摘要
目的:建立人血浆中伊立替康(CPT-11)及其代谢物7-乙基-10羟基喜树碱(SN-38)的浓度测定方法并进行方法学考证。方法:用Luna 5u CN100A(4.6 mm×150 mm,5μm)色谱柱,乙腈与醋酸铵缓冲溶液(50 mmol.L-1,pH4)为流动相梯度洗脱,CPT-11的检测波长为Ex/Em=368 nm/432 nm,SN-38的检测波长为Ex/Em=368 nm/535 nm。结果:CPT-11保留时间为(9.3±0.3)min,SN-38保留时间为(4.8±0.3)min。空白样品在CPT-11、SN-38及内标喜树碱出峰位置均无干扰。CPT-11在46.9～6 000.0nmol.L-1的范围内线性良好,SN-38在2.0～250.0nmol.L-1的范围内线性良好,r值均为0.998。低浓度点RSD均在20%内,其余浓度点的RSD均在15%内,准确度均在85%～115%之间。血浆样品长期冻存稳定性良好,反复冻融3次及提取后室温放置24 h条件下,样品浓度均无显著变化。结论:使用高效液相色谱-荧光检测方法简便,准确,灵敏,适用于伊立替康及其活性代谢物SN-38的血药浓度检测。
OBJECtIVE To develop and validate a method for simultaneous determination of irinotecan（CPT-11 ） and its metabolite SN-38 in human plasma by HPLC-FLD. METHODS Chromatographic separation was performed on a Luna 5u CN 100A column（4. 6 mm × 151） mm, 5 μm）, using acetonitrile： ammonium acetate（50 mmol· L^-1, pH = 4） as mobile phase. The detection wavelength of CPT-11 was Ex/Em = 368 nm/432 nm,and the detection wavelength of SN-38 was Ex/Em = 368 nm/ 535 nm. RESULTS The retention time of CPT-11 and SN-38 was （9. 3 ±0. 3）min and （4. 8 ±0. 3）min, respectively. No interfering peaks were observed at the retention time of both targets and interna standards.The assay was linear at the range 46.9 - 6 000. 0 nmol· L^-1 for CPT-11 （r= 0. 998） and 2. 0 - 250.0 nmol· L^-1 for SN-38 （r= 0. 998）. Both the inter- and intra batch precisions were less than 20% at the low concentration and less than 15 ~ at the middle and high concentrations. The in ter- and intra-accuracies were all between 85% and 115%. The spiked plasma samples were stable at -20℃ in a long-term stability test. After three freeze-thaw cycles of the spiked samples, the changes in levels of SN-38 and CPT-11 were both with- in 15%. The changes in levels of SN-38 and CPT-11 in 24 h after extraction were not obvious. CONCLUSION HPLC-FLD method for detection of concentrations of irinotecan（CPT-11 ） and its active metabolites SN-38 in human plasma is simple, accu rate and sensitive.
出处
《中国医院药学杂志》
CAS
CSCD
北大核心
2012年第1期17-19,共3页
Chinese Journal of Hospital Pharmacy
