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      Study on chemical fingerprinting of crude and processed Atractylodes macrocephala from different locations in Zhejiang province by reversed-phase high-performance liquid chromatography coupled with hierarchical cluster analysis

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          Abstract

          Background:

          In China, Atractylodes macrocephala is mainly distributed in Zhejiang province. It is, therefore, desirable to determine a reliable and accurate methodology to differentiate the samples collected from Zhejiang province. Although some studies on the fingerprints of Atractylodes macrocephala using HPLC have been published, none of them compared the processed product of Atractylodes macrocephala from different areas of Zhejiang province.

          Materials and Methods:

          Reversed-phase high-performance liquid chromatography (RP-HPLC) coupled with hierarchical cluster analysis was employed in the fingerprint analysis of Atractylodes macrocephala from Zhejiang province, China. The LC assay was performed on a reversed-phase C 18 column with linear gradient elution using water and acetonitrile. The LC data showed considerable variation of chemical constituents among Atractylodes macrocephala populations.

          Results:

          21 and 22 characteristic peaks in the 14 grants of Atractylodes macrocephala and its processed product were determined in samples from different habitats of Zhejiang province, respectively. Their chromatographic patterns were generally consistent although their contents of chemical compositions were greatly different. The results of hierarchical cluster analysis showed that the samples could be divided into four groups; it was able to select excellent resources from the groups.

          Conclusion:

          This was the first report of hierarchical cluster analysis of crude and processed Atractylodes macrocephala according to their chemical fingerprints and could be applied to the intrinsic quality control of crude and processed Atractylodes macrocephala. The processing technique of Atractylodes macrocephala through the pilot-scale experiment was first studied and is simple and suitable for prepared yin pian of Atractylodes macrocephala industrial manufacture.

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          Most cited references27

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          Atractylenolide I and atractylenolide III inhibit Lipopolysaccharide-induced TNF-alpha and NO production in macrophages.

          In order to clarify the mechanism involved in the antiinflammatory activity of atractylenolide I and atractylenolide III from the rhizomes of Atractylodes macrocephala Koidz, their effects on tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) production in peritoneal macrophages were examined. Atractylenolide I and atractylenolide III decreased the TNF-alpha level in LPS-stimulated peritoneal macrophages in a dose-dependent manner, their IC(50) values were 23.1 microm and 56.3 microm, respectively. RT-PCR analysis indicated that they inhibited TNF-alpha mRNA expression. Furthermore, they inhibited NO production in LPS-activated peritoneal macrophages, the IC(50) value of atractylenolide I was 41.0 microm, and the inhibition ratio of 100 microm of atractylenolide III was 45.1% +/- 6.2%. The activity analysis of inducible nitric oxide synthase (iNOS) indicated that they could inhibit the activity of iNOS, their IC(50) values were 67.3 microm and 76.1 microm, respectively. Western blot analysis showed that atractylenolide I and atractylenolide III attenuated LPS-induced synthesis of iNOS protein in the macrophages, in parallel. These results imply that the antiinflammatory mechanism of atractylenolide I and atractylenolide III may be explained at least in part, by the inhibition of TNF-alpha and NO production. Atractylenolide I showed more potent inhibition than atractylenolide III in the production of TNF-alpha and NO in LPS-activated peritoneal macrophages. So, atractylenolide I could be a candidate for the development of new drugs to treat inflammatory diseases accompanied by the overproduction of TNF-alpha and NO.
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            Anti-inflammatory components isolated from Atractylodes macrocephala Koidz.

            The petroleum ether-ether (1 : 1) extract of Atractylodis macrocephalae was screened by cell membrane chromatography (CMC) and subsequently separated by column chromatography (CC) and high performance liquid chromatography (HPLC). Five components were isolated and identified as atractylenolide III 1, atractylenolide I 2, 14-acetoxy-12-senecioyloxytetradeca-2E,8E,10E-trien-4,6-diyn-1-ol 3, 14-acetoxy-12-alpha-methylbutyl-2E,8E,10E-trien-4,6-diyn-1-ol 4 and 14-acetoxy-12-beta-methylbutyl-2E,8E,10E-trien-4,6-diyn-1-ol 5 by routine spectrometric methods. The data of 5 and (13)C-NMR data of 3 and 4 were reported for the first time. Further in vivo experiments showed that the five components exhibited significant inhibiting effects both on the ear edema induced by xylene and on the peritoneal capillary permeability induced by acetic acid in mice.
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              5-Lipoxygenase and cyclooxygenase-1 inhibitory active compounds from Atractylodes lancea.

              Lipophilic extracts of Atractylodes lancea rhizomes exhibited potent inhibitory activities in 5-lipoxygenase [IC50 (5-LOX) = 2.9 micrograms/mL (n-hexane extract)] and cyclooxygenase-1 [IC50 (COX-1) = 30.5 micrograms/mL (n-hexane extract)] enzymatic assays. Bioactivity-guided fractionation of the n-hexane extract led to the isolation of a new compound atractylochromene (1), a potent inhibitor in both test systems [IC50 (5-LOX) = 0.6 microM, IC50 (COX-1) = 3.3 microM]. Also obtained was 2-[(2E)-3,7-dimethyl-2,6-octadienyl]-6-methyl-2,5-cyclohexadiene-1 ,4-dione (2), which showed a selective inhibitory activity against 5-LOX [IC50 (5-LOX) 0.2 microM, IC50 (COX-1) 64.3 microM]. The sesquiterpene atractylon (3) and the coumarin osthol (4) turned out to be moderate but selective 5-lipoxygenase inhibitors. Atractylenolides I (5), II (6), and III (7) showed no significant inhibitory effects for either enzyme. Structures were established by spectral data interpretation.
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                Author and article information

                Journal
                Pharmacogn Mag
                Pharmacogn Mag
                PM
                Pharmacognosy Magazine
                Medknow Publications & Media Pvt Ltd (India )
                0973-1296
                0976-4062
                Oct-Dec 2012
                : 8
                : 32
                : 300-307
                Affiliations
                [1 ] Department of Chinese Materia Medica, College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing - 210046, People’s Republic of China
                [2 ] Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing - 210029, People’s Republic of China
                [3 ] Research Center of TCM Processing Technology, Zhejiang Chinese Medical University, Hangzhou - 310053, People’s Republic of China
                [4 ] National First-Class Key Discipline for Science of Chinese Materia Medica, Nanjing University of Chinese Medicine, Nanjing - 210046, People’s Republic of China
                Author notes
                Address for correspondence: Prof. Dr. Baochang Cai, Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing - 210029, People’s Republic of China. E-mail: bccai@ 123456126.com
                Article
                PM-8-300
                10.4103/0973-1296.103659
                3785168
                24082634
                50e85c24-89aa-4181-98a7-f8ccd449edaf
                Copyright: © Pharmacognosy Magazine

                This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 30 September 2011
                : 12 October 2011
                : 22 November 2012
                Categories
                Original Article

                Pharmacology & Pharmaceutical medicine
                processed,chemical fingerprinting,hierarchical cluster analysis,atractylodes macrocephala,high-performance liquid chromatography

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