Detection of a mixed-backbone oligonucleotide (GEM 231) in liver and tumor tissues by capillary electrophoresis |
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| Institution: | 1. Bioanalytical Center, Department of Pharmacology, Georgetown University Medical Center, 3900 Reservoir Road NW, Washington, DC 20007, USA;2. Lombardi Cancer Center, Georgetown University Medical Center, Washington, DC 20007, USA;1. Department of Chemistry, University of North Dakota, 151 Cornell Street, Grand Forks, ND 58202, USA;2. School of Medicine and Health Sciences, University of North Dakota, 501 N Columbia Rd., Grand Forks, ND 58203, USA;1. National Key Laboratory for Precision Hot Processing of Metals, Harbin Institute of Technology, Harbin 150001, PR China;2. State Key Laboratory of Advanced Welding and Joining, Harbin Institute of Technology, Harbin 150001, PR China;1. MOE Key Laboratory of Resources and Environmental Systems Optimization, North China Electric Power University, Beijing 102206, PR China;2. Hebei Key Lab of Power Plant Flue Gas Multi-Pollutants Control, Department of Environmental Science and Engineering, North China Electric Power University, Baoding, 071003, PR China |
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| Abstract: | A simple, rapid and sensitive method has been developed and validated for the analysis of a mixed-backbone oligonucleotide (GEM 231) in tumor tissues. The analysis was performed using a capillary electrophoresis (CE) system with UV detection. An extended light path (bubble cell) capillary column of 64.5 cm (effective length 56 cm)×50 μm I.D. is used as the separation column. The optimized chromatographic conditions were background electrolyte: sodium borate buffer (60 mM, pH 9.1), electrokinetic injection: 10 s, applied voltage: 30 kV, detection at λ=210 nm. A linear relationship was observed between the peak area and the amount of GEM 231 in the range of 1.0–1000 μg/ml. The lower detection limit of the drug was 100 pg with an average recovery of about 75±5%. The inter-day and intra-day relative standard deviations were <10%. Assay validation studies revealed that CE method is reproducible and specific for the determination of GEM 231 in tissue homogenates with a run time of less than 5 min. |
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