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Simultaneous analysis of monosaccharides using ultra high performance liquid chromatography-high resolution mass spectrometry without derivatization for validation of certified reference materials.
Journal of Chromatography B ( IF 2.8 ) Pub Date : 2020-09-06 , DOI: 10.1016/j.jchromb.2020.122370
Boyoung Han 1 , Jin Woo Park 2 , Minjeong Kang 3 , Byungjoo Kim 4 , Ji-Seon Jeong 5 , Oh-Seung Kwon 3 , Junghyun Son 3
Affiliation  

Monosaccharide composition of biological samples can reflect an individual’s health status. Monitoring the concentration of individual monosaccharides in human serum requires a technique for the simultaneous analysis of multiple monosaccharide molecules. Furthermore, certified reference materials (CRMs) for overall monosaccharide composition of human serum are required in order to validate the performance of clinical laboratory instruments. In the present study, we present a novel method for the simultaneous analysis of numerous monosaccharide molecules without the need for derivatization or post-column treatment. We utilized ultra-high-performance liquid chromatography (UHPLC)-quadrupole/orbitrap mass spectrometry incorporating a hydrophilic interaction chromatography (HILIC) column. We optimized the precursor ions, product ions, mobile phase composition and gradient program, flow rate, and column temperature. Seven monosaccharides (D-Ribose, L-Arabinose, D-Xylose, D-Fructose, D-Mannose, D-Galactose and D-Glucose) were able to be separated and quantified. We validated the method and the seven molecules showed favorable limits of detection and quantification, recovery rates, carry-over effects, intra- and inter-day accuracy and precision, resolution, and measurement uncertainty. We analyzed human serum samples using the method. To avoid ion suppression and D-d2-Glucose peak interference, compounds present at concentrations outside of the calibration range were analyzed from diluted samples. Quantification of serum samples corroborated some previous clinical research, in that increased D-Glucose concentration was associated with increased concentrations of D-Mannose and D-Ribose. We also validated the CRMs, and expect these to have utility as standards for serum monosaccharide profiling, thus contributing to public health.



中文翻译:

使用超高效液相色谱-高分辨率质谱法同时分析单糖,无需衍生化即可验证认证的参考物质。

生物样品中的单糖成分可以反映个人的健康状况。监测人血清中单个单糖的浓度需要同时分析多个单糖分子的技术。此外,为了验证临床实验室仪器的性能,需要用于人血清总单糖组成的认证参考材料(CRM)。在本研究中,我们提出了一种同时分析众多单糖分子而无需衍生化或柱后处理的新颖方法。我们利用了结合了亲水相互作用色谱(HILIC)柱的超高效液相色谱(UHPLC)-四极杆/轨道质谱。我们优化了前体离子,产物离子,流动相组成和梯度程序,流速和柱温。可以分离和定量七个单糖(D-核糖,L-阿拉伯糖,D-木糖,D-果糖,D-甘露糖,D-半乳糖和D-葡萄糖)。我们验证了该方法,七个分子显示出良好的检出限和定量限,回收率,残留效应,日内和日间准确度和精密度,分离度以及测量不确定度。我们使用该方法分析了人血清样品。为了避免离子抑制和D- 回收率,残留效应,日内和日间精度和精密度,分辨率以及测量不确定度。我们使用该方法分析了人血清样品。为了避免离子抑制和D- 回收率,残留效应,日内和日间精度和精密度,分辨率以及测量不确定度。我们使用该方法分析了人血清样品。为了避免离子抑制和D-d 2-葡萄糖峰干扰,从稀释样品中分析浓度超出校准范围的化合物。血清样品的定量证实了以前的一些临床研究,因为D-葡萄糖浓度的增加与D-甘露糖和D-核糖浓度的增加有关。我们还对CRM进行了验证,并期望它们可以作为血清单糖谱分析的标准,从而有助于公共健康。

更新日期:2020-09-16
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