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Simultaneous Determination of Furanocoumarins in Cultivated Changium smyrnioides Wollf and Its in Vitro Cultures by HPLC

??OBJECTIVE To determine simultaneously the contents of xanthotoxin,bergaptol, and bergapten in cultivated Changium smyrnioides Wollf and its in vitro cultures by HPLC.METHODS Agilent-C₁₈ column (4.6 mm??250 mm,5 ??m) was used for chromatographic separation and PAD was applied as detector. The flow rate was 1.0 mL??min^-1with a mobile phase of methanol-water in gradient elution mode. The detection wavelength was set at 314 nm while the injection volume was 10 ??L.RESULTS The linear regression equations of xanthotoxin, bergaptol, and bergapten were Y=17 057??-87.689 (r=1.000 0), Y=23 828??-380.44 (r=0.999 9) and Y=37 123??-441.16(r=0.999 9), respectively. The contents of xanthotoxin and bergapten in cultivated and regenerated specimens were obviously higher than those in calli and suspension cells, while bergaptol was only detected in the latter two samples. A larger amount of furanocoumarins accumulated in the cells from leaves and petioles. CONCLUSION The established method is simple and effective with high sensitivity and good repeatability. It can be adopted in studies on the utilization of Changium smyrnioides, especially the regulation and induction of furanocoumarins.