超高效液相色谱-串联质谱法同时测定人乳中的 胆碱、L-肉碱、乙酰基-L-肉碱和牛磺酸

Determination of Choline, L-Carnitine, Ace-L-Carnitine and Taurine in Human Milk using UPLC-MS/MS

中文关键词:  胆碱  L-肉碱  乙酰基-L-肉碱  牛磺酸  人乳

英文关键词:choline  L-carnitine  acetyl-L-Carnitine  taurine  human milk

中文摘要:

      目的 建立一种简单、快速、灵敏的可以同时测定人乳中胆碱、L-肉碱、乙酰基-L-肉碱和牛磺酸的超高效液相色谱-串联质谱法。方法 样品经甲醇沉淀蛋白、乙醚去除非极性杂质, UPLC BEH Amide色谱柱分离后采用保留时间和多反应离子检测(MRM)定性、定量, 同位素内标校正。结果 牛磺酸的定量下限(LOQ)为0.02 mg/100 g, 胆碱、L-肉碱和乙酰基-L-肉碱的LOQ均为0.003 mg/100 g。标准加入法测定所有被测物的高、中、低三个浓度加标水平的回收率在84.8%~90.3%之间, 相对标准偏差(RSD)在1.7%~5.3%之间。4种被测物在各自浓度范围内线性良好, R2均大于0.999。结论 本文建立了同时测定人乳中胆碱、L-肉碱、乙酰基-L-肉碱和牛磺酸的超高效液相色谱-串联质谱法。方法前处理简单、快速, 样品需要量少; 采用质谱检测器同位素内标一对一校正定量, 检测结果准确、灵敏。方法可以为人乳中相关营养素含量的调查、研究和为配方奶粉人乳化提供技术支持。

英文摘要:

      Objective To establish a rapid and sensitive method which can determine choline, L-carnitine, acetyl-L-carnitine and taurine in human milk by ultra-performance liquid-chromatography tandem mass spectrometry (UPLC-MS/MS). Methods The non-polar substances and protein were removed from samples by ether and methanol respectively. The sample extracts were separated by a UPLC BEH Amide column. The detection was performed with mass spectrometry under multiple reaction monitoring (MRM) mode with stable isotopedilution method. Results LOQs of choline, L-carnitine, acetyl-L-carnitine and taurine were between 0.003 mg/100 g and 0.02 mg/100 g. The spiked recoveries were 84.8%~90.3%. The precision RSD were below 5.3%. The lineal range was with high correlation coefficients (R2>0.999). Conclusion A simultaneous quantification of choline, L-carnitine, acetyl-L-carnitine and taurine in human milk was developed by using a UHPLC-MS/MS method. The sample preparation was simple, rapid and required small volume of samples. Four stable isotope standards which provide high precision and accurate were used for each analyte. The developed method was suitable for the study of four kinds of analytes which were concentrated in human milk and provided the technical support for the infant formula.