β-环糊精-荧光包合色谱法测定氧氟哌酸、甲氟哌酸和司帕氟哌酸

Simultaneous Determination of Trace Ofloxacin, Pefloxacin and Sparfloxacin by Inclusion Thin Layer Chromatography-Fluorescence Detection

采用荧光包合色谱法测定痕量的氧氟哌酸(ofloxacin,OFLX)、甲氟哌酸(pefloxacin,PFLX)和司帕氟哌酸(sparfloxacin,SPLX)。实验采用聚酰胺膜为固定相,β环糊精(βcyclodextrin,βCD)水溶液为流动相,并在流动相中直接加入EDTA消除金属离子的干扰,使混合物得到较好的分离。实验表明,氟哌酸化合物在βCD和EDTA的浓度比为0.005∶0.1(mol/L)时最为适宜。在此条件下,OFLX、PFLX、和SPLX的Rf值分别为0.74、0.60和0.25。试样斑点用薄层荧光色谱法扫描测定,激发波长为280nm(OFLX)、278nm(PFLX)和282(SPLX)。各化合物的线性范围分别0～110ng/斑点(OFLX)、0～90ng/斑点(PFLX)和0～93ng/斑点(SPLX);相对标准偏差为2.7%～6.9%;在血样和尿样中的回收率为98.2%～104.6%。

Simultaneous determination of ofloxacin (OFLX), peflxacin (PFLX) and sparfloxacin (SPLX) was realized by inclusion thin layer chromatography-fluorescence detection using beta-cyclodextrin (beta-CD) solutions as mobile phases, EDTA was used here for the mask of some transition metal ions to avoid the formation of norfloxacian-metal complexes. The experimental results show that the optimal molar ratio of beta-CD solutions to EDTA is 0.005:0.1. On the polyamide thin layer sheet, OFLX, PFLX and SPLX were separated from each other, and the corresponding R-f values were 0.74, 0.61 and 0.25, respectively. Fluorescence measurement was done using 278 nm(PFLX), 280 nm(OFLX) and 282 ran (SPLX) as the excitation wavelength and the cut-off filter was set at 400 nm. The respective linear ranges correspondingly fell in the concentration of 0 similar to 110 ng/spot for OFLX, 0 similar to 90 ng/spot for PFLX, and 0 similar to 93 ng/spot for SPLX. For all the three components, the relative standard deviations were in the range of 2.7% similar to 6.9% and the recoveries were found to be 98.2% similar to 104.6% in urine and serum samples.