Successive cartridge clean-up method for the simultaneous determination of matrine and oxymatrine in biopesticides containing Sophora flavescens extract was developed and validated by UPLC. The clean-up method was established with ENVI-Carb (0.5 g) and C18 SPE (0.5 g) cartridges for the bioactive alkaloid in biopesticides from S. flavescens, and the eluate was analyzed to quantify the matrine and oxymatrine by UPLC. The developed method was validated, and the recovery and LOQ of both materials were 105.0% and 103.6%, and 0.050 and 0.684 mg·kg −1 , respectively. Of the twenty one samples, the total content of matrines were analyzed by using the developed method and the result showed
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