Comparison of extraction methods for secologanin and the quantitative analysis of secologanin from Symphoricarpos albus using 1H-NMR.

In order to develop an efficient large-scale extraction of secologanin from Symphoricarpos albus, different methods have been compared. Ultrasonication with organic solvents and water, microwave-assisted extraction and hot water extraction methods were evaluated for their efficiencies. Among the methods tested, ultrasonication with methanol showed the highest yield of secologanin (3.35 +/- 0.24 mg/g fresh weight). For reliable quantification of secologanin a 1H-NMR method was developed. The experiment was performed by the analysis of the integral of the signal of H-9, which was well separated in the range delta 7.4-7.5 in the 1H-NMR spectrum. The quantity of the compound was calculated from the relative ratio of intensity of the target peak to the known amount of internal standard, 200 microg of gallic acid. This method allows rapid and simple quantification of secologanin in 5 min without any pre-purification steps.