High performance liquid chromatography and capillary electrophoresis determination of sanguinarine in biological matrices

HPLC and CE methods were employed to determine the quaternary benzo[c]phenanthridine alkaloid sanguinarine,in biological matrices (rat hepatocytes, human gingival fibroblasts, feed, porcine faeces, body fluids and tissues). HPLC was carried out on a C-18 column using gradient elution and ion pairing techniques with 1-heptanesulfonic acid as ion pairing agent under acidic conditions. The detection limit for fluorimetric detection at lambda(ex) = 327 nm and lambda(em) = 577 nm was 3 nM sanguinarine. CE analyses were performed in 50 mM phosphate-Na buffer pH 2.5, with 150 mM SDS used for pre-concentration by the sweeping effect. This experimental configuration allows injecting the total capillary length with sanguinarine sample. The detection limit for UV detection at 285 nm was 12 nM. Both methods are suitable for analysing submicromolar quantities of sanguinarine in biological materials. The HPLC method is more sensitive than CE because it uses fluorescence detection.