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Abstract
Objective: To investigate and compare the quantitative determination of naphthodianthrones of Hypericum perforatum L.by UV spectrophotometry and HPLC method.Methods: Hypericin was used as the reference compound for UV spectrophotometry,and the detection wavelength was 590 nm.HPLC was performed on a Diamonsil C18 analytical column(150 mm×4.6 mm,5 μm)with mobile phase consisted of CH3OH-10 mmol/L CH3COONH4(90∶10,pH 6.2) at the flow rate of 1.0 ml/min.The detection wavelength was 590 nm and column temperature was 30 ℃.The two peaks in HPLC chromatograms of herba Hypericum perforatum were identified as hypericin and pseudohypericin by UV spectra and negative ion mass spectra.The content of naphthodianthrones of Hypericum perforatum herba was determinated and estimated based on the sum of hypericin and pseudohypericin.Results: The linear range of hypericin was 2.44-19.52 μg/ml for UV spectra(r=0.999 6,n=3).The content of naphthodianthrones of Hypericum perforatum herba calculated with reference to hypericin was(0.990 2±0.006 0) μg/mg.For HPLC analysis,pseudohypericin had good linearity in the range of(0.402-8.032) μg/ml(r=0.999 5,n=3) and hypericin also had good linearity in the range of(0.488-9.760) μg/ml(r=0.999 7,n=3).The contents of pseudohypericin and hypericin from Hypericum perforatum herba calculated with its standard references were(0.387 2±0.001 4) μg/mg and(0.220 2±0.000 7) μg/mg,respectively.The total content of naphthodianthrones of Hypericum perforatum herba determined by HPLC with references to pseudohypericin and hypericin was(0.605 5±0.001 2) μg/mg.Conclusion: The contents of naphthodianthrones from Hypericum perforatum herba determined by UV spectrophotometry and HPLC are significantly different.This suggests that the accuracy of UV spectrophotometry is suspectable of determing the content of naphthodianthrones in Hypericum perforatum although the chemical structure and UV spectra are similar.
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