Palette C, Cordonnier P, Tillequin F, Koch M, Advenier C, Pays M
Department of Biochemistry, Pharmacology and Toxicology, Centre Hospitalier de Versailles, Le Chesnay, France.
Biomed Chromatogr. 1994 Mar-Apr;8(2):77-84. doi: 10.1002/bmc.1130080207.
The determination of the concentrations of antipyrine metabolites in biological fluids is hampered by the difficulty in obtaining pure conjugated compounds to be used as standards. Most authors have proposed determination of total forms by high performance liquid chromatography (HPLC) after deconjugation of these metabolites using chemical or enzymatic hydrolysis. Up to now there is no satisfactory hydrolysis method for the study of all antipyrine metabolites. The situation is further complicated by the fact that the deconjugated metabolites are highly unstable whichever technique is being used. Because of the lack of stability of all these molecules it has been necessary to isolate the glucuroconjugated compounds from urine. We describe a method which allows us to obtain highly purified glucuroconjugated metabolites of antipyrine. Sulphoconjugated compounds have been synthesized previously. We are thus able to propose a chromatographic procedure which allows us to determine simultaneously all stable phase I and phase II metabolites of antipyrine in biological fluids without any step of extraction. This analytical technique allows us to study the activity of the different isoenzymes implicated in the metabolism of antipyrine.
生物体液中安替比林代谢物浓度的测定受到阻碍，因为难以获得用作标准品的纯共轭化合物。大多数作者提议在使用化学或酶促水解使这些代谢物解共轭后，通过高效液相色谱法（HPLC）测定总形式。到目前为止，还没有一种令人满意的水解方法可用于研究所有安替比林代谢物。无论使用何种技术，解共轭后的代谢物都高度不稳定，这一事实使情况更加复杂。由于所有这些分子缺乏稳定性，因此有必要从尿液中分离出葡萄糖醛酸共轭化合物。我们描述了一种方法，该方法使我们能够获得高度纯化的安替比林葡萄糖醛酸共轭代谢物。硫酸共轭化合物先前已合成。因此，我们能够提出一种色谱程序，该程序使我们能够在不进行任何提取步骤的情况下，同时测定生物体液中安替比林所有稳定的I相和II相代谢物。这种分析技术使我们能够研究参与安替比林代谢的不同同工酶的活性。
