Abstract:

The method was established to determine the level of isosorbide-5-nitrate in human plasma by reversed-phase high performance liquid chromatography (RP-HPLC). The sample preparation was carried by alkalizing the plasma sample followed by extraction with dichloromethane. The HPLC analysis was performed under the conditions as follows: a mixture of an aqueous buffer (pH adjusted to 7.8 by 0.03 mol/L ammonia water) and acetonitrile (80∶20, v/v) as mobile phase, paracetamol as the internal standard, and the detection at 230 nm. The linear range was 20-1000 μg/L using the ratio of peak areas; the detection limit was 12 μg/L; the average recovery was (97.11±2.45)%-（104.34±2.17）%， the intra-day relative standard deviations (RSDs) were less than 2.52%, and inter-day RSDs were less than 5.21%.

Key words: isosorbide-5-nitrate, plasma , reversed-phase high performance liquid chromatography