Abstract
A method for the determination of pravastatin sodium (PS), a cholesterol–lowering agent, in plasma was developed by using an immobilized antibody column extraction followed by high‐performance liquid chromatography (HPLC). The analyte was monitored by a laser‐induced fluorescence detector after fluorogenic derivatization. The PS antibody was coupled to Sepharose 4B and used as an extraction phase for sample cleanup and extraction of the drug. A plasma sample was applied to the column and washed with water, and the drug was eluted with methanol.N‐Dansylethylenediamine (DNS‐ED) was coupled to the carboxyl moiety of the drug in the presence of diethyl phosphorocyanidate (DEPC) and triethylamine (TEA) in dioxane. Derivatization was completed in 5 min at room temperature. A column‐switching technique was utilized to remove excess reagents and byproducts. A He–Cd laser‐induced fluorescence detector was applied to achieve an ultrasensitive determination. The detection limit was 2 pg/injection of PS, which was 20 times more sensitive than the conventional fluorescence detection. The limit of quantitation was 100 pg/mL when 1 mL of plasma sample was available. An average coefficient of variations of the overall method were less than 8% at the concentration range of 1–100 ng/mL. A single oral dose of PS in rats (20 mg/kg) and dogs (5 mg/kg) resulted in average maximum concentrations of 142 and 310 ng/mL, respectively.
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