Pham Minh Hien, Menet Marie-Claude, Dugay Annabelle, Regazzetti Anne, Dauzonne Daniel, Auzeil Nicolas, Scherman Daniel, Chabot Guy G
INSERM, U640, CNRS, UMR8151, Université Paris Descartes, Faculté de Pharmacie, Ecole Nationale Supérieure de Chimie, Laboratoire de pharmacologie chimique et génétique, Paris, F-75006 France.
Rapid Commun Mass Spectrom. 2007;21(20):3373-86. doi: 10.1002/rcm.3226.
The experimental anticancer agent flavone-8-acetic acid (FAA) is metabolized into several monohydroxylated derivatives using mouse microsomes. Because these metabolites could be involved in the biological effects of FAA, the aim of this study was to characterize all its possible monohydroxylated derivatives. To do so, we have developed a methodology using reversed-phase high-performance liquid chromatography (RP-HPLC) coupled with ultraviolet (UV) detection and mass spectrometry (MS) to analyze and identify FAA derivatives hydroxylated at the 2', 3', 4', 3, 5, 6, or 7 position. In RP-HPLC, 4'-, 3'-, 2'-, 6-, and 7-OH-FAA eluted before FAA, whereas 3- and 5-OH-FAA eluted after FAA. UV spectra showed a bathochromic shift of band I for all derivatives and of band II for 5- and 6-OH-FAA. In addition, the position of the OH group could be determined by the presence of certain product ions in MS. Ions at m/z 133 and 151 were specific for 2'-, 3'-, 4'-, and 3-OH-FAA, whereas the ion at m/z 177 was specific for 3-OH-FAA only. The ions m/z 133, 151 and 167 were specific for 2'-OH-FAA. Ions at m/z 149 were specific for the presence of the OH group on cycle A only (i.e., 5-, 6- or 7-OH-FAA). The presence of both product ions m/z 149 and 179 were specific for 7-OH-FAA. Finally, ions at m/z 149 and several product ions of even m/z values were specific for 5-OH-FAA. In conclusion, the methodology described can be used to identify all possible monohydroxylated FAA derivatives.
实验性抗癌药物黄酮 - 8 - 乙酸（FAA）利用小鼠微粒体代谢生成多种单羟基化衍生物。由于这些代谢产物可能参与FAA的生物学效应，本研究的目的是对其所有可能的单羟基化衍生物进行表征。为此，我们开发了一种方法，使用反相高效液相色谱（RP - HPLC）结合紫外（UV）检测和质谱（MS）来分析和鉴定在2'、3'、4'、3、5、6或7位羟基化的FAA衍生物。在RP - HPLC中，4' - 、3' - 、2' - 、6 - 和7 - OH - FAA在FAA之前洗脱，而3 - 和5 - OH - FAA在FAA之后洗脱。紫外光谱显示所有衍生物的I带均有红移，5 - 和6 - OH - FAA的II带也有红移。此外，羟基的位置可通过MS中某些产物离子的存在来确定。m/z 133和151的离子对2' - 、3' - 、4' - 和3 - OH - FAA具有特异性，而m/z 177的离子仅对3 - OH - FAA具有特异性。离子m/z 133、151和167对2' - OH - FAA具有特异性。m/z 149的离子仅对A环上存在羟基（即5 - 、6 - 或7 - OH - FAA）具有特异性。m/z 149和179这两种产物离子的存在对7 - OH - FAA具有特异性。最后，m/z 149的离子和几个偶数m/z值的产物离子对5 - OH - FAA具有特异性。总之，所描述的方法可用于鉴定所有可能的单羟基化FAA衍生物。
