• Korean Journal of Food Science and Technology
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• v.32 no.5
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• pp.1198-1205
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• 2000

Effect of inhibitions of three kinds of Ginkgo biloba extracts(Ginkgo biloba extract, Ginkgolide A, and Ginkgolide B) on induction of reactive oxygen species and release of inflammation mediator arachidonic acid were tested. Three kinds of Ginkgo biloba extracts could not inhibit the pyrogallol auto-oxidation, but they showed the hydrogen atom donating activity in DPPH assay. When 10 ${\mu}M$ hydrogen peroxide and 400 ${\mu}g/mL$ of three kinds of Ginkgo biloba extracts were added to U937 monocytic macrophage, the induction of lipid peroxidation was not observed. The Ginkgo biloba extract showed the most powerful inhibition among the extracts. And only Ginkgolide A was good for the inhibition of the protein degradation. The release of inflammation mediator arachidonic acid was induced by adding TPA and calcimycin to U937. In this assay, even 10 ${\mu}g/mL$ of three different Ginkgo biloba extracts excellently blocked the release of arachidonic acid. Particularly, the inhibition efficiency of Ginkgolide B was about 11 times higher than that of induction, and was about 4 times higher than that of the control of noninduction. This result suggests that the release of arachidonic acid is not inhibited by the antioxidant activity of Ginkgo biloba extracts, but a pre-step of the release of arachidoinc acid is inhibited by Ginkgo biloba extracts.

• Korean Journal of Pharmacognosy
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• v.37 no.4 s.147
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• pp.278-282
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• 2006

In this study, we analyzed and quantified the contents of magnolol (1), honokiol (2), and obovatol (3) in the leaves of Magnolia species by HPLC. HPLC methods were developed and validated by ICH guide lines comparing the linearity, intraday precision, inter-day precision. The optimal conditions were as follows; wavelength: 288 nm, column: Zorbax Eclipse XDBC18 $(2.1\times150\;mm,\;5{\mu}m)$, mobile phase: Acetonitril - 0.1 % $H_3PO_4$ (45:55, v/v) buffer, flow rate: 0.5 ml/min. The contents of magnolol, honokiol and obovatol were 1.08%, 2.37%, and 0.57% in the leaves of M. officinalis, 0.50%, 0.03%, and 1.54% in the leaves of M. biloba, 0.37%, 0.03%, and 0.76% in the leaves of M. obovata. The contents of effective ingredients in the leaves of M. officinalis and M biloba were higher than that of M. obovata. Especially, the content of obovatol in the leaves of M. biloba was remarkably high. This result showed that the shapes of leaves have a certain relationship with the content of obovatol.

• Environmental Analysis Health and Toxicology
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• v.13 no.3_4
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• pp.105-115
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• 1998

Reactive oxygen species (ROS) are highly reactive molecules due to their unpaired electron. They have been suspected as one of the major tissue damage inducers in biological metabolic systems. Antioxidant enzymes, such as catalase and superoxide dismutase, could not repair all the oxidative damages resulting from those excessive toxic ROS. It is, therefore, urgent to develop effective antioxidants to relieve from the oxidatire damages. In this study antioxidative effects were investigated by using two flavonoids such as quercetin and naringenin and a flavonoid-rich extract, Ginkgo biloba extract in combination with paraquat that is known as a strong generator of oxygen radicals. The results are summeringed as follows: 1. To assess radical scavenging ability reduction concentrations (IC$_{50}$) of 1,1-diphenyl-2-picrylhydrazine (DPPH) within 15 minutes were measured. The values of the IC$_{50}$ of quercetin and Ginkgo biloba extract were 15.4 $\mu$M and 13.2$\mu$g/ml, respectively. Their radical removing activities showed concentration-dependent manners. 2. In the hydrogen peroxide assay by using PMS-NADH system, quercetin, naringenin and Ginkgo biloba extract led to removing hydrogen peroxide in concentrationdependent manner whose removing abilities at 100$\mu$M or 100 $\mu$g/ml were 75.6, 25.8 and 26.0%, respectively. 3. In the hydrogen peroxide-induced rat blood hemolysis assay all three compounds led to similar effects whose hemolysis inhibition ratios at 100$\mu$M or 100$\mu$g/ml were 68.0, 5.14 and 55.8%, respectively. 4. In the xanthinee oxidase assay by measuring degree of NADH oxidation in the presence of hypoxanthine and xanthinee oxidase, both quercetin and Ginkgo biloba extract showed excellent activities showing 42.8 and 24.2% inhibiting xanthine oxidase activity at 100$\mu$M or 100$\mu$g/ml concentrations, respectively.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.41 no.4
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• pp.383-389
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• 2015

In this study, we investigated the inhibitory effect on melanin synthesis of Ginkgo biloba seed oil. The results showed 9.96% inhibitory effect scavenging activity on DPPH and showed a value of 1.33 mM of $FeSO_4$ at a concentration of 0.06% in DMSO by using FRAP assay. G. biloba seed oil inhibited tyrosinase activity up tp 37.72% and suppressed the biosynthesis melanin up to 48.02% at 0.06% in B16/F10 mouse melanoma cell. In G. biloba seed oil treated group tyrosinase, TRP-1, TRP-2 and MITF gen expression levels significantly decreased compared to the contral group at a concentration of 0.04% and 0.06%. In conclusion, these results indicated that G. biloba seed oil extract have a good antimelanogenetic effects.

• Bulletin of the Korean Chemical Society
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• v.34 no.12
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• pp.3629-3634
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• 2013

In this study, an improved method for the quantitative analysis of ginkgolic acids (GAs) in Ginkgo biloba leaf extract was developed. The samples were extracted with a mixture of chloroform and 50 % ethanol, after which the chloroform extract was dried and reconstituted in methanol. GAs with 13:0, 15:1, and 17:1 in the extract were successfully separated within 40 min and determined with high throughput performance using an online column-switching HPLC method using an SP column C8 SG80 ($4.6{\times}150mm$, $5{\mu}m$) and a Cadenza 5CD C18 column ($4.6{\times}150mm$, $3{\mu}m$). The developed HPLC method was validated for Ginkgo biloba leaf extract. The validation parameters were specificity, linearity, precision, accuracy, and limits of detection and quantitation (LODs and LOQs, respectively). It was found that all of the calibration curves showed good linearity ($r^2$ > 0.9993) within the tested ranges. The LODs and LOQs were all lower than $0.04{\mu}g/mL$. The established method was found to be simple, rapid, and high throughput for the quantitative analysis of GAs in ten commercial Ginkgo biloba leaf extract and dietary supplements. The samples were also analyzed in LC-electrospray ionization (ESI) tandem mass spectrometry (MS/MS) - multiple-ion reaction monitoring (MRM) mode to confirm the identification results that were obtained by the column switching HPLC-DAD method. The developed method is considered to be suitable for the routine quality control and safety assurance of Ginkgo biloba leaf extract.

• Korean Journal of Organic Agriculture
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• v.29 no.1
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• pp.97-109
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• 2021

Plant parasitic nematodes are causing significant damage in crop production. There is a need to develop eco-friendly nematicide that reduces the damage of nematode and has little effect on the environment and human. In this study, we have isolated a substance having nematicidal activity from Ginkgo biloba L. outer seedcoat. Studies of G. biloba L. outer seedcoat are insufficient compared to the seed and leaves due to their odor and toxicity. The dried G. biloba L. outer seedcoat was extracted with dichloromethane:methanol (1:1) and fractionated into hexane, ethyl acetate and H₂O. Four steps TLC were performed from EtOAc fraction to purely isolate GB4-3 with nematicidal activity. To compare nematicidal activity, G. biloba L. seedcoat methanol extract and purified GB4-3 were investigated in terms of treatment concentration and time. As a result, the nematicidal activity increased with concentration and time. In the place treated with 20 ㎍/mL of crude G. biloba L. seedcoat MeOH extract, strong activity appeared after 12 hours, and 46% nematicidal activity shown after 18 hours. About 69% of nematicidal activity was confirmed in the place where GB4-3 purified from outer seedcoat was treated with 20 ㎍/mL, and the possibility of development as nematicide was very high. This study could be used as a basic data for the development of a nematode preparation from G. biloba L. outer seedcoat.

• Korean Journal of Pharmacognosy
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• v.30 no.3
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• pp.306-313
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• 1999

Polyphenol Oxidase(PPO) was purified from an extract of Ginkgo biloba leaves by ammonium sulfate fractionation followed by sephadex G-150 column chromatography, which resulted in a 18-fold increase in specific activity. The enzyme was most active at pH 8.5 and the temperature optimum for the PPO catechol oxidation reaction was $45^{\circ}C$. Heat inactivation studies showed that heating for 7, 9 and 48 min, at 80, 70 and $60^{\circ}C$ respectively caused a 50% loss in enzymatic activity and that the enzyme was completely inactivated after heat treatment at $90^{\circ}C$ for 60 min. Km values of the PPO for catechol, hydroquinone and 4-methylcatechol derived from Lineweaver-Burk plots were $6.06\;{\times}\;10^{-4}M,\;1.02\;{\times}\;10^{-3}M,\;1.41\;{\times}\;10^{-3}M$ respectively. Of the substrates tested, 4-methylcatechol was oxidized most readily and the enzyme did not oxidize monophenols. The enzyme datalyzed browning reaction was completely inhibited in the presence of reducing reagents, namely ascorbic acid, cysteine, glutathione, 2-mercaptoethanol, potassium metabisulfite at 0.5 mM level. Sodium chloride showed very little inhibition effect on Ginkgo biloba leaves PPO. Lineweaver-Burk analysis of inhibition data revealed that the inhibition by cysteine, 2-mercaptoethanol, potassium cyanide was competitive with ki values of $1.1\;{\times}\;10^{-5}M,\;2.4\;{\times}\;10^{-5}M,\;8\;{\times}\;10^{-5}M$, respectively. Among the divalent cations, $Cu^{2+}ion$ was a strong activator on PPO and $Mn^{2+}ion$ was little or no effect on PPO activity $Ni^{2+}ion$ was an inhibitor on PPO.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.1
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• pp.7-13
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• 2011

This study was conducted to investigate the antimicrobial activity of Ginkgo biloba L. leaves, seed and outer seedcoat against bacteria. Antimicrobial effects of Ginkgo biloba L. leaves (GBL), seed (GBS) and outer seedcoat (GBO) were examined by paper disc method and optical density method to determine minimum inhibition concentration (MIC), and observed by scanning electron microscope (SEM) to figure out the morphological change on the surface when Ginkgo biloba leaves extract was treated. The extracts of GBL, GBS and GBO were extracted by solvents such as methanol, ethanol, water. The methanol extract of GBL and GBO showed the highest antimicrobial activity against Bacillus cereus, Bacillus subtilis, Klebsiella pneumoniae, Listeria monocytogenes, Salmonella Typhimurium, Staphylococcus aureus, Yersinia enterocolitica except Escherichia coli and thus was further fractionated. The MICs of the chloroform fraction of GBL methanol extract were $125{\mu}g$/mL against B. subtilis, and L. monocytogenes; GBO methanol extract were $62.5{\mu}g$/mL against B. cereus and $125{\mu}g$/mL against B. subtilis, and L. monocytogenes. The microorganisms were treated with chloroform extracts ($2000{\mu}g$/mL) of GBL and GBO methanol extracts. It was observed by scanning electron microscope (SEM). The cells were expanded and a part of cell wall was completely destructed by GBL and GBO. Thus Ginkgo biloba L. leaves and outer seedcoat could be further developed into a natural antimicrobial agent.

• Korean Journal of Pharmacognosy
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• v.31 no.2
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• pp.240-244
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• 2000

Antimutagenicity profiles of the enzymatic browning reaction products(EBRP) were investigated. The rec-assay with Bacillus subtilis strains $H17(rec^+)$ and $M45(rec^-)$ was carried out using their spores. The biological activities were evaluated for seven different enzymatic browning reaction products, which resulted from the reactions of seven polyphenols with polyphenol oxidase isolated from Ginkgo biloba leaves. In the spore $rec^-$ assay, most of the polyphenolic compounds tested were positive, whereas their enzymatic browning reaction products were tested negative. The mutagenicity of enzymic browning mixtures of the polyphenols and the enzymes obtained from Ginkgo biloba leaves showed negative results in the mutagenicity test using Bacillus subtilis strains $H17(rec^+)$ and $M45(rec^-)$. In the case where polyphenol oxidase inhibitors were added in the enzymatic reaction mixtures with polyphenols, the polyphenols showed mutagenic effect in the spore $rec^-$ assay. This suggests that the activity of polyphenol oxidase is decreased.

• Journal of Korean Neurosurgical Society
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• v.49 no.1
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• pp.13-19
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• 2011

Objective: The purpose of this study is to investigate the combined effects of ginkgo biloba extract, ginkgolide A and B and aspirin on SK-N-MC, human neuroblastoma cell viability and mRNA expression of growth associated protein43 (GAP43), Microtubule-associated protein 2 (MAP2), B-cell lymphoma2 (Bcl2) and protein53 (p53) gene in hypoxia and reperfusion condition. Methods: SK-N-MC cells were cultured with Dulbecco's Modified Eagle's Medium (DMEM) media in $37^{\circ}C$, 5% $CO_2$ incubator. The cells were cultured for 8 hours in non-glucose media and hypoxic condition and for 12 hours in normal media and $O_2$ concentration. Cell survival rate was measured with Cell Counting Kit-8 (CCK-8) reagent assay. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to estimate mRNA levels of GAP43, MAP2, Bcl2, and p53 genes. Results: The ginkgolide A and B increased viable cell number decreased in hypoxic and reperfused condition. The co-treatment of ginkgolide B with aspirin also increased the number of viable cells, however, there was no additive effect. Although there was no increase of mRNA expression of GAP43, MAP2, and Bcl2 in SK-N-MC cells with individual treatment of ginkgolide A, B or aspirin in hypoxic and reperfused condition, the co-treatment of ginkgolide A or B with aspirin significantly increased GAP43 and Bcl2 mRNA levels. In MAP2, only the co-treatment of ginkgolide A and aspirin showed increasing effect. The mRNA expression of p53 had no change in all treating conditions. Conclusion: This study suggests that the combined treatments of Ginkgo biloba extracts and aspirin increase the regeneration of neuroblastoma cells injured by hypoxia and reperfusion.