Fangdi Hu

Synthesis of a biocompatible gelatin functionalized graphene nanosheets and its application for drug delivery

A simple and environmentally friendly synthetic route for the preparation of gelatin functionalized graphene nanosheets (gelatin-GNS) was reported by using exfoliated graphene oxide as a precursor, in which gelatin acted as not only a reducing reagent but also a functionalization reagent to guarantee good dispersibility and stability of the GNS in distilled water and various physiological solutions. The obtained biocompatible gelatin-GNS attaching methotrexate (MTX) via strong π-π stacking interaction, exhibited a high drug loading capacity of MTX and excellent ability for controlled drug release. The pH-dependent release behavior of MTX from MTX@gelatin-GNS showed that the release amount under acid conditions is much higher than that under neutral conditions, which experienced a gelatin-mediated sustained release process. From the cytotoxicity assay, we can see that the MTX@gelatin-GNS showed remarkable toxicity while the gelatin-GNS showed nontoxic at appropriate concentration, both of them might be taken up by A549 cells through a nonspecific endocytosis process. The prepared nanohybrids system offers a novel formulation that combines the unique properties of a biodegradable material, gelatin, and graphene for biomedical applications. Therefore, the gelatin-GNS with good stability and biocompatibility can be selected as an ideal drug carrier to be applied in biomedicine studies.

Structural characterization and antitumor activity of a pectic polysaccharide from Codonopsis pilosula

A pectic polysaccharide (CPP1b) was at first isolated from Codonopsis pilosula. Sugar analysis revealed that CPP1b is composed of rhamnose (Rha), arabinose (Ara), galactose (Gal) and galacturonic acid (GalA) with a molar ratio of 0.25:0.12:0.13:2.51. The result of esterification assay showed that about 46.7±0.4% of carboxylic groups in GalA residues existed as methyl ester. Combined with chemical and spectroscopic analyses, a preliminary structure of CPP1b was proposed as follows: 1,4-linked α-D-GalpA and 1,4-linked α-D-GalpA6Me interspersed with rare 1,2-linked β-L-Rhap, 1,2,6-linked α-D-Galp and terminal α-L-Arap. CPP1b had an average molar mass and root-mean square radius (RMS) of 1.45×10(5) Da and 29.7 nm, respectively, and presented a linear random coil conformation in 0.9% NaCl. The ultrastructure of CPP1b was further investigated by transmission electron microscope (TEM) and scanning electron microscope (SEM). CPP1b exhibited obvious cytotoxicity to human lung adenocarcinoma A549 cells in a dose- and time-dependent manner.

Electrochemically reduced graphene oxide-based electrochemical sensor for the sensitive determination of ferulic acid in A. sinensis and biological samples

An electrochemically reduced graphene oxide (ERGO) modified glassy carbon electrode (GCE) was used as a new voltammetric sensor for the determination of ferulic acid (FA). The morphology and microstructure of the modified electrodes were characterized by scanning electron microscopy (SEM) and Raman spectroscopy analysis, and the electrochemical effective surface areas of the modified electrodes were also calculated by chronocoulometry method. Sensing properties of the electrochemical sensor were investigated by means of cyclic voltammetry (CV) and differential pulse voltammetry (DPV). It was found that ERGO was electrodeposited on the surface of GCE by using potentiostatic method. The proposed electrode exhibited electrocatalytic activity to the redox of FA because of excellent electrochemical properties of ERGO. The transfer electron number (n), electrode reaction rate constant (ks) and electron-transfer coefficient (α) were calculated as 1.12, 1.24s(-1), and 0.40, respectively. Under the optimized conditions, the oxidation peak current was proportional to FA concentration at 8.49 × 10(-8)mol L(-1) to 3.89 × 10(-5)mol L(-1) with detection limit of 2.06 × 10(-8)mol L(-1). This fabricated sensor also displayed acceptable reproducibility, long-term stability, and high selectivity with negligible interferences from common interfering species. The voltammetric sensor was successfully applied to detect FA in A. sinensis and biological samples with recovery values in the range of 99.91%-101.91%.

Investigate electrochemical immunosensor of cortisol based on gold nanoparticles/magnetic functionalized reduced graphene oxide.

A sensitively competitive electrochemical immunosensor for the detection of cortisol was successfully developed based on gold nanoparticles and magnetic functionalized reduced graphene oxide (AuNPs/MrGO). In order to construct the base of the immunosensor, the MrGO was initially fabricated by chemical cross-linking and used to modify the nafion pretreated glassy carbon electrode. Subsequently, the surface of electrode was modified by AuNPs via electrochemical deposition. A variety of cortisol (Cor) can be firmly loaded in the AuNPs/MrGO with large specific surface area and good bioactivity to construct the basic electrode (Cor/AuNPs/MrGO/Nafion@GCE), which was characterized by the cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), respectively. Due to the cortisol on the surface of basic electrode and samples can competitively combine with the cortisol antibody labelled by horseradish peroxidase (HRP-Strept-Biotin-Ab). Finally, the detection signal of electrochemical immunosensor (HRP-Strept-Biotin-Ab-Cor/AuNPs/MrGO/Nafion@GCE) in the test liquid had negative correlations with the concentration of cortisol in samples. The AuNPs/MrGO with excellent electrical conductivity being applied, the electrochemical response of the immunosensor was immensely amplified. The immunosensor displayed excellent analytical performance for the detection of cortisol range from 0.1 to 1000ng/mL with a detection limit of 0.05ng/mL at 3σ. Moreover, compared the developed immunoassay with commercially available enzyme linked immunosorbent assay, the proposed method showed good precision, acceptable stability and reproducibility, indicating the immunosensor could be used for the sensitive, efficient and real-time detection of cortisol in real samples. Therefore, the present strategy provides a novel and convenient method for clinical determination of cortisol.

Structure and antioxidant activity study of sulfated acetamido-polysaccharide from Radix Hedysari

A new sulfated acetamido-heteropolysaccharide, HPS4-2A, was obtained by aqueous extraction followed by precipitation with ethanol and fractionation with DEAE column chromatography from Radix Hedysari. It was composed of rhamnose, arabinose, glucose, galactose and 2-acetamido-2-deoxy-d-galactose in the molar ratio of 10.09%:25.90%:25.90%:25.0%:12.30%. Elemental analysis indicated that HPS4-2A was a sulfated polysaccharide containing small amount of sulfate groups (1.87%). Partial acid hydrolysis, GC, GC-MS, 1D and 2D NMR spectroscopy analysis of the HPS4-2A revealed a predominance of glucose, galactose and 2-acetamido-2-deoxy-D-galactose linked in a highly-branched structure. The molecular weight of HPS4-2A was determined by HPSEC and HPSEC-MALLS. AFM study indicated that HPS4-2A took a highly branched conformation, which in consistent with the result studied by SEC-MALLS. Structural features of HPS4-2A were also investigated by SEM and TEM. Antioxidant assays demonstrated that HPS4-2A possessed of strong DPPH and hydroxyl radicals scavenging activities, suggesting that HPS4-2A could potentially be used as natural antioxidant.

Optimization of selenylation conditions for a pectic polysaccharide and its structural characteristic.

CPP1b is a pectic polysaccharide isolated from Codonopsis pilosula, and it possesses potent antitumor activity. In this study, a HNO3-Na2SeO3 method was performed to synthesize selenium-CPP1b (sCPP1b). The effects of BaCl2 usage amount and HNO3 concentration on the yield and selenium content of sCPP1b were investigated by single-factor experiments. Reaction time, reaction temperature and the ratio of polysaccharide to Na2SeO3 were optimized by orthogonal experimental design at three levels of each of the three factors (L9(3)(4)) based on antitumor activity, selenium content and yield of sCPP1b. Our results showed that 5h of reaction time, 60°C of reaction temperature, and 2:2 ratio of polysaccharide to Na2SeO3 were the optimal selenylation modification conditions. The validation experiments completed under the optimal conditions gave the mean selenium content and yield of sCPP1b were 478.17μg/g (RSD=5.7%) and 595mg/g (RSD=1.6%), respectively. Selenylation modification can significantly increase the antitumor activity of CPP1b in vitro. The structural characterization of sCPP1b was further characterized by Fourier-transform infrared spectroscopy, thermogravimetric analysis, and high-performance gel permeation chromatography coupled with multi-angle laser light scattering. These structure analysis results indicated that sCPP1b has been successfully selenylation modified with similar structure to polysaccharide of CPP1b.

Quantitative analysis of costunolide and dehydrocostuslactone in rat plasma by ultraperformance liquid chromatography-electrospray ionization-mass spectrometry.

Costunolide and dehydrocostuslactone are well-known sesquiterpene lactones contained in many plants used as popular herbs, such as Saussurea lappa and Laurus novocanariensis, and have been considered as potential candidates for the treatment of various types of tumor. In the present work, a sensitive UPLC-MS/MS for the quantification of costunolide and dehydrocostuslactone in biological matrices has been developed. The method is based on protein precipitation with acetonitrile followed by isocratic ultraperformance liquid chromatographic separation using methanol-formic acid (0.1% in water; 70:30, v/v) mobile phase. Detection was performed by ESI mass spectrometry in MRM mode with the precursor-to-product ion transitions m/z 233-187 and m/z 231-185, respectively. The calibration curves of analytes showed good linearity within the established range 0.19-760  ng/mL for costunolide and 0.23-908  ng/mL for dehydrocostuslactone. The lower limits of quantification of costunolide and dehydrocostuslactone were found to be 0.19 and 0.23  ng/mL, respectively. The intra-day and inter-day presicions of this method for the entire validation were less than coefficient of variation of 7% and the accuracy was within ±8% (relative error). The mean extraction recoveries were 73.8 and 75.3%, respectively. The method was found to be precise, accurate and specific during the study, and was successfully used to analyze the pharmacokinetics of costunolide and dehydrocostuslactone.

Ultrasensitive Detection of Ferulic Acid Using Poly(diallyldimethylammonium chloride) Functionalized Graphene-Based Electrochemical Sensor

The electrochemical redox of ferulic acid (FA) was investigated systematically by cyclic voltammetry (CV) with a poly(diallyldimethylammonium chloride) functionalized graphene-modified glassy carbon electrode (PDDA-G/GCE) as a working electrode. A simple and sensitive differential pulse voltammetry (DPV) technique was proposed for the direct quantitative determination of FA in Angelica sinensis and spiked human urine samples for the first time. The dependence of the intensities of currents and potentials on nature of the supporting electrolyte, pH, scan rate, and concentration was investigated. Under optimal conditions, the proposed sensor exhibited excellent electrochemical sensitivity to FA, and the oxidation peak current was proportional to FA concentration in the range of 8.95 × 10−8 M ~5.29 × 10−5 M, with a relatively low detection limit of 4.42 × 10−8 M. This fabricated sensor also displayed acceptable reproducibility, long-term stability, and high selectivity with negligible interferences from common interfering species. Besides, it was applied to detect FA in Angelica sinensis and biological samples with satisfactory results, making it a potential alternative tool for the quantitative detection of FA in pharmaceutical analysis.

Direct electrochemistry and electrocatalysis of lobetyolin via magnetic functionalized reduced graphene oxide film fabricated electrochemical sensor

A novel lobetyolin electrochemical sensor based on a magnetic functionalized reduced graphene oxide/Nafion nanohybrid film has been introduced in this work. The magnetic functionalized reduced graphene oxide was characterized by fourier transform infrared spectroscopy, atomic force microscope, X-ray diffraction, transmission electron microscopy and thermogravimetric analysis. The scanning electron microscopy characterized the morphology and microstructure of the prepared sensors, and the electrochemical effective surface areas of the prepared sensors were also calculated by chronocoulometry method. The electrochemical behavior of lobetyolin on the magnetic functionalized reduced graphene oxide/Nafion nanohybrid modified glassy carbon electrode was investigated by cyclic voltammetry and differential pulse voltammetry in a phosphate buffer solution of pH6.0. The electron-transfer coefficient (α), electron transfer number (n), and electrode reaction rate constant (Κs) were calculated as 0.78, 0.73, and 4.63s-1, respectively. Under the optimized conditions, the sensor based on magnetic functionalized reduced graphene oxide/Nafion showed a linear voltammetric response to the lobetyolin concentration at 1.0×10-7 to 1.0×10-4mol/L with detection limit (S/N=3)of 4.3×10-8mol/L. The proposed sensor also displayed acceptable reproducibility, long-term stability, and high selectivity, and performs well for analysis of lobetyolin in real samples. The voltammetric sensor was successfully applied to detect lobetyolin in Codonopsis pilosula with recovery values in the range of 96.12%-102.66%.

The pharmacokinetic study of rutin in rat plasma based on an electrochemically reduced graphene oxide modified sensor

An electrochemical method based on a directly electrochemically reduced graphene oxide (ERGO) film coated on a glassy carbon electrode (GCE) was developed for the rapid and convenient determination of rutin in plasma. ERGO was modified on the surface of GCE by one-step electro-deposition method. Electrochemical behavior of rutin on ERGO/GCE indicated that rutin underwent a surface-controlled quasi-reversible process and the electrochemical parameters such as charge transfer coefficient (α), electron transfer number (n) and electrode reaction standard rate constant (ks) were 0.53, 2 and 3.4 s−1, respectively. The electrochemical sensor for rutin in plasma provided a wide linear response range of 4.70×10−7−1.25×10−5 M with the detection limit (s/n=3) of 1.84×10−8 M. The assay was successfully used to the pharmacokinetic study of rutin. The pharmacokinetic parameters such as elimination rate half-life (t1/2), area under curve (AUC), and plasma clearance (CL) were calculated to be 3.345±0.647 min, 5750±656.0 µg min/mL, and 5.891±0.458 mL/min/kg, respectively. The proposed method utilized a small sample volume of 10 μL and had no complicated sample pretreatment (without deproteinization), which was simple, eco-friendly, and time- and cost-efficient for rutin pharmacokinetic studies.