Hao Qing-Xiu, Kang Li-Ping, Zhu Shou-Dong, Yu Yi, Hu Ming-Hua, Ma Fang-Li, Zhou Jie, Guo Lan-Ping
The State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijng 100700, China.
Infinitus (China) Company Ltd., Guangzhou 510663, China.
Zhongguo Zhong Yao Za Zhi. 2018 Mar;43(6):1201-1208. doi: 10.19540/j.cnki.cjcmm.2018.0046.
In this paper, an approach was applied for separation and identification of oligosaccharides in Morinda officinalis How by Ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) with collision energy. The separation was carried out on an ACQUITY UPLC BEH Amide C₁₈(2.1mm×100 mm,1.7 μm) with gradient elution using acetonitrile(A) and water(B) containing 0.1% ammonia as mobile phase at a flow rate of 0.2 mL·min⁻¹. The column temperature was maintained at 40 °C. The information of accurate mass and characteristic fragment ion were acquired by MSE in ESI negative mode in low and high collision energy. The chemical structures and formula of oligosaccharides were obtained and identified by the software of UNIFI and Masslynx 4.1 based on the accurate mass, fragment ions, neutral losses, mass error, reference substance, isotope information, the intensity of fragments, and retention time. A total of 19 inulin oligosaccharide structures were identified including D(+)-sucrose, 1-kestose, nystose, 1F-fructofuranosyl nystose and other inulin oligosaccharides (DP 5-18). This research provided important information about the inulin oligosaccharides in M. officinalis. The results would provide scientific basis for innovative utilization of M. officinalis.
本文采用超高效液相色谱/四极杆飞行时间质谱(UPLC-Q-TOF-MS)结合碰撞能量的方法,对巴戟天中的低聚糖进行分离和鉴定。分离在ACQUITY UPLC BEH酰胺C₁₈(2.1mm×100 mm,1.7μm)柱上进行,以含0.1%氨水的乙腈(A)和水(B)为流动相,梯度洗脱,流速为0.2 mL·min⁻¹。柱温保持在40℃。在低碰撞能量和高碰撞能量的ESI负离子模式下,通过MSE获取精确质量和特征碎片离子信息。基于精确质量、碎片离子、中性丢失、质量误差、对照品、同位素信息、碎片强度和保留时间,利用UNIFI和Masslynx 4.1软件获得并鉴定低聚糖的化学结构和分子式。共鉴定出19种菊粉型低聚糖结构,包括D(+)-蔗糖、1-蔗果三糖、蔗果四糖、1F-呋喃果糖基蔗果四糖和其他菊粉型低聚糖(聚合度5 - 18)。本研究为巴戟天中菊粉型低聚糖提供了重要信息。研究结果将为巴戟天的创新利用提供科学依据。