title: Simultaneous determination of the flavonoids robinin and kaempferol in human breast cancer cells by liquid chromatography-tandem mass spectrometry.
abstract: An accurate, precise and sensitive method was developed and validated for the simultaneous quantification of the flavonoid glycoside robinin, and its algycone kaempferol in human breast cancer MCF-7 cells. The application of liquid chromatography-tandem mass spectrometry (LC/MS/MS) with a TurboIonspray interface in negative mode under multiple reactions monitoring was investigated. Chromatographic separation was achieved on a C(18) column using a mobile phase consisting of (A) water with 0.025% formic acid and 1mM ammonium formate and (B) acetonitrile with 0.025% formic acid. Rutin was used as the internal standard for robinin and fisetin as the internal standard for kaempferol. The assay had a limit of detection of 0.1ng/ml for both compounds when present in cell lysate. The calibration curves were linear from 1 to 250ng/ml (r>0.999) for each compound. The intra- and inter-day coefficients of variation were less than 10% and intra- and inter-day accuracies were within 11%. This assay was successfully applied in a robinin cellular uptake study to determine the intracellular concentrations of robinin in MCF-7 cells.
authors.affiliation: Iovel Kutateladze Pharmacochemistry Institute, P. Sarajishvili st 36, Tbilisi 0159, Georgia.
authors.forename: Lia
authors.initials: L
authors.lastname: Tsiklauri
authors.forename: Guohua
authors.initials: G
authors.lastname: An
authors.forename: Donna M
authors.initials: DM
authors.lastname: Ruszaj
authors.forename: Mary
authors.initials: M
authors.lastname: Alaniya
authors.forename: Ether
authors.initials: E
authors.lastname: Kemertelidze
authors.forename: Marilyn E
authors.initials: ME
authors.lastname: Morris
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism / Q000493:pharmacokinetics; D001692:Biological Transport; D001943:Breast Neoplasms / Q000378:metabolism*; D002138:Calibration; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D005260:Female; D005419:Flavonoids / Q000378:metabolism* / Q000493:pharmacokinetics; D006801:Humans; D044949:Kaempferols / Q000378:metabolism*; D057230:Limit of Detection; D008832:Microchemistry / Q000379:methods*; D009994:Osmolar Concentration; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D005419:Flavonoids; D044949:Kaempferols; C006552:kaempferol; C005183:robinin