Oláh E, Bacsói Gy, Fekete J, Sharma V K
Budapest University of Technology and Economics, Department of Inorganic and Analytical Chemistry, Budapest, Hungary.
J Chromatogr Sci. 2012 Mar;50(3):253-8. doi: 10.1093/chromsci/bmr053.
This paper demonstrates the analysis of levetiracetam, a new chiral antiepileptic drug, at ng/mL levels using an ultra-high-performance liquid chromatography (UHPLC)-photodiode absorbance (PDA) method. Three different sample preparation methods, liquid-liquid extraction with Extrelut, solid phase extraction (SPE) with Oasis HLB and Oasis MAX SPE cartridges, and protein precipitation with organic solvents were carried out. The last preparatory method is the simplest and provides the best recoveries: between 97.1% and 100.4% with RSD value below 5%. The column for separation is BEH C18 column (1.7 µm particle size and 100 × 2.1 mm i.d.) and acetonitrile-phosphate buffer (pH = 6.6; 0.01 M) (10/90 v/v) is the mobile phase. The results obtained are compared to analysis conducted by the HPLC method. The UHPLC method was validated in the range of 2-100 µg/mL levetiracetam concentration (R(2) = 0.9997). LOD and LOQ are 10 ng/mL and 33 ng/mL, respectively. The developed UHPLC method was applied to plasma samples of patient with epilepsy.
本文展示了使用超高效液相色谱（UHPLC）-光电二极管吸光度（PDA）法在纳克/毫升水平分析新型手性抗癫痫药物左乙拉西坦。采用了三种不同的样品制备方法，即使用埃特拉路特进行液-液萃取、使用奥asis HLB和奥asis MAX固相萃取（SPE）柱进行固相萃取以及使用有机溶剂进行蛋白沉淀。最后一种制备方法最简单，回收率最佳：在97.1%至100.4%之间，相对标准偏差值低于5%。分离柱为BEH C18柱（粒径1.7 µm，内径100×2.1 mm），流动相为乙腈-磷酸盐缓冲液（pH = 6.6；0.01 M）（10/90 v/v）。将所得结果与通过HPLC法进行的分析进行比较。UHPLC法在左乙拉西坦浓度2-100 µg/mL范围内得到验证（R(2)=0.9997）。检测限和定量限分别为10 ng/mL和33 ng/mL。所开发的UHPLC法应用于癫痫患者的血浆样本。
