Hubei University of Science and Technology, Xianning, Hubei, China.
Phytochem Anal. 2022 Jun;33(4):577-589. doi: 10.1002/pca.3111. Epub 2022 Feb 6.
To establish a fast, sensitive and accurate high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for determining the monosaccharide content of Qingzhuan Dark Tea polysaccharides in different years (2 years, 5 years and 11 years).
The optimised chromatographic conditions were achieved on a C18 column (5.0 μm, 250 mm × 4.6 mm inner diameter). The mobile phase flow rate was 0.9 mL/min and the column temperature was set to 27°C. The aqueous phase A (5 mM aqueous ammonium acetate) and organic phase B (acetonitrile) were used to elute the target analyses isocratically (0-60 min: 18% B). The mass spectrometer detector was equipped with an electron spray ionisation (ESI)source, and multiple reaction monitoring (MRM) mode was used for the determination of 1-phenyl-3-methyl-5-pyrazolone (PMP) derived monosaccharides.
We carried out a comprehensive methodological validation of PMP derived monosaccharides, including linearity, precision, stability and repeatability. Nine monosaccharides (rhamnose, mannose, ribose, glucose, galacturonic acid, xylose, galactose, fucose and arabinose) of Qingzhuan Dark Tea polysaccharides were identified, in which ribose and fucose were reported for the first time. The results showed the contents of these nine monosaccharides differed significantly among different years.
The validated method is reliable, accurate, repeatable and can be applied to quality assessment of these monosaccharides.
建立一种快速、灵敏、准确的高效液相色谱-串联质谱（HPLC-MS/MS）法，用于测定不同年份（2 年、5 年和 11 年）青砖茶多糖中单糖的含量。
采用 C18 柱（5.0μm，250mm×4.6mm 内径）优化了色谱条件。流动相流速为 0.9mL/min，柱温设定为 27°C。水相 A（5mM 乙酸铵水溶液）和有机相 B（乙腈）等度洗脱目标分析物（0-60min：18%B）。质谱仪检测器配备电喷雾离子源（ESI），采用多反应监测（MRM）模式测定 1-苯基-3-甲基-5-吡唑啉酮（PMP）衍生的单糖。
对 PMP 衍生单糖进行了全面的方法学验证，包括线性、精密度、稳定性和重复性。鉴定出青砖茶多糖中的 9 种单糖（鼠李糖、甘露糖、核糖、葡萄糖、半乳糖醛酸、木糖、半乳糖、岩藻糖和阿拉伯糖），其中核糖和岩藻糖为首次报道。结果表明，这些单糖在不同年份的含量差异显著。
该验证方法可靠、准确、重复性好，可用于这些单糖的质量评价。
