title: Qualitative and quantitative analysis of the diuretic component ergone in Polyporus umbellatus by HPLC with fluorescence detection and HPLC-APCI-MS/MS.
abstract: 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.
authors.affiliation: Biomedicine Key Laboratory of Shaanxi Province, Northwest University, No.229 Taibai North Road, Xi'an 710069, China.
authors.forename: Ying-Yong
authors.initials: YY
authors.lastname: Zhao
authors.forename: Ye
authors.initials: Y
authors.lastname: Zhao
authors.forename: Yong-Min
authors.initials: YM
authors.lastname: Zhang
authors.forename: Rui-Chao
authors.initials: RC
authors.lastname: Lin
authors.forename: Wen-Ji
authors.initials: WJ
authors.lastname: Sun
mesh_terms: D002138:Calibration; D002783:Cholestenones; D002851:Chromatography, High Pressure Liquid; D004232:Diuretics / Q000032:analysis*; D004875:Ergosterol / Q000031:analogs & derivatives* / Q000032:analysis; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D055492:Polyporus / Q000737:chemistry*; D012015:Reference Standards; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence
chemical_list: D002783:Cholestenones; D004232:Diuretics; D007202:Indicators and Reagents; C015860:ergosta-4,6,8(14),22-tetraen-3-one; D004875:Ergosterol