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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 how 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 microm thick tissue cross section into a 50 microm i.d. 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 electrokinetic 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 microm 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 microM 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.

Direct Sampling from Human Liver Tissue Cross Sections for Electrophoretic Analysis of Doxorubicin