Winnik Bozena, Barr Dana B, Thiruchelvam Mona, Montesano M Angela, Richfield Eric K, Buckley Brian
Environmental and Occupational Health Sciences Institute, Rutgers University, 170 Frelinghuysen Rd, Piscataway, NJ 08854, USA.
Anal Bioanal Chem. 2009 Sep;395(1):195-201. doi: 10.1007/s00216-009-2929-z. Epub 2009 Jul 19.
Animal models, consistent with the hypothesis of direct interaction of paraquat (PQ) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) with specific areas of the central nervous system have been developed to study Parkinson's disease (PD) in mice. These models have necessitated the creation of an analytical method for unambiguous identification and quantitation of PQ and structurally similar MPTP and 1-methyl-4-phenylpyridinium ion (MPP+) in brain tissue. A method for determination of these compounds was developed using microwave-assisted solvent extraction (MASE) and liquid chromatography-mass spectrometry. Extraction solvent and microwave conditions such as power and time were optimized to produce recoveries of 90% for PQ 78% for MPTP and 97% for its metabolite MPP+. The chromatographic separation was performed on a C8, column and detection was carried out using an ion trap as an analyzer with electrospray ionization. Mass spectrometer parameters such as heated capillary temperature, spray voltage, capillary voltage and others were also optimized for each analyte. Analysis was done in selective ion-monitoring (SIM) mode using m/z 186 for PQ, m/z 174 for MPTP, and m/z 170 for MPP+. The method detection limit for paraquat in matrix was 100 pg, 40 pg for MPTP, and 20 pg MPP+.
为了在小鼠中研究帕金森病（PD），已经建立了与百草枯（PQ）和1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）与中枢神经系统特定区域直接相互作用假说相一致的动物模型。这些模型需要创建一种分析方法，用于明确鉴定和定量脑组织中的PQ以及结构相似的MPTP和1-甲基-4-苯基吡啶离子（MPP+）。使用微波辅助溶剂萃取（MASE）和液相色谱-质谱联用技术开发了一种测定这些化合物的方法。对萃取溶剂以及功率和时间等微波条件进行了优化，使得PQ的回收率达到90%，MPTP的回收率达到78%，其代谢物MPP+的回收率达到97%。在C8柱上进行色谱分离，并使用离子阱作为分析仪，采用电喷雾电离进行检测。还针对每种分析物对诸如加热毛细管温度、喷雾电压、毛细管电压等质谱仪参数进行了优化。分析在选择性离子监测（SIM）模式下进行，PQ使用m/z 186，MPTP使用m/z 174，MPP+使用m/z 170。基质中百草枯的方法检测限为100 pg，MPTP为40 pg，MPP+为20 pg。
