Liang Yu-Ting, Zhou Jun-Hui, Nan Tie-Gui, Qian Dan, Song Rui-Qi, Zhao Ya-Qiu, Yuan Yuan
School of Chinese Materia Medica, Capital Medical University, Beijing 100071, China.
National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2018 Nov;43(22):4469-4473. doi: 10.19540/j.cnki.cjcmm.20180724.004.
This study is to establish a pre-column derivatization procedure with 1-phenyl-3-methyl-5-pyrazolone (PMP) UPLC-MS/MS method for the determination of the monosaccharide composition of 12 polysaccharides. At the same time, the monosaccharide components of polysaccharides in Armillaria gallica were analyzed. The separation was performed on a ACQUITY ZORBAX RRHD Eclipse Plus C₁₈ column(2.1 mm×100 mm, 1.8 μm),using 95% acetonitrile (A) and ammonium acetate-5% acetonitrile-water (B) as mobile phase with gradient elution. The target components were detected in multiple-reaction monitoring (MRM) mode by mass spectrometry with electrospray ionization (ESI) source operated in ionization mode. The results showed that based on the monosaccharides detection method established by UPLC-MS/MS, the linearity of the 12 monosaccharides components were linear in their linear range (R²>0.990), and the recovery rate were 92.30%-105.6%. 11 monosaccharides such as fructose, mannose, and glucose were detected in A. gallica samples. The method established in this experiment is robust, highly reproducible and accurate, and is suitable for the determination of monosaccharide components such as A. gallica.
本研究旨在建立一种采用1-苯基-3-甲基-5-吡唑啉酮(PMP)的柱前衍生化超高效液相色谱-串联质谱(UPLC-MS/MS)法,用于测定12种多糖的单糖组成。同时,对蜜环菌中多糖的单糖成分进行了分析。分离在ACQUITY ZORBAX RRHD Eclipse Plus C₁₈柱(2.1 mm×100 mm,1.8 μm)上进行,以95%乙腈(A)和乙酸铵-5%乙腈-水(B)为流动相进行梯度洗脱。目标成分通过电喷雾电离(ESI)源在电离模式下运行的质谱仪,在多反应监测(MRM)模式下进行检测。结果表明,基于UPLC-MS/MS建立的单糖检测方法,12种单糖成分在其线性范围内呈线性(R²>0.990),回收率为92.30%-105.6%。在蜜环菌样品中检测到了果糖、甘露糖和葡萄糖等11种单糖。本实验建立的方法稳健、重现性高且准确,适用于蜜环菌等单糖成分的测定。