摘要
以α-氨基丁酸为内标,6-氨基喹啉基-N-羟基琥珀酰亚氨基甲酸酯为柱前衍生试剂,用高效Nova-PakC18柱,乙酸钠溶液(pH5.70)和60%乙腈为流动相,采用二元梯度洗脱,紫外检测器在248nm波长检测,建立了一种新的柱前衍生反相HPLC法同时测定动物细胞培养基中谷氨酰胺在内19种氨基酸。方法重现性好,精密度高。氨基酸的峰面积—浓度线性范围为20～500μmol/L,线性相关系数均大于0.99,峰面积的精密度为1.55～10.8%。在该色谱条件下获得了良好的商用细胞培养基DMEM和F12中氨基酸的色谱图,为动物细胞生长代谢研究提供了有利的实验手段。
A reversed phase high performance liquid chromatographic method for simultaneous determination of 19 amino acids in animal cell culture medium was established. The amino acid samples were mixed with α -aminobutylic acid as the internal standard, derivatized with 6-aminoquinolyl-N-hvdroxysuccinimldyl carbamate （AQC）, and then separated on a high-performauee Nova-Pak C 18 column using sodium acetate solution （pH5.70） and 60% acetonitrile solution as mobile phases with gradient elution technique. The ultraviolet detector at 248 nm was utilized for amino acid quantification. All 19 amino acid derivatives were separated well when an optimized elution condition was applied. Consequently, good reproducibility, high sensitivity and high precision of the method were achieved. When the amino acid concentration was set at the range of 25-500μ mol/L, the linear correlation coefficients of the amino acids were above 0.99 and the precision of each peak area was 1.55-10.8%. Satisfactory chromatograms of amino acids from the commercial cell media DMEM and F12 were obtained. The method provids a positive way for exploring the growth and metabolism of animal cells.
出处
《生命科学仪器》
2007年第4期23-25,共3页
Life Science Instruments
