Liu Yanhong, Zhang Weihua, Yang Yuhui
Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, PR China.
Talanta. 2008 Oct 19;77(1):412-21. doi: 10.1016/j.talanta.2008.07.004. Epub 2008 Jul 10.
A hydrophilic interaction high performance liquid chromatography-tandem mass spectrometric method has been developed and validated for simultaneous quantification of dacarbazine (DTIC) and its terminal metabolite, 5-amino-4-imidazole-carboxamide (AIC) in human plasma. The plasma samples are first extracted by a C8+SCX mixed-mode 96-well plate to extend the extraction stability of DTIC and AIC. The extracted residues are further cleaned by a primary and secondary amine (PSA) adsorbent for minimization of matrix effect. Analyses are done on an Amide-80 HPLC column coupled to a tandem mass spectrometer fitted with an atmospheric pressure turbo ion spray ionization interface in the positive-ion mode. Both DTIC and AIC have reproducible retention times on the Amide-80 HPLC column. This type of column not only has an excellent column life (over 4000 injections), but also has zero carryover effect. The injection volume should be limited at 10 microL or less to avoid the peak splitting. The validated concentration ranges are from 0.5 to 500 ng/mL for DTIC and from 2.0 to 500 ng/mL for AIC. The validated method has been successfully applied to determine the pharmacokinetic profiles for human patients receiving DTIC infusions.
已开发并验证了一种亲水作用高效液相色谱-串联质谱法，用于同时定量测定人血浆中的达卡巴嗪（DTIC）及其终末代谢产物5-氨基-4-咪唑甲酰胺（AIC）。血浆样品首先通过C8 + SCX混合模式96孔板进行萃取，以延长DTIC和AIC的萃取稳定性。萃取后的残留物用伯仲胺（PSA）吸附剂进一步净化，以最小化基质效应。分析在配备有大气压涡轮离子喷雾电离接口的串联质谱仪的酰胺-80高效液相色谱柱上进行，采用正离子模式。DTIC和AIC在酰胺-80高效液相色谱柱上均具有可重现的保留时间。这种类型的色谱柱不仅具有出色的柱寿命（超过4000次进样），而且具有零残留效应。进样体积应限制在10微升或更少，以避免峰分裂。DTIC的验证浓度范围为0.5至500 ng/mL，AIC的验证浓度范围为2.0至500 ng/mL。该验证方法已成功应用于测定接受DTIC输注的人类患者的药代动力学特征。
