Institute of Analytical Chemistry of the CAS, v. v. i., Veveří 97, 602 00, Brno, Czech Republic.
Institute of Analytical Chemistry of the CAS, v. v. i., Veveří 97, 602 00, Brno, Czech Republic.
Anal Chim Acta. 2019 Apr 11;1053:162-168. doi: 10.1016/j.aca.2018.12.010. Epub 2018 Dec 18.
Amphotericin B (AmB) is still, despite its severe nephrotoxicity, the first-line agent in the management of serious systemic fungal infections. A sensitive and reliable method is therefore required to control AmB concentration in body fluids of a patient. This study demonstrates the potential of the off-line combination of preparative isoelectric focusing (IEF) with capillary isoelectric focusing (CIEF) or capillary zone electrophoresis (CZE) in the determination of AmB in human blood serum. The required value of the isoelectric point of AmB was determined to be 6.1 using the CIEF technique. Preparative IEF served as a pre-separation and concentration technique. The pH gradient was traced by colored low molecular pI markers. The collected fraction with AmB was easily processed and then analyzed by CIEF and CZE. Tens of picograms of AmB in human blood serum sample can be determined by a combination of preparative IEF with CZE. The method was linear in the AmB concentration range of 0.3-600 ng mL. The recovery ranged from 93% to 98%.
两性霉素 B（AmB）尽管具有严重的肾毒性，但仍是治疗严重系统性真菌感染的一线药物。因此，需要一种灵敏、可靠的方法来控制患者体液中的 AmB 浓度。本研究证明了离线组合制备等电聚焦（IEF）与毛细管等电聚焦（CIEF）或毛细管区带电泳（CZE）在人血清中测定 AmB 的潜力。使用 CIEF 技术确定 AmB 的等电点值为 6.1。制备IEF 用作预分离和浓缩技术。通过有色低分子量 pI 标记物追踪 pH 梯度。收集含有 AmB 的馏分，然后通过 CIEF 和 CZE 进行分析。通过制备IEF 与 CZE 的组合，可以检测人血清样品中数十皮克的 AmB。该方法在 AmB 浓度范围为 0.3-600ng/mL 时呈线性。回收率在 93%至 98%之间。
