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Quantification of raf antisense oligonucleotide (rafAON) in biological matrices by LC‐MS/MS to support pharmacokinetics of a liposome‐entrapped rafAON formulation
Author(s) -
Johnson Jenifer L.,
Guo Wei,
Zang June,
Khan Sumsullah,
Bardin Sofia,
Ahmad Ateeq,
Duggan Jeffrey X.,
Ahmad Imran
Publication year - 2005
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.450
Subject(s) - chemistry , chromatography , pharmacokinetics , protein precipitation , liposome , standard curve , detection limit , oligonucleotide , tandem mass spectrometry , solid phase extraction , mass spectrometry , pharmacology , dna , biochemistry , biology
Abstract An LC‐MS/MS method was developed to quantify an antisense oligonucleotide against Raf‐1 expression (rafAON) in monkey and mouse plasma and in mouse tissue homogenates from animals dosed with a liposome‐entrapped rafAON easy‐to‐use formulation (LErafAON‐ETU) intended for use as an antineoplastic agent. RafAON was extracted from mouse and monkey plasma using solid‐phase extraction. Tissues were homogenized and sample cleanup was achieved by protein precipitation. RafAON and the internal standard (IS) were separated on a Hamilton PRP‐1 column and quantied by tandem mass spectrometry using an electrospray source in negative ion mode. The total run time was 4.0 min. The peak areas of two rafAON transitions were summed and plotted against the peak area of an IS transition to generate the standard curve. In monkey plasma the linear range was 50–10,000 ng/mL, and in mouse plasma it was 25–5000 ng/mL. The lower limit of quantication was 500 ng/mL (10 µg/g tissue) in heart, kidney, liver, lung and spleen homogenates, and the standard curve was linear up to 10,000 ng/mL. Accuracy, precision and stability were evaluated and found to be acceptable in all three matrices. The assay was used to support pharmacokinetics and tissue distribution studies of LErafAON‐ETU in mice and monkeys. Copyright © 2004 John Wiley & Sons, Ltd.