Kazemi Moloud, Emami Jaber, Hasanzadeh Farshid, Minaiyan Mohsen, Mirian Mina, Lavasanifar Afsaneh
Department of Pharmaceutics, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.
Department of Medical Chemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.
Res Pharm Sci. 2020 Feb 20;15(1):1-13. doi: 10.4103/1735-5362.278710. eCollection 2020 Feb.
A simple, rapid, and sensitive reversed-phase high performance liquid chromatography (RP-HPLC) method based on liquid-liquid extraction was developed and validated for determination of docetaxel (DTX) in plasma and homogenate tissues of tumor-bearing mice.
Samples were spiked with celecoxib as the internal standard and separation was achieved on a μ-Bondapak C18 HPLC column. The mobile phase consisted of a mixture of acetonitrile/water (40/60 v/v) at flow rate of 1.2 mL/min and the effluent was monitored at 230 nm.
Calibration curves were linear over the concentration range of 0.1-10 μg/mL of DTX in plasma and 0.25-50 μg/mL in tissue homogenates with acceptable precision and accuracy. The mean recoveries of the drug from plasma extraction was 94.6 ± 1.44% while those of tissue homogenates ranged from 73.5 ± 3.2 to 85.3 ± 2.8% depending on the type of tissues examined. DTX was stable in biological samples with no evidence of degradation during 3 freeze-thaw cycles and two months of storage at -70 ± 15 °C. The developed HPLC method was applied to quantify DTX in the mouse plasma and tissues after intravenous administration of 7.5 mg equivalent DTX/kg dose of DTX-loaded folic acid-polyethylene glycol-heparin-tocopherol (FA-PEG-HEP-CA-TOC) micelle formulation to female Balb/c mice.
A simple, sensitive, rapid, accurate, and prudent RP-HPLC method was developed, validated, and applied for DTX determination in plasma and tissues.
建立并验证了一种基于液-液萃取的简单、快速、灵敏的反相高效液相色谱（RP-HPLC）方法，用于测定荷瘤小鼠血浆和组织匀浆中的多西他赛（DTX）。
样品加入塞来昔布作为内标，在μ-Bondapak C18高效液相色谱柱上进行分离。流动相由乙腈/水（40/60 v/v）混合而成，流速为1.2 mL/min，在230 nm处监测流出物。
血浆中DTX浓度范围为0.1-10 μg/mL，组织匀浆中浓度范围为0.25-50 μg/mL时，校准曲线呈线性，精密度和准确度均可接受。血浆萃取药物的平均回收率为94.6±1.44%，而组织匀浆的平均回收率根据所检测组织类型在73.5±3.2%至85.3±2.8%之间。DTX在生物样品中稳定，在3次冻融循环以及在-70±15℃储存两个月期间无降解迹象。将所建立的高效液相色谱方法应用于雌性Balb/c小鼠静脉注射7.5 mg当量DTX/kg剂量的载DTX叶酸-聚乙二醇-肝素-生育酚（FA-PEG-HEP-CA-TOC）胶束制剂后小鼠血浆和组织中DTX的定量分析。
建立、验证并应用了一种简单,灵敏、快速、准确且可靠的反相高效液相色谱方法用于血浆和组织中DTX的测定。
