Permitted daily exposure from preclinical studies of Ginkgo biloba L. dry extract

Abstract Resolution 658/2022 of the Brazilian Regulatory Agency requires the determination of the permitted daily exposure (PDE) of pharmaceutical agents. Ginkgo biloba L. is used therapeutically to treat memory deficits and other brain diseases. However, published results indicate that more studies are needed to confirm the safety of Ginkgo biloba. This study aimed to evaluate the dry extract of Ginkgo biloba L. leaves PDE as an ingredient in an oral pharmaceutical product in preclinical studies using mice. Acute oral toxicity and repeated dose experiments were performed based on OECD guidelines, as well as genotoxicity tests. The results indicate that Ginkgo biloba L. has low acute toxicity, no liver toxicity, and does not alter blood glucose levels. No changes in weight gain were observed, but food intake decreased in males during the first week of treatment at the highest dose. Hematological parameters were not altered in males, whereas females presented lower leukocyte and lymphocyte counts and higher neutrophil counts at the highest dose. The lipid profile was not altered in males, whereas total cholesterol was increased in females. The estimated PDE was 0.1 mg/day and, when related to the maximum residual concentration, indicates that the cleaning process used is safe and does not require reassessment.

Effect of ginkgo biloba L. extract on oxidative stress and activity of protein kinase C in spleen tissue of mice with irradiation damage / 吉林大学学报(医学版)

Objective: To investigate the influence of Ginkgo biloba L. extract (GBE) in the expressions of oxidative damage-related proteins in the spleen tissue of the mice with radiation damage, and to clarify its mechanism. Methods: Sixty ICR mice were randomly divided into normal control (N O group, irradiation control (IC) group (4. 0 Gy y-ray radiation), low dose of GBE (IC+GBEL) group (4. 0 Gy y-ray+5. 0 mg • kg-1 GBE), medium dose of GBE (IC + GBEM) group (4.0 Gy y-ray+10. 0 mg • kg-1 GBE) and high dose of GBE (IC + GBEH) group (4. 0 Gy y-ray+20. 0 mg • kg_ 1GBE) (n = 1 2) . After continuous administration of GBE for 14 d, all mice received the whole body radiation of y-ray with the total dose of 4. 0 Gy on the 15th day except for the mice in NC group. The levels of malonaldehyde (MDA), the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-px) in the spleen tissue of the mice in various groups were measured by biochemical method 24 h after radiation. The expression levels of 8-hydroxy-2 deoxyguanosine 8-OHdG) in the spleen tissue of the mice in various groups were examined by immunohistochemistry. The activities of protein kinase C (PKC) in the spleen tissue of the mice in various groups were detected by ELISA. Results: Compared with NC group, the levels of MDA and 8-OHdG in the spleen tissue of the mice in IC and different doses of GBE groups were significantly increased (P < 0 . 05), and the activities of SOD, CAT, GSH-px and PKC were significantly decreased (P < 0 . 05). Compared with IC group, the levels of MDA and 8-OHdG in the spleen tissue of the mice in different doses of GBE groups were significantly decreased (P < 0 . 05), and the activities of SOD, CAT, GSH-px and PKC were significantly increased (P < 0 . 05). Conclusion: GBE can inhibit the oxidative stress and regulate the PKC activity in the spleen tissue of the mice with irradiation damage to play a protective role against the radiation damage.

Transcriptome sequencing analysis of gene expression of flavonoid biosynthesis in Ginkgo biloba / 中草药

Objective The transcriptome sequencing was used to analyze the expression of key genes of flavonoid biosynthesis in different stages of growth and development of Ginkgo biloba. Methods The leaves of young trees and adult trees of G. biloba in different periods were used as the test material. Transcriptome sequencing analysis was carried out by using Illumina HiSeq 2000, and analyses of gene functional annotation of Unigene and expression characteristics of key genes for biosynthesis of G. biloba flavonoids were also performed. Results A total of 43 073 Unigene were obtained by transcriptome sequencing, of which 35 179 were annotated and 5 117 genes were screened by differential gene expression. Fifty candidate genes were screened by analyzing KEGG pathway related to flavonoid synthesis. The expression patterns of 50 candidate genes were analyzed. It was found that the key genes of flavonoid synthesis were all highly expressed in young leaves of G. biloba, but there was no significant difference in the leaves between adult and young trees at same time. The 13 genes closely related to the synthesis of flavonoids were analyzed. Among them, the expression of C4H, CHS, ANS, ANR, and FOMT genes was high, and the expression of F3’H, F3’5’H, and FLS genes was relatively low. Conclusion Through transcriptome sequencing, we screened and analyzed the key genes of flavonoid biosynthesis of G. biloba and their expression characteristics, which provided the theoretical basis of molecular pharmacology for improving the yield of ginkgo flavonoids.

Advances research on chemical constituents, preparations and adverse reactions of Ginkgo biloba / 药物评价研究

The leaves and fruits of Ginkgo biloba L.are embodied in Pharmacopoeia of People's Republic of China,mainly contain ginkgo flavonoids,terpene lactones,phenolic acids,isopentenyl alcohol,steroids and other chemical compositions.The extract and active ingredients can be made into tablets,capsules,granules,oral liquid,injection,dripping pills,syrup,tincture and other formulations,clinically used for cardiovascular and cerebrovascular disease treatment.The adverse reactions are allergies,diarrhea,bleeding,liver and kidney toxicity.This review contains the chemical composition,dosage form and clinical adverse reactions of Ginkgo biloba in recent years,for providing a reference of further development and rational utilization of this plant resource.

Research progress on breeding and cultivation of Ginkgo biloba for leaf use in China / 中草药

Ginkgo biloba mainly contains many kinds of special flavonoids and terpenlactones in its leaves. The preparation of Ginkgo biloba extract is very good for curing patients suffering from cardiovascular and cerebrovascular diseases. In order to get more Ginkgo biloba leaves rich in flavonoids and terpenlactones, the rules are set to seek fine varieties including higher contents of flavonoids and terpenlactones, higher leaves yield than ordinary ones. Many fine varieties were selected according to these standards and applied in Ginkgo cultivation for leaf use. Factors affecting the contents of active ingredients and the amount of leaves covered Ginkgo varieties, sex, age, origin, environment, and harvest time, location in tree's canopy and branches, branch types and so on. The suitable application of fertilizer, trimming, pruning, and planting density will improve the yields of leaves and medicinal components. Future works should focus on two aspects, one is the germplasm resources collection, conservation, studying, and selection, another is directionally cultivation of leaf-used Ginkgo using improved varieties and standardized cultivation.

Comparison on antimicrobial activity of 40 endophytic fungi strains from Ginkgo biloba and preliminary analysis on ingredients with antimicrobial activity / 中草药

Objective: To compare the antimicrobial activity of 40 endophyte strains from Ginkgo biloba and analyze the ingredients with antimicrobial activity. Methods: Endophytic fungi inhabited in leaves, stems, and fruits of Ginkgo biloba were isolated. Bacillus subtilis, Saccharomyces cerevisiae, Aspergillus niger and Rhizopus were used as indicators to do the microbial inhibition experiment. Forty strains antimicrobial activity were investigated by using filter paper method and flat hole well diffusion method. Silica gel column chromatography, Sephadex LH-20 column chromatography, preparative HPLC, TLC, and HPLC-MS were used to isolate, purificate and identify the antimicrobial components from the broth of Penicillium oxalicum YXY009. Results: Eighteen strains produced sterile mycelia on PDA plates. Among the other strains, Penicillium spp. and Aspergillus spp. accounted for 17.5% and 15% of the total fungi. The strain with the highest antimicrobial activity was determined to be Penicillium oxalicum YXY009. The inhibiting zone diameters of the YXY009 strain reached (13.47 ± 0.46), (22.35 ± 1.32), and (23.60 ± 0.83) mm against Bacillus subtilis, Saccharomyces cerevisiae, and Rhizopus, respectively. Conclusion: An active compound POM was isolated from the fermentation broth of the strain P. oxalicum YXY009 and identified as 5-hydroxy-2-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl) oxan-2-yl] oxybenzoate for the first time.

Potential mechanism of Ginkgo Folium extract used in treatment of gout based on network pharmacology / 中草药

Objective: To screen the active compounds of Ginkgo Folium extrat (GFE) used in the treatment of gout to build network of compounds-target-pathway, and to elaborate the potential mechanism of GFE in the treatment of gout. Methods: The G. biloba compound-target interaction model was established by random forest algorithm, which was then applied to predict the potential targets interacted with compounds of G. biloba. The complicated compound-target-disease network of G. biloba was then constructed and analyzed. Results: Among the 158 compounds found in G. biloba, with (drug-likeness) DL ≥ 0.18 and (oral bioavailability) OB ≥ 30 as a filter condition, 39 active compounds were screened, including 11 flavonoids and 11 terpenes. There are 24 active compounds in G. biloba, which correspond to the 49 pathogenic genegenes of gout and reveal the molecular mechanism of G. biloba in the active ingredient and related targets. Conclusion: Active compounds in G. biloba may cure gout through the following three ways: First, to minimize the production of uric acid; Secondly, to increase the excretion of uric acid; Thirdly, to inhibit disorders induced by NALP3 inflammation.

Ginkgo biloba leaves history: A model of research and development for Chinese materia medica/phytomedicine / 中草药

Summary in this paper, the active chemical constituents from Ginkgo biloba leaves (GBL), their pharmacological effects, toxicity studies, clinical application, the comparison with the quality standard of the Pharmacopoeia, the history of medicinal development as well as the research and development of GBL in China were described, analyzed, and discussed in detail. GBL began to be recorded the medicinal value from the Song Dynasty in China. The scientists in research and development of GBL in Japanese and German have made significant pioneering contributions. The history of medicinal development of GBL has been fully embodied in the innovation, and has become a model for the research and development of Chinese materia medica and phytomedicine. The innovative drug research of GBL in China has made considerable progress, the significant results have been obtained. But the quality standard of Chinese GBL extract and its preparations have a significant difference with the European and American industries. The industry concentration is low.

Insight into recently issued community herbal monograph on Ginkgo biloba L., folium and its impact on registration strategy of Ginkgo Folium products in EU / 中草药

In January, 2014, European Medicines Agency (EMEA) issued a draft for community herbal monograph on Ginkgo biloba L., folium. That means the community herbal monograph on Ginkgo biloba L., folium has been settled finally. This assay introduces the new Ginkgo monograph briefly, describes the two registering methods and requirements for herbal medicines in EU, and reads the new monograph in more detail. By analyzing the impact of the new monograph on Ginkgo Folium products, this monograph will offer a very important reference and basis for the herbal registration application of Ginkgo Folium products in EU.

Component analysis of nucleosides, amino acids, and mineral elements in pollen of Gingko biloba / 中草药

Objective: To analyze the nutritional ingredient in the pollen of Gingko biloba and provide the theoretical basis for the comprehensive utilization of pollen of G. biloba. Methods: The nucleosides and dissociative amino acids in the pollen of G. biloba were detected by UPLC-TQM. The content of nucleosides in pollen of G. biloba was calculated with the help of UPLC-TQ/MS, and the mineral elements in the pollen of G. biloba were determined by ICP-AES. Results: In pollen of G. biloba there were 24 abundant dissociative amino acids accounting for 62.316 mg/g totally. In this number, dissociative essential amino acids accounted for 6.23 mg/g with the highest content of 17.11 mg/g aspartic acid (ASP). Meanwhile, a small quantity of taurine, approximately 0.24 mg/g, has been tested out. Furthermore, there were 16 kinds of nucleosides in the pollen of G. biloba. Among those nucleosides, guanosine made up for the highest percent with 251.99 μg/g while other nucleosides accounted for a low percent. In pollen of G. biloba, K is the most abundant with 26.301 mg/g, followed by Ca with 7.076 mg/g, Mg with 5.361 mg/g, and Fe with 2.545 mg/g. As to the heavy mental elements, as and Cd were not found in it. Conclusion: From above analysis, pollen of G. biloba can provide the theoretical basis for the development of health-care food with pollen of G. biloba.

Chemical constituents from root barks of Ginkgo biloba (I) / 中草药

Objective: To study the chemical constituents from the root barks of Ginkgo biloba. Methods: The constituents were isolated by column chromatography and their structures were elucidated by physicochemical properties and spectroscopic analyses. Results: Thirteen compounds were isolated including four fatty acids, palmitinic acid (1), stearic acid (2), behenic acid (3), and lignoceric acid (4); one fatty alcohol, 1-heptacosanol (5); two sterides, β-sitosterol (6) and daucosterol (7); two flavonoids, genkwanin (8) and apigenin (9); and four ginkgolides, bilobalide (10), ginkgolide A (11), ginkgolide B (12), and ginkgolide C (13). Conclusion: Among them, compounds 3-5 are isolated from this plant for the first time. The results are helpful for the comprehensive utilization of the resources of the root barks of G. biloba.

Optimization on preparation of Ginkgo biloba extract-β-cyclodextrin inclusion complex by central composite design and response surface / 中草药

Objective: To optimize the preparation of Ginkgo biloba extract-β-cyclodextrin inclusion (GBE50-β-CD inclusion) complex by central composite design and response surface methodology. Methods: Saturated aqueous solution method was used to prepare GBE50-β-CD inclusion complex. The preparation was optimized by central composite design. With the mass ratio of β-CD/GBE50, inclusion time and temperature as the independent variables and with the inclusion rate as the dependent variable, the binomial and linear equations were fitted to the data of overall desirabilities, and the resulting equation was used to produce 3-D response surface graphs, through which optimal formulation was predicted, the best prescription process was chosen to verification test. Results: It showed that the correlation coefficient of second-order quadratic model was higher and the prediction was better (r = 0.966); The optimum conditions for the preparation of GBE50-β-CD inclusion complex were as follows: the mass ratio of β-CD/GBE50 3:1, inclusion time 6 h, inclusion temperature at 60 °C. The deviation between, the result of the best craft verification test and the binomial fitting equation forecast value is 1.07%. Conclusion: Central composite design and response surface methodology is successfully used to optimize the preparation of GBE50-β-CD inclusion complex. The optimized process is reliable, stable, and available for the industrial production.

Atividade antioxidante dos extratos de Ginkgo biloba L / Antioxidant activity of extracts of Ginkgo biloba L

Atualmente, inúmeros ensaios têm demonstrado que os compostos bioativos encontrados em extratos vegetais, possuem eficácia no controle de reações oxidativas. Deste modo, este trabalho avaliou a efetividade de extratos obtidos das folhas de Ginkgo biloba L, como antioxidantes in vitro. Para tanto, elaboraram-se três extratos (um aquoso e dois purificados). As análises da atividade antioxidante, fenóis totais e redução de potência, foram realizadas para avaliar a ação antioxidante dos extratos. (...) O extrato aquoso apresentou um maior rendimento no conteúdo de fenóis totais, comparando-se com os extratos purificados, entretanto, a sua atividade antioxidante foi inferior, comprovando-se, deste modo, a maior eficácia da extração por fracionamento na seleção dos compostos ativos do Ginkgo biloba L.

A new ginkgolide from Ginkgo biloba / 中国药科大学学报

Aim: To study new chemical constituents of the leaves of Ginkgo biloba L. Methods: Isolation and purification were carried out by several chromatographic methods. The structures of the compounds were elucidated by detailed analysis of their UV, IR, MS, ~1H NMR and ~(13)C NMR spectra Results: A new ginkgolide, ginkgolide N( 1, 7, 10-trihydroxy-3,14- dehydroginkgolide, Ⅱ), along with a known compound, was isolated from the leaves of G. biloba. The structure of the known one was elucidated as ginkgolide L(Ⅰ). Conclusion: Compound Ⅱ was a new compound. The complete spectroscopic data of compound Ⅰ were reported for the first time.

Avaliação da qualidade de amostras comerciais de boldo (Peumus boldus Molina), pata-de-vaca (Bauhinia spp.) e ginco (Ginkgo biloba L.) / Control quality evaluation of commercial samples of boldo (Peumus boldus Molina), pata-de-vaca (Bauhinia spp.) and ginkgo (Ginkgo biloba L.)

O presente trabalho procurou avaliar a qualidade de produtos comercializados à base de boldo, pata-de-vaca e ginco, através dos parâmetros contidos na Farmacopéia Brasileira e na literatura específica. Foram analisadas oito amostras de boldo, nove de pata-de-vaca e sete de ginco, adquiridas em farmácias na cidade do Recife. A metodologia consistiu em avaliar: os rótulos e bulas dos produtos verificando se estavam de acordo com a RDC n o 17 de 24/02/2000 e a portaria 110/97 da ANVISA; realizar análise sensorial; verificar a autenticidade das amostras e sua pureza. Todas as embalagens de boldo, pata-de-vaca e seis de ginco continham erros ou ausência de informações científicas, além da falta de bula. Na verificação de impurezas, todas as de boldo, cinco de pata-de-vaca, e uma amostra de ginco foram reprovadas. Todos os produtos analisados apresentaram alguma irregularidade segundo os códigos oficiais, sendo necessário uma maior intensificação na vigilância de produtos à base de plantas medicinais no Brasil.

This study aimed to evaluate the quality of commercial products prepared with boldo (Peumus boldus Molina), pata-de-vaca (Bauhinia spp.) and ginkgo (Ginkgo biloba L.) by using parameters from the Brazilian Pharmacopoeia and specific literature. Eight samples of "boldo", nine of "pata-de-vaca", and seven of ginkgo were analyzed, all bought from pharmacies in Recife (Pernambuco, Brazil). The methodology consisted in evaluating the products’ labels and instructions to verify their accordance to RDC n o 17 of 02/24/2000 and ANVISA (National Sanitary Surveillance Agency) decree 110/97, undertaking a sensorial analysis, and verifying the authenticity and purity of the samples. All of the packages of "boldo" and "pata-de-vaca" and six of the packages of ginkgo contained mistakes or lacked scientific information; instructions were also missing. After analyzing for impurities, all samples of "boldo", five of "pata-de-vaca", and one of ginkgo were rejected. All of the products analyzed had some sort of irregularity in relation to the official codes, making it necessary to intensify the inspection of medicinal plant products in Brazil.

Extraction of flavones from leaves of Ginkgo biloba L. by microwave technique and optimized by experimental design and optimization / 第三军医大学学报

Objective To select the best condition for microwave assisted extraction (MAE) of flavones from leaves of Ginkgo biloba L . and compare this method with the most conventional extraction way. Methods Both microwave assisted extraction and theat maceration were adopted for flavones from leaves of Ginkgo biloba L. , and the total content was determined by spectrophotometry. Results Under appropriate MAE conditions, both the recovery and purity of total flavones obtained from the experiments by uniform design and the orthogonal design were very similar. The optimal recovery and purity by orthogonal experimental design were 212 4 mg/g and 61 9%, respectively. The optimal results by orthogonal experimental design were 216 2 mg/g and 57 1%, respectively. However, the results by maceration were only 114 6 mg/g and 40 1%, respectively. Conclusion MAE is a more suitable method for the extraction of flavones from leaves of Ginkgo biloba L . because of its higher extraction rate and purity.

Cell immobilization culture of Ginkgo biloba and determination of their affecting factors / 中草药

Object To investigate the application of cell immobilization culture to producing ginkgolides in Ginkgo biloba L. Methods To immobilize the cells by polyaminoresin foam as support materials, and determine the effects of different factors. Results The results showed that 71% of immobilization ratio and 22 mg/cm 3 of cell density were obtained using P29 of high density, small bore diameter with pieces of 0.5 cm ? 0.5 cm ? 0.5 cm, 0.72 g of support matrices per bottle in 65 mL of MS medium solution supplemented with 2,4-D 8.0 mg/L+KT 0.04 mg/L+NAA 0.4 mg/L and the incoulum quantity was 200 g/L. Conclusion It shows that present method has some advantages, such as simpleness, low cost, high immobilization ratio.

Quality analysis of seedling leaves of Ginkgo biloba Ⅱ / 中草药

Object To stuay the quality of seedling leaves cultured from improved variety of Ginkgo biloba L.. Methods The content of terpene lactones in seedling leaves of G. biloba were determined by RP-HPLC-ELSD.Results The content of total terpene lactone producted in Jun. was more than that in Aug., and it was obviously lower in Nov.. The content of terpene lactone in grafting was more than that in seedling. The content of terpene lactone in male trees was more than that in female trees.Conclusion The content of the active components was related to the season of growth, age of tree, raising modes, environment of growth, geographic conditions and others. The evaluation of the content of the main active components BB and GB showed that the quality of seedling leaves cultured from improved variety of G. biloba in Jinagxi is better than that reported by literature.

Quality and quantity analysis of terpene lactones from Ginkgo biloba leaves by GC-MS / 中草药

Object To develop a capillary GC-MS analytical method for identification and deter- mination of ginkgolide A, B, C and bilobalide (GA, GB, GC and BB) in Ginkgo biloba L. leaves. Methods The leave samples were extracted in ultrasonic bath with ethanol-water (20∶80). The extract was purified by liquid-liquid extraction with ethyl acetate followed by solid-phase extraction on a column mixed with acid Al 2O 3, active carbon and celite. The terpenes were trimethylsilylated by BSTFA (with 1% TMCS) for 60 min at 100 ℃ and determined by GC-MS with HP-5 MS capillary column in the selected-ion monitoring mode. The intense fragment ions were chosen as monitoring ions for quantitative analysis. Cholesterol was used as an internal standard. Column temperature gradient: initial temperature 180 ℃, maintained 1 min, and then increased at 20 ℃/min to 260 ℃, and finally at 2 ℃/min up to 300 ℃, maintained 2 min. Results The retention times of GA, GB, GC and BB were 13.7,14.3,15.3 and 6.8 min, the major fragmentation ions (monitoring) were at m/z 537, 625, 713 and 455 (299), the average recoveries of GA, GB, GC and BB were 102.0%, 99.4%, 96.0%, 96.3%, RSD were 0.54%, 2.40%, 1.98% and 2.43%, respectively. Conclusion This method is repeatable, specific, accurate and easy to operate. It is adoptable for quality and quantity analysis of terpene lactones from G. biloba leaves.

Study on changing rules for total flavonoids and total terpene lactones in Ginkgo biloba leaves at different ages of tree and collecting seasons / 中草药

Object To define suitable gathering season of Ginkgo biloba L. leaves. Methods The content of total flavonoids and total terpene lactones in G. biloba leaves were determined by HPLC DAD and HPLC ELSD. Results The content of total flavonoids and total terpene lactones in G. biloba were distinct in different ages of tree and collecting seasons. Conclusion The content of total flavonoids and total terpene lactones in G. biloba are the highest in 2 3 ages of tree. The total flavonoid is the highest in May and total terpene lactone is the highest in September.