| Purpose:To set up a rapid method to separate and determine the enantiomers of ofloxacin in serum by solid—phase extraction and high performance liquid chromatography. Method:To extract the ofloxacin in serum by solid phase extraction with C18 SPE column , then to separate and determine the enantiomers of ofloxacin in serum by high performance liquid chromatography with chiral mobile phase additive, using L–isoleucine as the chiral additive,Cu2+ as the ligand ion. The chromatography column used was a ODS—BP column (5μm,250mm×4.6mm). And the mobile phase was made up of the chiral additive solution (including L–isoleucine (1.31g/L) and CuSO4 (0.80g/L))and methanol(80:20 ,V:V).Ciprofloxacin was used as the internal standard.The excitation and emission wavelengths were 330nm and 504nm respectively. The flow rate was 0.8mL/min.ResμLts: In the selected chromatography conditions,the enantiomers of ofloxacin coμLd be baseline separated,and the resolution was higher than 1.5. Both the linear range of S-(-)-and R-(+)-ofloxacin were 0.0820 μg/mL ; r values are 0.9945 and 0.9947 respectively;and both the detection limits of them were 4ng/ml(S:N=3:1);and the mean extraction recoveries of them were 89.77% and 88.07%;and the mean method recoveries of them were 97.19 % and 98.39%; and the within-day RSDs were 2.9% and 3.0% respectively;and the between-day RSDs were 2.0% and2.2% .Conclusion: The method is simple,rapid, sensitive and precise.And it can be applied to determinations of S-(-)-and R-(+)-ofloxacin in serum respectively. |
PDF Full Text Request