High-Performance Liquid Chromatography (HPLC) Quantification of Liposome-Delivered Doxorubicin in Arthritic Joints of Collagen-Induced Arthritis Rats

Abstract

BackgroundNeoangiogenesis occurring in inflamed articular synovium in early rheumatoid arthritis (RA) is characterized by enhanced vascular permeability that allows nanoparticle agents, including liposomes, to deliver encapsulated drugs to arthritic joints and subsequently improve therapeutic efficacy and reduce adverse effects. However, the targeting distribution of liposomes in arthritic joints during RA has not been quantitatively demonstrated. We performed this study to evaluate the targeting distribution of PEGylated doxorubicin liposomes in the arthritic joints of collagen-induced arthritis (CIA) rats by high-performance liquid chromatography (HPLC).Material/MethodsTwo doxorubicin formulations were administered to CIA rats via tail intravenous injection at a single dose (50 mg/m2). CIA rats were sacrificed and the tissues of the inflamed ankle joints were collected. The content of doxorubicin in the arthritic joints was analyzed by a validated and reproducible HPLC method. A two-way ANOVA for 2×5 factorial design was used for statistical analysis.ResultsThe developed HPLC method was sensitive, precise, and reproducible. The method was successfully applied to quantify doxorubicin content in arthritic tissues. At each time point (6, 12, 24, 48, and 72 h), doxorubicin content in the arthritic joints of the doxorubicin liposome group (DOX-LIP group) was higher than in the free doxorubicin group (DOX group) (P<0.05). In the DOX-LIP group, doxorubicin levels in the arthritic joints increased gradually and significantly in the interval of 6–72 h post-administration.ConclusionsPEGylated doxorubicin liposomes were targeted to, accumulated, and retained in the arthritic joints of CIA rats. The present study indicates that liposome encapsulation increases the therapeutic efficacy of antirheumatic drugs, presenting a promising therapeutic strategy for RA.

Liquid Chromatography–Tandem Mass Spectrometry for the Simultaneous Determination of Doxorubicin and its Metabolites Doxorubicinol, Doxorubicinone, Doxorubicinolone, and 7-Deoxydoxorubicinone in Mouse Plasma

“…The matrix effects for doxorubicin, doxorubicinol, doxorubicinone, doxorubicinolone, 7-deoxydoxorubicinone, and daunorubicin (IS) were 112.9-119.7%, 94.8-109.6%, 105.1-117.9%, 99.0-111.0%, 98.3-108.3%, and 88.0%, respectively, with RSD ≤ 14.2% at low, medium, and high QC levels (Table 2), indicating the presence of a small matrix effect. Liquid−liquid extraction using chloroform:methanol (4:1, v/v) resulted in higher sensitivity and smaller matrix effects for doxorubicin and its four metabolites compared with the protein precipitation method using methanol, acetonitrile, or acetone [13][14][15]18,19,21,22,[25][26][27]31,32,34,35]. Our method yielded LLOQ levels of doxorubicin (0.5 ng/mL) and its major four metabolites (0.1-0.01 ng/mL) that were better than those reported using the least plasma volume of 10 µL.…”

“…Doxorubicinol, a major alcohol metabolite of doxorubicin formed by carbonyl reductases 1 and 3, is implicated in off-target cardiotoxicity of doxorubicin-treated patients [3,[7][8][9][10]. High-performance liquid chromatography (HPLC) with fluorescence [11][12][13][14][15][16][17][18][19] or ultraviolet (UV) [20,21] detection, LC with mass spectrometry (LC-MS) [22,23] or tandem mass spectrometry (LC-MS/MS) [24][25][26][27][28][29][30][31][32][33][34][35][36], and capillary electrophoresis [37][38][39] methods have been used to analyze doxorubicin alone or with its metabolite doxorubicinol in various biological matrices, such as blood, serum, plasma, cells, and tissues. Protein precipitation with methanol, acetonitrile, or acetone [13][14][15]18,19,21,22,[25][26][27]31,…”

Dual redox-responsive PEG–PPS–cRGD self-crosslinked nanocapsules for targeted chemotherapy of squamous cell carcinoma

“…The ow rate was 0.30 mL min À1 , and the injection volume was 5 mL. The column and sample temperature were maintained at 40 C and 4 C, 47,48 respectively. An Thermo Q EXACTIVE mass spectrometer equipped with an electro-spray ionization (ESI) source was used for mass spectrometric detection.…”