3~6.262μg·mL^(-1)的范围内线性关系良好(r=0.999 9);精密度试验的RSD值均小于0.2%;重复性试验的RSD值均小于2.0%;环-(甘氨酰谷氨酰胺)、焦谷氨酸和环-(甘氨酰酪氨酸)定量限分别为3.15、14.0和7.5 ng,检测限分别为0.95、7.0和2.5 ng;加样回收率分别为103.0%、101.5%和103.3%(n=9),RSD值分别为1.0%、0.9%和0.9%。9批样品环-(甘氨酰谷氨酰胺)的量为383~492 mg·L^(-1),焦谷氨酸的量为191~315 mg·L^(-1),环-(甘氨酰酪氨酸)的量为39~49 mg·L^(-1)。结论:该方法专属性强,结果准确,可用于复方氨基酸(15)双肽(2)注射液中有关物质的测定,能有效地反应药品质量。
Objective:To establish an HPLC method for the determination of related substances in compound amino acids (15)and dipeptides (2)injection.Methods :The test was performed in Ultimate AQ-C18 column (4.6 min× 250 mm,5 μm)under the elution of 0.01 mol·L^-1 diammonium hydrogen phosphate buffer solution (pH 2.0)-methanol (98:2).The flow rate was 0.7 mL ~rain-1,the detection wavelength was set at 205 nmand the injection volume was 50 μL.Results:The peaks of cyclo-(Gly-Gln),pyroglutamic acid and cyclo-(Gly-Tyr)were effectively separated from their adjacent chromatographic peaks under the developed chromatographic condition.The linear ranges of cyclo-(Gly-Gln ),pyroglutamic acid and cyclo-(Gly-Tyr)fell into 7.863 -52.42,5.256 - 35.04,0.939 3 -6.262 μg·mL^-1 with r =0.999 9,respectively.The RSDs of precision test were less than 0.2%and the RSDs of repeatability test were less than 2.0%.The LOQ values of cyclo-(Gly-Gln),pyroglutamic acid and cyclo-(Gly-Tyr)were 3.15,14.0 and 7.5 ng,respectively,LOD values of cyclo-(Gly-Gln),pyroglutam- ic acid and cyclo-(Gly-Tyr)were 0.95,7.0 and 2.5 ng,respectively.The average spiked recoveries of cyclo-(Gly- Gln),pyroglutamic acid and cyclo-(Gly-Tyr)were 103.0%,101.5 % and 103.3 %(n =9)with RSDs as 1.0%, 0.9%and 0.9%,respectively.The contents of cyclo-(Gly-Gln),pyroglutamic acid and cyclo-(Gly-Tyr)in 9 batches of samples ranged from 383 -492,191 -315,and 39 -49 μg·mL^-1,respectively.Conclusion:The established method has good specificity and accuracy.It can be used for the determination of the related substances in compound amino acids (15)and dipeptides (2)injection.
作者
陈宇堃
蒋玉辉
梁蔚阳
CHEN Yukun;JIANG Yuhui;LIANG Weiyang(Guangdong Institute for Drug Control,Guangzhou 510663,China)
出处
《中国药品标准》
CAS
2019年第1期25-30,共6页
Drug Standards of China
基金
广东省医学科学技术研究基金项目(B2018130
B2018220)
广东省省级科技计划项目(2015A030402004)