Mass spectrometric characterization and HPLC determination of the main urinary metabolites of nimesulide in man
Mass spectrometric characterization and HPLC determination of the main urinary metabolites of nimesulide in man
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DOI：
10.1016/s0731-7085(98)00172-1
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发表时间：
1998-10-01
影响因子：
3.4
通讯作者：
Facino, RM
中科院分区：
文献类型：
--
作者：
Carini, M;Aldini, G;Facino, RM
进行了一项研究，以表征和定量测定非甾体抗炎药（NSAID）尼美舒利（4-硝基-2-苯氧基-甲磺酰苯胺）单次口服给药（200 mg）后的主要尿液代谢产物。在给药后12、24、48、72和96 h采集6名健康志愿者的尿液，并在酶水解前（游离代谢物）和酶水解后（结合代谢物）进行液-液提取。通过TLG分离法分离的代谢物的结构通过质谱法（电子碰撞电离）进行了解析，并通过合成进行了确认。鉴别了5种代谢产物：它们产生于羟基化为苯氧基核（M1 =羟肟美舒利）;硝基还原为氨基衍生物（M2）;伴随的羟基化和还原（M3）; M2（M4）和M3（M5）代谢物的N-乙酰化。通过反相高效液相色谱法（Supelcosil LC-18 DB柱;移动的相：磷酸钠缓冲液（pH 3.0，50 mM）-乙腈（梯度洗脱）;流速：1 min（-1）; UV检测，230 nm）进行定量，该方法允许在单次色谱运行中同时测定原型药物及其代谢物。在总体96 h间隔内，药物和代谢产物（游离+结合）的尿液排泄约占给药剂量的40%：17.55 +/- 3.6% M1; 0.72 +/- 0.43% M2; 2.45 +/- 1.22% M3; 19.07 +/- 4.3% M5。大部分代谢产物为结合形式。原型药物和M4代谢产物的排泄率低于0.5%。所描述的方法适用于特异性和定量测量尼美舒利和代谢物在人尿中具有可接受的精密度和准确度。(C)1998 Elsevier Science B. V.保留所有权利。
A study was undertaken for the characterization and quantitative determination of the main urinary metabolites of the non-steroidal anti-inflammatory drug (NSAID) nimesulide (4-nitro-2-phenoxy-methanesulfonanilide) in man following single oral administration (200 mg). Urines were collected from six healthy volunteers at 12, 24, 48, 72 and 96 h post-administration and submitted to liquid-liquid extraction before (free metabolites) and after enzymatic hydrolysis (conjugated metabolites). The structure of the metabolites, isolated by TLG separation, was elucidated by mass spectrometry (electron impact ionization) and confirmed by synthesis. Five metabolites were identified: they arise from hydroxylation to the phenoxy nucleus (M1 = hydroxynimesulide); reduction of the nitro group to an amino derivative (M2); concomitant hydroxylation and reduction (M3); N-acetylation of the M2 (M4) and of the M3 (M5) metabolites. Quantitation was by reverse phase high performance liquid chromatography (Supelcosil LC-18 DB column; mobile phase: sodium phosphate buffer (pH 3.0, 50 mM)-acetonitrile (gradient elution); flow rate: 1 mi min(-1); UV detection, 230 nm), procedure which allows in a single chromatographic run the simultaneous determination of the unchanged drug and of its metabolites. The urinary excretion of the drug and metabolites (free + conjugated) in the overall 96 h-interval accounts for approximately 40% of the administered dose: 17.55 +/- 3.6% M1; 0.72 +/- 0.43% M2; 2.45 +/- 1.22% M3; 19.07 +/- 4.3% M5. The bulk of the metabolites was in conjugated form. Percentages excretion of the unchanged drug and of M4 metabolite were below 0.5%. The described method is suited to specifically and quantitatively measure nimesulide and metabolites in human urine with acceptable precision and accuracy. (C) 1998 Elsevier Science B.V. All rights reserved.