UPLC法测定CHO细胞培养液中17种氨基酸的含量及其变化规律  

Determination and Variation of Amino Acid in CHO Cell Culture Medium by UPLC

作　　者：周杰[1] 蔡俊[1] ZHOU Jie;CAI Jun(Hubei University of Technology,Wuhan 430068,China)

机构地区：[1]湖北工业大学,湖北武汉430068

出　　处：《化学与生物工程》2025年第4期64-68,共5页Chemistry & Bioengineering

基　　金：国家自然科学基金项目(31401807)。

摘　　要：采用超高效液相色谱(UPLC)法对CHO细胞培养液中17种氨基酸进行定量分析。结果表明,17种氨基酸色谱分离度均大于1.5,半胱氨酸在2~20μg·mL^(-1)浓度范围内相关系数R^(2)为0.9956,其他16种氨基酸在0.1~20μg·mL^(-1)浓度范围内相关系数R^(2)均不低于0.9987;方法检出限为0.02~1μg·mL^(-1),定量限为0.1~2μg·mL^(-1);精密度、重复性和稳定性相对标准偏差(RSD)均小于4%,适用于CHO细胞培养液中氨基酸的定量分析。根据CHO细胞培养液中氨基酸的含量变化,可推断酪氨酸、半胱氨酸可能为细胞生长的营养限制性成分,对CHO细胞基础培养基或补料培养基中氨基酸种类及含量的调控具有一定的指导意义。We quantitatively analyzed 17 amino acids in CHO cell culture medium by UPLC method.The results show that the chromatographic resolutions of 17 amino acids are all greater than 1.5.Specifically,for cysteine within the concentration range of 2-20μg·mL^(-1),the correlation coefficient R^(2)is 0.9956,as well as for the remaining 16 amino acids within the concentration range of 0.1-20μg·mL^(-1),the correlation coefficient R^(2)is not less than 0.9987.The detection limit and quantification limit of the method are 0.02-1μg·mL^(-1)and 0.1-2μg·mL^(-1),respectively.The relative standard deviation(RSD)of precision,repeatability,and stability are all less than 4%,indicating that the method is suitable for the quantitative analysis of amino acids in CHO cell culture medium.Based on the content change of amino acid in CHO cell culture medium,it can be inferred that tyrosine and cysteine may be nutrient-limiting components for cell growth,which has a certain guiding significance for the regulation of amino acid types and contents in basic or supplementary CHO cell culture media.

关 键 词：超高效液相色谱(UPLC) 氨基酸 CHO细胞培养液 

分 类 号：O657.7[理学—分析化学] R917[理学—化学]