Abstract：
INTRODUCTION:The aim of this investigation was to evaluate methods for increasing Caco-2 cell throughput for assessing drug intestinal absorption. The use of 6-, 12-, and 24-well membranes and the effect of membrane size on permeability and the integrity of the Caco-2 cell monolayer were assessed. In an effort to optimize the assessment of drug permeability, increased throughput was investigated by testing compounds singly or as mixtures of analytes. METHOD:The transepithelial electrical resistance (TEER) of cell monolayers was measured on 0.33, 1.0, and 4.7 cm2 polycarbonate membranes using EVOM, over a 25-day period. Absorptive transport was determined on all compounds tested using LC-MS/MS assays, or liquid scintillation spectrometry. RESULTS:The effect of multiple compounds in one well compared to single compounds was assessed with atenolol, nadolol, metoprolol, and propranolol for mixtures of four compounds and with RWJ-53308, atenolol, terbutaline, propranolol, naproxen, piroxicam, topiramate, and furosemide for mixtures of eight compounds. The apparent permeability (Papp) values correlated well between single analytes and mixtures of four and eight analytes in each well. Drug permeability decreased slightly with an increase in well size. The TEER value increased with the number of days in culture for each of the 6-, 12-, and 24-well sizes. DISCUSSION:It was demonstrated that the 24-well format system is ideal for high-throughput assessment. Furthermore, the approach of mixing four or eight analytes in each well to further increase throughput was also demonstrated to be valid.
journal_name
J Pharmacol Toxicol Methodsjournal_title
Journal of pharmacological and toxicological methodsauthors
Markowska M,Oberle R,Juzwin S,Hsu CP,Gryszkiewicz M,Streeter AJdoi
10.1016/s1056-8719(01)00161-7keywords:
subject
Has Abstractpub_date
2001-07-01 00:00:00pages
51-5issue
1eissn
1056-8719issn
1873-488Xpii
S1056-8719(01)00161-7journal_volume
46pub_type
杂志文章abstract：:For the rapid assay of norepinephrine (NE) and its major metabolite, 3-methoxy-4-hydroxyphenylglycol (MHPG) in the mouse brain, we developed a simple method using isocratic HPLC with coulometric detection. This method permits NE and MHPG assay within 5 min in one chromatographic run. Within-run coefficients of variati...
journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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doi：10.1016/j.vascn.2008.06.001
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journal_title：Journal of pharmacological and toxicological methods
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doi：10.1016/j.vascn.2009.04.196
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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journal_title：Journal of pharmacological and toxicological methods
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