The Key Laboratory of Forest Plant Ecology, Northeast Forestry University, Ministry of Education, Harbin 150040, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Mar 1;879(7-8):461-6. doi: 10.1016/j.jchromb.2011.01.001. Epub 2011 Jan 11.
A new, simple, sensitive and specific reversed-phase high performance liquid chromatographic (HPLC) method using tandem mass spectrometry detection was initially developed and validated for the analysis of 10-(2-pyrazolyl-ethoxy)-(20S)-camptothecin (CPT13) in rat plasma. Pretreatment of the sample obtained from plasma involved a single protein precipitation step with using acetonitrile containing 0.1% formic acid. An aliquot of 20 μl was injected into a C-18 column. The chromatographic separation was achieved using the mobile phase consisting of acetonitrile:water (35:65) at a flow rate of 1.0 mL/min. The total run time for each sample was 10 min, and camptothecin (CPT, IS) and CPT13 were well separated with retention times of 5.1 min and 5.6 min, respectively. Detection was performed using a triple quadrupole tandem mass spectrometer in multiple reaction monitoring (MRM) mode via an electrospray ionization (ESI) source. The calibration curve was linear (r² = 0.9998) over the concentration range of 1-1000 ng/mL, with a LLOQ of 1 ng/mL for CPT13. The inter- and intra-day precision (%R.S.D.) were <2.58% and 6.28%, respectively, and the accuracies (%) were within the range of 97.34-110.67%. CPT13 in rat plasma was stable when stored at -20 °C or 4 °C for three freeze-thaw cycles, The method was employed for the first time during pharmacokinetic studies of CPT13 in rats following a single intravenous dose (0.1 mg/kg) and three different oral doses (50 mg/kg, 30 mg/kg, and 10 mg/kg). This fully validated method was successfully applied to a pharmacokinetic study of CPT13 in rats.
建立并验证了一种新的、简单的、灵敏的和特异的反相高效液相色谱-串联质谱法，用于分析大鼠血浆中的 10-（2-吡唑基-乙氧基）-（20S）喜树碱（CPT13）。从血浆中得到的样品预处理包括用含 0.1%甲酸的乙腈进行单次蛋白沉淀。取 20 μl 进样到 C18 柱上。使用乙腈：水（35:65）作为流动相，流速为 1.0 mL/min 进行色谱分离。每个样品的总运行时间为 10 min，喜树碱（CPT，IS）和 CPT13 得到了很好的分离，保留时间分别为 5.1 min 和 5.6 min。检测采用电喷雾电离（ESI）源的三重四极杆串联质谱仪在多反应监测（MRM）模式下进行。CPT13 的校准曲线在 1-1000 ng/mL 浓度范围内呈线性（r²=0.9998），LOQ 为 1 ng/mL。日内和日间精密度（%R.S.D.）分别小于 2.58%和 6.28%，准确度（%）在 97.34-110.67%范围内。CPT13 在大鼠血浆中经过三次冻融循环，在-20°C 或 4°C 下储存时稳定。该方法首次应用于大鼠单次静脉注射（0.1 mg/kg）和三种不同口服剂量（50 mg/kg、30 mg/kg 和 10 mg/kg）后 CPT13 的药代动力学研究。该完全验证的方法成功地应用于大鼠 CPT13 的药代动力学研究。
