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      Metabolite profiles of ginsenosides Rk1 and Rg5 in zebrafish using ultraperformance liquid chromatography/quadrupole–time-of-flight MS

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          Abstract

          Background

          In the present study, metabolite profiles of ginsenosides Rk1 and Rg5 from red ginseng or red notoginseng in zebrafish were qualitatively analyzed with ultraperformance liquid chromatography/quadrupole–time-of-flight MS, and the possible metabolic were pathways proposed.

          Methods

          After exposing to zebrafish for 24 h, we determined the metabolites of ginsenosides Rk1 and Rg5. The chromatography was accomplished on UPLC BEH C18 column using a binary gradient elution of 0.1% formic acetonitrile–0.1% formic acid water. The quasimolecular ions of compounds were analyzed in the negative mode. With reference to quasimolecular ions and MS2 spectra, by comparing with reference standards and matching the empirical molecular formula with that of known published compounds, and then the potential structures of metabolites of ginsenosides Rk1 and Rg5 were acquired.

          Results

          Four and seven metabolites of ginsenoside Rk1 and ginsenoside Rg5, respectively, were identified in zebrafish. The mechanisms involved were further deduced to be desugarization, glucuronidation, sulfation, and dehydroxymethylation pathways. Dehydroxylation and loss of C-17 residue were also metabolic pathways of ginsenoside Rg5 in zebrafish.

          Conclusion

          Loss of glucose at position C-3 and glucuronidation at position C-12 in zebrafish were regarded as the primary physiological processes of ginsenosides Rk1 and Rg5.

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          Most cited references21

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          Chemical assessment of roots of Panax notoginseng in China: regional and seasonal variations in its active constituents.

          Root of Panax notoginseng (Radix Notoginseng, Sanqi) is a well-known traditional Chinese medicine and is mainly cultivated in Wenshan of Yunnan, China. The active constituents include saponin, dencichine, flavonoid, and polysaccharide; however, the levels of these components vary in different geographical regions of growth and also show a seasonal variation. By using high-performance liquid chromatography and spectrophotometry, the contents of notoginsenoside R1, ginsenoside R(g1), R(b1), R(d), dencichine, flavonoid, and polysaccharide were determined and compared with Radix Notoginseng collected from different regions of growth in China, as well as from different seasons of harvest and market grades. Using the contents of these active constituents as markers, the best quality of Radix Notoginseng is found in the southwestern parts of Wenshan, and the best season for the harvest is September to October. In addition, the unseeded plants produced a better quality of Radix Notoginseng. The current results provide useful information for the quality control of Radix Notoginseng and its further development in establishing the good agriculture practice standard of P. notoginseng in China.
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            Zebrafish for drug toxicity screening: bridging the in vitro cell-based models and in vivo mammalian models.

            Over the past decade, zebrafish have been tasked to play important roles from modeling human diseases to finding cures for them. Inadvertently, these fish now find themselves swimming along the drug development pipeline. A number of studies have been conducted to see if these small fish are up to the task of drug toxicity testing, an important rite of passage along the pharmaceutical pipeline. This review covers the recent publications (2008 - 2010) on the state-of-the-art applications that couple advanced technologies with the unique advantages of zebrafish for drug toxicity screening. The paper looks at the several automated high-throughput platforms that have been developed for zebrafish teratogenicity, cardiotoxicity and neuro-sensory organ toxicity assays over the past 3 years as well as the important studies related to metabolism and biotransformation of selected drugs that have been initiated. This paper also reviews their mechanistic and predictive omics applications. While there have been a number of developments over the past 3 years and indeed over the last 10 years, challenges and limitations still exist, which, unless overcome, will prevent zebrafish from truly reaching their full potential as a drug toxicological model. That being said, recent developments have suggested that zebrafish could play a role in bridging the gap between in vitro cell-based models and in vivo mammalian models.
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              Ultra-high performance liquid chromatography/time-of-flight mass spectrometry (UHPLC/TOFMS) for time-dependent profiling of raw and steamed Panax notoginseng.

              The metabolic profiles of Panax notoginseng and its associated therapeutic values are critically affected by the duration of steaming. The time-dependent steaming effect of P. notoginseng is not well-characterized and there is also no official guideline on its duration of steaming. In this paper, a UHPLC/TOFMS-based metabolomic platform was developed for the qualitative profiling of multiparametric metabolic changes of raw P. notoginseng during the steaming process. Our method was successful in discriminating the differentially processed herbs. Both the unsupervised principal component analysis (PCA) score plot (R(2)X=0.664, Q(2) (cum)=0.622, and PCs=2) and the supervised partial least square-data analysis (PLS-DA) model (R(2)X=0.708, R(2)Y=0.461, and Q(2)Y=0.271) demonstrated strong classification and clear trajectory patterns with regard to the duration of steaming. The PLS-DA model was validated for its robustness via a prediction set, confirming that the UHPLC/TOFMS metabolic profiles of the raw and differentially steamed P. notoginseng samples were highly reproducible. Based on our method, the minimum durations of steaming for the maximum production of bioactive ginsenosides such as Rg3 and Rh2 were also predicted. Our novel time-dependent metabolic profiling approach represents the paradigm shift in the quality control of P. notoginseng products. Copyright 2009 Elsevier B.V. All rights reserved.
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                Author and article information

                Contributors
                Journal
                J Ginseng Res
                J Ginseng Res
                Journal of Ginseng Research
                Elsevier
                1226-8453
                2093-4947
                13 January 2016
                January 2017
                13 January 2016
                : 41
                : 1
                : 78-84
                Affiliations
                [1 ]Key Laboratory of Chinese Medicine Delivery System of State Administration of Traditional Chinese Medicine, Jiangsu Provincial Academy of Chinese Medicine, Nanjing, China
                [2 ]Key Laboratory of New Drug and Clinical Application, Xuzhou Medical College, Xuzhou, Jiangsu, China
                Author notes
                []Corresponding author. Key Laboratory of Chinese Medicine Delivery System of State Administration of Traditional Chinese Medicine, Jiangsu Provincial Academy of Chinese Medicine, 100 Shizi Road, Hongshan Street, Nanjing, 210028, China. jeromechen@ 123456126.com
                Article
                S1226-8453(15)00134-7
                10.1016/j.jgr.2015.12.010
                5223078
                28123325
                836a107c-b1a9-44c9-b758-ac09f8a8c1ec
                Copyright © 2016, The Korean Society of Ginseng, Published by Elsevier.

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 24 August 2015
                : 24 December 2015
                : 30 December 2015
                Categories
                Research Article

                ginsenoside rg5,ginsenoside rk1,metabolism,ultraperformance liquid chromatography/quadrupole–time-of-flight ms,zebrafish

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