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首页> 外文期刊>Biomedical Chromatography: An International Journal Devoted to Research in Chromatographic Methodologies and Their Applications in the Biosciences >Validated LC-MS/MS method for simultaneous determination of doxorubicin and curcumin in polymeric micelles in subcellular compartments of MCF-7/Adr cells by protein precipitation-ultrasonic breaking method
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Validated LC-MS/MS method for simultaneous determination of doxorubicin and curcumin in polymeric micelles in subcellular compartments of MCF-7/Adr cells by protein precipitation-ultrasonic breaking method

机译：通过蛋白质沉淀 - 超声波破碎法同时测定在MCF-7 / ADR细胞亚细胞间在聚合物胶束中同时测定多柔比蛋白和姜黄素的验证的LC-MS / MS方法

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A specific and sensitive LC-MS/MS with protein precipitation- ultrasonic breaking method has been developed and validated for simultaneous determination of doxorubicin (DOX) and curcumin (Cur) in DOX and Cur co-loaded hyaluronic acid-vitamin E succinatemicelles [(DOX+Cur)-polymeric micelles (PMs)] in subcellular compartments of resistant MCF-7/Adr cells. Sequential extraction of four subcellular protein fractions (cytosolic, membrane/organelle, nucleic and cytoskeleton) was performed directly from MCF-7/Adr cells after incubation with (DOX+Cur)-PMs. An ultrasonic breaking-methanol precipitation method was used for extraction of the fractions, and the micelle breaking efficiency with methanol was 98.1 and 97.6% for DOX and Cur, respectively. The analytes were analyzed using positive electrospray ionization coupled with multiple reaction monitoring. The calibration curves were linear over a concentration range of 0.5-400ng/mL for DOX and 2-2000ng/mL for Cur, and the recovery for the two analytes were >85% with negligible matrix effect. The intra-day and inter-day precision was <10.80% and relative error was within +/- 7.70%. The developed method was successfully applied for subcellular determination of DOX and Cur in MCF-7/Adr cells. Moreover, Cur and (DOX+Cur)-PMs had a marked promoting effect on the distribution of DOX in the nucleic protein fraction.
机译：已经开发并验证了具有蛋白质沉淀 - 超声波破碎方法的特异性和敏感的LC-MS / MS，用于同时测定Dox和Cur Co-Loaded透明质酸 - 维生素E琥珀酸中的多柔比星（DOX）和姜黄素（CUR）[（DOX + Cur） - 聚合物胶束（PMS）]在耐药MCF-7 / ADR细胞的亚细胞室中。在与（DOX + CUR）-PM一起孵育后，直接从MCF-7 / ADR细胞中直接从MCF-7 / ADR细胞进行四个亚细胞蛋白级分（细胞溶质，膜/细胞器。使用超声波破碎 - 甲醇沉淀法用于萃取级分，分别用甲醇的胶束破碎效率分别为108.1和97.6％。使用阳性电喷雾电离与多重反应监测相结合分析分析物。校准曲线在0.5-400ng / ml的浓度范围内为DOX和2-2000ng / ml的CUR的浓度范围，两种分析物的回收率为85％，基质效应可忽略不计。日内和日间精度<10.80％，相对误差在+/- 7.70％内。开发方法已成功应用于MCF-7 / ADR细胞中DOX和Cur的亚细胞测定。此外，CU和（DOX + CU）-PMS对核蛋白级分中DOX分布有明显的促进作用。

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