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血清中游离胆碱和甜菜碱的高效液相色谱串联质谱法测定及稳定性研究
引用本文:王佳佳,张伟,谭炳炎,王娴,郑琳,朱惠莲.血清中游离胆碱和甜菜碱的高效液相色谱串联质谱法测定及稳定性研究[J].营养学报,2011,33(6).
作者姓名:王佳佳  张伟  谭炳炎  王娴  郑琳  朱惠莲
作者单位:1. 中山大学公共卫生学院卫生检验与检疫专业
2. 广州市越秀区疾控中心
3. 中山大学公共卫生学院实验教学中心
4. 中山大学公共卫生学院营养学系,广州,510080
基金项目:国家自然科学基金(No.30872099)
摘    要:目的建立一种高效液相色谱串联质谱法(HPLC-MS/MS)同时测定血清中游离胆碱和甜菜碱的方法,并观察血清中两种物质的稳定性。方法以乙腈为除蛋白剂,d9-氯化胆碱、d9-甜菜碱为内标,SiO2柱分离。流动相为乙腈-甲酸铵缓冲液(15 mmol/L,pH=3.5),等度洗脱,质谱检测。结果血清中游离胆碱和甜菜碱得到良好分离,最低定量检测限均为0.62 mol/L,线性范围均为0.62~400 mol/L,日内变异系数为2.50%~7.23%,日间变异系数为4.49%~14.79%。游离胆碱和甜菜碱的回收率均在88.64%~98.23%。稳定性研究中,血清保存温度、时间对游离胆碱和甜菜碱浓度均有影响,反复冻融只对甜菜碱的影响明显。结论高效液相色谱串联质谱法同时检测血清中的游离胆碱和甜菜碱具有较好的准确度和精密性,为临床和科研提供了一种简单快速的检测方法。建议在血清分离后3 h内完成测定或可在3 h内经乙腈处理后放置于4℃,24 h内测定完毕。

关 键 词:胆碱  甜菜碱  稳定性  高效液相色谱串联质谱

DETERMINATION AND STABILITY STUDY OF FREE CHOLINE AND BETAINE IN SERUM BY LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY
WANG Jia-jia , ZHANG Wei , TAN Bing-yan , WANG Xian , ZHENG Lin , ZHU Hui-lian.DETERMINATION AND STABILITY STUDY OF FREE CHOLINE AND BETAINE IN SERUM BY LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY[J].Acta Nutrimenta Sinica,2011,33(6).
Authors:WANG Jia-jia  ZHANG Wei  TAN Bing-yan  WANG Xian  ZHENG Lin  ZHU Hui-lian
Institution:WANG Jia-jia,ZHANG Wei1,TAN Bing-yan2,WANG Xian3,ZHENG Lin2,ZHU Hui-lian3(Dept.of Inspection and Quarantine,School of Public Health,Sun Yat-Sen University,1 Centers for Disease Control of Yuexiu District,Guangzhou,2Experimental Center,3 Faculty of Nutrition,Guangzhou 510080,China)
Abstract:Objective To develop a method for determination of free choline and betaine in serum simultaneously by liquid chromatography tandem mass spectrometry and to observe its stability.Method Acetonitrile was added into serum to precipitate protein and d9-choline and d9-betaine as internal standards.A normal-phase silica column was used to separate choline and betaine with acetonitrile / deionized water(60:40v/v,containing 15 mmol/L formic aicd in the water phase) under isocratic elution.Mass spectrometry was use...
Keywords:choline  betaine  stability  HPLC-MS/MS  
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