摘要
建立了液相色谱-质谱联用(LC-MS)测定血浆中稳定同位素^(15)N标记的甘氨酸、谷氨酸、天冬氨酸和尿素同位素丰度的方法。待测动物血浆样品经丙酮提取,氮吹浓缩后,用液相色谱电喷雾质谱(LC-ESI-MS)进行检测。以乙腈和质量分数为0.2%甲酸水溶液为流动相进行洗脱,经亲水色谱柱分离,在选择离子模式(SIM)下监测特定质荷比的上述四种待测物,依据测得的峰强度计算每种待测物的同位素丰度。通过对已知同位素丰度的甘氨酸-^(15)N、谷氨酸-^(15)N、天冬氨酸-^(15)N和尿素-15N2进行验证,相对偏差在0.7%以内。方法具有样品前处理简单、检测耗时少、表征结果准确,一次进样可同时测定血浆中稳定同位素标记甘氨酸-^(15)N、谷氨酸-^(15)N、天冬氨酸-^(15)N和尿素-15N2的同位素丰度等优点。
A liquid chromatography-mass spectrometry（LC-MS）method was developed for the determination of isotopic abundance of glutamic acid-^15N, glycine-^15N , aspartic acid-^15N and urea-^15N2. The blood sample was extracted with acetone to get the proteins. The compound was separated on Cosmosil Hilic column with acetonitrile and 0.2% formic acid aqueous solution as mobile phase. The detection of glutamic acid-^15N , glycine-^15N, aspartic acid-^15N and urea-^15N2 was carried out by LC-MS with electrospray ionization under selected ion monitoring mode. The known isotopic abundance of ^1glutamic acid-^15N , glycine-^15N, aspartic acid lS N and urea-^15N2 was checked by the developed method. The relative deviation between the known isotope abundance and the calculated result was below 0.7%. This method with simple pretreatment of blood sample, short characterizing time and high aecuracy was suitable to measure the isotopic abundance of glutamic acid-^15N , glyeine-^15N, aspartic acid-^15N and urea-^15N2 in animal blood.
出处
《化学世界》
CAS
CSCD
2016年第8期475-480,共6页
Chemical World
基金
上海市科技人才计划博士后基金(14R21421800)
科研院所技术开发研究专项资金(2014EG116258)
关键词
^15N-氨基酸
同位素丰度
液相色谱-质谱联用
^15N-labeled amino acids
isotope abundance
liquid chromatography-mass spectrometry
