Determination by liquid chromatography with fluorescence detection of total 7-ethyl-10-hydroxy-camptothecin (SN-38) in beagle dog plasma after intravenous administration of liposome-based SN-38 (LE-SN38).

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Guo W, Ahmad A, Khan S, Dahhani F, Wang YF, Ahmad I

Determination by liquid chromatography with fluorescence detection of total 7-ethyl-10-hydroxy-camptothecin (SN-38) in beagle dog plasma after intravenous administration of liposome-based SN-38 (LE-SN38).

J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Jul 5;791(1-2):85-92.

PubMed ID
12798168 [ View in PubMed
]
Abstract

An HPLC- fluorescence method to quantitate total 7-ethyl-10-hydroxy-camptothecin (SN-38) in beagle dog plasma spiked with liposome based formulation of SN-38 (LE-SN38) and using camptothecin (CPT) as the internal standard (I.S.) was developed and validated to support pharmacokinetics/toxicokinetics studies. Sample preparation was done by protein precipitation using acetonitrile with 0.5% acetic acid. The supernatant was evaporated, and reconstituted in acetonitrile-20 mM ammonium acetate, pH 3.5 (20:80, v/v). When injected onto a Zorbax SB-C(18) HPLC column SN-38 as well as I.S. were detected by fluorescence using an excitation at 368 nm and emission at 515 nm. The SN-38 concentrations in samples were calculated from a standard curve of peak area ratios of SN-38 to the I.S. using weighted linear regression. The sensitivity limit for SN-38 was 1.00 ng/ml in beagle dog plasma with a precision (expressed as relative standard deviation) of 12.4% and an accuracy (expressed as analytical recovery) of 104%. The assay was linear within the standard curve range of 1-750 ng/ml. Acceptable precision and accuracy were also obtained for concentrations over the balance of the standard curve range from between-run and within-run calculations.

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