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Refined extracts of Ginkgo biloba L. leaves can protect brain tissue from diverse types of tissue damage. Preliminary experiments identified 6-hydroxykynurenic acid (6-HKA) as a central contributor to this effect, and thus as a potential active marker for drug quality analysis. In contrast, genistein is a negative quality marker for this drug, as high contents of this compound in G. biloba leaves samples are in most cases caused by adulteration with Sophora flavescens Aiton, which is added to the extract by unlawful producers in order to boost its content in flavones, which serve as a quality marker. Some publications even dispute the natural occurrence of genistein in G. biloba leaves altogether, thus claiming that its presence even in trace amounts would prove adulteration. The present study aims to resolve the influence of harvesting time and location on the contents of genistein and 6-HKA - HPLC: Agilent SB-C18 column (5 μm, 4.6×250 mm); phosphoric acid pH2.5, acetonitrile, methanol, gradient; 1 mL/min; 25°C; UV 350 nm - in G. biloba leaves. The lowest contents of 6-HKA were observed in July and August, whereas the maximum was reached in May and October. In the case of genistein - its natural occurrence in G. biloba leaves could be demonstrated under controlled conditions for the first time - the lowest contents were found in May and June, whereas the highest were detected in September and October. These results show that harvesting of G. biloba leaves for pharmaceutical use should be avoided in July and August because of the reduced 6-HKA content, as well as in September and October, the season of highest genistein production. Due to both favorable 6-HKA production and a relative lack of genistein, May was identified as the ideal harvesting month for G. biloba leaves.

Seasonal Variability of Genistein and 6-Hydroxykynurenic Acid Contents in Ginkgo biloba Leaves from Different Areas of China