Simultaneous determination of ten flavonoids from Viscum coloratum grown on different host species and different sources by LC-MS.

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 C(18) 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.