Determination of trace amounts of ginkgolic acids in Ginkgo biloba L. leaf extracts and phytopharmaceuticals by liquid chromatography–electrospray mass spectrometry

Abstract

Ginkgolic acids (GAs) are toxic phenolic compounds present in the fruits and leaves of Ginkgo biloba L. (Ginkgoacae). Their maximum level in phytopharmaceuticals containing ginkgo extracts has been recently restricted to 5 μg/g by the Commission E of the former Federal German Health Authority. In order to detect ginkgolic acids at these low levels, a sensitive and selective analytical method, based on liquid chromatography–electrospray mass spectrometry (LC–ES-MS) has been developed. The three main phenolic acids ( 1– 3) of the chloroform fruit extract were isolated and used as standards for quantification. In the LC–ES-MS negative ion mode, calibration curves with good linearities ( r=0.9973, n=6) were obtained in the range of 0.5–10 μg/g for compounds 1, 2 and between 0.1 and 7.5 μg/g ( r=0.9949, n=6) for ginkgolic acid 3. The detection limits at a S/ N ratio of 3 were 0.1 ( 3) and 0.25 μg/g ( 1, 2). Recoveries were around 101% at 5 μg/g for the substances detected in the leaf extracts. Good precision was achieved with relative standard deviations of less than 4% ( n=6). The optimised method was applied to verify whether the amount of gingkolic acids was below 5 μg/g in a standardised leaf extract which is a constituent of a phytopreparation.