Yuanxing Wang

Quality control and original discrimination of Ganoderma lucidum based on high-performance liquid chromatographic fingerprints and combined chemometrics methods

In this paper, the feasibility and advantages of employing high-performance liquid chromatographic (HPLC) fingerprints combined with chemometrics methods for quality control of the cultured fruiting bodies of Ganoderma lucidum were investigated and demonstrated for the first time. In order to compare the HPLC fingerprints chromatograms between G. lucidum from different origins, the similarities of all the 60 samples and relative peak areas of 19 characteristic compounds were firstly calculated respectively. Then different pattern recognition procedures, including hierarchical cluster analysis (HCA), principal component analysis (PCA), partial least squares-discrimination analysis (PLS-DA) and soft independent modeling of class analogy (SIMCA) were applied to classify the G. lucidum samples according to their cultivated origins. Consistent results were obtained to show that G. lucidum samples could be successfully grouped in accordance with the province of origin. Furthermore, four marker constituents were screened out to be the most discriminant variables, which could be applied to accurate discrimination and quality control of G. lucidum by quantitative analysis. Finally, the chemical properties of those samples were also investigated to find out the differences of quality between them. Ranked in decreasing order, the quality of the G. lucidum can be arranged as Jinzhai/Huangshan, Shandong followed by Zhejiang samples. Our results revealed that the developed method has potential perspective for the original discrimination and quality control of G. lucidum.

Ultrasonic-assisted extraction, antimicrobial and antioxidant activities of Cyclocarya paliurus (Batal.) Iljinskaja polysaccharides

Recently, renewed interest has grown in the use of ultrasonic-assisted extraction as an alternative approach to the traditional extraction methods. In this study, this novel extraction method was used to isolate polysaccharides from Cyclocarya paliurus (Batal.) Iljinskaja, and response surface methodology (RSM) was employed to optimize the extraction conditions. The optimal conditions for the extraction of polysaccharides were determined to be the ratio of liquid to solid of 8, extraction time of 59 min and extraction temperature of 58 °C. Under these optimal conditions, the yield of polysaccharides obtained was 4.91 ± 0.11%, which was well matched with the value predicted by the model. In vitro antioxidant assays showed that the polysaccharides exhibited strong DPPH radicals (92.09% at 0.25 mg/ml) and self-oxidation of 1,2,3-phentriol (37.22% at 1 mg/ml) scavenging activities, moderate hydroxyl radicals (43.18% at 1 mg/ml) scavenging activity and lipid peroxidation inhibitory effect (31.66% at 1 mg/ml). In addition, the polysaccharides showed moderate antimicrobial activity.

Discrimination of Ganoderma lucidum according to geographical origin with near infrared diffuse reflectance spectroscopy and pattern recognition techniques

A rapid and nondestructive near infrared (NIR) method combined with chemometrics was used to discriminate Ganoderma lucidum according to cultivation area. Raw, first, and second derivative NIR spectra were compared to develop a robust classification rule. The chemical properties of G. lucidum samples were also investigated to find out the difference between samples from six varied origins. It could be found that the amount of polysaccharides and triterpenoid saponins in G. lucidum samples was considerably different based on cultivation area. These differences make NIR spectroscopic method viable. Principal component analysis (PCA), discriminant partial least-squares (DPLS) and discriminant analysis (DA) were applied to classify the geographical origins of those samples. The results showed that excellent classification could be obtained after optimizing spectral pre-treatment. For the discriminating of samples from three different provinces, DPLS provided 100% correct classifications. Moreover, for samples from six different locations, the correct classifications of the calibration as well as the validation data set were 96.6% using the DA method after the SNV first derivative spectral pre-treatment. Overall, NIR diffuse reflectance spectroscopy using pattern recognition was shown to have significant potential as a rapid and accurate method for the identification of herbal medicines.

Acetylation and carboxymethylation of the polysaccharide from Ganoderma atrum and their antioxidant and immunomodulating activities.

A water-soluble polysaccharide extracted from Ganoderma atrum was chemically modified to obtain its acetyled and carboxymethylated derivatives. The results of chemical analysis, Fourier-transform infrared and (13)C nuclear magnetic resonance spectroscopy showed that these modifications were successful, although the molecular weight of these derivatives decreased due to slight degradation during the reaction. The antioxidant and immunomodulating activities of these derivatives were then investigated to determine the structure-bioactivity relationship. Results showed that the acetyled derivative with appropriate degree of substitution and lower molecular weight exhibited stronger antioxidant abilities on scavenging DPPH radical, and inhibitory effects in β-carotene-linoleic acid systems compared with the native polysaccharide. In addition, it also enhanced the macrophage phagocytosis capacity and tumor necrosis factor-α secretion, whereas the carboxymethylated derivative was shown to be slightly less effective. These results indicated that the type of substitution group and their degree of substitution play a decisive role in the bioactivities of the derivatives.

Enhancement of Cyclophosphamide-Induced Antitumor Effect by a Novel Polysaccharide from Ganoderma atrum in Sarcoma 180-Bearing Mice

The aim of this study was to investigate the enhancement of Ganoderma atrum polysaccharide (PSG-1) on cyclophosphamide (CTX)-induced antitumor effect in sarcoma 180 (S-180)-bearing mice. Results showed that both CTX and PSG-1 delayed tumor growth and resulted in tumor apoptosis. The combined regimen was superior to either modality alone. Moreover, the combined treatment-induced apoptosis was mediated via mitochondrial pathway, as evidenced by alterations of Bcl-2 family proteins, loss of mitochondrial membrane potential (Δψ(m)), cytochrome c release, and caspases activation. Our results also showed that thymus and spleen indexes, lymphocytes proliferation, and concentrations of cytokine in the CTX group were decreased, which were alleviated by PSG-1. Additionally, the combined treatment ameliorated oxidative stress as compared with CTX alone. Taken together, we conclude that PSG-1 improved the antitumor effect of CTX, possibly in part mediated by enhancing the induction of apoptosis via mitochondrial pathways, activating host immune function, and modifying the redox system in S-180-bearing mice.

Sulfated modification of the polysaccharides from Ganoderma atrum and their antioxidant and immunomodulating activities

In this study, three chemically sulfated polysaccharides (S-PSG) with different degree of sulfate substitution (DS) were derived from Ganoderma atrum. FT-IR and (13)C NMR spectra indicated that the substitution was predominantly on C-6 position compared with the C-2 position. The antioxidant and immunomodulatory activities of S-PSG were further examined to determine the relationship between its structure and bioactivity. The results showed that compared with the native polysaccharide, the S-PSG-2 with moderate DS and molecular weight exhibited the highest immunomodulatory activity by increasing the macrophage phagocytosis capacity and TNF-α production. The DPPH radical scavenging activity was also greatly enhanced by S-PSG in all the groups compared with PSG. However, the reducing power and the antioxidant activities in β-carotene-linoleic acid assay were decreased. These results indicated that except for the DS, some other structural characteristics such as molecular weight, substitution position and chain conformation were also important factors affecting their bioactivities. And the S-PSG-2 we derived could be served as immunomodulator and free-radical inhibitors.

Analysis of the monosaccharide composition of purified polysaccharides in Ganoderma atrum by capillary gas chromatography

INTRODUCTION Ganoderma, one of the best-known traditional Chinese medicines, has attracted considerable attention owing to the fact that dozens of polysaccharides isolated from it have shown diverse and potentially significant pharmacological activities. However, no work has been reported on the analysis of monosaccharide composition of polysaccharide isolated from the aqueous extract of Ganoderma atrum yet. OBJECTIVE To develop a simple and sensitive GC-based method for the analysis of monosaccharide composition of purified polysaccharides in Ganoderma atrum. METHODOLOGY The polysaccharide was first hydrolysed to give the constituent monosaccharides, which were subsequently derived into acetylated aldononitriles and analysed by gas chromatography using a capillary column packed with a (5%phenyl) methylpolysiloxane stationary phase with the addition of acetyl inositol as the inner standard. High-performance liquid chromatography was also used for comparison. RESULTS The stable derivatives of the most common monosaccharides could be separated and reproducibly determined with high sensitivity. The limits of detection and quantification were 0.013 and 0.043 mg/mL, respectively. The intermediary precision values (expressed as the RSD) were less than 10%. The mean recovery of the method was 100 + or - 3%, with RSD values of less than 5%. The results obtained from GC and HPLC methods were found to be close to each other within acceptable error ranges. CONCLUSION This study demonstrated that the developed method could be applied as an accurate method for the compositional analysis of monosaccharides in the field of biological and biochemical study.

Sulfated modification, characterization and property of a water-insoluble polysaccharide from Ganoderma atrum.

Sulfated modification was carried out to modify a water-insoluble polysaccharide from Ganoderma atrum (AGAP). The effects of sulfation on structure, physicochemical and functional properties of AGAP were investigated. Three sulfated derivatives were prepared, designated as S-1, S-2 and S-3 with degree of substitution (DS) of 0.35, 0.74 and 1.14, respectively. AGAP was elucidated as an α-(1→3)-glucan with few branches terminated by single mannose or xylose residues. The molecular weight (Mw) and radius of gyration (Rg) were estimated to be 1665 kDa and 65.49 nm, respectively. After sulfated modification, non-selective sulfation occurred preferably at O-6, partially at O-2 and O-4 positions of the glucosyl residues. The water-solubility of the derivatives was significantly improved in a DS-dependent manner. Mw of the derivatives showed a sharp decrease, and the chain conformation was estimated to be expanded stiff in phosphate buffer. In vitro tests showed that sulfated modification improved its antioxidant activities and anti-proliferative ability against S-180 tumor cells. This study suggested that sulfated modification was an effective approach to improve the water-solubility and functional properties of insoluble polysaccharides.

An effective method for deproteinization of bioactive polysaccharides extracted from lingzhi (Ganoderma atrum)

Deproteinization procedure is a fundamental step for analyzing polysaccharide from natural plants. In this study, in the course of refining bioactive polysaccharides from lingzhi (Ganoderma atrum), an effective deproteinization method using lead acetate solution was established by comparing with other available methods. The percentages of deproteinization, polysaccharide loss, and its antioxidant activities loss were used as the index to evaluate and optimize the precipitation experimental conditions. The results showed that the modified method, precipitation with the addition of 0.4–0.52%(w/v) lead acetate, was superior to the others, as evidenced by the highest deproteinization efficiency (88%), as well as the lowest polysaccharide loss (17%). And notably its antioxidant activity also remained good (loss 15%). It provides a simple prefractionation step for the analysis of polysaccharide from natural plants. Polysaccharide isolated by this method is in the native state. Our method may offer a rapid method for removing protein from plant polysaccharides in large scale.

Study on discrimination of white tea and albino tea based on near-infrared spectroscopy and chemometrics.

BACKGROUND White tea and albino tea have their own nutritional characteristics, but from the appearance they are quite similar to each other. It is not easy to distinguish them with existing analytical tools or by visual inspection. The current study proposed a rapid method to discriminate them based on near-infrared (NIR) spectroscopy associated with supervised pattern recognition methods. RESULTS For this purpose, discriminant partial least-squares (DPLS) and discriminant analysis (DA) were employed to build classification models on the basis of a reduced subset of wavenumbers and different pretreatment methods. A completely independent validation set was also used to test the model performance. The results of the DA model showed that with the SNV Karl Norris derivative spectral pre-treatment samples from the two different origins could be 100% correctly discriminated. Similarly, for the DPLS model, the best classification results were obtained with the multiplicative scattering correction (MSC) + first derivative spectral pre-treatments; the accuracy of identification was 98.48% for the calibration set and 100% for the validation set. CONCLUSION The overall results demonstrated that NIR spectroscopy with pattern recognition could be successfully applied to discriminate white tea and albino tea quickly and non-destructively without the need for various analytical determinations.