Simultaneous Determination of Ten Flavonoids from Viscum coloratum Grown on Different Host Species and Different Sources by LC-MS

抄録

A high-performance liquid chromatographic-mass spectrometric method was developed for the simultaneous determination of 10 flavonoids in Viscum coloratum obtained from different host species and different sources. Viscum coloratum was extracted with 50% methanol. The extracts were separated on a C₁₈ column with a gradient of 0.1% (v/v) formic acid and methanol. The flavonoids in the extracts were detected by negative electrospray ionization mass spectrometry in selective ion monitoring mode. The calibration curves showed good linearity (r>0.998) within the test ranges (homoeriodictyol: 0.149—8.940 μg/ml, homoeriodictyol-7-O-β-D-glycoside: 0.230—13.80 μg/ml, homoeriodictyol-7-O-β-D-apiose (1→2)-β-D-glycoside: 5.000—300.0 μg/ml, homoeriodictyol-7-O-β-D-apiose (1→5)-β-D-apiose (1→2)-β-D-glycoside: 0.835—125.3 μg/ml, rhamnazin-3-O-β-D-glucoside: 0.064—3.840 μg/ml, rhamnazin-3-O-β-D-(6″-β-hydroxy-β-methyglutaryl)-glucoside: 1.435—86.10 μg/ml, isorhamnetin-3-O-β-D-glucoside: 0.930—55.80 μg/ml, 5-hydroxy-3,7,3′-trimethoxyflavone-4′-O-β-D-glucoside: 0.067—4.020 μg/ml, 5,7,4′-trihydroxy-3,3′-dimethoxyflavone: 0.270—16.20 μg/ml, pachypodol: 0.110—6.600 μg/ml). The limits of quantification were between 0.006—0.720 μg/ml. The assay was reproducible and the overall intra- and inter-day variations were less than 4.6%. The recoveries varied from 93.4 to 103.9% at three different concentration levels. The validation method was used to determine the contents of 10 flavonoids in Viscum coloratum. A one-way analysis of variance was applied to evaluate Viscum coloratum-host-source interactions. Compared with the host species, the sample source had a significant impact on the sample content.