Sarcotesta Of Ginkgo Biloba Research Articles

Extraction, Purification, and Elucidation of Six Ginkgol Homologs from Ginkgo biloba Sarcotesta and Evaluation of Their Anticancer Activities.

Ginkgols are active constituents from Ginkgo biloba L. (GB) and have pharmacological activities, such as antibacterial and antioxidant activities. In our previous report, only five ginkgols were separated. However, ginkgol C17:1 had two isomers, for which their separation, identification, and bioactivities have not yet been investigated. Hence, this research reports the successful isolation of six ginkgol homologs with alkyl substituents-C17:1-Δ12, C15:1-Δ8, C13:0, C17:2, C17:1-Δ10, and C15:0-for the first time using HPLC. This was followed by the identification of their chemical structures using Fourier transform infrared (FTIR), ultraviolet (UV), gas chromatography and mass spectrometry (GC-MS), carbon-13 nuclear magnetic resonance (13C-NMR), and proton nuclear magnetic resonance (1H-NMR) analysis. The results showed that two ginkgol isomers, C17:1-Δ12 and C17:1-Δ10, were obtained simultaneously from the ginkgol C17:1 mixture and identified entirely for the first time. That aside, the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay showed that the six ginkgol homologs possessed significant antiproliferation effects against HGC and HepG2 cells. Furthermore, the ginkgols with unsaturated side chains (C17:2, C15:1-Δ8, C17:1-Δ12, and C17:1-Δ10) exhibited more potent inhibitory effects than ginkgols with saturated side chains (C13:0, C15:0). In addition, unsaturated ginkgol C15:1-Δ8 showed the most potent cytotoxicity on HepG2 and HGC cells, of which the half-maximal inhibition concentrations (IC50) were 18.84 ± 2.58 and 13.15 ± 2.91 μM, respectively. The IC50 for HepG2 and HGC cells for the three unsaturated ginkgols (C17:1-Δ10, C17:2 and C17:1-Δ12) were ~59.97, ~60.82, and ~68.97 μM for HepG2 and ~30.97, ~33.81, and ~34.55 μM for HGC cells, respectively. Comparing the ginkgols' structure-activity relations, the findings revealed that the position and number of the double bonds of the ginkgols with 17 side chain carbons in length had no significant difference in anticancer activity.

Antibacterial, Antibiofilm, and Antioxidant Activity of Polysaccharides Obtained from Fresh Sarcotesta of Ginkgo biloba: Bioactive Polysaccharide that Can Be Exploited as a Novel Biocontrol Agent.

Staphylococcus aureus (S. aureus) biofilm plays an important role in the persistence of chronic infection due to its resistance to antibiotics. Because of their functional diversity, active polysaccharide is increasingly being applied as a biocontrol agent to inhibit the formation of biofilm by pathogens. In this study, a new polysaccharide, GBSPII-1, isolated from the fresh sarcotesta of Ginkgo biloba L. (G. biloba) was characterized and its effect on antibiofilm formation of S. aureus was examined in vitro. High-Performance Liquid Chromatography (HPLC) analysis showed that GBSPII-1 is an acidic heteropolysaccharide composed of mannose, rhamnose, glucose, glucuronic acid, and galacturonic acid. GBSPII-1 demonstrated a molecular weight of 34 kDa and may affect the accumulation of polysaccharide intercellular adhesion (PIA) by inhibiting icaA, icaB, icaC, and icaD gene expression at subinhibitory concentrations. Under 10 g/L, GBSPII-1 showed an antioxidant effect on the inhibition rate of H2O2-induced erythrocyte hemolysis and the scavenging rate of DPPH radicals was 76.5 ± 0.5% and 89.2 ± 0.26%, respectively. The findings obtained in this study indicate that GBSPII-1 has antibacterial effect, is a possible source of natural antioxidants, and may be a potential biocontrol agent for the design of new therapeutic strategies for biofilm-related S. aureus infections.

Application of coordination agent in high-speed counter-current chromatography for the preparative separation and isolation ginkgolic acids from the sarcotesta of Ginkgo biloba L.

An efficient coordination high-speed counter-current chromatography method for the preparative separation of ginkgolic acids from the sarcotesta of Ginkgo biloba L was developed. The type, concentration, and mechanism of the coordination agent were investigated. Following the use of four types of metal salts including silver nitrate, copper chloride, ferric chloride, and aluminium nitrate, n-heptane/ethyl acetate/methanol/acetic acid 5:4:1:1, v/v with 0.20mol/L silver nitrate as the coordination agent was chosen as the optimum two-phase solvent system. Five main ginkgolic acids including C13:0, C15:0, C15:1, C17:1, and C17:2 were successfully separated with purities greater than 98%. The sample loading was 500mg, the flow-rate was 2.0mL/min, rotation speed was 800 rpm and temperature was 20°C. The structures of the separated ginkgolic acids were identified by comparison with standard samples and electrospray ionization mass spectrometry. The introduction of coordination chemistry in high-speed counter-current chromatography is novel and effective for the preparative separation and isolation of ginkgolic acids from the sarcotesta of Ginkgo biloba L and could also be applied to separate compounds which form coordination bonds in other complex natural products.

An efficient method for the preparative separation and isolation of ginkgolic acids from the sarcotesta of Ginkgo biloba L by pH-zone-refining counter-current chromatography coupled with inner-recycling mode

An efficient method for the preparative separation and isolation of ginkgolic acids (GAs) from the sarcotesta of Ginkgo biloba L was developed by combining pH-zone-refining counter-current chromatography (PZRCCC) with inner-recycling mode. Two grams of GAs extract were separated by PZRCCC using a two-phase solvent system of n-heptane–ethyl acetate–methanol–water (2:1:3:1, v/v) with 10 mM HCl added to the upper phase as the retention acid and 10 mM triethylamine (TEA) in the lower phase as the elution base. In a one-step PZRCCC separation, two GA compounds, C15:1 and C17:2, were obtained with purities over 98% along with two mixtures. The mixtures were further separated by inner-recycling CCC mode with n-heptane–ethyl acetate–methanol–acetic acid (5:4:1:1, v/v) as the solvent system. Five main GAs were successfully separated with purities over 98%. The structures of the separated GAs were determined based on standard samples coupled with Electrospray Ionization Mass Spectrometry (ESI-MS) results. This separation method of coupling PZRCCC with inner-recycling CCC is effective for the preparation of GAs from the sarcotesta of G. biloba and could be further applied to separate organic acids of low polarity in other natural products.

Airborne contact dermatitis caused by the sarcotesta of Ginkgo biloba.

Airborne contact dermatitis caused by the sarcotesta of Ginkgo biloba.

Physiological Activities of Ginkgo biloba Sarcotesta Extract with Heat Treatment

Abstract This study was performed to investigate the physiological activities of Ginkgo biloba sarcotesta extracts before and after heat treatment. G. biloba sarcotesta was heated at 130 for 2 h and extracted with water, 70% ethanol and 80% methanol. ABTS and DPPH radical scavenging activities increased after heating in the water (14.95 mg AAE/g and 7.36 mg TE/g) and ethanol extracts (12.20 mg AAE/g and 6.23 mg TE/g). α-Glucosidase inhibitory activity decreased after heating in all but the water extract. Angiotensin converting enzyme I inhibitory activities decreased after heating in all extracts. Nitric oxide production inhibitory activity increased from 12.40~44.55% of the raw sample to 40.76~72.39% of the heated sample at a concentration of 200 μg/mL. Lipid accumulation inhibitory activities were similar before and after heat treatment. The highest antiproliferative effects on MCF-7 human breast cancer cell lines were observed in 80% methanol extract in the heated sample. Cell viability at concentrations of 25, 50, 100, and 200 μg/mL measured 34.88, 17.58, 8.44 and 10.48%, respectively. From the results, the antioxidant and antiproliferative activities of G. biloba sarcotesta extracts increased with heat treatment, and research on the identification of the structure for the active compounds are needed in further studies. Key words: Ginkgo biloba sarcotesta, heat treatment, physiological activity

Two new nonacosanetriols from Ginkgo biloba sarcotesta

Two new fatty alcohols named as (7S,8R,11S)-nonacosanetriol (1) and (10R,12R,15S)-nonacosanetriol (2), along with eight known compounds including ginkgolic acid (3), hydroginkgolic acid (4), sciadopitysin (5), ginkgetin (6), isoginkgetin (7), ginkgolide A (8), ginkgolide B (9) and ginkgolide C (10) have been isolated from the petroleum ether extract of Ginkgo biloba sarcotesta. Their structures were elucidated by means of chemical and extensive spectroscopic analysis. The absolute stereochemistry of compounds 1 and 2 was elucidated on the spectroscopic analysis of the R- and S-MTPA esters. Compounds 1 and 2 exhibited slight activity of antithrombin and moderate activity of antiplatelet aggregation in vitro. This was the first report regarding the anticoagulative activities of biflavonoids in G. biloba, and isoginkgetin (7) showed significant antithrombin and antiplatelet aggregation activity.

Effects of the molluscicidal agent GA-C13:0, a natural occurring ginkgolic acid, on snail mitochondria

Ginkgolic acids (GAs) from the leaves and sarcotesta of Ginkgo biloba L. represent a new kind of molluscicide agent. To date, the mechanism(s) for the observed molluscicidal activity remains largely unknown. Since GA-C13:0 has effectively inhibited snail mobility, we examined the effects of the compound on mitochondrial function and gene expression as compared to niclosamide. Snail mitochondrial damage induced by GAs was tested using transmission electron microscopy (TEM) and gene-expression profiling of five mitochondrial enzymes using real-time PCR. GA-C13:0 was found to have a pronounced effect on snail mitochondria with gross ultrastructural changes. In addition, GA-C13:0 was also found to inhibit the gene expression of four mitochondrial enzymes including cytochrome c oxidase, adenosine triphosphate (ATP) synthase, cytochrome b and dihydronicotinamide adenine dinucleotide (NADH) dehydrogenase. In contrast, niclosamide did not show such effects on mitochondrial function and gene expression, suggesting that the molluscicidal activity of GA-C13:0 and niclosamide differed. Our results imply that snail mitochondria are a potential target for the molluscicidal activity of ginkgolic acids.

Characterization of Alkylphenol Components in Ginkgo biloba Sarcotesta by Thermochemolysis–Gas Chromatography/Mass Spectrometry in the Presence of Trimethylsulfonium Hydroxide

A method to determine alkylphenols in Ginkgo biloba sarcotesta, especially focusing on polyunsaturated ginkgolic acid components, was developed on the basis of thermochemolysis–gas chromatography/mass spectrometry with a DB-23 column in the presence of trimethylsulfonium hydroxide [(CH3)3SOH]. By using this technique, powder sample of Ginkgo biloba sarcotesta can be directly analyzed without any tedious and time-consuming pretreatment. On the resulting chromatograms, seven kinds of ginkgolic acids including polyunsaturated ginkgolic acid GA17:2 and two kinds of ginkgols were clearly observed as their methyl derivative. Based on the peak areas, the ginkgolic acid compositions are in good agreement with these obtained by a high-performance liquid chromatography method dedicated to analysis of ginkgolic acid.

Isolation, purification and in vitro anti-tumor activity of polysaccharide from Ginkgo biloba sarcotesta

Isolation and purification of polysaccharide from Ginkgo biloba sarcotesta by cellulose DEAE-52 and Sephadex G-100 column chromatography were performed in this study. Four water-soluble polysaccharides named GBEPP11, GBEPP22, GBEPP33 and GBEPP44 were obtained and their anti-tumor activities in vitro were tested by MTT assay. GBEPP44 with molecular weight (MW) 9.5×105Da, GBEPP33 with MW 3.1×105Da and GBEPP22 with MW 4.8×104Da were composed of rhamnose, glucose, galactose with a molar ratio of 16:10:0.96, 2.64:1:1.43 and 37.01:1:8.46, respectively. GBEPP11 with MW 3.4×103Da was composed of rhamnose, glucose with a molar ratio of 1.9:1. MTT assay revealed that the polysaccharides had anti-tumor activity in a dose-dependent manner. The inhibitory rate of the polysaccharides on U937 was the highest among the three cancer cells, which increased as the MW decreased. Furthermore, the activity of acetylated GBEPP11 was higher than that of GBEPP11 and the inhibitory rate reached 78.97% at the concentration of 640μg/mL.

Antibacterial activity of hydroxyalkenyl salicylic acids from sarcotesta of Ginkgo biloba against vancomycin-resistant Enterococcus

A chloroform fraction prepared from the sarcotesta of Ginkgo biloba showed potent inhibitory activity against vancomycin-resistant Enterococcus (VRE). The active compounds were elucidated to be 2-hydroxy-6-(8-pentadecenyl) salicylic acid (1) and 2-hydroxy-6-(10-heptadecenyl) salicylic acid (2) based on their spectral analysis. Compounds 1 and 2 showed significant antibacterial activities against VRE.

Anti- Toxoplasma gondii activity of GAS in vitro

Ethnopharmacological relevance The sarcotesta of Ginkgo biloba is a Chinese herbal medicine used for treating toxoplasmosis, a serious disease requiring treatment with antibiotics that can have serious side effects. Aim of the study To investigate the anti- Toxoplasma gondii activity of ginkgolic acids (GAs) isolated from the Ginkgo biloba sarcotesta in Toxoplasma gondii-infected human foreskin fibroblast (HFF) cells in vitro. Materials and methods The safe concentration of GAs for HFF cells was determined by methyl thiazolyl tetrazolium (MTT) cell proliferation assay. The presence of Toxoplasma gondii was measured by [ 3H]-thymine deoxyriboside ([ 3H]-TdR) and [ 3H]-leucine ([ 3H]-Leu) incorporation, as well as Giemsa staining. The positive control was the commonly used and highly effective antibiotic azithromycin. Results Light microscopy revealed that most HFF cells were infected after 4 h of exposure to Toxoplasma gondii. After 48 h of exposure to either GAs or azithromycin, Toxoplasma gondii DNA and protein synthesis were minimal, there were no visible parasites in HFF cells, and the HFF cells had no significant morphological changes. Conclusions These results demonstrate that GAs have significant anti- Toxoplasma activity with low toxicity to HFF cells, suggesting that GAs could be an alternative treatment for toxoplasmosis.

Comparative molluscicidal action of extract of Ginko biloba sarcotesta, arecoline and niclosamide on snail hosts of Schistosoma japonicum

In search for new local plant molluscicides for the control of the vectors of schistosomiasis, we compared the molluscicidal action of the extract of Ginkgo biloba sarcotesta by benzinum (EGSB) to that of arecoline (ARE) and niclosamide (NIC) against Oncomelania hupensis snails. NIC showed the highest toxicity on snails with 24 h LC 50 vales of 0.12 mg/L and LC 90 of 0.98 mg/L, while the LC 50 and LC 90 of EGSB were much lower than that of ARE. Sublethal in vivo 24 h exposure to 40% and 80% LC 50 of NIC, EGSB and ARE altered the activities of different enzymes in different body tissues of snails. EGSB could significantly inhibit Choline esterase (ChE), Alanine aminotransferase (ALT), Alkaline phosphatase (ALP) and Malic dehydrogenase (MDH) activities both in the cephalopodium and liver. ARE could significantly cause a reduction in ChE, ALP activities in the cephalopodium and ChE, ALT, ALP, Succinodehydrogenase (SDH), MDH activities in the liver. NIC significantly altered activities of ChE, ALT, ALP, SDH, and MDH in the cephalopodium and ChE, ALT, ALP, SDH activities in the liver. All molluscicides could not affect Lactate dehydrogenase (LDH) activity in the cephalopodium and the liver. Maximum inhibition of ALT and MDH activities was found in the cephalopodium and liver of snails treated with 80% of 24 h LC 50 of EGSB. However, NIC and ARE caused maximum reduction in ALP and SDH activities, respectively. The results indicated that molluscicidal action of EGSB was different to that of ARE and NIC in some extent.