LC-MS/MS法测定肿瘤患者恶性腹水中紫杉醇的含量

Determination of Paclitaxel in Malignant Ascites of Tumor Patients by LC-MS/MS

目的:建立测定肿瘤患者恶性腹水中紫杉醇含量的方法。方法:采用液相色谱-质谱联用(LC-MS/MS)技术,以文多灵为内标,测定肿瘤患者给予紫杉醇后恶性腹水中紫杉醇的含量,色谱柱为Zorbax SB-C18,流动相为水溶液(含0.1%甲酸和10 mmol/L醋酸铵)-乙腈溶液(40∶60,V/V),流速为0.25 m L/min,柱温为30℃,进样量为5μL,离子源为电喷雾离子源,检测模式为多重反应离子监测正离子模式,干燥气温度为350℃,干燥气流速为10 L/min,毛细管电压为4 000 V,紫杉醇和内标在正离子模式下质荷比(m/z)分别为876.5→308.0、457.3→188.1,碎片电压分别为250、150 V,碰撞电压分别为30、20 eV。结果:紫杉醇检测质量浓度的线性范围为25~2 500 ng/mL(r^2=0.996 5,n=7);定量下限为25 ng/mL;日内、日间精密度试验的RSD为0.61%~3.62%(n=5、3);准确度为95.34%~98.76%(n=5、3);提取回收率的RSD为3.19%~3.72%(n=3);基质效应的CV为1.52%~2.93%(n=3);稳定性试验的RE均小于3%(n=3)。结论:该方法简便、准确,可用于测定肿瘤患者恶性腹水中紫杉醇的含量。

OBJECTIVE:To establish a method for content determination of paclitaxel in malignant ascites of tumor patients.METHODS:LC-MS/MS method was adopted. Using vindoline as internal standard,the content of paclitaxel in ascites of tumor patients was determined. The separation was performed on Zorbax SB-C18 column with mobile phase consisted of aqueous solution(containing 0.1% formic acid and 10 mmol/L ammonium acetate)-acetonitrile(40∶60,V/V)at the flow rate of 0.25 mL/min. the column temperature was 30 ℃,and sample size was 5 μL. The ion source was electrospray ion source,and the detection mode was multiple ion monitoring positive ion mode. MS parameters were set as following as dry gas temperature 350 ℃,dry gas flow rate 10 L/min,capillary voltage 4 000 V. Quantitative determination was operated in the multiple reaction monitoring(MRM)mode,with the ion transitions m/z 876.5→308.0 for paclitaxel and m/z 457.3→188.1 for the internal standard. The fragment voltage/collision energy for paclitaxel and the internal standard were 250 V/30 e V,and 150 V/20 eV,respectively. RESULTS:The linear range of paclitaxel were 25-2 500 ng/mL(r^2=0.996 5,n=7). The lowest limit of quantitation was 25 ng/mL. RSDs of inter-day and intra-day precision tests were 0.61%-3.62%(n=5,3). Accuracies were 95.34%-98.76%(n=5,3). RSDs of extraction recovery were 3.19%-3.72%(n=3). CV of matrix effect were 1.52%-2.93%(n=3). RE of stability tests were lower than 3%(n=3). CONCLUSIONS:The method is simple,accurate and suitable for the content determination of paclitaxel in malignant ascites of tumor patients.