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Chen Y.,Nanjing University | Chen J.-W.,Nanjing University | Chen J.-W.,Jiangsu Key Laboratory for TCM Formulae Research | Liu S.-J.,Nanjing University of Traditional Chinese Medicine | And 6 more authors.
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences | Year: 2012

A liquid chromatography-mass spectrometry method has been developed and validated for the quantification of bullatacin, a bistetrahydrofuran annonaceous acetogenin, in rat plasma. Squamostatin-A was selected as the internal standard. Analytes were extracted from rat plasma by liquid/liquid extraction using ethyl acetate with high efficiency. The chromatographical separation was performed on an Agilent Zorbax SB-C 18 column (150mm×2.1mm, 5μm). The mobile phase consisted of methanol and deionized water (95:5, v/v) containing 0.01% (v/v) formic acid. The chromatographic run time was 7min per injection and flow rate was 0.2mL/min. The retention time was 3.22 and 5.23min for internal standard and bullatacin, respectively. The elutes were detected under positive electrospray ionization and the target analytes quantified by selected ion monitoring mode (645.9m/z for bullatacin and 661.9m/z for squamostatin-A). The method was sensitive with the limit of quantitation at 0.5ng/mL in 100μL of rat plasma. Good linearity (r 2=0.9998) was obtained covering the concentration of 0.5-2000ng/mL. The intra- and inter-day assay precision ranged from 3.2 to 8.7% and 2.7 to 9.2%, respectively. In addition, the stability, extraction recovery and matrix effect involved in the method were also validated. This method was applied to measure the plasma bullatacin concentrations after a single tail vein intravenous administration of bullatacin in rats. © 2012 Elsevier B.V.

Yuan F.,Nanjing University | Bai G.,Nanjing University | Chen Y.,Nanjing University | Miao Y.,Nanjing University | And 4 more authors.
Bioorganic and Medicinal Chemistry Letters | Year: 2015

Fifteen annonaceous acetogenins (ACGs) with different stereochemical structures and configuration, representing three main classes of bis-adjacent-tetrahydrofuran (THF), bis-nonadjacent-THF, and mono-THF ACGs, were selected to tested for their inhibition activity on A549/Taxol cell line, which is multidrug resistant (MDR). The present study showed that some tested compounds showed significant activity toward A549/Taxol cells, and were more potent than the positive control Verapamil. For example, squamostatin-D (14) (IC50 value = 16.19 μM) was 7.8 times more active than Verapamil (IC50 value = 127.09 μM). Those ACGs with more carbons between the THF ring and the γ-unsaturated lactone were more potent. Moreover, ACGs with stereochemical arrangement of erythro were more active than those of threo, the compounds with THF ring configuration of cis seemed to be superior to those of trans. However, if all other structural features were identical, the ACGs with more hydroxyls on the aliphatic chain were not more potent towards A549/Taxol, which was not in accordance with previous studies. Furthermore, bis-nonadjacent-THF ACGs whose molecular weight is 622, with three hydroxyl groups located at carbon 16, 19, 24 and stereochemical arrangement of erythro possibly produced notable cytotoxicity. Based on the above conclusions, we proposed a compound model that may be a promising anti-MDR cancer candidate drug in the future clinical trial. © 2015 Elsevier Ltd. All rights reserved.

Bai G.-G.,Nanjing University | Yuan F.,Nanjing University | Mao K.-J.,Nanjing University | Li F.-Q.,Nanjing University | And 4 more authors.
Chinese Traditional and Herbal Drugs | Year: 2014

Objective: To study the chemical constituents from the root bark of Changium smyrnioides. Methods: Compounds were isolated by various kinds of column chromatographies on silica gel, Sephadex LH-20, and recycling preparative HPLC from the ethanol extract in the root bark of C. smyrnioides, and their structures were elucidated by the physicochemical characteristics and spectral analyses. Results: Fifteen chemical constituents were obtained and identified as imperatorin (1), phellopterin (2), xanthotoxol (3), 5-hydroxy-8-methoxy-psoralen (4), vanillic acid (5), alloimperatorin (6), psoralen (7), bergapten (8), 8-O-β-D-glucopyranosyl- 5-methoxylpsoralen (9), isopimpinellin (10), caffeic acid (11), aurantiamide acetate (12), vaginatin (13), β-sitosterol (14), and succinic acid (15). Conclusion: Compounds 6-13 are isolated from the plants in Changium Wolff for the first time.

Yao X.,Nanjing University | Bu D.,Nanjing University | Chen J.-W.,Nanjing University | Chen J.-W.,Jiangsu Key Laboratory for TCM Formulae Research | And 3 more authors.
Chinese Traditional and Herbal Drugs | Year: 2014

Objective: To investigate some factors which could influence the tissue culture of callus from Taxus chinensis var. mairei such as the culturing callus time, callus induction rate, callus development rate, growth rate, and browning degree. Then, the five taxane diterpenoids including paclitaxel in callus were studied and compared. Methods: The factors which influence the T. chinensis var. mairei callus induction and growth rate were studied by single factor analysis and orthogonal test design. The five taxane diterpenoids including paclitaxel in callus from different sources were determined by HPLC analysis. The weighted score for each of the above factors mentioned would be taken account to optimise the tissue culture conditions. Results: Among the explants, the stems intacted leaves would exhibit the best ability of dedifferentiation to form callus. The enrichment dark culture was conducive to paclitaxel and other five kinds of terpene constituents. The most optimum culture medium of the callus induction was B5 + 2, 4-D 3.0 mg/L + 6-BA 1.0 mg/L + NAA 1.0 mg/L + KT 0.5 mg/L. Conclusion: The callus culture conditions for T. chinensis var. mairei have indicated greater association with different types of the explantation, nutrient medium, and light illumination. The five kinds of enrichment taxane paclitaxel diterpenoids would be significantly affected by the type and concentration of the plant growth regulators. This study has shown some references value for T. chinensis var. mairei tissue culture and for the screening of high-yielding cell lines. ©, 2014, Editorial Office of Chinese Traditional and Herbal Drugs. All right reserved.

Xu C.-L.,Nanjing University | Chen J.-W.,Nanjing University | Chen J.-W.,Jiangsu Key Laboratory for TCM Formulae Research | Ju W.-Z.,Nanjing University of Traditional Chinese Medicine | And 7 more authors.
Biomedical Chromatography | Year: 2012

A rapid, sensitive and specific method using liquid chromatography with tandem mass spectrometric detection (LC-MS) was developed for the analysis of sauchinone in rat plasma. Di-O-methyltetrahydrofuriguaiacin B was used as internal standard (IS). Analytes were extracted from rat plasma by liquid-liquid extraction using ethyl acetate. A 2.1mm i.d. × 150mm, 5μm, Agilent Zorbax SB-C18 column was used to perform the chromatographic analysis. The mobile phase was methanol-deionized water (80:20, v/v). The chromatographic run time was 7min per injection and the flow-rate was 0.2mL/min. The tandem mass spectrometric detection mode was achieved with electrospray ionization interface in positive-ion mode (ESI+). The m/z ratios [M+Na]+, m/z 379.4 for sauchinone and m/z 395.4 for IS were recorded simultaneously. Calibration curve were linear over the range of 0.01-5μg/mL. The lowest limit of quantification was 0.01μg/mL. The intra-day and inter-day precision and accuracy of the quality control samples were 2.94-9.42% and 95.79-108.05%, respectively. The matrix effect was 64.20-67.34% and the extraction recovery was 93.28-95.98%. This method was simple and sensitive enough to be used in pharmacokinetic research for determination of sauchinone in rat plasma. © 2011 John Wiley & Sons, Ltd.

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