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Sun Y.-X.,Beijing Union University | Sun Y.-X.,CAS Lanzhou Cold and Arid Regions Environmental and Engineering Research Institute | Tang Y.,Peking Union Medical College | Wu A.-L.,Peking Union Medical College | And 4 more authors.
Journal of Asian Natural Products Research | Year: 2010

Our present study was conducted to investigate whether liquiritin (7-hydroxy-2-[4-[3,4,5-trihydroxy-6-(hydroxymethyl) oxan-2-yl] oxyphenyl]-chroman-4-one, 1), an active component of Glycyrrhiza uralensis Fisch., exerts a neuroprotective effect against focal cerebral ischemia/reperfusion (I/R) in male Institute of Cancer Research (ICR) mice. On the establishment of mice with middle cerebral artery occlusion (MCAO) for 2h and reperfusion for 22h, liquiritin at the doses of 40, 20, and 10mg/kg was administered before MCAO once a day intragastrically for a subsequent 3 days. Neurological deficits and infarct volume were measured, respectively. The levels of malondialdehyde (MDA) and carbonyl, activities of superoxide anion (SOD), catalase (CAT) and glutathion peroxidase (GSH-Px) and reduced glutathione/oxidized disulfide (GSH/GSSG) ratio in brain were estimated spectrophotometrically. 8-Hydroxy-2′-deoxyguanosine (8-OHdG) and terminal deoxynucleotidyl transferase-mediated DuTP-biotin nick end labeling (TUNEL)-positive cells were detected by immunohistochemical analysis. Our results showed that the neurological deficits, infarct volume, and the levels of MDA and carbonyl decreased, the ratio of GSH/GSSG and the activities of SOD, CAT, and GSH-Px were compensatorily up-regulated, and 8-OHdG and TUNEL-positive cells decreased after 22h of reperfusion in liquiritin-treated groups. These findings suggest that liquiritin might be a potential agent against cerebral I/R injury in mice by its antioxidant and antiapoptosis properties. © 2010 Taylor & Francis.


Liu Z.,Peking Union Medical College | Liu Z.,Beijing Institute for Drug Control | Feng Z.,Peking Union Medical College | Yang Y.,Peking Union Medical College | And 2 more authors.
Fitoterapia | Year: 2014

Eleven new acyl quinic acid derivatives, 4-O-caffeoyl-3-O-syringoylquinic acid methyl ester (1), 4-O-caffeoyl-3-O-vanilloylquinic acid (2), 4-O-caffeoyl-3-O-vanilloylquinic acid methyl ester (3), 5-O-caffeoyl-3-O-vanilloylquinic acid (4), 5-O-caffeoyl-3-O-vanilloylquinic acid methyl ester (5), 5-O-caffeoyl-3-O-sinapoylquinic acid (6), 5-O-caffeoyl-4-O-vanilloylquinic acid (7), 4-O-(7S, 8R)-glycosmisoyl-5-O-caffeoylquinic acid methyl ester (8), 4-O-(7S, 8-glycosmisoyl-5-O-caffeoylquinic acid (9), 3-O-(7S, 8R)-glycosmisoyl-4-O-caffeoylquinic acid (10), and 3-O-(7S, 8R)-glycosmisoyl-4-O-caffeoylquinic acid methyl ester (11), have been isolated from the stems of Erycibe obtusifolia together with eight known compounds (12-19). Their structures were elucidated on the basis of spectroscopic data analysis (UV, IR, HRESIMS, CD, and 1D and 2D NMR) and chemical evidence. In in vitro assay, compounds 7 and 16-18 exhibited significant neuroprotective effects against rotenone induced PC12 cell damage at 10 μM. © 2014 Elsevier B.V. All rights reserved.


Liu Y.,Beijing University of Chinese Medicine | Zhu X.-Q.,Beijing University of Chinese Medicine | Li W.-D.,Beijing Institute for Drug Control | Wen H.,Beijing University of Chinese Medicine | Liu C.-S.,Beijing University of Chinese Medicine
Chinese Journal of Natural Medicines | Year: 2015

The present study was designed to determine the effects of copy number variations (CNVs) of squalene synthase 1(SQS1) gene on the mevalonate (MVA) pathway. SQS1 gene from G. uralensis (GuSQS1) was cloned and over-expressed in Pichia pastoris GS115. Six recombinant P. pastoris strains containing different copy number of GuSQS1 were constructed. HPLC was used to assay the level of ergosterol in all transgenic P. pastoris strains containing GuSQS1. HPLC analysis showed that the contents of ergosterol in all of the transgenic P. pastoris containing GuSQS1 were higher than that in the negative control. And with the increase of copy number of GuSQS1, the content of ergosterol showed an increasing-decreasing-increasing pattern. The contents of ergosterol in 10-copy- GuSQS1 P. pastoris and 47-copy- GuSQS1 P. pastoris were significantly higher than that in the rest recombinant P. pastoris strains. In conclusion, the CNVs of GuSQS1 influence the content of secondary metabolites in the MVA pathway. The present study provides a basis for over-expressing GuSQS1 and increasing the content of glycyrrhizin in G. uralensis cultivars. © 2015 China Pharmaceutical University.


Zhang W.,Beijing Institute for Drug Control
Chinese Journal of Biologicals | Year: 2014

Objective: To determine the citrate ion content in human coagulation factor VIII (FVIII) by high performance anion exchange chromatography (HPAEC). Methods: The condition for HPAEC was as follows: chromatographic column: IonPac AS11-HC(4 mm x 250 mm); mobile phase: 20 mmol/L sodium hydroxide solution; time for isocratic elution: 20 min; flow rate; 1.0 ml/min; inhibitor: ASRS 4 mm anion inhibitor; inhibition current: 120 mA; detector: electrical conductivity detector; detector cell temperature: 30°C; sample loading: 25 μl. The developed method was determined for linear range and detection limit, verified for accuracy and precision, and applied preliminarily. Results: The chromatographic peak area of reference showed good linear relationship to citrate ion content at a range of 0.1-2.0 μg/ml (r = 0.999 0). Serving the signal-to-noise (S/N) ratio as 3, the detection limit of the developed method was 0.01 μg/ml. The total mean recovery of citrate ion in accuracy test was 97.80%, with a RSD of 0.69%. The RSD of 6 test results of one sample of FVIII was 0.08%. The determination results of three batches of FVIII by HPAEC, colorimetry and cation exchange chromatography were basically in agreement, which met the requirements in Chinese Pharmacopoeia(Volume III, 2010 edition). Conclusion: HPAEC may be used for determination of citrate ion content in FVIII, which was simple and rapid, with high accuracy and precision.


Fang X.,Shandong Agricultural University | Fang X.,State Key Laboratory of Crop Biology | Wang J.,Shandong Agricultural University | Wang J.,State Key Laboratory of Crop Biology | And 3 more authors.
Journal of Separation Science | Year: 2010

An optimized microwave-assisted extraction (MAE) method and an efficient HPLC analysis method were developed for fast extraction and simultaneous determination of oleanolic acid and ursolic acid in the fruit of Chaenomeles sinensis. The open vessel MAE process was optimized by using a central composite experimental design. The optimal conditions identified were microwave power 600 W, temperature 52°C, solvent to material ratio 32 mL/g and extraction time 7 min. The results showed that MAE is a more rapid extraction method with higher yield and lower solvent consumption. The HPLC-photodiode array detection analysis method was validated to have good linearity, precision, reproduction and accuracy. Compared with conventional extraction and analysis methods, MAE-HPLC-photodiode array detection is a faster, convenient and appropriate method for determination of oleanolic acid and ursolic acid in the fruits of C. sinensis. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA.


Zhao Y.-Y.,Beijing Institute for Drug Control | Wang J.-H.,Beijing Institute for Drug Control | Fu X.-T.,Beijing Institute for Drug Control | Chen Y.-G.,Beijing Institute for Drug Control | Guo H.-Z.,Beijing Institute for Drug Control
Yaoxue Xuebao | Year: 2013

A new UPLC method was developed for the simultaneous determination of eleven characteristic flavonoid glycosides in Ginkgo biloba leaves. The natural occurrence of flavonoid glycosides in Ginkgo biloba leaves within one vegetative season was investigated for the first time. The analysis was performed on an Agilent ZORBAX Eclipse Plus C18 column (50 mm x 4.6 mm, 1.8 μm), the mobile phase A was acetonitrile, the mobile phase B was 0.4% phosphate aqueous solution in a gradient elution at a flow rate of 0.6 mL·min -1, the detection was carried out at 360 nm. The result showed that eleven flavonoid glycosides had good linearity with good average recovery, separately. The method was proved to be accurate, rapid and good reproducible for the quality evaluation of Ginkgo biloba leaves, and provide an easy and rapid means for the quantitative analysis of flavonoid glycosides and their content fluctuation with seasons.


Zhao H.-Y.,Chinese Institute of Materia Medica | Fan M.-X.,Chinese Institute of Materia Medica | Fan M.-X.,Beijing Institute for Drug Control | Wu X.,Chinese Institute of Materia Medica | And 4 more authors.
Journal of Chromatographic Science | Year: 2013

An approach was established to analyze the chemical profiling of Xiao-Cheng-Qi Decoction (XCQD) using liquid chromatography coupled with electrospray ionization tandem mass spectrometry. XCQD consisted of three herbal medicines (Rhubarb, Fructus Aurantii Immaturus and Cortex Magnoliae Officinalis). The traditional water extractive method was applied in the sample preparation, which was identical with clinical use. The characteristic fragmentation pathways of 17 reference compounds were comprehensively studied, including precursors of tannins, flavonones, anthraquinones and lignan. In total, 71 constituents were identified or tentatively characterized based on their mass spectrometry fragmentations and chromatographic behaviors. By comparing their relative contents, flavanones and anthraquinones were supposed to be used together for the quality control of XCQD. Further pharmacology and pharmacokinetics investigations should be performed on the basis of the present chemical profiling study. © The Author [2012]. Published by Oxford University Press. All rights reserved.


PubMed | University of Sichuan and Beijing Institute for Drug Control
Type: | Journal: Journal of chromatographic science | Year: 2016

In this study, a selective and sensitive liquid chromatography-electrospray ionization-tandem mass spectrometric method was developed and validated for the determination of lycobetaine in rat plasma. Berberine was selected as the internal standard, and rat plasma samples were pretreated via protein precipitation and further separated on a diamonsil octadecyl-silylated silica column using 0.2% (v/v) aqueous formic acid and methanol as the mobile phase. Selected reaction monitoring was performed using the transitions m/z 266.1 208.1 and m/z 336.1 320.0 to determine the concentrations of lycobetaine and internal standard, respectively. The injection volume was 1 L, and the calibration curve was linear (R


PubMed | University of Sichuan and Beijing Institute for Drug Control
Type: | Journal: Colloids and surfaces. B, Biointerfaces | Year: 2016

Paclitaxel (PTX) is a widely used antineoplastic drug in clinic. Due to poor aqueous solubility, it is administrated by intravenous infusion of cremophor EL containing formulation with serious adverse effects. The low oral bioavailability is a great challenge for oral formulation development. In addition, P-gp mediated multidrug resistance limit its clinical use in various resistant cancers. In this study, a novel super-antiresistant PTX micelle formulation for oral administration was developed. A P-gp inhibitor, bromotetrandrine (W198) was co-encapsulated in the micelle. The micelles were composed of Solutol HS 15 and d-a-tocopheryl polyethylene glycol succinate to avoid Cremophor EL induced toxicity. The micelles were round with a mean particle size of 13nm and an encapsulation efficiency of 90%. A series of in vitro evaluations were performed in non-resistant MCF-7 cells and resistant MCF-7/Adr cells. The super-antiresistant PTX micelles showed higher cell uptake efficiency, significantly increased cytotoxicity and antimitotic effect in drug resistant MCF-7/Adr cells when compared with Taxol and other PTX micelle formulations. Compared with Taxol, the super-antiresistant PTX micelles significantly improved bioavailability after oral administration in rats, and inhibited tumor growth in multidrug resistance xenografted MCF-7/Adr nude mice. In summary, the noval super-antiresistant PTX micelles showed a great potential for oral delivery of PTX against resistant breast cancer.


PubMed | University of Sichuan and Beijing Institute for Drug Control
Type: | Journal: Journal of pharmaceutical and biomedical analysis | Year: 2015

A sensitive, selective and rapid liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-MS/MS) method has been developed for the quantification of the novel transforming growth factor- (TGF-) inhibitor SB-505124 in rat plasma and then validated. Plasma samples were prepared by simple protein precipitation. Separation was performed on a Diamonsil ODS chromatography column using a mobile phase of acetonitrile and 0.1% (v/v) aqueous formic acid. SB-505124 and the internal standard doxorubicin were detected in the positive ion mode using multiple reaction monitoring of the transitions at m/z 336.2320.1 and 544.2397.2, respectively. Calibration curve was linear (r>0.9996) over a concentration range of 10-5000 ng/mL with the lower quantification limit of 10 ng/mL. Both intra- and inter-day precision were within 6.5% and trueness were not more than 3.1%. Extraction recovery and matrix effect were within acceptable limits. Stability tests showed that SB-505124 and the IS remained stable throughout the analytical procedure. The validated LC-MS/MS method was then used to analyze the pharmacokinetics of SB-505124 administered to rats intravenously (8 mg/kg) or orally (10 mg/kg). Oral bioavailability of SB-505124 was calculated as 76.4%, indicating the potential of SB-505124 as an orally administered drug.

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