title: A Rapid and Sensitive HPLC Method for Quantitation of Paclitaxel in Biological Samples using Liquid-Liquid Extraction and UV Detection: Application to Pharmacokinetics and Tissues Distribution Study of Paclitaxel Loaded Targeted Polymeric Micelles in Tumor Bearing Mice.
abstract: A simple, rapid, and sensitive reversed-phase HPLC method was developed and validated for determination of paclitaxel (PTX) in plasma, various organs and tumor tissues of tumor-bearing mice. Tissue specimens of liver, kidneys, spleen, lungs, heart and tumor were separately homogenized in normal saline. Plasma or tissue homogenate (250 µl) containing PTX and internal standard (diazepam) were extracted by diethyl ether (6 ml). The separation was achieved on a µ-Bondapak C18 HPLC column using sodium acetate buffer solution (0.01 M)/acetonitrile (58/42 v/v) at pH 5 ± 0.1 and flow rate of 1.9 mL/min. The effluent was monitored at 227 nm and column temperature was adjusted at 58ºC. The internal standard and PTX were eluted at 4.2 and 5.2 min, respectively and no interfering peaks were observed. Calibration curves were linear over the concentration range of 0.25-10 µg/ml of PTX in plasma and 0.3-20 µg/ml PTX in tissue homogenates with acceptable precision and accuracy (<15%). The mean recoveries of the drug after plasma extraction was 87.4% ± 3.6 while those of tissue homogenates ranged from 62.1± 4.5 to 75.5± 3.2 depending on the type of tissues studied. PTX was stable in samples with no evidence of degradation during 3 freeze-thaw cycles and 3 months storage at -70 °C. The developed HPLC method was applied to quantify PTX in the mouse plasma and tissues after intravenous administration of 10 mg equivalent PTX/Kg dose of PTX-loaded tocopherol succinate-chitosan-polyethylene glycol-folate (TS-CS-PEG-FA) micelles formulation or Anzatax® (Cremophor® EL- based formulation of PTX) to female Balb/c mice.
authors.affiliation: Department of Pharmaceutics, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran...
authors.forename: Mahboubeh
authors.initials: M
authors.lastname: Rezazadeh
authors.forename: Jaber
authors.initials: J
authors.lastname: Emami
authors.forename: Abolfazl
authors.initials: A
authors.lastname: Mostafavi
authors.forename: Mahboubeh
authors.initials: M
authors.lastname: Rostami
authors.forename: Farshid
authors.initials: F
authors.lastname: Hassanzadeh
authors.forename: Hojjat
authors.initials: H
authors.lastname: Sadeghi
authors.forename: Mohsen
authors.initials: M
authors.lastname: Minaiyan
authors.forename: Afsaneh
authors.initials: A
authors.lastname: Lavasanifar
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000032:analysis* / Q000493:pharmacokinetics*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D016503:Drug Delivery Systems; D005260:Female; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction; D051379:Mice; D008807:Mice, Inbred BALB C; D008823:Micelles; D009374:Neoplasms, Experimental / Q000378:metabolism; D017239:Paclitaxel / Q000032:analysis* / Q000493:pharmacokinetics*; D012015:Reference Standards; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet; D014018:Tissue Distribution
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D008823:Micelles; D017239:Paclitaxel