Ahn Eun Jeong, Kim Hanna, Chung Bong Chul, Moon Myeong Hee
Department of Chemistry, Yonsei University, Seoul, Korea.
J Sep Sci. 2007 Nov;30(16):2598-604. doi: 10.1002/jssc.200700231.
A quantitative method was developed for the determination of phosphatidylcholine (PC) species concentration using nanoflow LC-ESI-MS/MS. In this study, a calibration method is developed to determine the effect of PC carbon chain length on MS peak intensity. Using the multiple standard addition method, a relationship between the peak intensities of different PC species from nanoflow LC-MS and carbon chain length is established first using different injection amounts of PC standards. From this relationship, a calibration curve for each carbon chain length can be obtained for the concentration calculation. It was found that the MS peak area of PC species analyzed by nanoflow LC-MS linearly decreased with increased acyl carbon numbers, and that the effect of the degree of acyl chain unsaturation on MS peak intensity was minimized when the injection amount was maintained at less than 1 pmol. The method was applied for the quantitative calculation of 34 PC species from rat liver, which were identified from data-dependent MS/MS analysis during nanoflow LC separation.
开发了一种使用纳流液相色谱-电喷雾串联质谱（nanoflow LC-ESI-MS/MS）测定磷脂酰胆碱（PC）种类浓度的定量方法。在本研究中，开发了一种校准方法来确定PC碳链长度对质谱峰强度的影响。使用多标准加入法，首先通过不同进样量的PC标准品建立纳流液相色谱-质谱中不同PC种类的峰强度与碳链长度之间的关系。根据这种关系，可以得到每条碳链长度的校准曲线用于浓度计算。结果发现，通过纳流液相色谱-质谱分析的PC种类的质谱峰面积随酰基碳数增加呈线性下降，并且当进样量保持在小于1皮摩尔时，酰基链不饱和度对质谱峰强度的影响最小化。该方法用于大鼠肝脏中34种PC种类的定量计算，这些PC种类是在纳流液相色谱分离过程中通过数据依赖型串联质谱分析鉴定出来的。
