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首页> 美国卫生研究院文献>Pharmaceutics >Paclitaxel-Loaded Silk Fibroin Nanoparticles: Method Validation by UHPLC-MS/MS to Assess an Exogenous Approach to Load Cytotoxic Drugs
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Paclitaxel-Loaded Silk Fibroin Nanoparticles: Method Validation by UHPLC-MS/MS to Assess an Exogenous Approach to Load Cytotoxic Drugs

机译：紫杉醇负载的丝素蛋白纳米颗粒：通过UHPLC-MS / MS进行方法验证以评估负载细胞毒性药物的外源性方法

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The aim of this work was to load an anticancer drug, paclitaxel (PTX), on Silk Fibroin Nanoparticles (SFNs) by using an exogenous approach. SFNs were produced, freeze-dried and then loaded with PTX. An exogenous method allowed us to reduce both drug loss and environmental impact. In order to quantify PTX loaded in SFNs, a simple and reliable method using reversed phase liquid chromatography coupled to tandem mass spectrometry (rp-UHPLC-MS/MS) was developed. This methodology was validated by the determination of spiked QC samples in three consecutive days. Good accuracy and precision of the method were obtained, while the intra-day and inter-day precisions were less than 10.3%. For PTX, the limit of quantitation (LOQ) was 5.0 ng/mL. Recovery from the matrix (SFNs-PTX pellets) was calculated (81.2% at LOQ value) as PTX was entrapped in a new matrix like the polymer silk fibroin-based. This method was successfully applied to determine the encapsulation efficiency (1.00 ± 0.19%) and the nanoparticle loading (0.12 ± 0.02% w/w). The in vitro anticancer activity of SFNs-PTX was tested against CFPAC-1 cancer cells; results demonstrated a very high cytotoxic activity of SFNs-PTX, with a dose dependent inhibition of CFPAC-1 proliferation, confirmed by the IC50 value of 3450 ± 750 ng/mL.
机译：这项工作的目的是通过使用外源性方法将抗癌药紫杉醇（PTX）负载在丝素蛋白纳米颗粒（SFN）上。生产SFN，冷冻干燥，然后装载PTX。一种外源方法使我们能够减少药物损失和环境影响。为了量化SFN中负载的PTX，开发了一种简单可靠的方法，该方法使用了反相液相色谱-串联质谱（rp-UHPLC-MS / MS）。通过连续三天测定加标的QC样品验证了该方法。该方法具有较高的准确性和精密度，而日内和日间精度均小于10.3％。对于PTX，定量限（LOQ）为5.0 ng / mL。当将PTX截留在新的基质（如基于聚合物丝素蛋白的基质）中时，计算出从基质（SFNs-PTX颗粒）的回收率（LOQ值为81.2％）。该方法已成功应用于确定包封效率（1.00±0.19％）和纳米颗粒负载量（0.12±0.02％w / w）。测试了SFNs-PTX对CFPAC-1癌细胞的体外抗癌活性；结果表明，SFNs-PTX具有极高的细胞毒活性，并具有剂量依赖性的CFPAC-1增殖抑制作用，IC50值为3450±750 ng / mL。

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