Frigerio E, Magi N, Benecchi A, Pianezzola E, Strolin Benedetti M
Department of Pharmacokinetics and Metabolism, Pharmacia S.p.A., Milan, Italy.
J Chromatogr A. 1996 Apr 5;729(1-2):237-42. doi: 10.1016/0021-9673(95)00976-0.
A sensitive and selective ion-pair high-performance liquid chromatographic method for the determination of 7,7'-carbonylbis[imino-N-methyl-4, 2-pyrrolecarbonylimino(N-methyl-4,2-pyrrole)carbonylimino]¿- bis(1,3-naphthalenedisulphonic acid), tetrasodium salt in monkey plasma has been developed. The compound and internal standard (bromphenol blue) were extracted from plasma samples were methylene chloride (twice) after deproteination with acetonitrile and addition of the ion-pairing agent (tetrabutylammonium hydroxide). The combined organic phases were dried, the residue dissolved in the mobile phase and then analysed by reversed-phase ion-pair liquid chromatography under isocratic conditions. The HPLC analysis time was about 20 min. Quantitation was achieved by UV detection at 323 nm. The linearity, precision and accuracy of the method were evaluated. The limit of quantitation was 0.3 microgram/ml plasma. No interference from blank monkey, mouse, rat, dog and human plasma was observed. The suitability of the method for in vivo samples was checked by analysis of plasma samples drawn from three male cynomolgus monkeys that had received a 20 mg/kg single i.v. dose of the test compound.
已开发出一种灵敏且具选择性的离子对高效液相色谱法，用于测定猴血浆中的7,7'-羰基双[亚氨基-N-甲基-4,2-吡咯羰基亚氨基（N-甲基-4,2-吡咯）羰基亚氨基]-双（1,3-萘二磺酸）四钠盐。该化合物和内标（溴酚蓝）在用乙腈进行蛋白沉淀并加入离子对试剂（氢氧化四丁铵）后，从血浆样品中用二氯甲烷萃取两次。合并的有机相经干燥后，残渣溶解于流动相中，然后在等度条件下通过反相离子对液相色谱法进行分析。高效液相色谱分析时间约为20分钟。通过在323nm处进行紫外检测实现定量。评估了该方法的线性、精密度和准确度。定量限为0.3微克/毫升血浆。未观察到来自空白猴、小鼠、大鼠、狗和人血浆的干扰。通过分析三只接受20毫克/千克单次静脉注射受试化合物的雄性食蟹猴采集的血浆样品，检验了该方法对体内样品的适用性。
