摘要
以AQC为柱前衍生剂,采用氨基酸专用分析柱、四元梯度洗脱、紫外检测的方法测定CCM1细胞培养液在不同温度(4℃和37℃)存贮的条件下,17种游离氨基酸的浓度。结果表明,该法分离细胞培养液中游离氨基酸的效果较好,在50～1000μmol.L-1范围内线性关系良好,相关系数R2在0.9718～0.9991之间,日内精密度RSD≤0.0917%,日间精密度RSD≤0.0981%。CCM1培养液在不同存贮条件下,随着时间的延长,其中部分游离氨基酸的浓度发生了明显改变:在4℃下存贮28 d,Asp、Ser、His的浓度分别下降了44.66%、53.79%和53.44%,其余游离氨基酸浓度下降率≤20.25%;在37℃、5%CO2的环境中存贮6 d,17种游离氨基酸浓度下降率≤19.57%。提示对于细胞培养要求较高的实验过程,应配制新鲜培养液,并及时进行细胞换液。
The effects of different storage temperature（4℃ and 37℃ ） on the concentration of free amino acids in CCM1 cell culture medium were investigated in the article. The microdialysis fluids were derived with 6- aminoquinolyl-N-hydroxysuccinimidyl carbamate （AQC） reagent and the derivatives were separated by a four gradient elution and detected by UV detector. The intraday and daylight RSD was less than 0. 0917% and 0. 0981% respectively. The good linear relationship of peak area and concentration of free amino acids was in the range of 50- 1000 μmol · L^-1 and the regression coefficient of standard calibration curves was from 0. 9718 to 0. 9991. After being preserved at 4℃ for 28 days, Asp, Ser, His concentration in CCM1 cell culture medium decreased by 44.66% , 53. 79% and 53.44% , respectively, and the rest of free amino acids concentration decreased by less than 20.25 %. After being preserved at 37℃ with 5 % CO2 for 6 days, 17 kinds of free amino acid concentration decreased by less than 19.57 %. The concentrations of some free amino acids in CCM1 culture medium preserved under different conditions changed significantly with the extension of the time. It was suggested that the medium had better be prepared freshly and the cell culture medium should be replaced in a timely manner.
出处
《化学与生物工程》
CAS
2009年第2期69-72,共4页
Chemistry & Bioengineering
关键词
游离氨基酸
培养液
高效液相色谱
free amino acid
culture medium
high performance liquid chromatography （HPLC）
