State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
Molecules. 2023 Jul 2;28(13):5166. doi: 10.3390/molecules28135166.
A rapid, precise, and dependable method for quantifying flavonoids in the fruiting bodies of was established using ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS). Separation was achieved using a ZORBAX Eclipse Plus C18 column (1.8 μm, 3.0 mm × 100 mm) with a 15 min gradient of a mobile phase consisting of 0.01% aqueous formic acid and 2 mm/L ammonium formate (mobile phase A), and 0.01% formic acid and 2 mm/L ammonium formate in methanol (mobile phase B). A mass spectrometry analysis was performed using the multiple reaction monitoring (MRM) mode with an electrospray ion source. This method enabled the simultaneous detection of 10 flavonoids (sakuranetin, quercitrin, myricitrin, kaempferol, luteolin, rutin, hyperoside, kaempferol-3--rutinoside, catechin, and catechin gallate) in the fruiting bodies of . Additionally, we applied this method to analyze the flavonoid content in fruiting bodies of various species. The results revealed substantial variations in flavonoid content, up to a 100-fold difference, among different species, with myricitrin, hyperoside, and rutin identified as the most abundant flavonoids. This protocol serves as a valuable tool for quantifying flavonoid compounds in different species or under diverse cultivation conditions, particularly for identifying species with high levels of specific flavonoid compounds.
建立了一种使用超高效液相色谱-三重四极杆串联质谱法（UHPLC-QQQ-MS/MS）快速、准确、可靠地测定蛹虫草子实体中类黄酮的方法。采用 ZORBAX Eclipse Plus C18 柱（1.8μm，3.0mm×100mm），以 15min 梯度洗脱的方式分离，流动相由 0.01%甲酸水溶液和 2mm/L 甲酸铵（流动相 A）和 0.01%甲酸和 2mm/L 甲酸铵甲醇溶液（流动相 B）组成。采用电喷雾离子源多反应监测（MRM）模式进行质谱分析。该方法能够同时检测 10 种类黄酮（樱花素、槲皮苷、杨梅素、山奈酚、木樨草素、芦丁、圣草酚、山奈酚-3-O-芸香糖苷、儿茶素和儿茶素没食子酸酯）在蛹虫草子实体中的含量。此外，我们还应用该方法分析了不同蛹虫草品种子实体中的类黄酮含量。结果表明，不同种间类黄酮含量差异很大，最高可达 100 倍，其中杨梅素、圣草酚和芦丁含量最高。该方法为定量测定不同蛹虫草品种或不同栽培条件下类黄酮化合物提供了一种有价值的工具，特别是用于鉴定特定类黄酮化合物含量高的品种。
