柱前衍生化-高效液相色谱法测定大鼠灌服朱砂后血浆中活性硫的药动学特征

Pharmacokinetic Features of Reactive Sulfide in Plasma of Rats After Oral Administration of Cinnabaris by Using HPLC Coupled with Pre-column Derivatization

目的测定朱砂给药后活性硫的药动学特征。方法建立单溴二胺(MBB)柱前衍生化-高效液相色谱法测定朱砂灌胃后大鼠血浆中活性硫水平的方法,并将该法用于大鼠灌服朱砂后血浆中活性硫的药动学研究。结果大鼠血浆中活性硫在0.25~15μmol/L范围内线性关系良好(r>0.99),定量限和检测限分别为0.1μmol/L和0.02μmol/L,方法的日内、日间精密度分别小于4.4%、3.5%,准确度为-9.9%~6.0%,平均回收率为74.9%。灌胃给予大鼠朱砂0.6 g/kg,血浆中活性硫C_(max)为(1.33±0.13)μmol/L,t_(max)为(150±34)min,t_(1/2)为(323±62)min,AUC_(0-∞)为(5743±297)ng/m L·h。结论本研究建立了灵敏、专属、准确的柱前衍生化-高效液相色谱法测定血浆中活性硫含量的方法,可用于朱砂灌胃大鼠血浆中活性硫的动力学特征研究。

Objective To study the pharmacokinetic features of reactive sulfide in rats after oral administration of Cinnabaris. Methods An HPLC coupled with precolumn derivatization method was developed for the pharmacokinetic features study on reactive sulfide in rats after oral administration of Cinnabaris. Results Good linearity （r〉0.99） was found for reactive sulfide in plasma in the concentration range of 0.25–15 μmol/L （r〉0.99）. The LOQ and LOD of the method were 0.1 μmol/L and 0.02 μmol/L, respectively. The intra- and inter-day precision was less than 4.4% and 3.5% respectively, and the accuracy was -9.9%–6.0%. The average recovery rate was 74.9%. 0.6 g/kg Cinnabaris was given the rats for gavage, and the time-course pharmacokinetics parameters were as follows： Cmax （1.33±0.13） μmol/L, tmax （150±34） min, t1/2 （323±62） min, AUC0-∞ （5743±297） ng/mL？h. Conclusion A sensitive, robust and accurate precolumn derivatization-HPLC method for the determination of plasma reactive sulfide is developed and validated. The method is successfully applied in the pharmacokinetic features study on reactive sulfide in plasma of rats after administration of Cinnabaris.