摘要
目的建立一种简单、灵敏、专属的反相高效液相色谱荧光检测(HPLCFD)方法,测定大鼠体内十二个组织样本中的羟基喜树碱分布。方法以喜树碱为内标,乙腈01%三乙胺溶液(pH30)(25∶75,v/v)为流动相,采用C18色谱柱(250mm×45mm,5μm),柱温35℃,流速为10ml/min,荧光检测的激发波长(Ex)为382nm,发射波长(Em)为528nm。结果羟基喜树碱(HCPT)的保留时间约为99min,内标物喜树碱(CPT)的保留时间约为198min。组织匀浆中HCPT的最低定量限为01ng/ml(S/N>10)。HCPT在各组织中一定浓度范围内线性关系良好,各组织中低、中、高浓度的回收率及日间精密度均符合方法学要求。结论本文建立的HPLCFD法可准确测定大鼠各组织样本中的羟基喜树碱分布。
AIM:To established an HPLC-FD method for determination of tissue distribution of 10-hydroxycamptothecin in rats.METHOD:Camptothecin is used as an internal standard.The liquid chromatograph is equipped with a fluorescence detector,whose excitation wavelength is set at 382 nm and emission wavelength is set at 528 nm,and a reversed-phase column(250 mm×4.6 mm),whose temperature is maintained at 35 ℃ is used.The flow rate is about 1.0 ml/min.The mobile phase is composed of CH 3 CN-triethylamine solution(0.1%,pH 3.0)(25∶75,v/v).RESULT:The retention time of 10-hydroxycamptothecin and camptothecin is 9.9 min and 19.8 min respectively. The limit of quantification is 0.1ng/ml of 10-hydroxycamptothecin in the tissues(S/N>10).The results of extraction recovery and inter-day precision of each tissue meet the requirement.CONCLUSIONS:The method proves to be suitable for determination of 10-hydroxycamptothecin in tissues of rats.
出处
《中国药科大学学报》
CAS
CSCD
北大核心
2004年第6期540-544,共5页
Journal of China Pharmaceutical University
