Shou Dan, Dong Yu, Wang Nani, Li Hongyu, Zhang Yang, Zhu Yan
Department of Chemistry, Zhejiang University, Tianmushan Road 148, Hangzhou 310028, China.
Department of Medicine, Zhejiang Academy of Traditional Chinese Medicine, Tianmushan Road 132, Hangzhou 310007, China.
PLoS One. 2016 Sep 30;11(9):e0163797. doi: 10.1371/journal.pone.0163797. eCollection 2016.
Natural antioxidants are widely used in the life sciences. Phellinus igniarius is a historically used natural antioxidant containing a variety of active compounds. Phenols, particularly Inoscavin A and Hypholomine B, are found in the high concentrations. Better quantitative methods are needed to perform quality control in order to support further research of this mushroom. An ultra-performance liquid chromatography method coupled to photodiode-array detection and an electrospray ionization tandem mass spectrometry method (UPLC-PAD-MS) was developed to simultaneously quantify Inoscavin A and Hypholomine B levels in the medicinal fungus Phellinus igniarius. The two compounds were quantified using UPLC-PAD and UPLC-MS. The methods were accurate (mean accuracy for spiked matrix ranged from 101.5% to 105.8%), sensitive (limit of detection ranged from 0.28 to 1.14 mg L-1) and precise (the relative standard deviations ranged from 0.13 to 2.8%). Inoscavin A and Hypholomine B were purified using high-speed counter-current chromatography (HSCCC), structural evaluated to meet the request of standard substances. UPLC separation was performed on a reversed-phase C18 column using gradient elution with acetonitrile and 0.1% formic acid over 10 min. The developed method was successfully applied to determine Inoscavin A and Hypholomine B in twelve Phellinus igniarius samples of different origins and the results showed that it was suitable for the analysis of these active components in Phellinus igniarius samples.
天然抗氧化剂在生命科学中被广泛应用。桑黄是一种历史悠久的天然抗氧化剂，含有多种活性化合物。其中酚类物质，特别是桑黄菌素A和亚侧耳素B的含量较高。为了支持对这种蘑菇的进一步研究，需要更好的定量方法来进行质量控制。开发了一种超高效液相色谱法结合光电二极管阵列检测和电喷雾电离串联质谱法（UPLC-PAD-MS），用于同时定量药用真菌桑黄中桑黄菌素A和亚侧耳素B的含量。使用UPLC-PAD和UPLC-MS对这两种化合物进行定量。这些方法准确（加标基质的平均回收率在101.5%至105.8%之间）、灵敏（检测限在0.28至1.14 mg L-1之间）且精密（相对标准偏差在0.13至2.8%之间）。采用高速逆流色谱法（HSCCC）对桑黄菌素A和亚侧耳素B进行纯化，并对其结构进行评估以满足标准物质的要求。在反相C18柱上进行UPLC分离，使用乙腈和0.1%甲酸在10分钟内进行梯度洗脱。所建立的方法成功应用于测定12个不同产地桑黄样品中的桑黄菌素A和亚侧耳素B，结果表明该方法适用于桑黄样品中这些活性成分的分析。
