D w During the last decade. Preparation of the sample generally comprises either a single protein or F Llungsschritt solid phase extraction or liquidextraction liquid prior to injection into the chromatograph. The separation of DOX and its metabolites, daunorubicin, was used as internal standard at different station Later phases. Detection methods are UV, tandem mass spectrometry, electrochemistry, chemiluminescence and fluorescence. Fluorescence detection is by far 5-HT Receptor the most hours Ufigsten used method due to the high fluorescence yield of Dox. To our knowledge, only two methods have been developed with the determination of DOX in biological samples from animals re To reach underground injections of Dox PACA. These methods have not been documentation of the mice in the reporting on the pharmacokinetics and biodistribution of Dox at M After birth with the first generation of Dox PACA described. Data validation methods were not reported and lacked details on the extraction methods. The aim of this study was to develop a rapid, sensitive and specific HPLC with fluorescence detection for quantification of Dox and the detection of its major metabolites was: doxorubicinon and doxorubicinol in plasma and tissues of rats treated with various formulations of Dox PACA. A rapid method was developed for LLE sample preparation of biological samples with Dox or free Dox PACA developed. Second Materials and methods 2.1. Doxorubicin was purchased from Chemo-reactive GmbH. Doxorubicinol, doxorubicinon and idarubicin as an internal standard were purchased from Chemical Research in Toronto. Tris, trichloroacetic Acid, sodium dodecyl sulfate and dextran were from Sigma. Isobutyl was a gift from Henkel Biomedical.
All L Solvents were HPLC quality, and were purchased from Carlo Erba. Dox PACA nanoparticles were prepared as described above, by redox radical polymerization or anionic emulsion polymerization. Dox concentrations in suspensions of nanoparticles were 1 mg / ml and 0.78 mg / ml Dox Dox RREP and AEP, respectively. 2.2. HPLC Instrumentation HPLC analysis was performed on a Waters LC Module 1 HPLC system with a C18-S Column, which filled at 30 ° C and by a pilot Column with the same station Ren phase. The mobile phase consisted of a mixture of pH 2.5 0.05 M acetic Trichloroacetic acid and acetonitrile. The fluorimetric detection was performed with a Waters 470 Scanning Fluorescence Hedgehog Pathway detector. Fluorescence detection device at a wavelength Length of 480 nm excitation and Emissionswellenl length Conducted of 558 nm. Azure software was used for HPLC monitoring and data collection. To qualify, the sampling device automatically, the accuracy and linearity t be regular Ig Strips with Standardl Controlled measurements of methyl paraben. 2.3. Animal experiments were conducted g of male pattern Wistar rats weighing from 250 20th All experiments were carried out in accordance with the guidelines of the Europ European Community. The ethics of the protocol were institutionally recognized. The rats were housed individually in K Provisional housed and allowed to 1 week prior to treatment to hnen weight. Nine rats were Feeder Llig into three groups of three rats were intravenously F s with Dox or Dox RREP or AEP Dox a single dose of 6 mg of Dox Equivalents per kg Administered body weight divided.