Analytical and Bioanalytical Chemistry ( IF 4.3 ) Pub Date : 2018-01-23 , DOI: 10.1007/s00216-018-0872-6 Lingling Yu , Chao Wen , Xing Li , Shiqi Fang , Lichuan Yang , Tony Wang , Kaifeng Hu
Quantification of endogenous and exogenous plasma glucose can help more comprehensively evaluate the glucose metabolic status. A ratio-based approach using isotope dilution liquid chromatography tandem mass spectrometry (ID LC-MS/MS) with indirect multiple reaction monitoring (MRM) of the derivative tag was developed to simultaneously quantify endo-/exogenous plasma glucose. Using diluted D-[13C6] glucose as tracer of exogenous glucose, 12C6/13C6 glucoses were first derivatized and then data were acquired in MRM mode. The metabolism of exogenous glucose can be tracked and the concentration ratio of endo/exo-genous glucose can be measured by calculating the endo-/exo-genous glucose concentrations from peak area ratio of specific daughter ions. Joint application of selective derivatization and MRM analysis not only improves the sensitivity but also minimizes the interference from the background of plasma, which warrants the accuracy and reproducibility. Good agreement between the theoretical and calculated concentration ratios was obtained with a linear correlation coefficient (R) of 0.9969 in the range of D-glucose from 0.5 to 20.0 mM, which covers the healthy and diabetic physiological scenarios. Satisfactory reproducibility was obtained by evaluation of the intra- and inter-day precisions with relative standard deviations (RSDs) less than 5.16%, and relative recoveries of 85.96 to 95.92% were obtained at low, medium, and high concentration, respectively. The method was successfully applied to simultaneous determination of the endo-/exogenous glucose concentration in plasma of non-diabetic and type II diabetic cynomolgus monkeys.
中文翻译:
通过同位素稀释LC-MS / MS与衍生标签的间接MRM同时定量内源性和外源性血浆葡萄糖
内源性和外源性血浆葡萄糖的定量可以帮助更全面地评估葡萄糖的代谢状态。开发了一种基于比例的方法,该方法使用同位素稀释液相色谱串联质谱(ID LC-MS / MS)和衍生标签的间接多反应监测(MRM)来同时量化内源/外源血浆葡萄糖。使用稀释D- [ 13 C ^ 6 ]葡萄糖作为外源性葡萄糖示踪剂,12 Ç 6 / 13 Ç 6首先将葡萄糖衍生化,然后以MRM模式获取数据。通过从特定子离子的峰面积比计算内源/外源葡萄糖浓度,可以跟踪外源葡萄糖的代谢,并且可以测量内源/外源葡萄糖的浓度比。选择性衍生化和MRM分析的联合应用不仅可以提高灵敏度,而且还可以将血浆背景的干扰降到最低,从而保证了准确性和可重复性。理论浓度比和计算浓度比之间具有良好的一致性,并且具有线性相关系数(RD葡萄糖从0.5到20.0 mM的范围内为0.9969),涵盖了健康和糖尿病的生理情况。通过评估日内和日间精度的相对标准偏差(RSD)小于5.16%,可以获得令人满意的重现性,并且在低,中和高浓度下的相对回收率分别为85.96至95.92%。该方法已成功地用于同时测定非糖尿病和II型糖尿病食蟹猴血浆中的内源/外源葡萄糖浓度。