Odontiadis John, MacKenzie Erin M, Natesan Sridhar, Mamo David, Kapur Shitij, Baker Glen B
Bebensee Schizophrenia Research Unit and Neurochemical Research Unit, Department of Psychiatry, University of Alberta, Edmonton AB, Canada.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 May 1;850(1-2):544-7. doi: 10.1016/j.jchromb.2006.11.039. Epub 2006 Dec 11.
A sensitive and reliable assay for the quantification of l-stepholidine (SPD) in rat plasma and brain was developed using high performance liquid chromatography (HPLC) combined with fluorescence detection. Brain regions (prefrontal cortex, striatum, and cerebellum) and plasma from rats treated with SPD (10 mg/kg s.c.) 20, 40, 60, or 90 min prior to euthanasia were analyzed for SPD levels. Brain samples were homogenized in ice-cold 0.1M perchloric acid and centrifuged to remove proteins. The supernatants and diluted plasma samples, to which O-desmethylvenlafaxine was added as a process standard, were basified and extracted with ethyl acetate. The organic phase was taken to dryness and the residue taken up in mobile phase. The samples were then injected into an HPLC equipped with a fluorescence detector (excitation and emission wavelengths set at 280 and 320 nm, respectively). The mean recovery of SPD was 74.6%, and reliability studies confirmed the reproducibility of the assay (intra- and inter-assay coefficients of variation of 4.8% and 5.3%, respectively). The assay was readily applicable to the brain and plasma samples obtained from rats injected with SPD as described above; the levels and patterns of disappearance of SPD in brain regions and plasma are shown.
采用高效液相色谱（HPLC）结合荧光检测技术，建立了一种灵敏可靠的大鼠血浆和脑组织中左旋千金藤啶碱（SPD）定量检测方法。对安乐死之前20、40、60或90分钟经皮下注射SPD（10 mg/kg）的大鼠的脑区（前额叶皮质、纹状体和小脑）及血浆进行SPD水平分析。脑样本在冰冷的0.1M高氯酸中匀浆，然后离心去除蛋白质。向上清液和稀释的血浆样本中加入O-去甲基文拉法辛作为过程标准品，碱化后用乙酸乙酯萃取。将有机相蒸干，残渣用流动相复溶。然后将样品注入配备荧光检测器的HPLC中（激发波长和发射波长分别设定为280和320 nm）。SPD的平均回收率为74.6%，可靠性研究证实了该检测方法的可重复性（批内和批间变异系数分别为4.8%和5.3%）。该检测方法可轻松应用于上述注射SPD的大鼠所获得的脑和血浆样本；展示了SPD在脑区和血浆中的水平及消失模式。
