Programa de Pós-Graduação em Saúde Pública e Meio Ambiente, Escola Nacional de Saúde Pública Sergio Arouca, FIOCRUZ, Rio de Janeiro, RJ, Brazil; Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Jun 1;1146:122127. doi: 10.1016/j.jchromb.2020.122127. Epub 2020 Apr 24.
Ultrafiltration (UF) is used to separate unbound drugs; however, non-specific binding (NSB) may be a limiting factor of this technique. Pretreatment of UF devices has been suggested to reduce NSB. Therefore, the pretreatment methodologies for UF devices were evaluated in order to test their effectiveness in reducing NSB of protease inhibitors (PIs).
Two PIs (lopinavir-LPV and ritonavir-RTV) were tested. UF devices were pretreated with ultrapure water, Tween-20 or Tween-80. To evaluate the NSB, after UF devices being pretreated, ultrafiltrate solutions containing the analytes at two concentrations (low and high) were used. Samples were quantified by LC-MS/MS.
UF devices pretreated with Tween-5% had the lowest NSB for both analytes. NSB values varied between 7 and 11% at low concentration 16-34% at high LPV concentration, respectively. For RTV, NSB was approximately 6% for low concentration and 18% for high concentration. Failure to completely remove Tween in UF devices could results in an overestimation of NSB.
Pretreatment of UF device with Tween and subsequent removal proved to be effective in reducing NSB of PI.
超滤（UF）用于分离未结合的药物；然而，非特异性结合（NSB）可能是该技术的限制因素。有人建议对 UF 装置进行预处理以减少 NSB。因此，评估了 UF 装置的预处理方法，以测试它们在减少蛋白酶抑制剂（PI）的 NSB 方面的有效性。
测试了两种 PI（洛匹那韦[LPV]和利托那韦[RTV]）。用超纯水、吐温-20 或吐温-80 对 UF 装置进行预处理。为了评估 NSB，在对 UF 装置进行预处理后，使用含有两种浓度（低和高）分析物的超滤滤液溶液。通过 LC-MS/MS 对样品进行定量。
用 5%吐温预处理的 UF 装置对两种分析物的 NSB 最低。在低浓度时，NSB 值在 7%至 11%之间，在高 LPV 浓度时，NSB 值分别在 16%至 34%之间。对于 RTV，低浓度时的 NSB 约为 6%，高浓度时的 NSB 约为 18%。如果不能完全去除 UF 装置中的吐温，可能会导致 NSB 高估。
用吐温对 UF 装置进行预处理并随后去除，被证明能有效减少 PI 的 NSB。
