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Core-shell magnetic zeolite imidazolate framework-8 (Fe3O4@PAA@ZIF-8) was successfully synthesized and first employed as adsorbent of magnetic solid-phase extraction (MSPE) for the determination of brucine and strychnine in human urine sample coupled with high performance liquid chromatography. The as-prepared Fe3O4@PAA@ZIF-8 was characterized by transmission electron microscopy, Fourier-transform infrared spectrometry, X-ray diffraction, vibrating sample magnetometer and zeta potential analysis. Main parameters affecting the MSPE efficiency, including amount of adsorbent, sample solution pH, extraction time, ionic strength, desorption solvent, desorption time and desorption volume were further optimized. Under the optimized conditions, the proposed method provided good linearity (5.0-1000.0 µg L-1) with determination coefficients between 1.0000 and 0.9998, low limits of detection in the range of 1.1-1.2 µg L-1, and excellent reproducibility with relative standard deviations of less than 7.7%. The intra-day and inter-day precision were 1.5-3.2% and 2.1-7.2%, respectively. Satisfactory spiked recoveries were between97.2% and 115.4% with the relative standard deviations less than 6.3%. The results demonstrated that Fe3O4@PAA@ZIF-8 composite was a promising magnetic adsorbent for the preconcentration of brucine and strychnine in human urine sample.

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The traditional Chinese medicine of Radix Hedysari plays an important role in invigorating gas for ascending, benefiting blood for promoting production of fluid, and promoting circulation for removing obstruction in collaterals, which is consistent with the principle of treatment for osteoporosis. This study is designed to investigate the bioactive components on increasing peak bone mass (PBM) by exploring the spectrum-effect relationship between chromatography fingerprints and effect. Multiple indicators are selected to evaluate the pharmacological activity. In fingerprints, 21 common peaks are obtained, five of which are identified. Furthermore, gray relational analysis (GRA) is a quantitative method of gray system theory and is used to describe the correlation degree of common peaks and pharmacological activities with relational value. 21 components are then divided into three different regions, of which ononin and calycosin play an extremely significant role in increasing PBM. In addition, factor analysis and hierarchical cluster analysis (HCA) are used to screen the optimal producing area for Radix Hedysari. This provides a comprehensive and efficient method to improve the quality evaluation of Radix Hedysari, confirming the bioactive components for PBM-enhancement and further develop its medicinal value.

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Drug metabolism is significantly affected under hypoxia environment with changes of pharmacokinetics, expression and function of drug-metabolizing enzymes and transporters. Studies have shown that hypoxia increases the release of a series of inflammatory cytokines which can modulate drug metabolism. Besides, both hypoxia inducible factor 1α (HIF-1α) and microRNA-mediated pathways play a role in regulating drug metabolism. This article reviewed the impact and single-factor modulating mechanisms of drug metabolism under hypoxia, and put forward the speculation and prospects of multi-factor modulating mechanisms.

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In this study, complex enzymes combined with ultrasonic extraction technology（MC） were used, to select optimal extraction combinations by single factor and orthogonal test, with Hedysarum polysaccharides yield and content as the comprehensive indexes. The components, physicochemical properties and antioxidant activity of Hedysarum polysaccharides from complex enzyme combined with ultrasonic extraction（HPS-MC）and the Hedysarum polysaccharides from hot water extraction（HPS-R）were analyzed. The results showed that：complex enzymes had significant effect on the yield and content of Hedysarum polysaccharides, and the ultrasonic power could significantly improve the content of Hedysarum polysaccharides. The optimum technological parameters were as follows: complex enzyme ratio 1:1, ultrasonic power 105 W, ultrasonic time 60 min, and enzymatic hydrolysis pH 5, achieving （14.01±0.64）% and （92.45±1.47）% respectively for the yield and content of Polysaccharides. As compared with HPS-R, the molecular weight, absolute viscosity and protein content of HPS-MC were decreased, while the content of uronic acid was increased. In the antioxidant system, the concentration of polysaccharide was within the range of 1-7 g·L⁻¹; the antioxidant activity of HPS-MC was higher than that of HPS-R, and HPS-MC (80%) with the lowest molecular weight showed a significant dose effect relationship with the increase of the experimental concentration. In conclusion, MC is a simple, convenient, economical and environmentally friendly extraction technology, and the Hedysarum polysaccharides extracted by this method have obvious antioxidant activity.

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The purpose of this study was to compare the effects of different chromatographic columns for the separation of seven flavonoids. Four different stationary phases are available, including bridged ethyl hybrid, BEH and the same hybrid phase modified with 2-ethylpyridine, CSH fluorophenyl, and HSS C18 SB. The analytes included calycosin, genistein, medicarpin, calycosin-7-O-ß-d-glucoside, formononetin, formononetin-7-O-ß-d-glucoside, and liquiritigenin. The CSH fluorophenyl column was determined to be the most suitable and provided the fastest separation within 17 min using gradient elution with carbon dioxide as the mobile phase and methanol as the co-solvent. Good peak shapes were obtained, and the values of the peak asymmetry were close to 1.0 for all of the flavonoids. The resolution was more than 1.41 for all of the separated peaks. Baseline separation on the optimal columns was achieved by changing the co-solvent type and adjusting the temperature and pressure. Quantitative performance was evaluated under optimized conditions, and method validation was accomplished. The validation parameters, such as linearity, sensitivity, precision, and accuracy, were satisfactory. Good repeatability of both peak area (relative standard deviation <1.02%) and retention time (relative standard deviation <0.88%) was observed. The optimized chromatographic methods were successfully used for the determination of seven flavonoids in Radix astragali. The sensitivity was sufficient for the analysis of real samples.

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A rapid and simple ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS) method for the determination of astragaloside III was developed and used in a pharmacokinetic and tissue distribution study in rats following the oral administration 95% ethanol extraction of Zhenqi Fuzheng capsules. Although astragaloside III and astragaloside IV have the same molecular weight and very similar structures, they were successfully separated using this method. Quantification was performed using low-energy collision tandem mass spectrometry (CID-MS-MS) with the multiple reaction monitoring scan mode of the following precursor ion → product ion atm/z807.61→335.22 for astragaloside III and atm/z633.18→331.18 for the internal standard (hesperidin). Both astragaloside III and astragaloside IV in rat plasma were best fit to a two-compartment model. The tissue distribution study showed the overall trend of disposition of astragaloside III were C thymus > C spleen > C stomach > C liver > C heart > C kidney > C lung > C testicle The high levels of astragaloside III in thymus and spleen indicated an accumulation in organs involved in immune responses and showed that these organs are major target sites in vivo The results in the article will provide valuable information for use in clinical applications of astragaloside III.

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An HPLC method was established for the determination of adenosine, γ-aminobutyric acid (GABA) and six flavonoids (calycosin-7-glucoside, ononin, calycosin, isoliquiritigenin, formononetin and medicarpin） in Radix Hedysari. The samples were extracted with methanol by refluxing for 4 h. The HPLC-DAD was performed on a Diamonsil C(18) column (250 mm × 4.6 mm, 5 µm) with acetonitrile-water as the mobile phase. The column temperature was at 40 ℃ and the flow rate was 1.0 m L·min(-1), while the temperature of drift tube was 110.5 ℃ and the nebulizing gas flow was 3.1 L·min-1 for the ELSD system. The results showed all the eight components had good linear relationships (r(2) =0.992 8-1.000 0) in the range of the test concentration. The RSD of precision, stability and repeatability were less than 2%.The average recovery rates were 96.78%-103.45%, and RSD were 0.29%-1.61%.The index component contents of Radix Hedysari form 24 different origins were determined and used as variable factors in clustering analysis. The results were classified into 2 groups basically in accordance with the regional cluster. And the consequence was in consistent with the results of principal component analysis. This HPLC method is simple, shows good sensitive and accurate, and provides the experimental basis for multi-index control of Radix Hedysari. Clustering analysis for Radix Hedysari quality control has a certain reliability and objectivity.

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Radix Astragali (Huangqi in Chinese) and Fructus Ligustri Lucidi (Nvzhenzi in Chinese) (2:1, w/w) are combined in an herbal formulation called Zhenqi Fuzheng capsules (ZFCs) for use in China to improve immunity, promote the recovery of normal functions after surgical operations, and as the most important adjuvant therapy in cancer. In this study, the tissue distribution profiles of the six major bio-active constituents (calycosin-7-O-ß-D-glucoside, ononin, calycosin, formononetin, astragaloside IV and astragaloside II) were examined after oral administration of ZFCs to rats. All six constituents in each tissue were detected simultaneously using UPLC-ESI-MS, and the concentration of each constituent per gram of each tissue was determined. Quantification was performed using low-energy collision tandem mass spectrometry (CID-MS/MS) in multiple reaction monitoring (MRM) scan mode for the following precursor ion→product ion transitions at m/z 447.21→285.30 for calycosin-7-O-ß-D-glucoside, m/z 285.29→270.38 for calycosin, m/z 431→269 for ononin, m/z 269→237 for formononetin, m/z 807.40→627.50 for astragaloside IV, m/z 849.60→669.65 for astragaloside II and m/z 633.18→331.18 for the internal standard (hesperidin). The results showed that in general the tissue concentrations for all six constituents were in the following order: spleen>stomach>thymus>lung>liver>kidney>heart>testicle. The high levels in the spleen and thymus indicated that all six compounds accumulated in organs involved in the immune response, consistent with the immunity effects of ZFC. The high levels in the stomach were consistent with the oral administration of ZFC. This study was the first to compare the tissue distribution of calycosin-7-O-ß-D-glucoside with that of calycosin or of ononin with that of formononetin in rats. It was also the first study to examine the tissue distribution of astragaloside II, calycosin and formononetin following oral administration of ZFC to rats.

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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.

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This study provided a comprehensive component analysis and structure identification of active substances for the anti-gastric ulcer effects of Radix Astragali. The data were generated by organically combining the results from in vivo pharmacodynamic experiments, a cell growth-promoting assay, structure identification, content determination, fingerprinting, and correlation analyses. The fingerprints from high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD) and from HPLC coupled with evaporative light scattering detectors (ELSD) from 95% ethanol extracts of Radix Astragali (ERA) were determined using HPLC-DAD-ELSD. The structures of 16 compounds were identified using ultra-pressure liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS). The contents of these 16 compounds were simultaneously determined in a single run using HPLC-DAD-ELSD. The strength of the anti-ulceration effect of each of the 16 compounds was correlated to its content in the HPLC spectrum using gray relation statistics. The sequence of the contribution from each of the 16 compounds to the anti-gastric ulcer effect was determined. The results showed that ononin, astragalosideIII, and astragalosideIV contributed most to the observed anti-gastric ulcer effects and that these three compounds also exhibited strong growth-promoting effects in cultured GES-1 cells. The results of this study can be used to evaluate the quality of Radix Astragali and to provide a theoretical foundation for its further study.

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A sensitive and reliable ultra-pressure liquid chromatography with tandem mass spectrometry (UPLC-MS) was developed and validated for simultaneous quantification of six main bioactive components, i.e., calycosin-7-O-ß-D-glucoside, ononin, calycosin, formononetin, astragaloside IV, and astragaloside II in rat plasma after oral administration of the 95 % ethanol extraction from Radix Astragali. Plasma samples were extracted with Waters Oasis(TM) HLB 1 cc (30 mg) Extraction Cartridges (SPE) separated on an UPLC™ BEH C18 column and detected by MS with electro spray ionization interface in positive selective ion monitoring mode. Calibration curves offered linear ranges of two orders of magnitude with r (2) > 0.99. The method had the lower limit quantification of 1.30, 0.73, 1.17, 2.33, 0.63, and 0.83 ng/mL for ononin, calycosin, calycosin-7-O-ß-D-glucoside, formononetin, astragaloside IV, and astragaloside II, respectively, with precision less than 10 %. The RSD of intra- and inter-day variations ranged from 1.66 to 6.46 and 3.39 to 6.58 %. This developed method was applied subsequently to pharmacokinetic studies of the six compounds in rats successfully. The proposed method was for the first time to compare the pharmacokinetic difference between calycosin-7-O-ß-D-glucoside and calycosin in rat plasma, so as between ononin and formononetin, and studied to the astragaloside II pharmacokinetics in rat plasma.

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HPS1-D, an active polysaccharide,was isolated and purified from Hedysarum polybotrys. HPS1-D was obtained after treated with Savage method and H2O2, and purified with DEAE-cellulose 52 and Sephadex G-100 gel filtration chromatography. Then physicochemical property analysis, GC, methylation, partial acid hydrolysis, and NMR method were used to study chemical structural of HPS1-D. The conformation was primarily analyzed with GPC-MALLS method and Congo red reaction. The anti-complementary activity of HPS1-D was evaluated with the hemolysis assay. HPS1-D was a heteropolysaccharide and consisted of D-glucose, L-arabinose, (7.2:1.3). HPS1-D proved to be a neutral sugar, with 1, 4-and 1, 4, 6-alpha-D-glucopyranosyl residues in backbone ,and 1, 5-and 1, 3, 5-alpha-L-arabinofuranosyl residues in branches. HPS1-D has a random coil state conformation with monodisperse mass distribution in 0.9% NaCl solution. And HPS1-D had triple-helix conformation in concentrate of NaOH solution. Anti-complementary activity of HPS1-D was closed to its positive control heparin.

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A water-soluble polysaccharide (HPS3aS) with a molecular mass of 1.22 × 10(4) Da was isolated from Hedysarum polybotrys using anion-exchange and gel-permeation chromatography. HPS3aS exhibits a globular-shaped conformation in 0.1 M NaNO3 by size exclusion chromatography with multi-angle laser light scattering (SEC-MALLS). The investigation of the structural features of this heteropolysaccharide through a combination of chemical and instrumental analyses revealed that the backbone of HPS3aS is composed of α-D-(1 → 4)-linked glucopyranose residues, which occasionally branched at O-6. The branches are composed of (1 → 4)-linked galactopyranose residues and terminated with glucopyranose residues. HPS3aS possesses good in vitro antioxidant activity, as evaluated by scavenging assays with 1,1-diphenyl-2-picrylhydrazyl, hydroxyl, and superoxide radicals, which suggests that HPS3aS could be a potential antioxidant.

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SHG was sulfated by chlorosulfonic acid-pyridine method, and six samples which we got were prepared in different reaction conditions. There is a characteristic absorption peak near 260 nm in UV spectra and there are two characteristic absorption peaks near 1240 cm(-1) and 810 cm(-1) in the FT-IR. Degree of sulfation (DS) was calculated by elemental analysis and turbidimetry. Under the same conditions the absorption peaks become strong with the DS increase. The anticoagulant activity of SHG and sulfated modification samples was evaluated by the classic coagulant assays of prothrombin time (PT), activated partial thrombin time (APTT) live enzymes, and plasma thrombin time (TT). Results show that sulfated SHG has a good anticoagulant activity in vitro, and DS increased activity within a certain range.

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Blood deficiency model of mice was copied by subcutaneous injection with 200, 100 and 100 mg x kg(-1) (0.01 mL x g(-1)) acetyl phenylhydrazine (APH) at the frist, fourth, and seventh days. Mice in each group were perfused with different extracted parts of Angelica sinensis (drug dosage was 2.4 g x kg(-1)) at the tenth day, once a day for 10 days. Then compare the influence of different extracted parts of Angelica sinensis to RBC, Hb, PLT and thymus, spleen and weight changes of blood deficiency mice. The peak areas of each common peak from HPLC fingerprint were associated with the date of replenishing blood pharmacodynamics efficacy by using gray relation statistic, which was used to research the chromatogram-pharmacodynamics relationship. The results showed that the part of DSC has the better effect in replenishing blood. The contribution degree of the DSC to replenishing blood of each component were determined by correlation size, and ferulic acid made the largest contribution, but contribution of other components should not be ignored. In this paper, we research the relationship of the HPLC fingerprint and spectrum activity relationship, determine the material basis of the DSC for replenishing blood, and provide effective way to represent the spectral correlation effect.

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OBJECTIVE: To analyze the relationship between high-performance liquid chromatography (HPLC) fingerprints of the chloroform extract fractions of Peucedanum harrysmithii var. subglabrum (PHS) and its phlegm-reducing effect, in order to establish "active component group for reducing phlegm". METHOD: HPLC was adopted to determine and analyze HPLC fingerprints of chloroform extract fractions of PHS. Phenol red expectorant experiment was used to observe the phlegm-reducing effect in mice. Mice were administered intragastrically with chloroform extract fractions for 6 days (1.4 g x kg(-1)), with acute bronchitis syrup as the positive control drug (12 mL x kg(-1)). The phenol red secretion in mice was determined by spectrophotometer. Then the grey relational analysis was used to study the spectrum-effect relationship. RESULT: The phlegm-reducing effect of the chloroform extract fractions of PHS were resulted from the combined effect of all of its chemical components. Its various characteristic peaks represented different chemical components, and the order of their contributions to the phlegm-reducing effect was (number of peaks) 13 > 12 > 16 > 18 > 19 > 6 > 20 > 14 > 1 > 11 > 15 > 10 > 17 > 2 > 5 > 4 > 7 > 3 > 8 > 9, in No. 1, 3, 4, 10, 13 and 16 characteristic peaks were identified as marmesin, psoralen, xanthotoxin, Pd-Ib, pteryxin and peuformosin. CONCLUSION: The chloroform extract fractions of PHS show strongly phlegm-reducing effect. There may be certain relationship between their HPLC fingerprint and phlegm-reducing effect.

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OBJECTIVE: To elaborate the correlation of chromatography fingerprint of Astragali Radix and the efficacy of improving immunity, and express the effective substances foundation. METHODS: The water extract was given to irrigation stomach of mice for carbon clearance experiment. Associated the peak area of each common peak from HPLC fingerprint and the date of improving immunity, and studied the correlation between fingerprint and efficacy and found out the effective substances foundation of improving immunity in the method of gray correlation analysis. RESULTS: Polysacharides was the main component of the water extract of Astragali Radix, which could improve immunity function. In carbon clearance experiment, the phagocytic index of the mice with water extract was the largest, and the ability to enhance immunity was the strongest. Grey correlation analysis was used to evaluate the correlation of the effect of improving immunity and the component of the water extract of Astragali Radix. CONCLUSION: It is effective to study the fingerprint and the correlation of fingerprint and the efficacy of traditional chinese medicine for expressing its correlation.

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A quantitative determination method of the total flavonoids in Olea europaea L. leaves using rutin as a standard substance was developed. Two coloration systems including Al(NO3)3-NaNO2-NaOH and AlCl3 were both used. The latter was selected as the optimal determination method by comparing with the results. The main interference of the matrix was discussed. The result showed that the AlCl3 coloration method is simple, accurate and reproducible with good linear relationship, and it can be looked as a reference method for the determination of total flavonoids in herbal materials or natural products.

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A simple and accurate high-performance liquid chromatography (HPLC) coupled with diode array detector (DAD) and evaporative light scattering detector (ELSD) was established for the determination of six bioactive compounds in Zhenqi Fuzheng preparation (ZFP). The monitoring wavelengths were 254, 275 and 328 nm. Under the optimum conditions, good separation was achieved, and the assay was fully validated in respect of precision, repeatability and accuracy. The proposed method was successfully applied to quantify the six ingredients in 31 batches of ZFP samples and evaluate the variation by hierarchical cluster analysis (HCA), which demonstrated significant variations on the content of these compounds in the samples from different manufacturers with different preparation procedures. The developed HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of this preparation.

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An approach was proposed to evaluate preparation technology by means of fingerprint-peak matching technology of high Performance liquid chromatography with diode array detector (HPLC-DAD). Similarity and hierarchical clustering analysis (HCA) were applied to identify the 15 batches of Xiaochaihu granules from different manufacturers and our laboratory, and peak pattern matching between the composite formulae and Radix Bupleuri Chinensis, which was one of the main ingredients of Xiaochaihu granules, was utilized to evaluate the preparation technology of Xiaochaihu granules via the indexes of the relative deviation of retention time (RT) and UV spectrum feature similarity of their corresponding peaks. Eleven matching peaks were found between Xiaochaihu granules and Radix Bupleuri Chinensis. However, the saikosaponin A and saikosaponin D, which are the important active components in Radix Bupleuri Chinensis, were not found in Xiaochaihu granules from any manufacturers. The peak areas of 11 characteristic peaks of Xiaochaihu granules samples formed a matrix of 11 × 15. The result of HCA showed that Xiaochaihu granules samples were divided into four kinds of category. Xiaochaihu granules samples from the same manufacturer were basically clustered of the same category. The results suggested that the saikosaponin A and saikosaponin D are prone to structural transformation under the condition of decoction and in the presence of the organic acidic components. These active components, existing in raw herb, might transform to a series of non-active secondary saikosaponin due to unfavourable preparation technology. So the conventional decoction-based preparation technology of Xiaochaihu granules might greatly affect its quality and therapeutic effectiveness. This study demonstrates that fingerprint-peak matching technology can not only be used for quality control of this composite formulae, but also provide some guidance for preparation technology of Xiaochaihu granules.