Isolation, purification, and structural characterization of polysaccharides from Codonopsis pilosula and its therapeutic effects on non-alcoholic fatty liver disease in vitro and in vivo.

Codonopsis pilosula is a famous edible and medicinal plants, in which polysaccharides are recognized as one of the important active ingredients. A neutral polysaccharide (CPP-1) was purified from C. pilosula. The structure was characterized by HPSEC-MALLS-RID, UV, FT-IR, GC-MS, methylation analysis, and NMR. The results showed that CPP-1 was a homogeneous pure polysaccharide, mainly containing fructose and glucose, and a small amount of arabinose. Methylation analysis showed that CPP-1 composed of →1)-Fruf-(2→, Fruf-(1→ and Glcp-(1→ residues. Combined the NMR results the structure of CPP-1 was confirmed as α-D-Glcp-(1 â†’ [2)-ß-D-Fruf-(1 â†’ 2)-ß-D-Fruf-(1]26 â†’ 2)-ß-D-Fruf with the molecular weight of 4.890 × 103 Da. The model of AML12 hepatocyte fat damage was established in vitro. The results showed that CPP-1 could increase the activity of SOD and CAT antioxidant enzymes and reduce the content of MDA, thus protecting cells from oxidative damage. Subsequently, the liver protective effect of CPP-1 was studied in the mouse model of nonalcoholic fatty liver disease (NAFLD) induced by the high-fat diet. The results showed that CPP-1 significantly reduced the body weight, liver index, and body fat index of NAFLD mice, and significantly improved liver function. Therefore, CPP-1 should be a potential candidate for the treatment of NAFLD.

Optimization of Three Extraction Methods and Their Effect on the Structure and Antioxidant Activity of Polysaccharides in Dendrobium huoshanense.

Dendrobium huoshanense is a famous edible and medicinal herb, and polysaccharides are the main bioactive component in it. In this study, response surface methodology (RSM) combined with a Box-Behnken design (BBD) was used to optimize the enzyme-assisted extraction (EAE), ultrasound-microwave-assisted extraction (UMAE), and hot water extraction (HWE) conditions and obtain the polysaccharides named DHP-E, DHP-UM, and DHP-H. The effects of different extraction methods on the physicochemical properties, structure characteristics, and bioactivity of polysaccharides were compared. The differential thermogravimetric curves indicated that DHP-E showed a broader temperature range during thermal degradation compared with DHP-UM and DHP-H. The SEM results showed that DHP-E displayed an irregular granular structure, but DHP-UM and DHP-H were sponge-like. The results of absolute molecular weight indicated that polysaccharides with higher molecular weight detected in DHP-H and DHP-UM did not appear in DHP-E due to enzymatic degradation. The monosaccharide composition showed that DHPs were all composed of Man, Glc, and Gal but with different proportions. Finally, the glycosidic bond types, which have a significant effect on bioactivity, were decoded with methylation analysis. The results showed that DHPs contained four glycosidic bond types, including Glcp-(1→, →4)-Manp-(1→, →4)-Glcp-(1→, and →4,6)-Manp-(1→ with different ratios. Furthermore, DHP-E exhibited better DPPH and ABTS radical scavenging activities. These findings could provide scientific foundations for selecting appropriate extraction methods to obtain desired bioactivities for applications in the pharmaceutical and functional food industries.

Contamination and Health Risk Assessment of Multiple Mycotoxins in Edible and Medicinal Plants.

Edible and medicinal plants (EMPs) are widely used but are easily infected by harmful fungi which produce mycotoxins. Herein, 127 samples from 11 provinces were collected to investigate 15 mycotoxins based on geographic, demographic, processing, and risk characteristics. A total of 13 mycotoxins were detected, and aflatoxin B1 (0.56~97.00 µg/kg), deoxynivalenol (9.41~1570.35 µg/kg), fumonisin B1 (8.25~1875.77 µg/kg), fumonisin B2 (2.74~543.01 µg/kg), ochratoxin A (0.62~19.30 µg/kg), and zearalenone (1.64~2376.58 µg/kg) occurred more frequently. Mycotoxin levels and species were significantly different by region, types of EMPs, and method of processing. The margin of exposure (MOE) values was well below the safe MOE (10,000). AFB1 exposure from Coix seed and malt consumption in China was of high health concern. The hazard Index (HI) method showed the range of 113.15~130.73% for malt, indicating a public health concern. In conclusion, EMPs should be concerned because of the cumulative effects of co-occurred mycotoxins, and safety management strategies should be developed in follow-up studies.

Comparison of two commercial methods with a UHPLC-MS/MS method for the determination of multiple mycotoxins in cereals.

Immunoassay-based techniques are important on-site screening tools for the detection of mycotoxins in cereals. This study aims to evaluate the trueness, precision, repeatability and cross-reactivity of commercially available test strips, ELISA kits and UHPLC-MS/MS on analyzing zearalenone, ochratoxin A, deoxynivalenol, T-2 toxin and fumonisin B1. The results showed that false negative rate (25.7 %-37.4 %) of all tested mycotoxins by test strips was higher than the false positive rate (0 %-31.0 %). The repeatability of ELISA kits at the declared LOD dispersed from -85.7 % to +98.4 %. ELISA kits were more accurate at 50 % of the maximum residue limit (MRL) of mycotoxins than 150 % and 200 %. All the tested deoxynivalenol/zearalenone derivatives had cross-reactivity with different level, and sample matrix could reinforce this overestimation of target mycotoxin. This study emphasized that higher-quality antibody screening and more analytical performance investigations are need to address for on-site detection of mycotoxins in the future.

Preparation, characterization and immunoregulatory activity of derivatives of polysaccharide from Atractylodes lancea (Thunb.) DC.

A polysaccharide (ALP-1) extracted from Atractylodes lancea (Thunb.) DC. was carboxymethylated (C-ALP-1), phosphorylated (P-ALP-1) and acetylated (A-ALP-1) to improve its physicochemical properties and bioactivities. The solubility of all derivatives was increased, and the solubility of A-ALP-1 increased to 137.5 mg/mL, which was much higher than the solubility of ALP-1 (15.0 mg/mL). The results of HPSEC-MALLS-RID showed that the molecular weight of polysaccharides was slightly increased after the modification, and the root mean square radius of rotation (Rz) and morphology of polysaccharides in solution were also changed. The scanning electron microscopy (SEM) and X-ray diffraction (XRD) results confirmed that the surface morphology of ALP-1 changed dramatically and the crystallinity decreased after structural modification. From thermal analysis results, the T50 of ALP-1, C-ALP-1, P-ALP-1 and A-ALP-1 were 281.34, 292.14, 333.75 and 298.70 °C, which showed that derivatives had stronger thermal stability than ALP-1. The immunomodulatory activity studies displayed that P-ALP-1 showed the best ability to stimulate RAW264.7 macrophages to release NO, and A-ALP-1 showed the best capacity to stimulate TNF-α and IL-6 releasing. These results indicated that chemical modification could enhance the solubility, the thermal stability and immunomodulatory activity of polysaccharides, which is beneficial for the development and utilization of natural polysaccharides.

A metal-organic framework@hydrogen-bond framework as a matrix for MALDI-TOF-MS analysis of small molecules.

A novel MOF@HOF composite that can serve as a matrix for analysis of small molecules by MALDI-TOF-MS was fabricated through a simple solvothermal method. Taking falvonoids as an example, this composite/matrix presents high desorption/ionization efficiency, low background interference, high salt tolerance, and satisfactory signal reproducibility for MALDI-TOF-MS analysis.

Exploring kinetics, removal mechanism and possible transformation products of tigecycline by Chlorella pyrenoidosa.

The accumulation of antibiotics in wastewater leads to broad antibiotic resistance, threating human health. Microalgae have been receiving attention due to their ability to remove antibiotics from wastewater. Tigecycline (TGC) is a broad-spectrum glycylcycline antibiotic. It has not been investigated for removal by microalgae. The removal kinetics of TGC by Chlorella pyrenoidosa were evaluated under different initial dry cell densities, TGC concentrations, temperatures and light intensity conditions. Approximately 90% of TGC could be removed when the TGC concentration was 10 mg∙L-1 and the initial dry cell density was more than 0.2 g∙L-1. A low value of TGC per g dry cell weight ratio led to a high removal efficiency of TGC. The initial dry cell density of microalgae was also critical for the removal of TGC. A high initial dry cell density is better than a low initial dry cell density to remove TGC when the ratio of the TGC concentration to dry cell weight are the same at the beginning of the cultivation. The removal mechanisms were investigated. Photolysis was a slow process that did not lead to removal at the beginning. Adsorption, hydrolysis, photolysis and biodegradation by microalgae were the main contributors to the removal of TGC. TGC was easily hydrolyzed under high -temperature conditions. Three transformation products of TGC by microalgae were identified. The stability of TGC was evaluated in water and salt solutions of citric acid, K2HPO4·3H2O and ferric ammonium citrate. TGC was stable in ultrapure water and citric acid solution. TGC was hydrolyzed in K2HPO4·3H2O and ferric ammonium citrate solutions.

Preparation, characterization, and bioactivity evaluation of oligosaccharides from Atractylodes lancea (Thunb.) DC.

Sixteen oligosaccharide monomers with the degree of polymerization 3 to 18 (DP 3 to DP 18) and three active fractions (DP 3-9, DP 8-11, and DP 11-17) were separated from Atractylodes lancea (Thunb.) DC. by optimized fast protein liquid chromatography coupled with refractive index detector (FPLC-RID) and preparation hydrophilic interaction chromatography (Pre-HILIC). Gas chromatography-mass spectrometer (GC-MS), liquid chromatography tandem mass spectrometry (LC-MS/MS), nuclear magnetic resonance (NMR) spectroscopy, and methylation analysis showed that the oligosaccharide in A. lancea was 1-kestose [ß-D-fructofuranosyl-(2 â†’ 1)-ß-D-fructofuranosyl-(2 â†’ 1)-α-D-glucopyranoside] (inulin-type fructooligosaccharides, FOS). Particularly, DP 3-9 showed the best capacity in stimulating phagocytic, NO, and cytokines production on RAW264.7 cells than any other purified oligosaccharide monomers and active fractions. It could also activate T-cells in Peyer's patch cells and enhance the production of colony stimulation factors. Besides, FPLC-RID showed a good capacity for large-scale preparation of DP 3-9 with the recovery of more than 93%. The bioactivity of sixteen FOS monomers (DP 3 to DP 18) and three FOS fractions (DP 3-9, DP 8-11, and DP 11-17) investigated in this study are beneficial for the utilization of FOS as a functional ingredient in novel product development.

ZrO2/CeO2/polyacrylic acid nanocomposites with alkaline phosphatase-like activity for sensing.

In the present work, we synthesized ZrO2/CeO2/polyacrylic acid (PAA) nanocomposites (nanozyme) with phosphatase-like activity. ZrO2 evenly distributed in CeO2 nanorods considered as lewis acids to enhance the phosphatase-like activity of CeO2 nanorods. Furthermore, PAA was used to coat ZrO2/CeO2/ nanorods and improve the dispersion, stability and robustness. The ZrO2/CeO2/PAA nanocomposites had 100% enhanced phosphatase-like activity compared with CeO2 nanorods and excellent adaptability in a wide pH range from 4.0 to 12.0. ZrO2/CeO2/PAA nanocomposites could hydrolyze methyl parathion (MP) to p-nitrophenol (p-NP) with bright yellow color for colorimetric detection. The developed colorimetric detection system showed a linear response from 7.60 × 10-11-7.60 × 10-8 M with a detection limit of 0.021 nM and was successfully applied for the determination of MP in corn samples.

Highly Sensitive Simultaneous Detection of Multiple Mycotoxins Using a Protein Microarray on a TiO2-Modified Porous Silicon Surface.

A new protein microarray method for multiplex mycotoxin detection in parallel has been established on a stable TiO2-modified porous silicon (PSi) surface. A typical competitive immunoassay microarray protocol has been developed for simultaneous detection of multiplex mycotoxins including aflatoxin B1 (AFB1), ochratoxin A (OTA), and fumonisin B1 (FB1) on the TiO2-PSi surface. The epoxy groups were selected to modify the surface of a TiO2-PSi wafer for the immobilization of artificial antigens of mycotoxins because of their high signal-to-noise ratios. Under optimal conditions, the developed method showed wide linear detection ranges of 0.01-1 ng/mL for OTA, 0.001-1 ng/mL for AFB1, and 0.01-1 ng/mL for FB1 and low limit of detections (LODs) of 0.433 ng/mL for OTA, 0.243 ng/mL for AFB1, and 0.093 ng/mL for FB1. The microarray method can specifically identify the three mycotoxins and their analogues. The recovery rates in real samples were within 75-120%, which were in agreement with that of the classical ELISA method. The new method has great application potential for rapid, sensitive, and high-throughput screening of multiplex mycotoxins and other target molecules.

Oligosaccharide-based quality evaluation of Atractylodis rhizome and a strategy for simplifying its quality control.

BACKGROUND: Atractylodis rhizoma, is the dried rhizomes of Atractylodes lancea (Thunb.) DC. or A. chinensis (DC.) Koidz. Both of two are pharmacologically and economically important, while with differences in efficacy. Therefore, an authentication system is vital for evaluation the quality and discrimination adulteration of Atractylodis rhizoma. Fructooligosaccharides (FOS), which are regarded as functional ingredients in Atractylodis rhizoma, have not been used for quality control of Atractylodis rhizoma for shortage of reference compounds. RESULTS: A HPLC-ELSD method was developed for the quantification of FOS in Atractylodis rhizoma. And chemometrics analysis showed that 2 markers including content of degree of polymerization (DP) 12 and total content of DP 3-15 could be used as the main distinctive elements for quality evaluation of Atractylodis rhizome. Actually, the separation and purification of high DP FOS, such as DP 12, is still a challenge because of high polarity. Then DP 5-based qualification evaluation was investigated for quality control of Atractylodis rhizoma. The results showed that A. lancea and A. chinensis could be clearly separated. CONCLUSIONS: DP 5-based quantification method was credible and effectively adopted for solving the shortage of reference compounds and improving the quality control of Atractylodis rhizoma.

Structural characterization and immunoregulatory activity of two polysaccharides from the rhizomes of Atractylodes lancea (Thunb.) DC.

A neutral polysaccharide (ALP-1) and an acidic polysaccharide (ALP-3) were purified from Atractylodes lancea (Thunb.) DC. ALP-1 exhibited a linear backbone composed of (2 → 1)-linked ß-d-fructofuranose. The backbone of ALP-3 was elucidated as →4)-GalAp-(1 → 3,4)-Rhap-(1→, and with branch chain substituted at O-3 position of →3,4)-Rhap-(1→. The branch chain consists of →3,5)-Araf-(1→, →5)-Araf-(1→, and Araf-(1→. Particularly, ALP-3 could significantly stimulate macrophage proliferation in a dose-dependent manner, and stimulate phagocytic, NO and cytokines production than ALP-1 on RAW264.7 cells. Besides, both ALP-1 and ALP-3 could activate T-cells in Peyer's patch cells and enhance the production of colony stimulation factors. While the ALP-3 also showed better intestinal immune system modulating activity than ALP-1. The result of this study indicated that the structure diversity of polysaccharide is crucial for its bioactivity. And also provided evidence for that carbohydrate polymers in A. lancea definitely contributed to its pharmaceutical effects.

Preparation and Application of Standardized Typical Volatile Components Fraction from Turmeric (Curcuma longa L.) by Supercritical Fluid Extraction and Step Molecular Distillation.

A green and reliable method using supercritical fluid extraction (SFE) and molecular distillation (MD) was optimized for the separation and purification of standardized typical volatile components fraction (STVCF) from turmeric to solve the shortage of reference compounds in quality control (QC) of volatile components. A high quality essential oil with 76.0% typical components of turmeric was extracted by SFE. A sequential distillation strategy was performed by MD. The total recovery and purity of prepared STVCF were 97.3% and 90.3%, respectively. Additionally, a strategy, i.e., STVCF-based qualification and quantitative evaluation of major bioactive analytes by multiple calibrated components, was proposed to easily and effectively control the quality of turmeric. Compared with the individual calibration curve method, the STVCF-based quantification method was demonstrated to be credible and was effectively adapted for solving the shortage of reference volatile compounds and improving the QC of typical volatile components in turmeric, especially its functional products.

Cordyceps collected from Bhutan, an appropriate alternative of Cordyceps sinensis.

Natural Cordyceps collected in Bhutan has been widely used as natural Cordyceps sinensis, an official species of Cordyceps used as Chinese medicines, around the world in recent years. However, whether Cordyceps from Bhutan could be really used as natural C. sinensis remains unknown. Therefore, DNA sequence, bioactive components including nucleosides and polysaccharides in twelve batches of Cordyceps from Bhutan were firstly investigated, and compared with natural C. sinensis. Results showed that the fungus of Cordyceps from Bhutan was C. sinensis and the host insect belonged to Hepialidae sp. In addition, nucleosides and their bases such as guanine, guanosine, hypoxanthine, uridine, inosine, thymidine, adenine, and adenosine, as well as compositional monosaccharides, partial acid or enzymatic hydrolysates, molecular weights and contents of polysaccharides in Cordyceps from Bhutan were all similar to those of natural C. sinensis. All data suggest that Cordyceps from Bhutan is a rational alternative of natural C. sinensis, which is beneficial for the improvement of their performance in health and medicinal food areas.

Optimization of microwave-assisted extraction of bioactive alkaloids from lotus plumule using response surface methodology.

In this work, a fast and efficient microwave-assisted extraction (MAE) method was developed to extract main bioactive alkaloids from lotus plumue. To optimize MAE conditions, three main factors were selected using univariate approach experiments, and then central composite design (CCD). The optimal extraction conditions were as follows: methanol concentration of 65%, microwave power of 200 W, and extraction time of 260 s. An high performance liquid chromatography-diode array detector (HPLC-DAD) method was established to quantitatively analyze these phytochemicals in different lotus plumule samples and in different part of lotus. Chromatographic separation was carried out on an Agilent Zorbax Extend-C18 column (4.6 mm×150 mm, 3.5 µm). Gradient elution was applied with the mobile phase constituted with 0.1% triethylamine in water (A) and acetonitrile (B): 40%-70% B at 0-8 min, 70%-100% B at 8-9 min, 100% B for 2 min, and then equilibrated with 40% B for 2 min.

Quality control of sweet medicines based on gas chromatography-mass spectrometry.

Sweet medicines are a relatively untapped source of new drugs. Their biological activities are closely correlated to their chemical characteristics. However, accurately defining the chemical characteristics of glycans is a challenge due to their chemical heterogeneity and diversity. Gas chromatography-mass spectrometry (GC-MS) is an excellent technique for the analysis of glycans even though the preparation of adequate derivatives is necessary. We reviewed and discussed the most important methodologies currently used for glycan analysis in sweet medicines based on GC-MS, including the derivatization for monosaccharide analysis, hydrolysis methods for polysaccharide analysis, glycosidic linkage analysis based on methylation, and pyrolysis gas chromatography in carbohydrate analysis. Finally a strategy for quality control of sweet medicines based on quantification analysis is proposed.

A comparative study on immunomodulatory activity of polysaccharides from two official species of Ganoderma (Lingzhi).

Two Ganoderma species, G. lucidum and G. sinense, are listed as Lingzhi in Chinese Pharmacopoeia and they are considered to have the same therapeutic effects. Polysaccharides were the main immunomodulatory and anticancer components in Ganoderma. In this study, the chemical characters and the effects of polysaccharides from G. lucidum (GLPS) and G. sinense (GSPS) on macrophage functions were investigated and compared. Chemical studies showed that GLPS and GSPS were different, displaying various molecular weight distribution and ratio of monosaccharide components. In vitro pharmacological studies showed that both GLPS and GSPS had potent effects on macrophage functions, such as promoting macrophage phagocytosis, increasing their release of nitric oxide and cytokines interleukin (IL)-1α, IL-6, IL-10, and tumor necrosis factor-α. Generally, GLPS was more powerful than GSPS. This study is helpful to elucidate the active components and pharmacological variation between the 2 Ganoderma species. The structure-activity relationship of polysaccharides from Ganoderma needs further study.

Determination of inulin-type fructooligosaccharides in edible plants by high-performance liquid chromatography with charged aerosol detector.

Fructooligosaccharides (FOS), which are regarded as functional ingredients, are commonly classified as dietary fibers in many countries. However, few analytical methods for separation and analysis of individual FOS in plants, crops, and food products have been developed. In this study, a simple, rapid, and sensitive high performance liquid chromatography with charged aerosol detector (HPLC-CAD) method was developed for simultaneous determination of 11 inulin-type FOS with degree of polymerization (DP) 3-13 in different samples. The separation was performed on a Waters XBridge Amide column (4.6 × 250 mm i.d., 3.5 µm) with gradient elution. All calibration curves for investigated analytes showed good linear regression (R(2) > 0.9962). Their limits of detection (LOD) and quantification (LOQ) were in the ranges 0.4-0.6 µg/mL and 1.4-2.3 µg/mL, respectively. The recoveries ranged from 94.0% to 114.4%. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was applied to qualitative analysis of FOS in different samples. The developed method was successfully applied to analysis of 11 FOS in different samples of plants from Compositae, Campanulaceae, and Rubiaceae families. The developed HPLC-CAD nethod with microwave-assisted extraction can be used for quantitative analysis of FOS and is helpful for quality control of plants containing FOS.

Chain conformation and immunomodulatory activity of a hyperbranched polysaccharide from Cordyceps sinensis.

A polysaccharide, named as cordysinan, extracted from natural Cordyceps sinensis, was identified as a hyperbranched heteropolysaccharide from the results of FT-IR, GC-MS, and carbohydrate analysis by carbohydrate gel electrophoresis analysis, as well as the degree of branching of cordysinan was 43.3%. The solution properties of cordysinan were investigated by using size exclusion chromatography coupled with multi-angle laser light scattering and triple detector array, respectively. The molecular weights, the radius of gyration and the intrinsic viscosity of cordysinan were determined as 22.45±0.26 kDa and 22.37 kDa, 15.4±2.4 nm and 1.41 mL/g, respectively. By applying the polymer solution theory, the exponent (ν and α) values of g1/2=kMwv and [η]=kMwα were calculated as 0.28 and 0.42, respectively, which firstly revealed that cordysinan existed as a globular shape in 0.9% NaCl aqueous solution. Moreover, the results showed that cordysinan could obviously stimulate macrophages functions.

Novel gas sensor combined active fiber loop ring-down and dual wavelengths differential absorption method.

A novel active fiber loop ring-down gas sensor combined with dual wavelengths differential absorption method is proposed. Two Distributed Feedback Laser Diodes (DFB LDs) with different wavelengths are employed. One LD whose wavelength covered with the absorption line of target gas is used for sensing. Another LD whose wavelength is centered outside the absorption line is used for reference. The gas absorption loss can be obtained by differencing the reference signal and sensing signal. Compared with traditional method of one wavelength employed, it can eliminate the influence of the cavity loss variety and photoelectric device drift in the system efficiently. An Erbium Doped Fiber Amplifier (EDFA) with Automatic Gain Control (AGC) is used to compensate the loss of the light in the ring-down cavity, which will increase the cavity round trips and improve the precision of gas detection. And two fiber Bragg gratings (FBGs) are employed to get rid of amplified spontaneous emission (ASE) spectrum noise as filters. The calibrating ethyne samples of different concentrations are measured with a 65 mm long gas cell in order to evaluate the effect of reference. The results show the relative deviation is found to be less than ± 0.4% of 0.1% ethyne when a certain additional loss from 0 to 1.2dB is introduced to the cavity and the relative deviation of measured concentration is less than ± 0.5% over 24 hours.