Quantitation of anthocyanins in elderberry fruit extracts and nutraceutical formulations with paper spray ionization mass spectrometry

The ability to rapidly identify and quantitate, over a wide range of concentrations, anthocyanins in food and therapeutic products is important to ensuring their presence at medicinally significant levels. Sensitive, yet mild, analysis conditions are required given their susceptibility to degradation and transformation. Paper spray ionization has been used to detect and quantify the levels of anthocyanin levels in extracts of fresh and dried elderberries, and elderberry stems, as well as 3 commercially available nutraceutical formulations. The component cyanidin glucosides, including cyanidin-3-sambubioside, cyanidin-3-glucoside, cyanidin-3,5-diglucoside, cyanidin-3-sambubioside-5-glucoside, and the aglycone cyanidin, were readily detected in a range of sources. Quantitation was achieved by establishing a calibration plot from dilutions of a stock solution of cyanidin-3,5-diglucoside containing malvidin-3,5-diglucoside as an internal standard at a fixed concentration. The same standard was used to quantify the anthocyanin content in the fruit and nutraceutical formulations. Wide 5-fold variations in anthocyanin concentration were detected in the nutraceutical formulations from different suppliers ranging from 1050 to 5430 mg/100 g. These concentrations compared with 500 to 2370 mg/100 g measured in the dried stems and fruit, respectively.