Simultaneous determination of in total 17 opium alkaloids and opioids in blood and urine by fast liquid chromatography–diode-array detection–fluorescence detection, after solid-phase extraction

Abstract

A fast liquid chromatographic method with tandem diode array–fluorescence detection for the simultaneous determination of in total 17 opium alkaloids and opioids is presented. Blank blood and urine samples (1 ml) were spiked with different concentrations of a standard mixture, as well as with the internal standard, butorphanol (2000 ng/ml). After solid-phase extraction, based on weak cation exchange (Bond Elut ® CBA SPE columns), the extracts were examined by HPLC–DAD–FL. By using a “high-speed” phenyl column (53×7.0 mm I.D., particle size 3 μm) eluted with a gradient system (A: water–methanol (90:10, v/v), B: methanol, both containing 25 m M triethylammoniumformate (pH A=4.5)) all compounds could be baseline separated within 12 min. The method was validated and its applicability was demonstrated by the analysis of real-time forensic cases.