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采用4-氟-7-硝基-2,1,3-苯并恶二唑进行柱前衍生化并结合荧光检测,通过高效液相色谱法测定大鼠血浆中的氟伏沙明。

Determination of fluvoxamine in rat plasma by HPLC with pre-column derivatization and fluorescence detection using 4-fluoro-7-nitro-2,1,3-benzoxadiazole.

作者信息

Higashi Yasuhiko, Matsumura Hiroki, Fujii Youichi

机构信息

Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Hokuriku University, Ho-3, Kanagawa-machi, Kanazawa 920-1181, Japan.

出版信息

Biomed Chromatogr. 2005 Dec;19(10):771-6. doi: 10.1002/bmc.514.

Abstract

A sensitive, simple and reliable method using high-performance liquid chromatographic (HPLC) assay of fluvoxamine (FLU), a selective serotonin reuptake inhibitor (SSRI), in rat plasma after pre-column derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) was developed in this study. Extracted plasma samples were mixed with NBD-F at 60 degrees C for 5 min and injected into HPLC. Retention times of FLU and an internal standard (propafenone) derivative were 15.5 and 13.5 min, respectively. The calibration curve was linear over the range 0.015-1.5 microg/mL (r2 = 0.9985) and the lower limits of detection and quantification of FLU were 0.008 and 0.015 microg/mL, respectively, in 100 microL of plasma. The derivative sample was stable at 4 degrees C for 1 day. The coefficients of variation for intra-day and inter-day assay of FLU were less than 8.3 and 9.6%, respectively. Other SSRIs and centrally acting drugs did not interfere with the peak of the FLU derivative. The method was applied for analysis of the plasma samples from rats treated with FLU. These results indicate that the method presented is useful to determine the FLU levels in rat plasma of volumes as small as 100 microL and can be applied to pharmacokinetic studies.

摘要

本研究建立了一种灵敏、简便且可靠的方法,用于在大鼠血浆中测定选择性5-羟色胺再摄取抑制剂(SSRI)氟伏沙明(FLU)。该方法采用高效液相色谱(HPLC)法,在柱前用4-氟-7-硝基-2,1,3-苯并恶二唑(NBD-F)进行衍生化。提取的血浆样品在60℃下与NBD-F混合5分钟后注入HPLC。氟伏沙明及其内标(普罗帕酮)衍生物的保留时间分别为15.5分钟和13.5分钟。在100微升血浆中,校准曲线在0.015-1.5微克/毫升范围内呈线性(r2 = 0.9985),氟伏沙明的检测限和定量下限分别为0.008和0.015微克/毫升。衍生化样品在4℃下可稳定保存1天。氟伏沙明日内和日间测定的变异系数分别小于8.3%和9.6%。其他SSRI和中枢作用药物不干扰氟伏沙明衍生物的峰。该方法用于分析经氟伏沙明处理的大鼠血浆样品。这些结果表明,所提出的方法可用于测定低至100微升大鼠血浆中的氟伏沙明水平,并可应用于药代动力学研究。

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