摘要
目的:建立复方氨基酸注射液(3AA)的主成分和有关物质测定的高效液相色谱方法。方法:采用反相高效液相色谱法并结合二维柱切换LC/MSn法对制剂中主要杂质进行分离和结构鉴定,建立复方氨基酸注射液(3AA)中缬氨酸、异亮氨酸和亮氨酸及其有关物质含量检测方法。采用Capcell PAK AQ C_(18)(250 mm×4.6 mm,3μm)色谱柱,以0.2 mol·L^(-1)磷酸二氢钠溶液(用磷酸调pH至2.8)-乙腈(98∶2)为流动相,流速1.0 mL·min^(-1),柱温40℃,检测波长210 nm,进样量20μL。采用Thermo Accucore AQ C18(100 mm×4.6 mm,2.6μm)色谱柱,以0.1%甲酸溶液为流动相A,以0.1%甲酸溶液-乙腈为流动相B,流速0.4 mL·min^(-1),柱温40℃;质谱条件采用ESI电离源,正离子扫描模式,扫描范围为m/z 100~1000,二级质谱采用数据依赖型扫描开展。结果:主峰和相邻杂质峰分离度良好,缬氨酸的线性范围为1.263~5.050 mg·mL^(-1),平均回收率(n=9)为99.0%;异亮氨酸的线性范围为1.350~5.402 mg·mL^(-1),平均回收率(n=9)为99.4%;亮氨酸的线性范围为1.647~6.588 mg·mL^(-1),平均回收率(n=9)为99.5%。3批样品中主要杂质均为原料引入的工艺杂质,其中甲硫氨酸含量分别为4.344、3.751、4.503μg·mL^(-1),苯丙氨酸含量分别为4.636、4.889、4.753μg·mL^(-1)。其他单个未知杂质分别为0.01%、0.02%、0.01%。结论:经方法学验证,本方法可用于复方氨基酸注射液(3AA)的质量控制。
Objective:To establish an HPLC method of the content and related substances of compound amino acid injection(3AA).Methods:RP-HPLC was adopted to determine compound amino acid injection(3AA),combining with the use of two-dimensional column switching-LC/MSn method was applied to separate and identify the impurities.The determination was performed on Capcell PAK AQ C_(18)(250 mm×4.6 mm,3μm)column with 0.2 mol·L^(-1)sodium dihydrogen phosphate solution(adjusted pH to 2.8 with phosphoric acid)-acetonitrile(98∶2)as mobile phase at the flow rate of 1.0 mL·min^(-1).The column temperature was 40℃,and the detection wavelength was 210 nm.And the injection volume was 20μL.The LC/MSn method was performed on a Thermo Accucore AQ C18(100 mm×4.6 mm,2.6μm)column with 0.1%formic acid solution as mobile phase A,0.1%formic acid solution-acetonitrile as mobile phase B,at a flow rate of 0.4 mL·min^(-1),and at a column temperature of 40℃.The mass spectrometry conditions were performed using an ESI ionisation source in the positive-ion scanning mode with a scan range of m/z 100-1000,and the secondary mass spectrum was carried out by data-dependent scanning.Results:The related substances were completely separated from the main constituents in RP-HPLC.The standard curve of valine was linear over the range of 1.263-5.050 mg·mL^(-1),with the average recovery of 99.0%(n=9).The standard curve of isoleucine was linear over the range of 1.350-5.402 mg·mL^(-1),with the average recovery of 99.4%(n=9).The standard curve of leucine was linear over the range of 1.647-6.588 mg·mL^(-1),with the average recovery of 99.5%(n=9).The main impurities in the three batches of samples were all process impurities introduced from the raw materials,with methionine content of 4.344μg·mL^(-1),3.751μg·mL^(-1),4.503μg·mL^(-1),respectively,phenylalanine content of 4.636μg·mL^(-1),4.889μg·mL^(-1),4.753μg·mL^(-1),respectively.The maximum single impurity contents were 0.01%,0.02%and 0.01%,respectively.Conclusion:The method is proved by the methodology validation that it can be used for the quality control of compound amino acid injection(3AA).
作者
左利民
茹仙古丽·依明
郭鑫
肖菁
徐士婕
赵婷
连晓芳
刘惠一
周怡
山广志
ZUO Li-min;Ruxianguli·Yiming;GUO Xin;XIAO Jing;XU Shi-jie;ZHAO Ting;LIAN Xiao-fang;LIU Hui-yi;ZHOU Yi;SHAN Guang-zhi(Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences,Beijing 100050,China;Xinjiang Institute of Materia Medica,Urumqi 830004,China;Hunan Institute for Drug Control,Changsha 410001,China;Chinese Pharmacopoeia Commission,Beijing 100061,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2024年第7期1161-1168,共8页
Chinese Journal of Pharmaceutical Analysis
基金
中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-070)
湖南省自然科学基金科药联合基金项目(2022JJ80071)。
关键词
复方氨基酸注射液(3AA)
含量测定
二维柱切换
串联质谱法
有关物质
结构鉴定
质量控制
compound amino acid injection(3AA)
content determination
two-dimensional column switching
tandem mass spectrometry
related substances
structural identification
control of drug quality