Weight. Was twenty-four hours after treatment, urine collected and animals were sacrificed under anesthesia by cardiac puncture for blood collection agencies. Blood was collected in heparinized R Hrchen S1P Receptors collected and centrifuged at 2000 g for 10 min. To obtain the degradation of Dox, all samples were at 20 ° C until analysis. 2.4. General Procedure for the preparation of samples for the analysis of doxorubicin in plasma and tissue samples of plasma or tissues were treated with 100 L IS. After addition of 100 l pH 8.8, 1.0 M Tris buffer-L was Measurement carried out the extraction of anthracyclines twice by adding 3 ml of chloroform / methanol and stirring for 3 min. After centrifugation, the organic phases were collected and evaporated to dryness at 30 ° C under a nitrogen stream. Chemical Residues walls Were from the plasma and tissue in 100 l or 1000 l of mobile phase gel St, respectively, and 50 l of the resulting L Solution was injected into the chromatograph. For plasma samples with Dox concentrations below detection limits were h Used here amounts of plasma and then extracted by adding 30 volumes of chloroform / methanol. The dried residue was then dissolved in 100 L of mobile phase before injection into the chromatograph gel St. 2.5. Production of standards and samples of the contr The quality of t of w Ssrigen L Measurements were made of Dox and Doxol Doxon and at 20 ° C until use. A w Ssrige L Sung by Ida was treated with 1 g / ml. For a first separation a mixture of DOX Doxol was Doxon, and Ida, manufactured to a final concentration of 1 g / ml was obtained for each compound. Stamml solutions used Of Dox for the calibration of the method were made three times. Dox Stamml Solution was diluted to a series of L Substack solutions with concentrations ranging from 0.5 g / ml to obtain up to 15 g / ml.
Standard L Solutions were solutions by appropriate dilution of Stamml In rat plasma prepared with untreated liver homogenates. The final concentrations of the calibration samples used to determine the calibration curves in plasma erm Aligned ranges from 0.05 to 1.5 g / ml controlled samples with premium quality were t prepared from seed Ant untreated rat plasma and tissue samples with Dox RREP nanoparticles give final concentrations of Dox 0.05, 0.3 and 1 g / mL for plasma samples, 0.5, 5.0 and 10 g / g tissue of the liver and 10 g / g for the spleen and heart tissue. To obtain the degradation of Dox, were all standard for calibration and quality Tszirkel and frozen at 20 ° C until analyzed on the HPLC system. 2.6. Method Validation The linearity t of the process in plasma was 0.05 to 1.5 g tested / ml and in tissues from 0.5 to 10 g / g with samples prepared as described in Section 2.5. The extraction yield of Dox was spiking samples from animals not treated with Dox to put twoconcentrations determined. internal standard used was less than 1 g / ml for Histamine Receptor samples of plasma and 10 g / ml for tissue samples. The extraction was carried out as described in Section 2.4. Recoveries were determined by comparison Peakfl Weeks after the extraction with those after direct injection into the chromatograph corresponding pure L Obtained measurements calculated. For plasma and liver, was the accuracy of the method by analyzing the intra and inter-day QCS samples analyzed on the same day and.