Direct Sampling from Human Liver Tissue Cross Sections for Electrophoretic Analysis of Doxorubicin
Direct Sampling from Human Liver Tissue Cross Sections for Electrophoretic Analysis of Doxorubicin
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DOI：
10.1021/ac802542e
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发表时间：
2009-05-01
影响因子：
7.4
通讯作者：
Arriaga, Edgar A.
中科院分区：
文献类型：
--
作者：
Wang, Yaohua;Hong, Jennifer;Arriaga, Edgar A.
在用阿霉素（Dox）对肝脏进行化疗栓塞后，该药物及其代谢产物在该器官中分布不均匀。分布不容易测量，使得难以评估药物在不同组织区域中的表现。在这里，我们报告了一种技术取样组织横截面，可以分析微米大小的区域的内容。组织横截面来自肝细胞癌患者的切除的肝脏。将样品从5 μ m厚的组织横截面直接抽吸到50 μ m W.毛细管，其中组织用含有十二烷基硫酸钠的分离缓冲液溶解。样品溶解后，Dox和天然荧光化合物分离，并通过胶束电动色谱与激光诱导荧光检测。使用10%（w/v）明胶作为组织模拟物评估采样重现性和回收率。从含有Dox的明胶切片中取样显示相对标准偏差为13%，这与从溶液中取样的相对标准偏差相当。Dox回收率为82% +/-16%（n = 5）。当对肿瘤和非肿瘤组织区域进行采样时，可以从相隔100 pm的同一区域采集样本。使用原子力显微镜测定每个样品的体积为8.4 +/-1.0 pL，这使得可以分别测定取自肿瘤和非肿瘤区域的样品的Dox浓度在0.4-1.3 μ M和0.3-0.5 μ M的范围内。结果表明，在微米尺寸的区域中采样、检测和定量Dox的可行性，这可能是用于分析高度异质性组织中Dox浓度和分布的有用资源。
After chemoembolization of the liver with doxorubicin (Dox), this drug and its metabolites are not homogeneously distributed in this organ. The distribution cannot be easily measured making it difficult to assess bow the drug performs in different tissue regions. Here we report a technique for sampling tissue cross sections that can analyze the contents of micrometer size regions. The tissue cross sections were from the explanted liver of a hepatocellular carcinoma patient. Samples were directly aspirated from a 5 mu m thick tissue cross section into a 50 pm W. capillary where the tissue was solubilized with a separation buffer containing sodium dodecyl sulfate. Upon sample dissolution, Dox and natively fluorescent compounds were separated and detected by micellar electro-kinetic chromatography with laser-induced fluorescence detection. Sampling reproducibility and recovery were assessed using 10% (w/v) gelatin as tissue mimic. Sampling from gelatin slices containing Dox revealed a relative standard deviation of 13%, which was; comparable to that of sampling from solution. Dox recovery was 82% +/- 16% (n = 5). When sampling tumor and nontumor tissue regions, samples could be taken from the same region 100 pm apart. Atomic force microscopy was used to determine that each sample was 8.4 +/- 1.0 pL in volume which made it possible to determine Dox concentrations in the ranges of 0.4-1.3 and 0.3-0.5 mu M for the samples taken from tumor and nontumor regions, respectively. The results demonstrated the feasibility of sampling, detection, and quantification of Dox in micrometer size regions, which could be a useful resource for analyzing the Dox concentration and distribution in highly heterogeneous tissues.