Before gavage administration to the rats AP | Proteasome Inhibitors Guide
One day before the pharmacokinetic study, each animal was operated with a cannula insert into the right jugular vein under anesthesia by intraperitoneal injection of pentobarbital sodium (50 mg/kg). A surgical incision was made on the ventral side of the neck of rats to expose the jugular vein. The jugular vein was then cannulated with a polyethylene tubing (0.5 mm ID, 1 mm OD, Portex Ltd., Hythe, Kent, England) that was led under the hatu peak and exteriorized at the back of the neck for blood sampling. 50 IU/ml of heparin sodium in normal saline was filled into the catheter to prevent the blood clotting. After the exposed areas were surgically sutured, the rats were placed individually in standard cages. The animals were allowed to recover for 24 h and were fasted overnight prior to administration (Qian et al., 2012). 2.8.2. Drug administration 2.8.3. Analysis of AP in rat plasma In this study, a modified HPLC/UV method was employed to determine the concentration of AP in rat plasma using a reversed phase HPLC (Shimadzu LC 10AD, Shimadzu Corporation, Kyoto, Japan) (Lou et al., 2009). Chromatographic amersham separation of AP was performed on a C18 column (Kromasil ODS column, 150 mm 4.6 mm, 5 μm) guarded with a C18 precolumn (Shimadzu). Mobile phase consisting of methanol, acetonitrile and 0.1% phosphoric acid aqueous solution in a volume ratio of 32/18/50 was run at 1.0 ml/min and monitored at 337 nm with the column temperature at 30 C.
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One day before the pharmacokinetic study, each animal was operated with a cannula insert into the right jugular vein under anesthesia by intraperitoneal injection of pentobarbital sodium (50 mg/kg). A surgical incision was made on the ventral side of the neck of rats to expose the jugular vein. The jugular vein was then cannulated with a polyethylene tubing (0.5 mm ID, 1 mm OD, Portex Ltd., Hythe, Kent, England) that was led under the hatu peak and exteriorized at the back of the neck for blood sampling. 50 IU/ml of heparin sodium in normal saline was filled into the catheter to prevent the blood clotting. After the exposed areas were surgically sutured, the rats were placed individually in standard cages. The animals were allowed to recover for 24 h and were fasted overnight prior to administration (Qian et al., 2012). 2.8.2. Drug administration 2.8.3. Analysis of AP in rat plasma In this study, a modified HPLC/UV method was employed to determine the concentration of AP in rat plasma using a reversed phase HPLC (Shimadzu LC 10AD, Shimadzu Corporation, Kyoto, Japan) (Lou et al., 2009). Chromatographic amersham separation of AP was performed on a C18 column (Kromasil ODS column, 150 mm 4.6 mm, 5 μm) guarded with a C18 precolumn (Shimadzu). Mobile phase consisting of methanol, acetonitrile and 0.1% phosphoric acid aqueous solution in a volume ratio of 32/18/50 was run at 1.0 ml/min and monitored at 337 nm with the column temperature at 30 C.
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