Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 779, Issue 2, Pages 259-269Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S1570-0232(02)00395-1
Keywords
doxorubicin; doxorubicinol
Funding
- NCI NIH HHS [R01 CA52168] Funding Source: Medline
- NCRR NIH HHS [M01 RR000070, M01 RR00070] Funding Source: Medline
- NIAID NIH HHS [R01 AI052168] Funding Source: Medline
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A high-performance liquid chromatographic method was developed for the quantification of doxorubicin derived from PEGrylated liposomal doxorubicin (Doxil) and its major metabolite in human plasma. This method utilizes Triton X-100 to disperse the liposome, followed by a protein precipitation step with 5-sulfosalicylic acid. Analytes in the resultant supernatant are separated on a Discovery RP amide C-16 column (250 x 3 min I.D., 5 mum) using an isocratic elution with a mobile phase consisting of 0.05 M sodium acetate (pH 4.0) and acetonitrile (72:28). The retention times for doxorubicin and the internal standard daunorubicin were 4.8 and 10.1 min, respectively. The column eluate was monitored by UV-visible detection at 487 mn. The determination of doxorubicin was found to be linear in the range of 1.0 ng/mL to 25 mug/mL, with intra-day and inter-day coefficients of variation and percent error less than or equal to10%. The recovery of doxorubicin from plasma was >69.3%, with a liposomal dispersion efficiency of >95.7%. Our analytical method for free and PEGylated doxorubicin in human plasma is rapid, avoids organic extractions, and maintains sensitivity for the parent compound and its major metabolite, doxorubicinol. (C) 2002 Elsevier Science B.V. All rights reserved.
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