Huaguo Chen

Anti-inflammatory, antiviral and quantitative study of quercetin-3-O-β-D-glucuronide in Polygonum perfoliatum L.

Quercetin-3-O-β-D-glucuronide, isolated from Polygonum perfoliatum L., was evaluated by antiviral efficacy against influenza A virus and anti-inflammatory activity in vivo in mouse, and it was used for quality evaluation of P. perfoliatum L.. In vivo study, oral administration of quercetin-3-O-β-D-glucuronide significantly suppressed ear edema induced by dimethyl benzene and peritoneal permeability induced by acetic acid in mice, and quercetin-3-O-β-D-glucuronide also showed to possess inhibitory activity against influenza A virus (FLUAV). In the present study, additionally, a rapid, simple and sensitive method for quantitative analysis of quercetin-3-O-β-D-glucuronide in P. perfoliatum L. was developed using high performance liquid chromatography (HPLC) coupled with photodiode array detection. The separation was carried out on a Lichrosher-C18 column (250 mm × 4.6mm, 5 μm) together with a C18 guard column at isocratic elution systems of methanol (A) and 0.05% aqueous phosphoric acid (B) (43:57, v/v) with detection wavelength at 258 nm and column temperature at 30°C. The method was validated for linearity, repeatability, limit of quantification (LOQ), precision and robustness. The contents of quercetin-3-O-β-D-glucuronide in 28 samples from different regions of China were between 0.06% and 2.09%. The developed analytical method was applied to investigate P. perfoliatum L. and for quality control of the herb.

Optimization of the microwave-assisted extraction and antioxidant activities of anthocyanins from blackberry using a response surface methodology

Blackberry contains high amounts of anthocyanins, whose extraction method is closely related with anthocyanin content and antioxidant activity. The extraction yield and antioxidant capacity acted as the comprehensive evaluation indexes, and a Box–Behnken design (BBD) of response surface methodology (RSM) was employed to further optimize the microwave-assisted extraction (MAE) conditions for blackberry anthocyanins (BBAC). A significant correlation was found between the double indexes extraction yield and the antioxidant capacity (P < 0.01). The results showed that the optimized extraction conditions included a microwave power of 469 W, a solvent concentration of 52%, a liquid–solid ratio of 25 g mL−1, and a microwave time of 4 min. Under these conditions, the mean experimental value of the extraction yield (2.18 ± 0.06 mg g−1), ABTS assay (32.18 ± 1.54 μM TEAC per g) and DPPH assay (27.18 ± 1.33 μM TEAC per g) corresponded well with the predicted values. Moreover, these mean experimental values were 120% higher than those obtained during ethanol leaching extraction.

Determination of dehydroevodiamine in Evodia rutaecarpa (Juss.) Benth by high performance liquid chromatography and classification of the samples by using hierarchical clustering analysis.

A simple, sensitive and accurate liquid chromatographic method with photodiode-array detection was developed for determination of dehydroevodiamine with detection wavelength at 368 nm and column temperature at 30 degrees C. The separation was carried out on an Agilent Zorbax SB-C(18) column (250 mm x 4.6 mm, 5 microm) together with a C(18) guard column. The mobile phase was acetonitrile-water (containing 30 mM sodium acetate trihydrate and 0.15% acetic acid) in the ratio of 30:70 (v/v) delivered at a flow rate of 1 mL/min. Excellent linear behavior was observed over the concentration range investigated, with correlation coefficient (R(2))=0.9998. This validated method was applied to determine the contents of dehydroevodiamine in 36 samples from different regions of China, and hierarchical clustering analysis was firstly used to classify and differentiate Evodia rutaecarpa samples. The analysis is specific and can be successfully applied to analyze E. rutaecarpa which is helpful for quality control of the herb.

Studies on Chromatographic Fingerprint and Fingerprinting Profile-Efficacy Relationship of Saxifraga stolonifera Meerb.

This work investigated the spectrum-effect relationships between high performance liquid chromatography (HPLC) fingerprints and the anti-benign prostatic hyperplasia activities of aqueous extracts from Saxifraga stolonifera. The fingerprints of S. stolonifera from various sources were established by HPLC and evaluated by similarity analysis (SA), hierarchical clustering analysis (HCA) and principal component analysis (PCA). Nine samples were obtained from these 24 batches of different origins, according to the results of SA, HCA and the common chromatographic peaks area. A testosterone-induced mouse model of benign prostatic hyperplasia (BPH) was used to establish the anti-benign prostatic hyperplasia activities of these nine S. stolonifera samples. The model was evaluated by analyzing prostatic index (PI), serum acid phosphatase (ACP) activity, concentrations of serum dihydrotestosterone (DHT), prostatic acid phosphatase (PACP) and type II 5α-reductase (SRD5A2). The spectrum-effect relationships between HPLC fingerprints and anti-benign prostatic hyperplasia activities were investigated using Grey Correlation Analysis (GRA) and partial least squares regression (PLSR). The results showed that a close correlation existed between the fingerprints and anti-benign prostatic hyperplasia activities, and peak 14 (chlorogenic acid), peak 17 (quercetin 5-O-β-d-glucopyranoside) and peak 18 (quercetin 3-O-β-l-rhamno-pyranoside) in the HPLC fingerprints might be the main active components against anti-benign prostatic hyperplasia. This work provides a general model for the study of spectrum-effect relationships of S. stolonifera by combing HPLC fingerprints with a testosterone-induced mouse model of BPH, which can be employed to discover the principle components of anti-benign prostatic hyperplasia bioactivity.

Characterizations and hepatoprotective effect of polysaccharides from Mori Fructus in rats with alcoholic-induced liver injury

Crude polysaccharides of Mori Fructus (MFPs) were found to have anti-inflammatory antioxidant, and immuno-enhancing activities. However, the structure of the polysaccharides was ambiguous and its holistic hepatic protection evaluation was defective. This study was conducted to illustrate the characterization of MFPs, and evaluate its hepatoprotective activities. The results found that MFPs contained 67.93±1.18% carbohydrates, 31.03±0.54% uronic acid, and little protein and sulfate. The average molecular weight was ranging from 112.2kDa to 181.9kDa. Monosaccharide component analysis indicated that MFPs was mainly composed of glucose, galacturonic acid, rhamnose and galactose. Both the acute and subacute alcoholic-induced liver injury animal models were adopted to evaluate the MFPss hepatoprotective activity. After administration of MFPs, both serological indexes (aspartate aminotransferase and alanine aminotransferase) and hepatic indicators (glutathione, superoxide dismutase, glutathione peroxidase and malondialdehyde) were improved by comparing with the non-MFPs group. The hepatic histopathology results also showed a prominent lipid degeneration and microvesicular steatosis attenuation in the MFPs groups. These outstanding hepatic protecting activities of MFPs might be related to its activation of ethanol dehydrogenase, elimination of free radicals and/or inhibition of lipid peroxidation capacities. MFPs could be important active substances for preventing and remedying liver injury.

A polyamide resin based method for adsorption of anthocyanins from blackberries

The present study deals with the downstream processing of anthocyanins from blackberries in order to obtain anthocyanins in a purified form. Adsorption was carried out employing four different mesh sizes of polyamide resin and among these, the 60–100 mesh polyamide resin showed the highest adsorption capacity (4.41 mg mL−1 of the adsorbent) and desorption capacity (3.29 mg mL−1 of the adsorbent). Adsorption results were found to correlate best using the Freundlich equation at all the temperatures studied. The second order kinetics model was found to be more appropriate to describe the adsorption of anthocyanins. Under all conditions, the polyamide resin showed optimal adsorption and desorption capacity towards anthocyanins at an initial concentration of 0.75 mg mL−1, an adsorption liquid pH of 1, an eluent pH of 3 and an ethanol concentration of 80%. The dynamic adsorption process parameters for the purification of anthocyanins using the 60–100 mesh polyamide resin were as follows: processing volume, 5 BV; the flow rate of adsorption, 5 BV per h; the temperature of adsorption, 15 °C; eluent volume, 3.5 BV and the flow rate of desorption, 5 BV per h. Polyamide resin adsorption resulted in a gram of concentrated blackberry extract that contained 432 mg of anthocyanins, which was superior to cation exchange resin adsorption (160 mg g−1) and macroporous adsorbent resin adsorption (176 mg g−1). This method provides an efficient and low-cost approach for anthocyanin purification for industrial applications.

Inhibition effects of chlorogenic acid on benign prostatic hyperplasia in mice

&NA; This study aimed to evaluate the inhibitory effects and explore mechanisms of chlorogenic acid against testosterone‐induced benign prostatic hyperplasia (BPH) in mice. Benign prostatic hyperplasia model was induced in experimental groups by daily subcutaneous injections of testosterone propionate (7.5 mg/kg/d) consecutively for 14 d. A total of 60 mice were randomly divided into six groups: (Group 1) normal control group, (Group 2) benign prostatic hyperplasia model control group, (Group 3) benign prostatic hyperplasia mice treated with finasteride at a dose of 1 mg/kg, (Group 4) benign prostatic hyperplasia mice treated with chlorogenic acid at dose levels of 0.8 mg/kg (low dose group), (Group 5) benign prostatic hyperplasia mice treated with chlorogenic acid at dose levels of 1.6 mg/kg (medium dose group) and (Group 6) benign prostatic hyperplasia mice treated with chlorogenic acid at dose levels of 3.2 mg/kg (high dose group). Animals were sacrificed on the scheduled termination, pick out the eyeball to get blood, then prostates were weighed and prostatic index were determined. Then the serum acid phosphatase (ACP), prostatic acid phosphatase (PACP) and typeII5‐alpha‐reductase (SRD5A2) levels were measured and observed morphological changes of the prostate. Comparing with benign prostatic hyperplasia model group, the high and medium dose of chlorogenic acid could significantly reduce prostate index and levels of acid phosphatase, prostatic acid phosphatase and typeII5‐alpha‐reductase (P<0.05 or P<0.01). These findings were supported by histopathological observations of prostate tissues. Histopathological examination also indicated that chlorogenic acid treatment at the high and medium doses inhibited testosterone‐induced prostatic hyperplasia. The results indicated that chlorogenic acid exhibited restraining effect on benign prostatic hyperplasia model animals, and its mechanism might be related to inhibit typeII5‐alpha reductase activity.

The Tissue Distribution and Urinary Excretion Study of Gallic Acid and Protocatechuic Acid after Oral Administration of Polygonum Capitatum Extract in Rats.

In the present study, we investigated the tissue distribution and urinary excretion of gallic acid (GA) and protocatechuic acid (PCA) after rat oral administration of aqueous extract of Polygonum capitatum (P. capitatum, named Herba Polygoni Capitati in China). An UHPLC-MS/MS analytical method was developed and adopted for quantification of GA and PCA in different tissue homogenate and urine samples. Interestingly, we found that GA and PCA showed a relatively targeted distribution in kidney tissue after dosing 60 mg/kg P. capitatum extract (equivalent to 12 mg/kg of GA and 0.9 mg/kg of PCA). The concentrations of GA and PCA in the kidney tissue reached 1218.62 ng/g and 43.98 ng/g, respectively, at one hour after oral administration. The results helped explain the empirical use of P. capitatum for kidney diseases in folk medicine. Further studies on urinary excretion of P. capitatum extract indicated that GA and PCA followed a concentrated elimination over a 4-h period. The predominant metabolites were putatively identified to be 4-methylgallic acid (4-OMeGA) and 4-methylprotocatechuic acid (4-OMePCA) by analyzing their precursor ions and characteristic fragment ions using tandem mass spectrometry. However, the amount of unchanged GA and PCA that survived the metabolism were about 14.60% and 15.72% of the total intake, respectively, which is reported for the first time in this study.

Optimized microwave-assistant extraction combined ultrasonic pretreatment of flavonoids from Periploca forrestii Schltr. and evaluation of its anti-allergic activity: General

Microwave extraction combined ultrasonic pretreatment of flavonoids from Periploca forrestii Schltr. was investigated in this study, extraction process was first performed in an ultrasonic cleaner, then treated by microwave irradiation. The optimum ultrasonic time of 25 min was selected by single‐factor experiments. A response surface methodology has been used to obtain a mathematical model that describes the process and analyzes the significant parameters ethanol concentration 59.92%, liquid to raw materials ratio 21.24 mL/g, microwave radiation time 209.53 s, and microwave power 274.14 w. In these optimum conditions, the yield of flavonoids from P. forrestii (TFPF) could be up to 9.11 ± 0.08%, which was increased by 14.30 and 19.86% compared microwave extraction and ultrasonic extraction, respectively. In vitro suppress hyaluronidase experimentation showed that TFPF purified using polyamide exhibited good anti‐hyaluronidase ability with IC50 value of 1.033 mg/mL, possessing certain anti‐antiallergic and potential application prospect in pharmaceutical production of treating inflammation and other related fields.

HPLC-DAD-ELSD Combined Pharmacodynamics and Serum Medicinal Chemistry for Quality Assessment of Huangqi Granule

Objective To more scientifically and reasonably control the quality of Huangqi Granules, preliminary studies on the pharmacodynamics and serum pharmacochemistry of this medicine were performed. DPPH and MTT experiments showed that water extracts of Huangqi Granules had good antioxidant activity and increased immunity. Timed blood samples collected 5 min, 15 min, and 30 min after oral administration of a set amount of Huangqi Granules were collected and tested using UPLC-ESI-MS/MS. As a result, calycosin-7-O-β-D-glucoside, ononin, calycosin, astragaloside IV, and formononetin were found to exist in rat blood after dosing, indicating that the five chemical compounds might have pharmacological activity, and based on this result, they were designated biomarkers for quality control of Huangqi Granules. Consequently, a simple, rapid and efficient method was developed in the present study for the simultaneous determination of the five characteristic compounds in Huangqi Granules using HPLC-DAD-ELSD. Materials and Methods The separation was performed using an Agilent Hypersil ODS column (4.6 × 250 mm, 5 μm) at 30 ℃. The mobile phase was composed of water (solvent A) and acetonitrile (solvent B) with a flow rate of 1 mL/min. The drift tube temperature of the ELSD system was set to 85 ℃, and the nitrogen pressure was 3.5 bar. Results All five characteristic compounds had good linear behavior with r2 values greater than 0.9972. The recoveries varied from 96.31% to 101.22%. Subsequently, the developed method was applied to evaluate the quality of Huangqi Granules from different batches, and hierarchical clustering analysis (HCA) was used to analyze the classification of the samples based on the values of the five compounds. Conclusion The established HPLC method combined with HCA proved to be effective to evaluate the quality of Huangqi Granules.