Determination of primaquine and hepatic targeting neoglycoalbumin-primaquine in whole blood and liver of mouse by normalphase high-performance liquid chromatography

We have developed a method to estimate primaquine (PQ) and hepatic targeting antimalarial agent neoglycoalbumin-primaquine (NGA PQ) in whole blood and liver of mice by sensitive and selective high-performance liquid chromatography. The primaquine was extracted out with ether from the basified biological samples in the PQ diphosphate group or from the samples which were acidic hydrolyzed then basified in NGA-PQ group. The extracts were evaporated under N₂ then dissolved in the mobile phase. A linear chain analogue of primaquine was used as the internal standard. The samples were injected into the normal phase silica column with chloroform:methanol:ammonia solution (86.8:12.5:0.7, V/V/V) as mobile phase and detected at 254 nm. The average recovery of the method was 100.09 ± 3.46 (x̄ ± s)%; the CV of within-day and between-day were 2.7%-6.13% (n = 5) and 8.32%-13.56% (n = 9) respectively; in PQ group and NGA-PQ group, the PQ concentrations ranged from 10-20000 ng/ml blood or lg. liver respectively, and they all yielded a good linear relationship with the peak height ratios. No endogeneous interference was found in chromatograms of biological samples.