Zhao Ying-Yong, Zhao Ye, Zhang Yong-Min, Lin Rui-Chao, Sun Wen-Ji
Biomedicine Key Laboratory of Shaanxi Province, Northwest University, No.229 Taibai North Road, Xi'an 710069, China.
Pharmazie. 2009 Jun;64(6):366-70.
Polyporus umbellatus is a widely used anti-aldosteronic diuretic in Traditional Chinese medicine (TCM). A new, sensitive and selective high-performance liquid chromatography-fluorescence detector (HPLC-FLD) and high-performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (HPLC-APCI-MS/MS) method for quantitative and qualitative determination of ergosta-4,6,8(14),22-tetraen-3-one(ergone), which is the main diuretic component, was provided for quality control of P. umbellatus crude drug. The ergone in the ethanolic extract of P. umbellatus was unambiguously characterized by HPLC-APCI, and further confirmed by comparing with a standard compound. The trace ergone was detected by the sensitive and selective HPLC-FLD. Linearity (r2 > 0.9998) and recoveries of low, medium and high concentration (100.5%, 100.2% and 100.4%) were consistent with the experimental criteria. The limit of detection (LOD) of ergone was around 0.2 microg/mL. Our results indicated that the content of ergone in P. umbellatus varied significantly from habitat to habitat with contents ranging from 2.13 +/- 0.02 to 59.17 +/- 0.05 microg/g. Comparison among HPLC-FLD and HPLC-UV or HPLC-APCI-MS/MS demonstrated that the HPLC-FLD and HPLC-APCI-MS/MS methods gave similar quantitative results for the selected herb samples, the HPLC-UV methods gave lower quantitative results than HPLC-FLD and HPLC-APCI-MS/MS methods. The established new HPLC-FLD method has the advantages of being rapid, simple, selective and sensitive, and could be used for the routine analysis of P. umbellatus crude drug.
猪苓是一种在传统中药中广泛使用的抗醛固酮利尿剂。建立了一种新的、灵敏且选择性高的高效液相色谱 - 荧光检测器（HPLC - FLD）和高效液相色谱 - 大气压化学电离 - 质谱联用（HPLC - APCI - MS/MS）方法，用于猪苓药材质量控制中主要利尿成分麦角甾 - 4,6,8(14),22 - 四烯 - 3 - 酮（麦角甾四烯酮）的定量和定性测定。通过HPLC - APCI对猪苓乙醇提取物中的麦角甾四烯酮进行了明确表征，并与标准化合物比较进一步确认。痕量的麦角甾四烯酮通过灵敏且选择性高的HPLC - FLD检测。线性关系（r2 > 0.9998）以及低、中、高浓度的回收率（100.5%、100.2%和100.4%）均符合实验标准。麦角甾四烯酮的检测限约为0.2μg/mL。我们的结果表明，猪苓中麦角甾四烯酮的含量因产地不同而有显著差异，含量范围为2.13±0.02至59.17±0.05μg/g。HPLC - FLD与HPLC - UV或HPLC - APCI - MS/MS的比较表明，对于所选药材样品，HPLC - FLD和HPLC - APCI - MS/MS方法给出的定量结果相似，HPLC - UV方法给出的定量结果低于HPLC - FLD和HPLC - APCI - MS/MS方法。所建立的新HPLC - FLD方法具有快速、简便、选择性高和灵敏的优点，可用于猪苓药材的常规分析。
