Document Detail

Measurement of intracellular accumulation of anthracyclines in cancerous cells by direct injection of cell lysate in MEKC/LIF detection.
MedLine Citation:
PMID:  20217860     Owner:  NLM     Status:  MEDLINE    
Anthracyclines are chemotherapeutic drugs that are broadly used in the treatment of various types of solid cancers and leukemia. Herein, we report on a novel analytical method for intracellular accumulation of anthracyclines using MEKC/LIF detection. An aqueous separation system permitted the injection of cell lysates directly into the capillary. The MEKC migrating solution was made up of borate buffer at pH 9.22 containing sodium taurodeoxycholate, (2-hydroxypropyl)-gamma-CD, and SDS. The anthracyclines, Doxorubicin (DOX) and epirubicin (EPI) were detected by LIF using a Nd:YAG laser (532 nm) or an argon ion laser (488 nm) for excitation. Two cell lines, human humerus tumor cells (RDES) and human lung tumor cells (A549), were treated with a mixture of the two anthracyclines for fixed periods of time, and then intracellular concentrations were determined by injecting cell lysates directly. Recovery values of 96.0-100.8% were obtained for DOX and EPI. Reproducibility quantified by RSD was less than 3.9% intraday and 6.7% interday at concentrations ranging between 50 and 500 nM. The uptake of EPI was found to be slightly less than that of DOX for A549, but higher levels of EPI were observed in RDES. Intracellular accumulation of anthracyclines was greater in RDES than in A549, but both types of cells excreted anthracyclines after 12 h. These results indicate that MEKC with an aqueous medium is useful for investigating intracellular uptake and accumulation of drugs, since cell lysates can be used directly with no pretreatment such as deproteination or solvent extraction of analytes.
Julius Mbuna; Takashi Kaneta; Totaro Imasaka
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Electrophoresis     Volume:  31     ISSN:  1522-2683     ISO Abbreviation:  Electrophoresis     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-04-27     Completed Date:  2010-07-28     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8204476     Medline TA:  Electrophoresis     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1396-404     Citation Subset:  IM    
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka, Japan.
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MeSH Terms
Cell Line, Tumor
Chromatography, Micellar Electrokinetic Capillary / methods*
Culture Media
Doxorubicin / analysis*,  chemistry,  pharmacokinetics
Epirubicin / analysis*,  chemistry,  pharmacokinetics
Hydrogen-Ion Concentration
Intracellular Space / chemistry
Linear Models
Neoplasms / chemistry*,  drug therapy,  metabolism
Reproducibility of Results
Spectrometry, Fluorescence / methods*
Reg. No./Substance:
0/Culture Media; 23214-92-8/Doxorubicin; 56420-45-2/Epirubicin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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