Ginkgo Seeds Research Articles

Artificial Neural Network Modeling of Drying Kinetics and Color Changes of Ginkgo Biloba Seeds during Microwave Drying Process

Ginkgo biloba seeds were dried in microwave drier under different microwave powers (200, 280, 460, and 640 W) to determinate the drying kinetics and color changes during drying process. Drying curves of all samples showed a long constant rate period and falling rate period along with a short heating period. The effective moisture diffusivities were found to be 3.318 × 10−9 to 1.073 × 10−8 m2/s within the range of microwave output levels and activation energy was 4.111 W/g. The L⁎ and b⁎ values of seeds decreased with drying time. However, a⁎ value decreased firstly and then increased with the increase of drying time. Artificial neural network (ANN) modeling was employed to predict the moisture ratio and color parameters (L⁎, a⁎, and b⁎). The ANN model was trained for finite iteration calculation with Levenberg-Marquardt algorithm as the training function and tansig-purelin as the network transfer function. Results showed that the ANN methodology could precisely predict experimental data with high correlation coefficient (0.9056–0.9834) and low mean square error (0.0014–2.2044). In addition, the established ANN models can be used for online prediction of moisture content and color changes of ginkgo biloba seeds during microwave drying process.

Limited proteolysis initiates massive degradation of ginnacin, storage 11S globulin from Ginkgo seeds

Limited proteolysis initiates massive degradation of ginnacin, storage 11S globulin from Ginkgo seeds

Elucidation of micromolecular phenylpropanoid and lignan glycosides as the main antioxidants of Ginkgo seeds

Ginkgo biloba L. is a widely used medicinal plant that has a high concentration of its bioactive components, flavonoids and ginkgolides, in its leaves. Gingko seeds are also consumed as a functional food and medicine throughout the world to slow the ageing process. However, there have been few phytochemical studies of the phenylpropanoids from Ginkgo seeds. To elucidate the active components, we studied the antioxidant activities and related main compounds in Ginkgo seeds. Four main phenylpropanoids and lignans were isolated and identified from the antioxidative fraction and their antioxidant activities were measured. An HPLC/DAD-Q-TOF-MS/MS method was developed to identify the other minor phenylpropanoid and lignan glycosides to elucidate the antioxidant profile of Gingko seeds. Four compounds were identified by NMR and mass data, three of which (8, 17, and 28) exhibited substantial antioxidant activities (IC50: 67–100 μM). Sixteen minor compounds were tentatively identified based on their tandem mass spectrometry data and the fragmentation of four isolated standards from antioxidative n-butanol fraction. Fifteen compounds were reported in Ginkgo seeds for the first time, and compound 16 was tentatively identified as a new chemical structure. Thus, understanding their antioxidant profiles will lay the foundation for research and development of the health benefits of Ginkgo seeds.

Streptomyces ginkgonis sp. nov., an endophyte from Ginkgo biloba.

A novel endophytic actinomycete strain, designated KM-1-2T, was isolated from seeds of Ginkgo biloba at Yangling, China. A polyphasic approach was used to study the taxonomy of strain KM-1-2T and it was found to show a range of phylogenetic and chemotaxonomic properties consistent with those of members of the genus Streptomyces. The diamino acid of the cell wall peptidoglycan was identified as LL-diaminopimelic acid. No diagnostic sugars were detected in whole cell hydrolysates. The predominant menaquinones were identified as MK-9(H6) and MK-9(H8). The diagnostic phospholipids were found to be phosphatidylethanolamine and phosphatidylcholine. The DNA G+C content of the novel strain was determined to be 72.9mol%. The predominant cellular fatty acids (>10.0 %) were identified as iso-C14 : 0, iso-C16 : 0, C16 : 0 and C17 : 0 cyclo. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that the strain is closely related to Streptomyces carpaticus JCM 6915T (99.3%), Streptomyces harbinensis DSM 42076T (98.9%) and Streptomyces cheonanensis JCM 14549T (98.5%). DNA-DNA hybridizations with these three close relatives gave similarity values of 39.1±1.9, 35.8±2.3, and 47.4±2.7%, respectively, which indicated that strain KM-1-2T represents a novel species of the genus Streptomyces. This is consistent with the morphological, physiological and chemotaxonomic data. Cumulatively, these data suggest that strain KM-1-2T represents a novel Streptomyces species, for which the name Streptomyces ginkgonis sp. nov. is proposed, with the type strain KM-1-2T (=CCTCC AA2016004T=KCTC 39801T).

Changes in 4'-O-methylpyridoxine (ginkgotoxin) and antioxidant activity in ginkgo biloba seeds in different cooking conditions

Changes in 4'-O-methylpyridoxine (ginkgotoxin) and antioxidant activity in ginkgo biloba seeds in different cooking conditions

Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba.

Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. Previous research pointed out that the protein allergen in Ginkgo biloba seeds is glycosylated, and the oligosaccharides conjugated to these proteins might also contribute to the allergy. The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. Results of monosaccharide composition and immunoblotting assays indicated the existence of N-glycans. Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In total, 14 out of 16 structures detected by UPLC were confirmed by MALDI-TOF-MS and tandem mass spectrometry, among which complex-type N-glycans bearing Lewis A determinants and high-mannose-type N-glycans were identified from Ginkgo seeds for the first time. Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation.

Changes in Seed Germination Ability, Lipid Peroxidation and Antioxidant Enzyme Activities of Ginkgo biloba Seed during Desiccation

With the aim of investigating the antioxidant system and germinability in response to the desiccation of Ginkgo biloba seeds, they were put in a drying room (25 ± 2% relative humidity, 25 °C) for 67 days. Results showed that the germination rate remained constant when seed moisture content (MC) decreased from 48% (fresh seeds) to 45.1%. However, when MC reached 40.1%, the germination percentage decreased from 92% to 50%. A significant positive correlation was observed between the MC and seed germination percentage (r = 0.910). The electrical conductivity was significantly increased during the initial desiccation (48–45.1%). Furthermore, both the superoxide dismutase (SOD) and peroxidase (POD) activity first reduced, then elevated to peak values before they declined again. POD activity rose earlier than SOD activity, indicating that the POD reaction was more desiccation-sensitive than the SOD. Significant negative correlations were observed between the MC and malondialdehyde (MDA) content (r = −0.619) and electrical conductivity (r = −0.745). Our collective results suggest that G. biloba seeds are highly sensitive to desiccation. Excessive desiccation could reduce the antioxidant enzyme activity of G. biloba seeds and intensify membrane lipid peroxidation, which causes the consequent reduction—or even the complete loss—of seed germinability.

Review of Ginkgo biloba-induced toxicity, from experimental studies to human case reports

ABSTRACTGinkgo biloba seeds and leaves have been used as a traditional herbal remedy for thousands of years, and its leaf extract has been consumed as a botanical dietary supplement for decades. Ginkgo biloba extract is a complex mixture with numerous components, including flavonol glycosides and terpene lactones, and is one of the most widely sold botanical dietary supplements worldwide. Concerns about potential health risks for the general population have been raised because of the widespread human exposure to Ginkgo biloba and its potential toxic and carcinogenic activities in rodents. The National Toxicology Program conducted 2-year gavage studies on one Ginkgo biloba leaf extract and concluded that there was clear evidence of carcinogenic activity of this extract in mice based on an increased incidence of hepatocellular carcinoma and hepatoblastoma. Recently, Ginkgo biloba leaf extract has been classified as a possible human carcinogen (Group 2B) by the International Agency for Research on Cancer. This review presents updated information on the toxicological effects from experimental studies both in vitro and in vivo to human case reports (caused by ginkgo seeds or leaves), and also summarizes the negative results from relatively large clinical trials.

Effect of heating on the content and composition of ginkgolic acids in ginkgo seeds

Ginkgolic acids (GAs) are generally identified as the allergen in ginkgo seeds. The effect of steaming and microwave treatment on GAs in ginkgo seeds was investigated in this paper. GAs were extracted with ethanol/water (85:15, v/v) and detected by high performance liquid chromatography (HPLC). The results indicated that GAs contents decreased significantly after heating treatments compared to the control. After steaming (115 °C, 75 min) and microwave treatment (1000 W, 6 min), GAs contents reduced with 54.5 and 27.0%, respectively. Three main GAs were detected by HPLC, and the alkyl side chains of which were C13:0, C15:1 and C17:1, respectively. Steaming and microwave treatment could effectively decrease GAs contents in ginkgo seeds, which could be used in the industrial production of ginkgo foods.

Effects of high hydrostatic pressure treatment on structural, allergenicity, and functional properties of proteins from ginkgo seeds

Abstract The effects of high hydrostatic pressure (HHP) treatment on the structural, allergenicity and functional properties of ginkgo seed protein (GSP) were investigated. GSP was treated under pressures of 100, 200, 300, 400, 500, 600 and 700 MPa. Western blotting and ELISA assay revealed that HHP treatment at pressure ranges from 300 up to 700 MPa can significantly reduce the allergenicity of GSP. Meanwhile, SDS-PAGE, MALDI-TOF-MS, CD spectra, fluorescence spectra and UV absorption spectra analysis revealed that, after HHP treatment, the GSP disintegrated into proteins with small molecular weights ranging from 4 to 30 kDa; the secondary structures of α-helices and β-sheet were largely destroyed and turned into random coil; and the UV absorption intensity, surface hydrophobicity and sulfhydryl group (SH) content of GSP were significantly increased. Moreover, the HHP-treated GSP showed markedly improved heat stability and emulsifying properties compared to the untreated GSP. Industrial relevance As a traditional food and medicine source, the seeds of Ginkgo biloba were used in china for several thousand years. Ginkgo seeds could be added to desserts, glazed fruit, beverages and tipple, also ginkgo seeds, as a traditional Chinese medicinal material, have been recorded in the Compendium of Materia Medical. Research shows that ginkgo seeds have a relatively high content (10%–15%) of proteins, which possess rich and reasonable composition of amino acids, and much higher content of essential amino acids than the recommended FAO/WHO standard, belonging to the high-quality protein. Ginkgo seeds proteins (GSP) have many biological activities, including anti-oxidation, anti-aging, anti-tumor, and anti-bacterial, so it has high value of nutrition and medicine. However, eating GSP could result in allergic reaction to humans, infant, or children are more sensitive to GSP, the clinical symptoms such as nausea, vomiting, bellyache, diarrhea, dysphoria, exanimation, convulsions, dyspnea, and mydriasis, and sometimes even death can occur. In the study, we used the method of high hydrostatic pressure (HHP) treatments, which can significantly improve the functional properties and reduce the antigenicity of GSP. After the HHP treatment, GSP can be transformed into proteins with small molecular weights ranging from 4 to 30 kDa, the antigenicity of GSP can be reduced by more than 95%, and the heat stability and emulsifying properties of GSP can be markedly improved. Thus, this study is very helpful for designing the hypoallergenic and improved functional GSP, which could have a relevant practical application for their use as base ingredients of food.

Effects of Enzymatic Hydrolysis Assisted by High Hydrostatic Pressure Processing on the Hydrolysis and Allergenicity of Proteins from Ginkgo Seeds

The purpose of this work was to study the combined effect of high hydrostatic pressure (HHP) and enzymatic hydrolysis treatment on the hydrolysis and allergenicity of ginkgo seed proteins (GSPs). Four food-grade proteases (papain, alcalase, pepsin, and neutrase) were used, and HHP (200, 300, and 400 MPa separately) was applied prior to hydrolysis. The extent of hydrolysis was measured with the o-phthaldialdehyde method, SDS-PAGE, and MALDI-TOF-MS, and the allergenicity was assessed with a Western blot and enzyme-linked immunosorbent assay (ELISA). The results showed that HHP could significantly improve the extent of proteolysis by papain, alcalase, or pepsin and reduce the antigenicity of GSP, whereas neutrase showed poor effects at any pressure. Papain and alcalase showed the highest proteolysis at 300 MPa, followed by pepsin at 400 MPa, and all of the obtained hydrolysates showed molecular weights lower than 10 kDa; furthermore, papain or alcalase at 300 MPa as well as pepsin at 400 MPa reduced antigenicity by more than 95 %, and all of the immunoreactive bands disappeared in the obtained hydrolysates. These results suggest that HHP can enhance the hydrolysis of GSP by certain enzymes and reduce the residual antigenicity of the hydrolysates. The obtained hypoallergenic hydrolysates could be used as a source of peptides for food ingredients.

β-Galactosidase from Ginkgo biloba seeds active against β-galactose-containing N-glycans: purification and characterization.

In this study, we purified an acidic β-galactosidase to homogeneity from Ginkgo biloba seeds (β-Gal'ase Gb-1) with approximately 270-fold purification. A molecular mass of the purified β-Gal'ase Gb-1 was estimated about 35 kDa by gel filtration and 32 kDa by SDS-PAGE under non-reducing condition, respectively. On the other hand, β-Gal'ase Gb-1 produced a single band with a molecular mass of 16 kDa by SDS-PAGE under reducing condition. The N-terminal amino acid sequences of 32 kDa and 16 kDa molecules were the same and identified as H-K-A-N-X-V-T-V-A-F-V-M-T-Q-H-, suggesting that β-Gal'ase Gb-1 may function as a homodimeric structure in vivo. When complex-type N-glycans containing β-galactosyl residues were used as substrates, β-Gal'ase Gb-1 showed substantial activity for β1-4 galactosyl residue and modest activity for β1-3 galactosyl residue with an optimum pH near 5.0. Based on these results, the involvement of β-Gal'ase Gb-1 in the degradation of plant complex-type N-glycans is discussed.

Decrease in pyridoxal-5′-phosphate concentration and increase in pyridoxal concentration in rat plasma by 4′-O-methylpyridoxine administration

Food poisoning from Ginkgo biloba seeds can cause epilepsy because of a decrease in γ-aminobutyric acid (GABA) concentrations in the brain. We previously demonstrated that 4′-O-methylpyridoxine (MPN) is responsible for this observed toxicity of G biloba seeds; however, the mechanism for the decrease in GABA and plasma concentration profile of MPN has not been clarified. Our hypothesis is that MPN induces a decrease in vitamin B6 concentrations, resulting in a decrease in GABA concentration. This study aimed to characterize the plasma concentration profile of MPN and intrinsic vitamin B6 concentrations (pyridoxal [PL], PL-5′-phosphate [PLP], and 4-pyridoxic acid) using a rat model. Plasma concentrations of B6 vitamers after intravenous MPN administration (5 mg/kg) were determined using high-performance liquid chromatography with a fluorescence detector. The half-life of MPN (0.91 ± 0.05 hours) was shorter in rats than the previously reported value in humans. We found a significant decrease in the plasma concentration of PLP, an active form of vitamin B6, after MPN administration. We also observed an increase in plasma PL and 4-pyridoxic acid concentrations; the increase in PL concentration may be caused by either metabolism of MPN to PL or by MPN-mediated inhibition of PL kinase. The present study is the first in vivo study showing relatively rapid elimination of MPN in rats and a decrease in plasma PLP concentration caused by MPN.

Novel analysis of maturation of murine bone-marrow-derived dendritic cells induced by Ginkgo Seed Polysaccharides

Our understanding of the mechanisms of effect of Ginkgo Seed Polysaccharides (GSPs) on the immune system remains unclear. The aim of this work was to investigate the effect of GSPs on the maturation and function of bone-marrow-derived dendritic cells (BMDCs). The results demonstrate that GSP could exert positive immune modulation on the maturation and functions of BMDCs. This effect was evidenced by decreased changes of phagosome number inside BMDCs, decreased activity of acidic phosphatase (ACP), decreased phagocytosis of BMDCs, and increased changes of key membrane molecules on BMDCs. Upregulated production of cytokines IL-12 and TNF-α also was confirmed. Therefore, it can be concluded that GSPs can efficiently induce the maturation of BMDCs. Our exploration provides direct data and a rationale for potential application of GSPs as an immune enhancer in improving immunity and as a potent adjuvant in the design of DC-based vaccines.

Study on physicochemical and in-vitro enzymatic hydrolysis properties of ginkgo (Ginkgo biloba) starch

Ginkgo biloba seed starch was isolated and its morphological, compositional, physicochemical, and structural characteristics, and in-vitro enzymatic hydrolysis, were investigated. Ginkgo starch granules were oval or spherical (5–20 μm) with a smooth surface. Compared with normal maize and potato starches, ginkgo starch had a significantly higher amylose content of 33.26%, a CA-type crystal pattern and a significantly higher crystallinity degree (40.2%). Because of its different chemical composition and structural characteristics, ginkgo starch exhibited a higher pasting temperature (80.95 °C), setback viscosity (1463 cP), gelatinization temperature (69.23 °C), lower swelling factor and gelatinization enthalpy (4.34 J/g). Moreover, ginkgo starch showed the lowest hydrolysis degree of the three starches in vitro, indicating its higher resistance to α-amylase digestibility in the case of retrogradation. The present study demonstrated that ginkgo starch is a robust and potentially useful nutritional source of enzyme-resistant starch, and provides useful information for food industries using ginkgo seeds.

Identification and quantification of biomarkers to confirm the poisoning by Ginkgo biloba seeds in a 2-year-old boy

Ginkgo seed poisoning was confirmed by measuring 4′-O-methyl pyridoxine (MPN) as a serum biomarkers. Picture shows the kinetic of MPN at 14 (B) and 20 (C) hours from intake.

Anti-inflammatory and PPAR Transactivational Effects of Components from Ginkgo biloba Seeds

Anti-inflammatory and PPAR Transactivational Effects of Components from Ginkgo biloba Seeds

An optimized ultrasound-assisted extraction and simultaneous quantification of 26 characteristic components with four structure types in functional foods from ginkgo seeds

An optimized method of ultrasound-assisted extraction followed by ultra-high-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UAE–UHPLC–TQ/MS2) was proposed for the simultaneous extraction and determination of 26 characteristic components covering four structure types (flavonoids, terpene lactones, ginkgolic acids and phenylpropanols) in ginkgo seeds (GSs). The UAE parameters (ultrasound power, time and solvent-to-material ratio) were optimized using a response surface methodology. This is the first report of the simultaneous analysis of 26 compounds in Ginkgo biloba using UHPLC–TQ/MS2; this analysis afforded good linearity, precision, repeatability and accuracy. UAE–UHPLC–TQ/MS2 was successfully applied to ginkgo seed samples, and the analysis showed that GSs are rich in terpene lactones and could be selected as a healthy food resource. The results suggest that UAE–UHPLC–TQ/MS2 might be able to be utilized as a tool for the quality assessment of samples from GSs or other related products using flavonoids, terpene lactones, ginkgolic acids and phenylpropanols as markers.

Effect of immersion in solutions with 6-benzylaminopurine on the germination and growth of seeds of Ginkgo biloba L.

Ginkgo biloba seed germination has hormonal inhibitors. Because of this, it has to be stratified in cold (4 °C) for 9 weeks before planting. This study evaluated other management by dipping prior to the sowing of seeds in solutions of 6-benzylaminopurine (BA) to obviate those nine-week waiting period and thus increase seedling growth period during the first year of cultivation. Dose experiments were performed combining BA and cold days. Treatments were two solutions with BA (2.5 ppm and 5 ppm), four periods of cold stratification (25, 31, 38 and 49 days) and two growth conditions:(a) in pots and (b) in soil without impedances. Germination (PG), dry weight partitioning, plant height and stem diameter at the neck (DAC) were evaluated. We observed a significant increase in seeds without pretreatment PG cold when treated with both dilutions of BA; there were no differences in dry weight partitioned, but the high and DAC variables show that BA-treated plants were less stable and presented fewer resources for later transplant field. We conclude that growers can choose seed immersion with 2.5 ppm of BA, no cold period, and thus sow nine weeks before with the consequent increase in biomass at the end of the first growing season; however, a higher susceptibility of most of the plants treated with BA to stress situations should be taken into account.

Hydrophilic interaction ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (HILIC-UPLC–TQ-MS/MS) in multiple-reaction monitoring (MRM) for the determination of nucleobases and nucleosides in ginkgo seeds

In this study, a rapid, simple and sensitive analytical method was developed for the quantitative determination of 20 nucleosides and nucleobases in functional foods at trace levels using hydrophilic interaction ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (HILIC-UPLC–TQ-MS/MS) in multiple-reaction monitoring (MRM) mode. Under optimised chromatographic conditions, good separation of 20 target compounds was achieved using a Waters Acquity UPLC BEH Amide column and gradient elution in 11min. The limits of detection (LODs) and quantification (LOQs) were between 0.02–42.54ng/mL and 0.05–98.18ng/mL for the 20 analytes, respectively. This is the first report about simultaneous analysis of nucleosides and nucleobases in functional foods using this method, which afforded good linearity, precision, repeatability and accuracy. The method developed was successfully applied to quantify target compounds in batches of ginkgo seeds. The method potentially could be used to determine polar and trace-level nucleosides and nucleobases in ginkgo seeds.