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Objective: To comprehensively evaluate the different passivation materials on growth and cadmium uptake and accumulation of Ophiopogon japonicus cultivated in cadmium contaminated soil of. Methods: Using one-year-old O. japonicus as experimental material, the effects of five different passivation materials (white marble Ar, straw biochar Br, fly ash Fh, bacterial residue Me, diatomite Dm) on the growth, physiological and biochemical indexes and cadmium absorption and accumulation of O. japonicus were studied by soil pot experiment. Results: The biomass, chlorophyll, soluble sugar and soluble protein of O. japonicus could be significantly increased in soils contaminated with two kinds of Cd under different passivation materials. Among them, Ar and Me were the most improved. In addition, the activities of antioxidant enzyme system of O. japonicus were significantly decreased, and the content of MDA had no noteworthy change. Different passivation materials remarkably increased the content of flavonoids in the underground part of O. japonicus, among which Ar and Fh treatments increased the most. The contents of heavy metal Cd in all parts of O. japonicus were decreased by different passivation materials, and the contents of Cd in aboveground and underground parts of O. japonicus were significantly lower than those of the control, and the effects of Ar, Br and Fh treatments were better than those of the control. Conclusion: Studies have shown that by combining and comparing treatments of different passivating materials, in terms of the biomass of O. japonicus treated with Me, the biomass improvement effect of Me was better than that of Ar and Br. Aiming at the effect of reducing Cd absorption and accumulation of O. japonicus and improving the effective component flavone of O. japonicus, it can be concluded that Ar and Fh have better effects in treating Cd polluted soil and improving the quality of O. japonicus.
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Objective: An efficient method was established using high content screening (HCS) for the hepatotoxicity evaluation of sulfur-fumigated Ophiopogonis Radix. Methods: Cytotoxicity of positive control group, negative control group, Ophiopogonis Radix extracts group and sulfur-fumigated Ophiopogonis Radix extracts group were tested based on HepG2 human hepatoma cells. HCS was applied to detect the cell number, DNA content, level of glutathione (GSH), reactive oxgyen species (ROS), and mitochondrial membrane potential (MMP). Results: Compared with the cells of Ophiopogonis Radix extracts group, GSH of sulfur-fumigated Ophiopogonis Radix extracts decreased significantly at the concentration of 50 mg/mL; The MMP of sulfur-fumigated Ophiopogonis Radix extracts changed signally at the concentration of 12.5 mg/mL. Conclusion: Ophiopogonis Radix showed pontential cytotoxicity after sulfur-fumigated. The hepatotoxicity of sulfur-fumigated Ophiopogonis Radix may be related to mitochondria-mediated apoptosis according to the influence of its MMP from the results.
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Ophiopogon japonicus is widely distributed in China and rich in resources, and is a kind of medicinal plants with great utilization value. It could nourish yin, promote body fluid production, moisten the lung, ease the mind, and clear away heart fire. Modern research showed that there are several secondary metabolites, such as steroidal saponins, homoisoflavonoids and polysaccharides. Its pharmacological research mainly focused on the hypoglycemic effect, cardiovascular protection, enhanced immunity, anti-aging, anti-inflammatory, and antitumor effect. In this paper, the chemical compositions and pharmacological activities of O. japonicas were summarized, in order to provide reference for further study and application of O. japonicas.
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Objective To investigate the rules of root tuberenlargement and medicinal materials formation, and lay the foundation for breeding a new Ophiopogon japonicus variety. Method The study was carried out with field cultivation. Samples were collected continuously for every interval 7-10 d. The rules of dry matter distribution and accumulation in each parts of O. japonicus was calculated based on the mean value of 15 plant samples. The number and size of root tuber were calculated based on the mean value of 15 samples. The fiber characteristics of root tuber of O. japonicus were observed by optical microscope. Characters of O. japonicus were observed according to the Chinese Pharmacopoeia 2015 edition and recorded the changes of medicinal characteristics. Results The root tuber of O. japonicus inflated starting from October to April in the next year in Sichuan province. Adventitious roots were initially formed into nutritive roots around caudex, and able to form into root tuber around caudex later. Tuber number of per plant is basic from 5 to 20, most in 8-10, very few more than 20. Adventitious roots expanded gradually into a fusiform root from one end of root tip, root tip can continue to grow to form adventitious roots after the formation of root tuber, the newly formed adventitious roots could be expanded for the second time, very few might appear swollen three times to form beads. Microscopic observation showed that root tubers were inflated primarily by the enlargement of cortical cells and the increase of cell layers, in which cortical cells enlargement played a leading role. While tubers started expanding, the accumulation of dry matter in fibrous roots and stem leaves quickly slowed to stop eventually, and the accumulation of root dry matter increased rapidly in the early stage, and it kept basically dynamic balance in mid and late March, and the whole behavior of medicinal materials was in the process from dry to full. Conclusion The results show the apparent change rules of O. japonicus root tuber in the expanding process, which proves the direction and goal for research on the mechanism of root tuber development of O. japonicus.
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Ultrafiltration is one of the most fascinating technologies, which makes it possible to improve the quality of traditional medicines for application in the pharmaceutical industry. However, researchers have paid little attention to the effect of ultrafiltration membrane on traditional medicines chemical constituents. In this work, Ophiopogon japonicus (L.f) Ker-Gawl. was used as an example to illuminate the influence of ultrafiltration with different material and molecular weight cut-off (MWCO) membrane on natural chemical constituents as measured by ultra-fast liquid chromatography coupled with ion trap time-of-flight mass spectrometry (UFLC-IT-TOF/MS). Our results indicated that ultrafiltration membrane significantly impacted homoisoflavonoids, especially homoisoflavonoids that were almost completely retained on the polyethersulfone (PES) membrane. We also found that the larger number of aglycone hydroxy and sugar moiety in steroid saponins, the higher the transmittance. Furthermore, the passage rate (%) of ophiogenin type saponins was higher than that of others. The possible adsorptive mechanisms were hydrogen bonding, hydrophobic interactions, and benzene ring interaction by π-π stacking. In conclusion, it is crucial to choose appropriate ultrafiltration membrane based on the characteristics of produce products for application of ultrafiltration technique.
Subject(s)
Chromatography, High Pressure Liquid , Methods , Chromatography, Liquid , Methods , Drugs, Chinese Herbal , Isoflavones , Molecular Structure , Molecular Weight , Ophiopogon , Chemistry , Plant Extracts , Chemistry , Polymers , Saponins , Spectrometry, Mass, Electrospray Ionization , Methods , Sulfones , Ultrafiltration , MethodsABSTRACT
Objective: To optimize the suitable technology of Ophiopogon japonicus (OJ) fumigation based on the changes of relative indexes after the different levels of sulfur fuming. Methods: The phenol-sulfuric acid method was used to determine the content of polysaccharide. HPLC-ELSD method was used to determine the contents of Ophiopogon saponins D (OPD) and D' (OPD') with the mobile phase of acetonitrile-water (50:50), drift tube temperature at 40℃, and nitrogen volumetric flow of 2.0 L/min. The fingerprints of OJ were determined by HPLC-UV method, the mobile phase was eluted with acetonitrile-0.1% phosphoric acid gradient, and the detection wavelength was 297 nm. The contents of ethylophiopogonanones A (MA) and B (MB) were determined using the same method. Using the contents of total polysaccharides of OJ, OPD and OPD', MA and MB, and the contents of other components in the fingerprints as control indexes, and the dosage of sulfur and the time of sulfur fumigation as factors, the central composite design-response surface methodology (CCD-RSM) was used to optimize sulfur fumigation process. Results: In the experimental design range, the content of polysaccharides increased first and then decreased with the increase of sulfur dosage and sulfur fumigation time, and the contents of OPD and OPD' increased slightly and then decreased. The contents of MA and MB were slightly reduced. The sulfur fuming process was determined as follows: sulfur dosage was 120-200 g/m3, sulfur fuming time was 0.6-1.5 h, binomial equation: Y = 0.574 + 0.054 3 X1-0.008 13 X2-0.518 X12 + 0.131X22 + 0.153 X1X2 (R-Sq = 0.971). Process validation: the absolute value of deviation was less than 3%. Conclusion: In this study, the changes of chemical composition of OJ were studied, and the sulfur smelting process suitable for OJ was established.
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Objective: To develop an LC/MS approach based on precise mass and tandem mass spectrometer to identify homoisoflavonoids from Ophiopogon japonicus alcohol extract. Methods: Separation was performed on an Alltima C18 reverse-phase column (250 mm x 4.6 mm, 5 μm) and an Alltech pre-column. The mobile phase consisted of water containing 1 mmol/L ammonium acetate-acetonitrile containing 0.01% acetic acid was used as gradient elution. The flow rate was 1.0 mL/min. The detection wavelength was 298 nm. A combination of time-of-flight mass spectrometer (TOF/MS) and ion trap tandem mass spectrometer (Trap/MSn) was applied for qualitative analysis under negative ion mode. Results: From the alcohol extract of O. japonicus, the molecular formulae of 11 components were screened by TOF/MS and 11 chemical components were identified by Trap/MSn. All of the peaks were attributed. Conclusion: This method can be used to identify medicinal ingredients quickly and provide an effective and reliable analytical model, as well as other complex system of traditional Chinese medicine compound.