Highly selective and sensitive assay for paclitaxel accumulation by tumor cells based on selective solid phase extraction and micro-flow liquid chromatography coupled to mass spectrometry.
Highly selective and sensitive assay for paclitaxel accumulation by tumor cells based on selective solid phase extraction and micro-flow liquid chromatography coupled to mass spectrometry.
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DOI：
10.1039/b806856a
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发表时间：
2008-12
期刊：
影响因子：
--
通讯作者：
Straubinger RM
中科院分区：
文献类型：
--
作者：
Gaspar JR;Qu J;Straubinger NL;Straubinger RM
紫杉烷是过去20年里批准临床使用的最重要的癌症化疗药物之一。紫杉醇被用作多种癌症的一线治疗方法，许多药物输送方法正在研究中，以提高其对肿瘤的选择性和有效性。一种策略是在肿瘤内产生持续、低水平的药物，以促进细胞凋亡和抑制血管生成。改变药物浓度/时间暴露曲线以改善治疗结果的兴趣产生了对细胞或组织中低浓度紫杉醇进行量化的必要性。建立了一种选择性固相萃取与毛细管液相色谱-串联质谱仪(μLC-MS/MS)联用的方法，用于定量常规LC-MS/MS无法分析的低浓度、治疗相关的紫杉醇。在优化的固相萃取洗脱条件下，从生物样品中选择性地提取紫杉醇，并去除大部分基质成分。色谱柱为15 0×0.5 mm内径为0.5 mm的ODS毛细管柱，流速为12μL/m in。样品萃取物集中在μLC柱的前面，然后用梯度洗脱。检测下限和定量下限分别为5和20pg/ml，允许在小组织样本或低药物浓度下的培养细胞中定量紫杉醇。定量线性范围为20~20000 pg/m L。定量这些低浓度紫杉醇的能力为研究这种重要药物的浓度依赖药理作用提供了重要的工具。
The taxanes are among the most important cancer chemotherapy drugs approved for clinical use in the last two decades. Paclitaxel is used as first-line therapy for a variety of cancers, and numerous drug delivery approaches are under investigation to enhance its selectivity and effectiveness against tumors. One strategy is to produce sustained, low drug levels within the tumor to enhance apoptosis and inhibit angiogenesis. The interest in altering drug concentration/time exposure profiles to improve therapeutic outcomes creates the necessity to quantify low concentrations of paclitaxel in cells or tissues. Here, a selective solid phase extraction (SPE) method, coupled with a capillary liquid chromatography-tandem mass spectrometry (μLC-MS/MS) method, was developed to quantify low, therapeutically relevant concentrations of paclitaxel that could not be analyzed using conventional LC-MS/MS. Under optimized SPE wash and elution conditions, paclitaxel was selectively extracted from biological samples, and most matrix components were removed. A 150×0.5 mm ID ODS capillary column was used for μLC separation and the flow rate was 12 μL/min. Sample extracts were focused at the front of the μLC column and then eluted with a gradient. The lower limits of detection and quantification were 5 and 20 pg/mL, respectively, permitting quantification of paclitaxel in small tissue samples or in cultured cells exposed to low drug concentrations. The quantitative linear range was 20–20,000 pg/mL. The ability to quantify these low concentrations of paclitaxel provides an important tool to study the concentration-dependent pharmacological effects of this important drug.
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影响因子：
11.2
作者：
Ng, SSW;Figg, WD;Sparreboom, A
通讯作者：
Sparreboom, A
影响因子：
15
作者：
WANI, MC;TAYLOR, HL;MCPHAIL, AT
通讯作者：
MCPHAIL, AT
影响因子：
2
作者：
Gardner, Erin R.;Liau, Chi-Ting;Sparreboom, Alex
通讯作者：
Sparreboom, Alex
影响因子：
2
作者：
Wang, LZ;Goh, BC;Lee, HS
通讯作者：
Lee, HS
影响因子：
4.8
作者：
Pushkarev, VM;Starenki, DV;Yamashita, S
通讯作者：
Yamashita, S