摘要
建立了一种基于超高效液相色谱-单四极杆质谱检测器(UPLC-QDA)定量血浆游离氨基酸谱的方法。血浆样本经甲醇去除蛋白后,所得到的上清液挥干后用6-氨基喹啉-N-羟基琥珀酰亚胺基氨基甲酸酯(AQC)进行衍生。反应液在Cortecs C18色谱柱上进行6 min梯度洗脱达到色谱分离,采用正离子模式下的选择离子扫描模式测定。结果表明,34种氨基酸的检出限与定量限分别在0.1~5.0μmol/L和0.2~10.0μmol/L之间,并具有较好的线性关系(R 2>0.991)和良好的准确度(80.4%~121.8%),较高的精密度(相对标准偏差<15%)以及较好的稳定性,低、中、高3个浓度加标回收率在78.1%~127.6%之间,基质效应在-21.9%~29.7%之间。本方法简单快速,稳定可靠,并成功地应用于正常人和小鼠血浆样本中氨基酸谱的定量。
A method was established for the quantification of serous free amino acids profiles based on ultra-high performance liquid chromatography-single quadrupole mass spectrometry detector(UPLC-QDA).After protein precipitation with methanol,the supernatant was evaporated to dryness,and then derived with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate(AQC).The chromatographic separation was achieved on a Cortecs C18 column using gradient elution within 6 minutes.Electrospray ionization with positive ion mode was used to detect under selected ion recording mode.As the results,the detection and quantification limits of 34 amino acids were between 0.1-5.0μmol/L and 0.2-10.0μmol/L,respectively.A good linear relationship(R 2>0.991),acceptable accuracy ranging from 80.4%to 121.8%,high precision and appropriate stabilities with relative standard deviation(RSD)<15%were obtained.The recoveries at low,medium and high standard concentrations were in the range of 78.1%-127.6%while the matrix effects ranged over-21.9%-29.7%.The developed method with the characteristics of simplicity,rapidity and stability was successfully applied to the quantification of the amino acids in the plasma of healthy people and mice,respectively.
作者
瞿纯
刘佳健
郑丹
郑晓皎
贾伟
赵爱华
QU Chun;LIU Jiajian;ZHENG Dan;ZHENG Xiaojiao;JIA Wei;ZHAO Aihua(Center for Translational Medicine,Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200233)
出处
《分析科学学报》
CAS
CSCD
北大核心
2023年第5期557-564,共8页
Journal of Analytical Science
基金
国家自然科学基金(82122012)
上海自然科学基金(21ZR1448800)。
