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A HPLC method for simultaneous determination of 5-aminoimidazole-4-carboxamide riboside and its active metabolite 5-aminoimidazole-4-carboxamide ribotide in tumor-bearing nude mice plasma and its application to pharmacokinetics study
- Cheng, Xiaoliang, Guo, Liping, Li, Zaiquan, Li, Liang, Zhou, Tianyan, Lu, Wei
- Journal of Chromatography B 2013 v.915-916 pp. 64-70
- ethyl ether, high performance liquid chromatography, metabolites, mice, pharmacokinetics, solid phase extraction, temperature, trichloroacetic acid, wavelengths
- A HPLC method with on-line solid phase extraction (SPE) and column switching was developed for simultaneous determination of 5-aminoimidazole-4-carboxamide riboside (AICA riboside) and its active metabolite 5-aminoimidazole-4-carboxamide ribotide (AICA ribotide) in nude mice plasma. Plasma sample was deproteinized by adding a half volume of 10% trichloroacetic acid (TCA), and the resulting supernatant was extracted with diethyl ether to remove TCA. 50μl aqueous fraction was injected onto a WAX-1 SPE column, and AICA ribotide was trapped on the SPE column, while AICA riboside was eluted from the SPE column. The chromatographic separation of AICA riboside was achieved on CG16 column, and separation of AICA ribotide was performed on HILIC-10 and WAX-1 column. The columns temperature was maintained at 40°C, and the optimal detection wavelength was 268nm for both AICA riboside and AICA ribotide. The total analytical run time was 40min. The proposed method was linear over the range of 0.1–500μg/ml for AICA riboside and 0.03–50μg/ml for AICA ribotide. The lower limit of quantification (LLOQ) was 100 and 30ng/ml for AICA riboside and AICA ribotide, respectively. The sensitivity, accuracy and precision of this method were within acceptable limits during validation period. The method was successfully applied to investigate the pharmacokinetics characteristics of AICA riboside and its active metabolite AICA ribotide in nude mice bearing MCF-7 cell xenografts.