Four Natural Compounds Separated from Folium Isatidis: Crystal Structures and Antibacterial Activity.

Four natural compounds were obtained by concentrating, separating and purifying from the Folium isatidis. These natural compounds have been characterized by elemental analysis, IR spectrum, NMR and single-crystal X-ray diffraction analysis. The results show that these natural compounds are 4(3H)-quinazolinone (I), 2,4(1H,3H)-quinazolinedione (II), methyl 3,4-dihydro-4-oxoquinazoline-2-carboxylate (III) and ethyl 3,4-dihydro-4-oxoquinazoline-2-carboxylate (IV). The antibacterial activity experiment showed that I and II had better activity against Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Salmonella than III, IV and other multiple components, because III and IV have long branches and steric hindrance effect. Compounds I and II have planar structure, which can more easily combine with these bacteria and kill them. The above results have good guiding significance for studying the antibacterial activity for single components or mixtures from natural origin.

n-Butanol extract from Folium isatidis inhibits lipopolysaccharide-induced inflammatory cytokine production in macrophages and protects mice against lipopolysaccharide-induced endotoxic shock.

Sepsis, which is caused by severe infection, is an important cause of mortality, but effective clinical treatment against sepsis is extremely limited. As the main component of the outer membrane of Gram-negative bacteria, lipopolysaccharide (LPS) plays a major role in inflammatory responses. Studies have shown beneficial pharmacological effects for Folium isatidis. The present study further illuminated the effects of n-butanol extract from Folium isatidis in LPS-induced septic shock and identified the main active chemical components. Our study showed that pretreatment with n-butanol extract from Folium isatidis not only significantly inhibited LPS-induced tumor necrosis factor-α and interleukin-6 production but also markedly and dose dependently enhanced the recruitment of MyD88, the phosphorylation of extracellular signal-regulated kinase, and the degradation of IκB-α. Additionally, the extract exhibited dramatic protective effects against lung injury and death in mice with septic shock. Eight main active compounds were identified, including organic acids, glycoside, indolinones, and flavonoids. These findings provide a perspective on the respiratory protection offered by n-butanol extract from Folium isatidis in LPS-induced sepsis and outline a novel therapeutic strategy for the treatment of sepsis.

Analysis of Volatile Constituents in Folium Isatidis by Head-space Solid Micro-extraction Coupled with GC-MS / 中国药师

Objective:To investigate the volatile constituents in Folium isatidis. Methods:The volatile constituents from Folium isatidis were analyzed by head-space solid micro-extraction coupled with GC-MS for the first time. Results: Thirty-five compounds (89. 95%) were identified from the volatile constituents in Folium isatidis. The main volatile constituents of Folium isatidis were 6, 10, 14-trimethyl-2-pentadecanone (6. 32%), nonanal (5. 99%), phenethyl isothiocyanate (5. 79%) and palmitic acid (5. 62%). Conclusion:Palmitic acid and benzyl alcohol may be the main effective constituents in Folium isatidis.

Antiviral Activity of the Effective Monomers from Folium Isatidis Against Influenza Virus in Vivo / 中国病毒学·英文版

In order to evaluate the anti-influenza virus activity of the effective monomer from Folium Isatidis (FI) in vivo, we established mice model with viral pneumonia and divided them into 3 different dose groups, then observed their lung indexes, pulmonary pathological changes, pulmonary virus hemagglitination titers, living time and death rates. The results showed that the monomer could reduce the pulmonary index from 2.64 to 1.93, 1.63 and 1.40 (P<0.01) and decrease the hemagglitination titer from 1.15 to 0.84, 0.70 and 0.59 (P<0.01). In addition,the living time from 5.1d to 6.5d, 8.4d and 8.9d respectively(P<0.01). The high dose (75 mg/kg/d) has the similar effect with 100 mg/kg/d dose of virazole(P>0.05), and more effective than 200 mg/kg/d dose of antiviral liquor (P<0.05).

Content Determination of organic Acid in Folium Isatidis from Differernt Habitats / 中国中医药信息杂志

Objective To compare the content of organic acid in Folium Isatidis from differernt Habitats. Methods According to ultraviolet absorption atlases characteristics of the three main organic acid in Folium Isatidis, DSA UV-2401 was used to determinate the acidcontent of Folium Isatidis from differernt Habitats, and carried on to inspect the methodology. Results The difference of the acidcontent of Folium Isatidis from differernt Habitats is notable. Conclusion The method is simple, accurate and duplicated good, and can be used as quantitative analysis method of organic acid in Folium Isatidis.

Precipitation of Folium Isatidis Extract with Chitosan and ZTC1+1-ⅡClarifiers / 中国药房

OBJECTIVE:To optimize the precipitation process of the extract of Folium Isatidis using chitosan and ZTC1+ 1-Ⅱ natural clarify agents.METHODS:Using the remaining rates of polysaccharide and indirubin and the defecation level as indexes,the optimum process of removing impurity was investigated with single-factor test.RESULTS:The optimum process conditions were as follows:chitosan:precipitating the extract solution which was condensed to 1∶10(herb:the herb solution) with the concentration of chitosan clarify agent at 8% at a heating temperature of 50 ℃;ZTC1+1-Ⅱ:precipitating the extract solution which was condensed to 1∶10(herb:the herb solution) with the concentration of chitosan clarify agent at 5% and the heating temperature at 70 ℃,and the adding sequence of ZTC 1+1-Ⅱ natural clarify agents was clarifier B before clarifier A.CONCLUSION:Chitosan and ZTC1+1-Ⅱ natural clarify agents can be used to effectively remove the impunity of extract of Folium Isatidis.

Research on the Extraction and Purification of Organic Acids from Folium Isatidis with Macroporous Adsorptive Resin / 中国药房

OBJECTIVE:To extract and purify organic acids with macroporous resins from Folium Isatidis.METHODS:Folium Isatidis was extracted with 70% aqueous ethanol(pH=2)by Soxhlet extraction.The process was optimized by employing orthogonal test design with the content of anthranilic acid as the index.RESULTS:The resin HPD100 had much better adsorption capacity and desorption ratio by using 3 BV 60% aqueous ethanol to extract organic acids in Folium Isatidis than other resins.The results showed that HPD100 possessed the better ecomic under the concentration of sample 0.18mg?mL-1,the condition of pH 3.5,and flowing rate of 2BV?h-1.CONCLUSION:Macroporous resin HPD100 may be value of isolating and purifying the organic acids from Folium Isatidis.

Content Determination of Amino Acids in Folium Isatidis by RP-HPLC Precolumn Derivatization Method / 中国药房

OBJECTIVE:To establish a RP-HPLC precolumn derivatization method for the content determination of argi-nine(Arg)and proline(Pro)in Folium Isatidis.METHOD:2,4-dinitro-fluorobenzene was undergone precolumn derivatization and the amino acids was determined directly under alkaline condition with Kromasil C 18 as chromatographic column,the mobile phase was composed of sodium acetate buffer solution(pH=6.4)-acetonitrile(850∶150)with detection wavelength at360nm,the content was calculated by external reference method.RESULTS:The linear ranges for Arg and Pro were0.627?g～5.016?g(r=0.9996)and0.874?g～7.000?g(r=0.9995),respectively.The average recovery was98.2%with RSD at2.3%and2.2%,respectively.CONCLUSION:The method is simple,accurate and reliable,and suitable for the assaying of amino acids in Folium Isatidis.

Extraction and seperation of indirubin in Folium Isatidis / 中成药

AIM: To study the suitable conditions of the extraction-separation process of the indirubin in Folium Isatidis with NIR. METHODS: The indirubin was chromatographed on an aluminium oxide column and chloroform-ethyl acetate as eluting agent,eluate was recrystalized by methanol-acetone. RESULTS: The indirubin extracted by this method was of high purity (97.78%).The sample purity was similar to reference substance one((98.01%)).The spectra of NMR and IR are the same. CONCLUSION: This method is proved to be fast,safe,lower cost,simple and has good results.

Determination of Paracetamol and Caffeine in New compound Folium Isatidis Tablets by HPLC / 中成药

Objective:To determine the contents of paraceltamol and caffeine in New compouad Folium Isatidis Tablets simultaneously by HPLC. Methods:The determination was carried out with C 18 chemical bonded silica gel as a solid phase, methanol-water (25∶75) as a mobile phase and UV deterction wavelength at 215 nm. Results: The average recovenies of the added sample were 99.6%( RSD=0.67, n=5) for paracetamol and 99.3%(RSD=0.58, n=5) for caffine, There was a good linear relationship between the concentration and absorption area value in the rang of 1.6?g～6.4?g for paractamol or 0.16?g～0.64?g for caffeine.Conclusions: The method is simple, quick and accurate.