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Objective: Fusarium oxysporum is a common pathogenic fungus in ginseng cultivation. Both pathogens and antagonistic fungi have been reported to induce plant resistance responses, thereby promoting the accumulation of secondary metabolites. The purpose of this experiment is to compare the advantages of one of the two fungi, in order to screen out more effective elicitors. The mechanism of fungal elicitor-induced plant resistance response is supplemented. Methods: A gradient dilution and the dural culture were carried out to screen strains. The test strain was identified by morphology and 18 s rDNA. The effect of different concentrations (0, 50, 100, 200, 400 mg/L) of Penicillium sp. YJM-2013 and F. oxysporum on fresh weight and ginsenosides accumulation were tested. Signal molecules transduction, expression of transcription factors and functional genes were investigated to study the induction mechanism of fungal elicitors. Results: Antagonistic fungi of F. oxysporum was identified as Penicillium sp. YJM-2013, which reduced root biomass. The total ginsenosides content of Panax ginseng adventitious roots reached the maximum (48.95 ± 0.97 mg/g) treated with Penicillium sp. YJM-2013 at 200 mg/L, higher than control by 2.59-fold, in which protopanoxadiol-type ginsenosides (PPD) were increased by 4.57 times. Moreover, Penicillium sp. YJM-2013 activated defense signaling molecules, up-regulated the expression of PgWRKY 1, 2, 3, 5, 7, 9 and functional genes in ginsenosides synthesis. Conclusion: Compared with the pathogenic fungi F. oxysporum, antagonistic fungi Penicillium sp. YJM-2013 was more conducive to the accumulation of ginsenosides in P. ginseng adventitious roots. Penicillium sp. YJM-2013 promoted the accumulation of ginsenosides by intensifying the generation of signal molecules, activating the expression of transcription factors and functional genes.
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ABSTRACT Passiflora caerulea L., P. alata Curtis and P. incarnata L. (synonym for P. edulis Sims), are the most popular representatives of the Passiflora genus in South America. In recent years, a growing attention is paid to the biological activity and phytochemical profiles of crude extracts from various species of Passiflora in worldwide. The aim of this study was to evaluate and to compare of anti-leukemic activity of the dry crude extracts from leaves of three Passiflora species from greenhouse of Poland in two human acute lymphoblastic leukemia cell lines: CCRF-CEM and its multidrug resistant variant. Two systems of liquid chromatography in order to assessment of phytochemical composition of extracts were applied. Extracts of P. alata and P. incarnata showed the potent inhibitory activity against human acute lymphoblastic leukemia CCRF-CEM, while P. caerulea not showed activity (or activity was poor). Despite similarities in quality phytochemical profile of extracts from P. caerulea and P. incarnata, differences in quantity of chemical compounds may determine their various pharmacological potency. For the activity of P. alata extract the highest content of terpenoids and a lack of flavones C-glycosides are believed to be crucial. Summarizing, the crude extract from P. alata leaves may be considered as a substance for complementary therapy for cancer patients.
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Through comprehensively characterizing components in blood after oral administration of Tang-Bi-Kang (TBK) granules by UPLC-ESI-MSn,this study was aimed to explain the pharmaceutical material basis of TBK initially.UPLC-LTQ-Orbitrap was used under both positive and negative ion modes of electrospray ionization.The blank serum and rat serum after oral administration of TBK were analyzed.Components in rat serum were identified and characterized based on ion fragment information,evenelectron law,nitrogen rule and so on.Reference data was used to establish the UPLC-ESI-MSn method.The results showed that after oral administration of TBK granules,15 components were detected in the serum,of which 13 components were taken as the prototype to blood and 2 metabolites.It was concluded that constituents of TBK granules in rat serum were generated from compatibility of all herbal medicines.In rat serum,most of the components had been absorbed by rat's metabolism;a few were absorbed as the prototype.This research provided references for pharmacodynamic material basis and metabolism of TBL granules in vivo.
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Objective: Qualitative analysis of chemical components in Paeoniae Alba Radix extracts (PARE) in rat serum and brain. Methods: After ig administration of PARE, rat blood and brain tissues were collected. The chemical components in serum and brain were analyzed by HPLC-ESI-MSn. Results: By comparing retention time, MS and MSn spectra with data in the literature and reference standards, a total of 34 compounds including eight new metabolites were identified in serum, and three components including one new metabolite were identified in rat brain. Conclusion: This research will contribute to elucidate the pharmacodynamic material basis of antidepressant effect of PARE.
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OBJECTIVE: To compare the chemical composition and therapeutic effect between Prunella vulgaris L stem leaf and ear, thus to provide evidence for judging whether the stem leaf of Prunella vulgaris L can substitute the ear as an herbal medicine. METHODS: Aqueous extracts of the stem leaf and ear of Prunella vulgaris L from different producing areas were analyzed with HPLC-ESI-MSn. The anti-inflammatory effects were observed by inflammatory models of ear edema induced by dimethylbenzene in mice and hind paw edema induced by carrageenan in rats.Enzyme-linked immunosorbent assay (ELISA) was used to detect the plasma level of TNF-α and antioxidant activities were detected by ABTS method. RESULTS: Prunella vulgaris L stem leaf and ear were not significantly different in chemical composition, both of which contained mainly triterpenoids, flavonoids, phenolic acids and other substances. Compared with the model group, Prunella vulgaris L ear significantly reduced the hind paw edema in rats induced by carrageenan from 1 h after oral administration (P<0.05), while the onset time of stem leaf was later than 1 h.Both groups could significantly reduce the ear edema in mice induced by dimethylbenzene(P<0.01). The TNF-α levels in the Prunella vulgaris L stem leaf and ear groups [(24.16±1.24) and (24.33±2.36 )ng·mL-1] were lower than that in the model group [(31.34±1.94) ng·mL-1] (P<0.01).Prunella vulgarisL stem leaf and ear groups showed strong antioxidant activities in the ABTS·+ scavenging test. CONCLUSION: The contents of the main constituents in Prunella vulgaris L stem leaf and ear have significance differences.The RESULTS of animal tests indicate that the aqueous extracts of Prunella vulgaris L stem leaf and ear have significant anti-inflammatory and antioxidant effects.
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Millettia nitida var.hirsutissima is a Chinese herbal medicine used for the treatment of gynecological diseases.An HPLC/DAD/ESI-MSn method was established for the rapid separation and characterization of bioactive flavonoids in M.nitida var.hirsutissima.A total of 32 flavonoids were detected,of which 14 compounds were unambiguously characterized by comparing their retention time,UV,and MS spectra with those of the reference standards,and the others were tentatively identified based on their tandem mass spectrometry fragmentation data obtained in the negative ionization mode on line.Nineteen of these compounds characterized were reported from this plant for the first time.
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OBJECTIVE: To identify flavone C-glycosides in Dendrobium officinale leaves by high performance liquid chromatography with DAD and electrospray ionization mass spectrometry (HPLC-DAD-ESI-MSn). METHODS: The chromatographic separation was carried out at 30°C on a XB C18 column (4.6 mm × 250 mm, 5 μm) eluted with gradient program. The mobile phase consisted of methanol and water containing 0.4% formic acid. The detection wavelength was 335 nm and the on-line UV spectra was recorded in the range of 190-400 nm. The mass spectra was obtained by Agilent ion trap mass spectrometer in the negative ion mode with capillary voltage 4 500 V, dry gas temperature 350°C, dry gas flow 12.0 L · min-1, nebulizer pressure 241 kPa; mass range recorded m/z 50-600. RESULTS: Eight flavone di-C-glycosides from D. officinale leaves, whose aglycone was apigenin and monosaccharide was connected with C-6 and C-8, were identified by HPLC-DAD-ESI-MSn for the first time. The study further proved the characteristics of fragmentation pattern of flavone C-glycosides in ESI-MSn. CONCLUSION: The method is simple and rapid for the identification of D. officinale leaves. The characteristics of fragmentation pattern of ESI-MSn in flavone di-C-glycosides provide a reference for rapid detection and identification of flavone C-glycosides.
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Objective: To establish an LC-MS method for identification of the main components in Xinyue Capsula. Methods The main components in Xinyue Capsula were detected with the LC-ESI-MS in negative monitoring mode. These components were further analyzed by LC-MS2iv and MS3 spectra, and by comparing to the corresponding standards and the references, these components were elucidated. Results: Eighteen major ginsenosides, which were belonging to protopanaxadiol, protopanaxatriol, and ocotillol type saponin, were found in Xinyue Capsula. With the corresponding reference substances, ten of the 18 components were domenstrated as ginsenoside Re, Rg1, pseudoginsenoside F11, ginsenoside Rg2, Rc, Rb 2, Rb3, Rd, F2, and 20(S)-ginsenoside Rg 3. Conclusion: The LC-MSn method established in the pre sent study could simultaneously analyze various kinds of ginsenosides, which include ocotillol, protopanaxadiol, and protopanaxtiol type ginsenoside. This method is sensitive, rapid, and simple for analyzing and identifying the ginsenosides in Xinyue Capsula and Panax quinque folius.
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Objective: To analyze the bioactive components of Acnoitum Kusnezoffii Reichb..Methods: To detect the water extraction of Aconitum kusnezoffii Reichb.by the method of ESI-MSn.Results: Three kinds of alkaloids were detected in the water extraction of Aconitum kusnezoffii Reichb.:monoester alkaloids,diester-diterpenoid aconitines and lipo-alkaloids.Conclusion: The effect components of processed and unprocessed Acnoitum Kusnezoffii Reichb.can be analyze rapidly by ESI-MSn,and only needs few sample.
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Objective To study the collision-induced dissociation fragmentation pathways of the jasminoidin and catalpol in ESI-Msn. Methods The samples were studied by using high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-Msn) both in positive and negative. Results The collision-induced dissociation fragmentation pathways of both compounds were described. In the spectrum of two compounds, the loss of glucose residue and whole glucose were observed in positive mode, while only the loss of glucose residue could be observed in negative mode. Furthermore, we found that as the effect of the substituting groups, the fragment ions of jasminoidin were more in positive mode and those of catalpol were more in negative mode. Conclusion Conclusion The established method could be used for the sensitive and rapid identification of this kind of iridoid glycosides。
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Objective To study how and why the lipo-alkaloids changed in decocting the flowers of Aconitum kusnezoffii (FAK). [WT5HZ]Methods The alkaloids in ethanol extract of FAK, FAK decoction, and the residues of decocted FAK were analyzed and compared by electrospray ionization tandem mass spectrometry (ESI-MSn). Results No lipo-alkaloids were detected in FAK decoction, however, triester-lipo-alkaloids become the dominant components in the residues of FAK decoction. Conclusion Besides hydrolysis reactions, ester-exchange reactions happen for diester-aconitines and triester-aconitines in the decocting of FAK. It is the first time to report that C_8-acetyl is displaced by long chain fatty acyl for triester-aconitines in the ester-exchange reactions.
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AIM: To apply the solid phase extraction (SPE) on salvianolic acids in Radix Salvia Miltiorrhizae by means of high performance liquid chromatography with mass spectroscopic detector (HPLC MSn). METHODS: A C 18 solid phase column and negative ion electrospray ionization (ESI) mode were used by orthogonal design to optimize four factors which might affect recoveries of samples, which included vacuum tightness, sample size, amount of eluting agent and eluting power. RESULTS: The optimized results were as follows: vacuum tightness was 0.002?106 Pa, sample size was 0.75 mL, amount of eluting agent was 15mL and eluting power was 1.0mL?s -1 . CONCLUSION: The established method is simple, reliable and suitable for the usual quality control method of salviamolic acids in Radix Salvia Miltiorrhizae.