Department of Pharmaceutical Sciences, University of Perugia, Via Fabretti 48, 06123 Perugia, Italy.
Center for Perinatal and Reproductive Medicine, University of Perugia, Santa Maria della Misericordia University Hospital, 06132 Perugia, Italy.
Molecules. 2021 Jan 27;26(3):665. doi: 10.3390/molecules26030665.
Amikacin (Amk) analysis and quantitation, for pharmacokinetics studies and other types of investigations, is conventionally performed after extraction from plasma. No report exists so far regarding drug extraction from whole blood (WB). This can represent an issue since quantification in plasma does not account for drug partitioning to the blood cell compartment, significantly underrating the drug fraction reaching the blood circulation. In the present work, the optimization of an extraction method of Amk from murine WB has been described. The extraction yield was measured by RP-HPLC-UV after derivatization with 1-fluoro-2,4-dinitrobenzene, which produced an appreciably stable derivative with a favorable UV/vis absorption. Several extraction conditions were tested: spiked Amk disulfate solution/acetonitrile/WB ratio; presence of organic acids and/or ammonium hydroxide and/or ammonium acetate in the extraction mixture; re-dissolution of the supernatant in water after a drying process under vacuum; treatment of the supernatant with a solution of inorganic salts. The use of 5% (by volume) of ammonium hydroxide in a hydro-organic solution with acetonitrile, allowed the almost quantitative (95%) extraction of the drug from WB.
阿米卡星（Amk）分析和定量，用于药代动力学研究和其他类型的研究，通常是在从血浆中提取后进行的。到目前为止，还没有关于从全血（WB）中提取药物的报告。这可能是一个问题，因为在血浆中的定量并不能说明药物向血细胞隔室的分配，这大大低估了到达血液循环的药物分数。在本工作中，描述了从鼠 WB 中提取 Amk 的优化方法。通过 1-氟-2,4-二硝基苯衍生化后，用反相高效液相色谱-紫外法（RP-HPLC-UV）测量提取产率，该方法产生了一种明显稳定的衍生物，具有良好的紫外/可见吸收。测试了几种提取条件：加标 Amk 二硫酸盐溶液/乙腈/WB 比；在提取混合物中存在有机酸和/或氨和/或乙酸铵；在真空下干燥过程后将上清液重新溶解在水中；用无机盐溶液处理上清液。在乙腈的水有机溶液中使用 5%（体积）的氨，可几乎定量（95%）地从 WB 中提取药物。
