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Simultaneous determination of free and total paclitaxel in blood in a three-phase laminar flow microchip.
Journal of Chromatography A ( IF 3.8 ) Pub Date : 2020-07-06 , DOI: 10.1016/j.chroma.2020.461391
Qidan Cai 1 , Huaidong Peng 2 , Jiang Meng 3 , Youshao Yan 1 , Yu Zeng 3 , Paul C H Li 4 , Yue Sun 3
Affiliation  

In this study, a three-phase laminar flow microfluidic chip (TPL chip) combined with HPLC was developed for monitoring free and total concentrations of paclitaxel (PTX) in blood simultaneously. A diluted whole blood sample (aqueous phase) was introduced into the chip, ethyl acetate (organic phase) was introduced into the chip for extraction, and an interphase was used to prevent the blood sample from coming into direct contact with the organic phase. Because only free drug can quantitatively diffuse into the organic extraction phase and the free drug fraction has a linear relationship with the dilution factor of blood, both the free and total drug concentrations can be obtained by detecting the concentration of paclitaxel in the organic extraction phase. The governing factor such as flow rate for extraction was optimized. Docetaxel was used as an internal standard. The reliability of the quantitative diffusion of molecules in the TPL chip was proved by the methodological investigation of PTX in PBS sample, which showed a good linearity in the concentration range of 0.5 - 100 µg/mL and a detection limit of 7 ng/mL. Good repeatibilities for retention time (RSD of PTX is 1.23%, docetaxel is 1.14%, n = 5) and peak area ratio of PTX to docetaxel (RSD is 4.38%) were obtained. For blood sample analysis, only 100 µL of sample was needed and whole pretreatment was finished in 35 min, and a recovery of 94~117% were obtained. The provided method showed advantages in fast analysis speed, minimum sample handing, and potential ability of automation, and integration.



中文翻译:

三相层流微芯片中血液中游离和总紫杉醇的同时测定。

在这项研究中,开发了一种三相层流微流控芯片(TPL芯片)与HPLC相结合,用于同时监测血液中紫杉醇(PTX)的游离浓度和总浓度。将稀释的全血样品(水相)引入芯片中,将乙酸乙酯(有机相)引入芯片中进行提取,并使用中间相来防止血液样品直接与有机相接触。因为只有游离药物才能定量地扩散到有机萃取相中,并且游离药物组分与血液的稀释因子具有线性关系,所以可以通过检测有机萃取相中紫杉醇的浓度来获得游离药物浓度和总药物浓度。优化了控制因素,例如提取流速。多西紫杉醇用作内标。通过PBS样品中PTX的方法学研究证明了TPL芯片中分子定量扩散的可靠性,该浓度范围为0.5-100 µg / mL时线性良好,检出限为7 ng / mL。保留时间的重复性好(PTX的RSSD为1.23%,多西他赛为1.14%,n  = 5),获得PTX与多西他赛的峰面积比(RSD为4.38%)。对于血液样品分析,仅需要100 µL样品,整个预处理过程在35分钟内即可完成,回收率达到94〜117%。所提供的方法具有分析速度快,样品处理量最少,潜在的自动化和集成能力等优点。

更新日期:2020-07-13
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