Journal
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
Volume 30, Issue 2, Pages 737-745Publisher
SPRINGER
DOI: 10.1007/s11274-013-1495-x
Keywords
DPVL; Antifungal; Plumbagin; Isolation; Quantification
Categories
Funding
- National Mega Project on Major Drug Development [2011ZX09401-302]
- National Natural Science Foundation of China [31301585]
- Commonweal Specialized Research Fund of China Agriculture [201103016]
- Key Program of Natural Science Foundation of Hubei Province of China [2010CBB02301]
- Fundamental Research Funds for the Central Universities [20103010101000185]
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Drosera peltata Smith var. lunata (Buch.-Ham.) C. B. Clarke (DPVL) fractions and plumbagin were tested via broth microdilution techniques on Rhizopus oryzae, Aspergillus flavus, Aspergillus niger, Aspergillus oryzae, Penicillium citrinum. All of the test substances [petroleum ether, chloroform, ethyl acetate, n-butanol fraction and aqueous residue (AR)] except for the AR were active against all the tested strains. The petroleum ether fraction (PEF) was the most active (MIC = 5.86-46.88 mu g/ml, MFC = 23.44-93.75 mu g/ml) of the five tested substances and therefore, was selected for further analysis. Based on antifungal activity, bioactivity-guided fractionation of the PEF led to the isolation of plumbagin. The structure of plumbagin was elucidated by H-1 and C-13 NMR. Using HPLC, DPVL was found to be a new source of plumbagin. Reversed-phase HPLC was performed using a mobile phase of water and methanol, and peaks were detected at 254 nm. Plumbagin showed a good linear relationship at concentrations ranging from 0.625 to 10 mu g/ml. Both the intraday and the interday precision showed that the method was precise, with RSDs of at least 3 % at different concentrations. Recovery rates ranging from 97.86 to 99.94 % were observed, which indicate that the method is accurate. The specificity of the method was established by checking the peak purity of plumbagin. For six independent measurements, the average plumbagin content in DPVL was 11.05 +/- A 0.31 mg/g of dried material. The validated HPLC method provides a new basis for assessing DPVL quality.
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