A simple and accurate high-performance thin-layer chromatography (HPTLC)-bioautographic method was developed for the quantitative analysis of magnolol and honokiol in the herbal medicine Magnoliae officinalis Cortex. The samples were separated on a silica gel HPTLC plate with a mixture solution of toluene-methanol (10:1, v/v) as the mobile phase. Spots were visualized by dipping in 2,2-diphenyl-1-picrylhydrazyl radical (DPPH*) reagent and measured at a wavelength of 550 nm in a reflection mode, scanning after derivatization for 40 min. The method had excellent linearity (r2 = 0.9939 for magnolol and r2 = 0.9989 for honokiol, respectively) in the concentration range of 0.16–0.97 mg spot−1 for both analytes. The recoveries were 94.5–105.9% for magnolol and 86.6–103.4% for honokiol, respectively. The established HPTLC-bioautographic method was evaluated comprehensively in quantitative and antioxidant activity analysis of magnolol and honokiol in Magnoliae officinalis Cortex and various plants.
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