Multi-Response Optimization of Ultrasonic Assisted Enzymatic Extraction Followed by Macroporous Resin Purification for Maximal Recovery of Flavonoids and Ginkgolides from Waste Ginkgo biloba Fallen Leaves.

Guisheng Zhou,Xinmin Wang,Jin-Ao Duan,Jing Zhang,Yuping Tang,Jiayan Ma

doi:10.3390/molecules23051029

Abstract

In the present study, the process of ultrasonic assisted enzymatic extraction (UAEE), followed by macroporous resin purification, was successfully developed to achieve maximal recovery of flavonoids and ginkgolides from Ginkgo biloba fallen leaves (GBFL). Three effective extracted factors, including UAE power, EtOH%, and the amount of cellulase were screened by Plackett–Burman design (PBD). The important variables were further optimized by rotatable central composite design (RCCD). After the combination of PBD and RCCD, the resulting optimal UAEE conditions were as follows: UAE power of 218 W; EtOH% of 68%; the amount of cellulase of 8.4 mg; UAE temperature of 40 °C; UAE time of 20 min; pH of 5.0; and, sample particle size of 40 mesh. Under the optimum conditions; the yields of flavonoids were 0.74 ± 0.05% (n = 3) and ginkgolides was 0.42 ± 0.06% (n = 3), which were close to the predicted values. Moreover, the further enriching flavonoids and ginkgolides from the obtained GBFL extracts using the above optimum UAEE condition was successfully achieved by macroporous resin DA-201. After column adsorption and desorption on DA-201; the percentage of total flavonoids was (25.36 ± 1.03)%; ginkgolides was (12.43 ± 0.85)% and alkylphenols was (0.003 ± 0.0005)% from the obtained dry extracts of GBFL which were complied with Chinese pharmacopoeias. Therefore, the present study provided a convenient and efficient method for extraction and purification of flavonoids and ginkgolides from waste GBFL.

Highlights

Ginkgo biloba L. (G. biloba), the only remaining member of the Ginkgoaceae family, was widely distributed in extratropical, warm and subtropical zones, especially in eastern China
This study reported the effect of extraction and purification of flavonoids and ginkgolides from waste Ginkgo biloba fallen leaves (GBFL) for the first time, which will provide a scientific basis for the comprehensive utilization and development of G. biloba resources
HPLC-DAD and HPLC-MS methods, we developed an ultra-high performance liquid chromatography coupled with diode array detector-Q/TOF mass spectrometry (UPLC-DAD-Q/TOF-MS) method for the identification and the determination of flavonoids and ginkgolides in GBFL

Summary

Introduction

Ginkgo biloba L. (G. biloba), the only remaining member of the Ginkgoaceae family, was widely distributed in extratropical, warm and subtropical zones, especially in eastern China. (G. biloba), the only remaining member of the Ginkgoaceae family, was widely distributed in extratropical, warm and subtropical zones, especially in eastern China. G. biloba has existed on the earth for 200 million years, but its true value has induced a range of attentions all around the world until the recent five decades. The extract from G. biloba leaves (EGb) was among the most commonly used herbal medicines and/or dietary supplements. G. biloba leaves was one of the most sold medicinal plants of estimates of worldwide annual sales, and EGb was standardized for the two fractions of main active ingredients [6], namely flavonoids and ginkgolides. EGb has been of great interest to the food and pharmaceutical industries due to their inimitable benefits [7]

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