超高效液相色谱-串联质谱法测定Beagle犬血浆中抗癌新药CC1007

目的：建立超高效液相色谱-串联质谱法（UHPLC-MS/MS）测定Beagle犬血浆中抗癌新药CC1007。方法：以甲醇为沉淀剂处理犬血浆样品。采用Phenomenex Synergi 4μ Polar-RP 80A分析柱（50 mm×2.00 mm，4 μm），预柱为Phenomenex Security Guard C₁₈ Cartridges Polar-RP（4 mm×3.0 mm），以甲醇-0.1 %甲酸水溶液为流动相，梯度洗脱（0～0.5 min，30%A;0.5～1 min，30%A→90%A，保持3 min;4.1～5 min，30%A），流速0.2 mL·min^-1，柱温40℃，5 min内实现快速分离。采用电喷雾负离子（ESI^-）模式电离，选择反应监测（SRM）模式检测，以P1作为内标物质进行定量。用于监测的离子分别为CC1007 m/z 402.0→384.0/213.2和P1 m/z 312.0→170.1/95.3。结果：犬血浆中CC1007的质量浓度在1～1000 μg·L^-1范围内线性良好（r=0.9974，权重系数W=1/X²）;血浆中CC1007的检出限（信噪比为3）为0.2 μg·L^-1;其平均回收率在85%～115%之间;日内及日间的RSD均小于15%，满足生物样品检测的要求。结论：该方法可用于犬口服灌胃、静脉与阴道给药后的血浆样品中CC1007的检测。该方法选择性强、灵敏度高、操作简便快速、重复性好，适用于CC1007药代动力学等方面的研究。

Objective: To establish an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the determination of CC1007 in Beagle dog plasma. Methods: Methanol was added in the dog plasma sample for deproteinization.A Phenomenex Synergi 4μ Polar-RP 80A column (50 mm×2.00 mm,4 μm) was employed with a guard column of Phenomenex SecurityGuard Cartridges Polar-RP(4 mm×3.0 mm) and the column temperature was set at 40℃.Methanol and water (containing 0.1%(v/v) formic acid) as the mobile phase was performed at a flow rate of 0.2 mL·min^-1 with the program of gradient elution (0-0.5 min,30%A;0.5-1 min,30%A→90%A,keep 3 min;4.1-5 min,30%A),and a rapid separation was completed in 5 min.The electrospray was operated in the negative ionization mode and the CC1007 and P1 were identified by selected reaction monitoring (SRM) mode,and the monitoring ions of them were m/z 402.0→384.0/213.2 and m/z 312.0→170.1/95.3 respectively. Results: The calibration curve showed good linearity within the concentrations of 1 to 1000 μg·L^-1(r=0.9974;weighing:1/X²);the limit of detection (S/N=3) was 0.2 μg·L^-1.The mean recoveries were from 85% to 115% at the spiked levels of 1,3,100 and 800 μg·L^-1;the relative standard deviations (RSDs) of intra-and inter-day of variation were both less than 15%,which met the determination requirements of biological samples. Conclusion: The method can be used for the determination of CC1007 in dog plasma after oral gavage (po),intravenous injection (iv) and vaginal suppository (vs) administration of CC1007.The result indicates that the method is selective,sensitive,convenient,rapid and reproducible for the determination of CC1007,and can be used for the pharmacokinetics research of CC1007 in plasma.