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Ichthyophthirius multifiliis is a holotrichous protozoan that invades the gills and skin surfaces of fish and can cause morbidity and high mortality in most species of freshwater fish worldwide. The present study was undertaken to investigate the antiparasitic activity of crude extracts and pure compounds from the leaves of Macleaya microcarpa. The chloroform extract showed a promising antiparasitic activity against I. multifiliis. Based on these finding, the chloroform extract was fractionated on silica gel column chromatography in a bioactivity-guided isolation affording two compounds showing potent activity. The structures of the two compounds were elucidated as dihydrosanguinarine and dihydrochelerythrine by hydrogen and carbon-13 nuclear magnetic resonance spectrum and electron ionization mass spectrometry. The in vivo tests revealed that dihydrosanguinarine and dihydrochelerythrine were effective against I. multifiliis with median effective concentration (EC(50)) values of 5.18 and 9.43 mg/l, respectively. The acute toxicities (LC(50)) of dihydrosanguinarine and dihydrochelerythrine for richadsin were 13.3 and 18.2mg/l, respectively. The overall results provided important information for the potential application of dihydrosanguinarine and dihydrochelerythrine in the therapy of serious infection caused by I. multifiliis., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Objective: To develop an HPLC method for the determination of a Tibetan medicine Meconopsis quintuplinervia., Method: A Hypersil-Keystone-C18 column (4.6 mm x 250 mm, 5 microm) was used with the isocratic elution of acetonitrile and 0.012% glacial acetic acid. The flow rate was 1.0 mL x min(-1), and the detection wavelength was set at 237 nm., Result: The linear range of 0-methylflavinantine was 0.2-2.4 microg (r = 0.999 7). The average recovery was 96.26%., Conclusion: The developed method was reliable, and can be used for the quality control of M. quintuplinervia Regel.

Transforming technology for semi-synthesized isocorydione from the natural product ofisocorydine was studied. The factors affecting on the reaction yield were investigated. UV spectrophotometry was used to indicate the semi-synthesized yield of isocorydione. The optimum reaction conditions were determined as following: reacting for 12 h in the solution of sodium dihydrogen phosphate at pH 10, the temperature was 25 degrees C and the ratio of isocorydine to Fremy's radical was 1 : 2. Under the optimum conditions, the yield could reach up to 50.0%. The molecular structure of isocorydione was elucidated by X-ray single-crystal diffraction analysis for the first time.

The substance archive of the laboratory of Johannes Gadamer (1867-1928), Marburg, Germany, was analyzed thoroughly with modern instrumental methods, with the samples purified when necessary, and the authenticity of the samples confirmed by historical and analytical evidence. Eight formerly unknown alkaloids of the benzylisoquinoline type were identified in the archive originally isolated from Corydalis cava or Fumaria vaillantii. This finding underscores the importance of the work of Johannes Gadamer and his group in stimulating overall progress in natural product chemistry. Several alkaloids were isolated by the group long before they were officially reported.

Objective: To establish the quantitative determination method and TLC identification of Meconopsis and study the Vis-UV spectrum character for fast identification., Methods: Meconopsis were identified by TLC, and the content of quercetin was determined by HPLC., Results: The acid hydrolyze condition was HCl-80% MeOH(8:25) 90 degrees C calefaction by water for 1.5 hour. The condition of HPLC: Hypersil-Keystone-C18 (250 mm x 4.6 mm, 5 microm) column, MeOH-Water (50: 50) as mobile phase, detection wavelength at 360nm, flow rate at 1.0 mL/min, column temperature at 35 degrees C, quercetin showed a good linear relationship at a range of 0.01 - 0.12 microg, r =0.9997. The average recovery was 98.83%, and RSD was 1.69% (n = 9). The contents of quercetin in Meconopsis integrifolia and Meconopsis quintuplinervia of Gansu were 0.1135 mg/g and 0.0842 mg/g respectively., Conclusion: The method is simple, accurate and specific. It can be used for the quality control and fast identification of Meconopsis.

Objective: To search for natural benzylisoquinoline alkaloids against tumor., Method: In this study, taken DNA as target, a method was developed for screening of natural benzylisoquinoline alkaloids against tumor from traditional Chinese medicine by the use of centrifugal ultrafiltration combined with HPLC technology. The anti-tumor activity for the compounds screened was evaluated in vitro., Result: Nine compounds interacted with DNA were discovered and identified from Macleaya cordata, Chelidonium majus, Coptis chinensis, and the proliferation of four types of human solid cancer cell lines was markedly inhibited by these compounds., Conclusion: The developed method was considered to be suitable for screening of natural benzylisoquinoline alkaloids against tumor from traditional Chinese medicine.

The antifungal activities of dihydrosanguinarine and dihydrochelerythrine, isolated from the leaves of Macleaya microcarpa, were evaluated on 12 plant pathogenic fungi; the two compounds exhibited the highest antifungal activity against Botrytis cinerea Pers. Among the 11 tested plant pathogenic fungi in vitro, the two compounds showed the highest antifungal activity against B. cinerea Pers, with 95.16% and 98.32% mycelial growth inhibition at 50 µg mL⁻¹, respectively. In addition, the two compounds inhibited spore germination in vitro in a concentration-dependent manner. They also showed potent protective and curative effects against Erysiphe graminis and B. cinerea in vivo. This is the first report on the antifungal activity of dihydrosanguinarine and dihydrochelerythrine against pathogenic plant fungi.

In this study, five derivatives of sanguinarine (1) and chelerythrine (2) were prepared, with 1 and 2 as starting materials, by reduction, oxidation and nucleophilic addition to the iminium bond C=N+. The structures of all compounds were elucidated on account of their MS, ¹H-NMR and ¹³C-NMR data. The antibacterial activities of all compounds were screened, using Staphylococcus aureus, Escherichia coli, Aeromonas hydrophila and Pasteurella multocida as test bacteria. The minimum bacteriostatic concentration and minimum bactericidal concentration of the active compounds were determined by the turbidity method. The structure-activity relationships of 1 and 2 were discussed. The results showed that 1, 2 and their pseudoalcoholates were found to be potent inhibitors to S. aureus, E. coli and A. hydrophila, while the other derivatives were found to be inactive. The pseudoalcoholates might be the prodrugs of 1 and 2. The iminium bond in the molecules of 1 or 2 was the determinant for antibacterial activity, and the substituents at the 7 and 8 positions influenced the antibacterial activities of 1 and 2 against different bacteria.

Macleaya microcarpa (Maxim.) Fedde belongs to the genus Macleaya, family Papaveraceae. Together with the better known and more frequently studied species M. cordata (Willd.) R. Br. it is a main source of quaternary benzo[c]phenanthridine alkaloids. Using HPLC we determined the content of eight isoquinoline alkaloids in the aerial and underground parts of 1-, 2-, 12- and 13-year old plants and followed their changes during the vegetative period. The dominant alkaloid of all samples collected in the end of this period was allocryptopine (3.8-13.6 mg/g for aerial parts, 24.2-48.9 mg/g for underground parts). Chelerythrine, sanguinarine and protopine were also present in both parts of the plant. Additionally, measurable concentrations of chelilutine (CL), chelirubine (CR), macarpine (MA) and sanguirubine (SR) were detected in underground parts. The most important finding was that contents of CR, CL, SR and MA in the 12- and 13-year old plant roots were significantly higher (approximately 3-fold for CR, 6-fold for CL, 5-fold for SR, and at least 14-fold for MA) than in 1- or 2-year old plants. The proportion of individual alkaloids in aerial and underground parts thus changed significantly during the vegetative period.

The human cytochrome P450 (CYP) enzymes play a major role in the metabolism of endobiotics and numerous xenobiotics including drugs. Therefore it is the standard procedure to test new drug candidates for interactions with CYP enzymes during the preclinical development phase. The purpose of this study was to determine in vitro CYP inhibition potencies of a set of isoquinoline alkaloids to gain insight into interactions of novel chemical structures with CYP enzymes. These alkaloids (n=36) consist of compounds isolated from the Papaveraceae family (n=20), synthetic analogs (n=15), and one commercial compound. Their inhibitory activity was determined towards all principal human drug metabolizing CYP enzymes: 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6 and 3A4. All alkaloids were assayed in vitro in a 96-well plate format using pro-fluorescent probe substrates and recombinant human CYP enzymes. Many of these alkaloids inhibited the CYP3A4 form, with 30/36 alkaloids inhibiting CYP3A4 with at least moderate potency (IC₅₀ < 10 μM) and 15/36 inhibiting CYP3A4 potently (IC₅₀ < 1 μM). Among them corydine, parfumine and 8-methyl-2,3,10,11-tetraethoxyberbine were potent and selective inhibitors for CYP3A4. CYP2D6 was inhibited with at least moderate potency by 26/34 alkaloids. CYP2C19 was inhibited by 15/36 alkaloids at least moderate potently, whereas CYP1A2, CYP2B6, CYP2C8, and CYP2C9 were inhibited to a lesser degree. CYP2A6 was not significantly inhibited by any of the alkaloids. The results provide initial structure-activity information about the interaction of isoquinoline alkaloids with major human xenobiotic-metabolizing CYP enzymes, and illustrate potential novel structures as CYP form-selective inhibitors., (Copyright © 2010 Elsevier GmbH. All rights reserved.)

Chelerythrine (a natural quaternary benzophenanthridine alkaloid) is an extract from the roots of Chelidonium majus with potential antimycobacterial activity. To reveal the possible mechanism of action of chelerythrine against Mycobacterium tuberculosis (M. tuberculosis), commercial oligonucleotide microarrays were used to analyze the genome-wide transcriptional changes triggered by treatment with subinhibitory concentrations of chelerythrine. Quantitative real-time RT-PCR was performed for selected genes to verify the microarray results. We interpreted our microarray data using Agilent software. Analysis of the microarray data revealed that a total of 759 genes were differentially regulated by chelerythrine. Of these, 372 genes were upregulated, and 387 genes were downregulated. Some of the important genes that were significantly regulated are related to different pathways (such as urease), methoxy-mycolic acid synthase, surface-exposed lipids, the heat shock response, and protein synthesis. This genome-wide transcriptomics approach produced the first insights into the response of M. tuberculosis to a chelerythrine challenge.

The aim of this experiment was to investigate whether a low-dietary application (15 mg/kg) of an alkaloid preparation containing quaternary benzo[c]phenanthridine alkaloids and obtained from Macleaya cordata (Sangrovit) influenced caecal metabolism, growth performance and long-chain fatty acid composition of breast meat. One-day-old broiler chicks (Cobb 500) were fed a diet without supplement or with a 15 mg/kg dose of Sangrovit (C and A groups respectively) for 5 weeks. Although the A treatment was not accompanied by an enhanced final body weight of broilers, the intake of a diet with Sangrovit influenced the caecal microflora activity. The addition of Sangrovit to a diet decreased potentially harmful β-glucuronidase and β-glucosidase activities (p<0.05 and p=0.075 respectively), and at the same time led to a significant increase in activities of bacterial glycolytic enzymes α-glucosidase, α-galactosidase, β-galactosidase in comparison to the control group. The concentration of total short-chain fatty acids in the caecal digesta was increased in the A treatment contributing to the tendency towards lower caecal pH (p=0.078). The analysis of breast meat fatty acids showed that the dietary application of Sangrovit evoked some changes in contents of monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) contents. The applied dosage of Sangrovit caused an increase in the sum of MUFA and the tendency towards lower PUFA sum (p<0.05 and p=0.062 respectively) as in relation with the C group. Although the nutritionally relevant n-6/n-3 PUFA and the (PUFA+MUFA)/saturated fatty acids ratios remained similar in both groups, further research is postulated to establish the effect of this preparation on meat quality. To sum up, despite of a lack of the improvement in final body weight, a low dose of dietary Sangrovit was found to exert positive effects on the caecal metabolism of the broilers., (© 2010 Blackwell Verlag GmbH.)

Objective: To analyze the effects of Eomecon chinanthe sanguinarine (SAN) on glucogen, enzyme activity and lipid peroxidation of Oncomelania hupensis liver so as to explore the mechanism of SAN against Oncomelania hupensis., Methods: SAN was extracted and purified from the dry powder of Eomecon chionantha. Oncomelania hupensis were immersed in 5 mg/L sanguinarine (50 Oncomelania hupensis per 500 ml solution) or clean water at 25 degrees C for 36 h, the livers were isolated from live snails. Total glucogen content, malondialdehyde (MDA) level, activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), acid phosphatase (ACP), alkaline phosphatase (AKP), superoxide dismutase (SOD), peroxidase (POD) were determined respectively and the data were analyzed by independent t test., Results: The glucogen content of snail livers in the SAN group and the control group were (12.151 +/- 0.204) and (18.113 +/- 0.163) mg/g respectively, the difference between the two groups was significant (P < 0.05); the MDA levels of the two groups were (5.298 +/- 0.441) and (4.351 +/- 0.197) nmol/mgprot respectively, and the difference was not significant (P > 0.05); the activities of ALT, AST, ACP, AKP, SOD in the SAN group were (2.760 +/- 0.076) U/mgprot, (68.723 +/- 2.295) U/mgprot, (407.949 +/-19.868) U/gprot, (191.287 +/- 0.771) U/ gprot and (48.452 +/- 0.193) U/mgprot respectively, the activities of these enzymes in the control group were (1.104 +/- 0.000) U/mgprot, (49.448 +/- 1.626) U/mgprot, (344.475 +/- 30.186) U/gprot, (121.905 +/- 3.127) U/gprot and (38.814 +/- 2.765) U/mgprot respectively, the activities of ALT, AST, ACP, AKP and SOD were significantly increased after immersed in 5 mg/L SAN for 36 h, the differences were significant (All P values < 0.05); yet the difference of POD between the SAN group [(22.170 +/- 0.018) U/mgprot] and the control group [(21.747 +/- 0.264) U/mgprot] was not significant (P > 0.05)., Conclusion: SAN can destroy liver functions of Oncomelania hupensis through decreasing glucogen content and changing activities of some important enzymes in snail liver.

Retention parameters of alkaloid standards were determined on different stationary phases, i.e., octadecyl silica, base-deactivated octadecyl silica, cyanopropyl silica, preconditioned cyanopropyl silica, and pentafluorophenyl, using different aqueous eluant systems: acetonitrile-water mixtures; buffered aqueous mobile phases at pH 3 or 7.8; and aqueous eluants containing ion-pairing reagents (octane-1-sulfonic acid sodium salt and pentane-1-sulfonic acid sodium salt) or silanol blockers (tetrabutyl ammonium chloride and diethylamine). Improved peak symmetry and separation selectivity for basic solutes was observed when basic buffer, ion-pairing reagents, and, especially, silanol blockers as mobile phase additives were applied. The best separation selectivity and most symmetric peaks for the investigated alkaloids were obtained in systems containing diethylamine in the mobile phase. The influence of acetonitrile concentration and kind and concentration of ion-pairing reagents or silanol blockers on retention, peak symmetry, and system efficiency was also examined. The most efficient and selective systems were used for separation of the investigated alkaloids and analysis of Fumaria officinalis and Glaucium flavum plant extracts.

Objective: To study the active ingredients of analgesic and anti-inflammatory from Meconopsis quintuplinervia, Tracking the active ingredients of such medicines., Methods: The compounds Meconopsis quintuplinervia,were separated with chromatography and its chemical structure was elucidated by means of MS, NMR spectroscopy methods respectively., Results: Five compounds were obtaine as O-methylflavinantine (I), flavinantin (II), tricin (III), quercitrin (IV) and methyl linoleate (V)., Conclusion: Compounds I is obtained from the plant for the first time.

Macleaya cordata (plume poppy) is a source of bioactive compounds, mainly isoquinoline alkaloids which are used in phytopreparations with anti-inflammatory and antimicrobial activities. In this study, the alkaloids sanguinarine, chelerythrine, their dihydro derivatives, protopine and allocryptopine and phenolics, gallic, protocatechuic, p-hydroxybenzoic, m-hydroxybenzoic, gentisic, p-coumaric, caffeic, ferulic and sinapic acids were determined in extracts prepared from M. cordata aerial part, seeds, and seed capsules using HPLC with UV detection and/or LC/MS with electrospray ionization. The highest content of sanguinarine and chelerythrine was found in capsules. Protopine and allocryptopine were major alkaloids in leaves including footstalks. The seed oil contained dihydrosanguinarine, dihydrochelerythrine and twelve fatty acids of which linoleic, oleic, palmitic and stearic acids predominated. In addition, sanguinarine reductase, a key enzyme in sanguinarine/dihydrosanguinarine equilibrium in plants, was found for the first time, in the soluble proteins of leaves. Finally, extracts were tested for antimicrobial activity using the microdilution method on standard reference bacterial strains., (Copyright © 2010 Elsevier B.V. All rights reserved.)

The ciliate Ichthyophthirius multifiliis is one of the most pathogenic parasites of fish maintained in captivity. In this study, effects of crude extracts, fractions, and compounds from the leaves of Macleaya cordata against I. multifiliis were investigated under in vitro conditions by bioactivity-guided isolation method. The dried ethanol extract of M. cordata was extracted successively in a separating funnel with petroleum ether, ethyl acetate, chloroform and n-butanol. Among them, only the chloroform extract showed promising activity and therefore, was subjected to further separation and purification using various chromatographic techniques. Four compounds were isolated from chloroform extract, but only one compound showed potent activity. The structure of the active compound was elucidated as sanguinarine by hydrogen, carbon-13 nuclear magnetic resonance spectrum and electron ionization mass spectrometry. In vitro antiparasitic efficacy tests exhibited that sanguinarine was 100% effective against I. multifiliis at a concentration of 0.7 mg l(-1), with LC(50) and LC(90) values of 0.437 and 0.853 mg l(-1) after 4 h of exposure. In vivo antiparasitic efficacy tests showed that the number of I. multifiliis on the gills in the treatment group (in 0.9 mg l(-1) sanguinarine) was reduced by 96.8%, in comparison to untreated group at 25°C for 48 h. Mortality of fish did not occur in the treatment group during the trail, although 40% of untreated fish died. Our results indicate that the studied plant extracts, as well as sanguinarine might be potential sources of new antiparasitic drug for the control of I. multifiliis.

The present study aims to evaluate the anthelmintic properties of aerial part of Macleaya microcarpa (Maxim) Fedde. Bioassay-guided fractionation and isolation of the compounds with anthelmintic activity were performed on the ethanolic extract of M. microcarpa yielding five bioactive alkaloids namely: sanguinarine, cryptopine, beta-allocryptopine, protopine and 6-methoxyl-dihydro-chelerythrine by comparing spectral data (UV, NMR, and EI-MS) with literature values. According to in vivo anthelmintic assays, they were found to be 100% effective at the concentrations of 0.7, 8.0, 8.0, 16.0 and 7.0 mgl(-1), and the median effective concentration (EC(50)) values for the five compounds were 0.37, 3.31, 4.64, 8.13 and 3.63 mgl(-1), respectively. Additionally, the acute toxicity on goldfish for the five active compounds was also investigated with median lethal concentrations (LC(50)) values of 1.13, 16.12, 15.88, 21.69 and 10.91 mgl(-1), respectively. The resulting therapeutic indices for sanguinarine, cryptopine, beta-allocryptopine, protopine and 6-methoxyl-dihydro-chelerythrine were 3.03, 4.82, 3.40, 2.66 and 2.99 correspondingly. Correlations analysis between the logP and EC(50), LC(50) of the five alkaloids revealed that the activity of the five alkaloids was well correlated with their hydrophobicity and r(2)=0.45 is for anthelmintic activity while r(2)=0.47 is for acute toxicity for goldfish, respectively. These results provided evidence that the studied plant extract, as well as the isolated compounds, especially sanguinarine, might be potential plant-based medicines for the treatment of D. intermedius infection., (Copyright 2010 Elsevier B.V. All rights reserved.)

In this review, the literature data on the phytochemical and biological investigations on the genus of Meconopsis are summarized from 49 references. Up to now, more than 95 compounds were isolated from 19 Meconopsis plant species. The chemical constituents are mostly alkaloids, flavonoids, phenols, steroids, and terpenes, together with minor constituents of essential oil, and others. The crude extracts and metabolites have been found to possess various bioactivities including antitumor activity, central action, cardiovascular system effects, antibiosis, antiviral activity, anti-inflammatory effects, and other biological activities.

Leishmanicidal activities of benzophenanthridine alkaloids isolated from fruits of Bocconia pearcei and their derivatives were examined. Seven benzophenanthridine compounds were isolated from the methanolic extracts of B. pearcei. Among them, dihydrosanguinarine showed the most potent leishmanicidal activities (IC(50) value: 0.014 microg/ml, respectively). To examine the structure-activity relationship of the benzophenanthridine skeleton, the leishmanicidal activities for 32 synthetic samples were examined. The existence of bulky groups at the C(7)-C(8) position was found to enhance the activity. On the other hand, the bulkiness at the C(2)-C(3) position on the D-ring, a carbonyl group at C-6, substitution at C-6 and cleavage or saturation of the C(5)-C(6) bond reduced activity. A methyl group on nitrogen of the C-ring was thought to be necessary for significant activity.

The purpose of the research was to investigate the multiple response optimizations for the extraction of protopine and allocryptopine from the stems of Macleaya cordata (Willd) R. Br. by using microwave-assisted extraction (MAE). A three-level, three-factor Box-Behnken design of response surface methodology was used to develop response model, and desirability function was employed to optimize the effects of main extraction parameters. Three variables, ethanol concentration (20-80%, v/v), extraction temperature (30-70 degrees C) and solvent/solid ratio (10:1 to 30:1, mL/g), were investigated in this study. The results showed that the optimum parameters of MAE were ethanol concentration of 45.2 % (v/v), extraction temperature of 54.7 degrees C and solvent/solid ratio of 20.4:1 (mL/g). Under these conditions, the extraction yields of protopine and allocryptopine were 89.4 and 102.0%, respectively, and the extracta sicca yield was 12.5%. The combination use of response surface methodology, Box-Behnken design and the appropriate desirability function could provide an insight into a lab-scale MAE process, and help to develop procedures for commercial production of active ingredients from medical plants.

Glaucium flavum collected in Sardinia was studied using a phytochemical approach in order to evaluate its alkaloid composition and obtain a comparison with the alkaloid contents of the same species in populations of other geographic proveniences. In fact, different chemoecotypes of G. flavum have been identified, on the basis of their particular content and composition in alkaloids, in accordance with the different distribution areas. The analysis showed that Sardinian G. flavum contains a homogeneous alkaloid pattern of aporphyne type, significantly different from those reported for populations from other parts of Europe.

A technique based on strip dispersion hybrid liquid membrane was developed for the separation and extraction of four main alkaloids from fruits of Macleaya cordata (Willd) R. Br. A microporous polypropylene membrane impregnated with an organic membrane solution comprised the heart of the strip dispersion hybrid liquid membrane system. The membrane solution was made by dissolving a cationic carrier, di-(2-ethylhexyl) phosphoric acid in an inexpensive, less toxic membrane solvent, kerosene. The transport of alkaloids from an aqueous feed solution through the membrane to a strip dispersion phase was driven by the concentration gradient of H(+) and facilitated by di-(2-ethylhexyl) phosphoric acid. The effects of the extraction time and reuse times of the membrane, the strip solution composition, the carrier concentration, the volume ratio of the aqueous strip solution to the organic membrane solution, and the flow rates of the feed solution and the strip dispersion phase on the transport of alkaloids were investigated. Under the optimal conditions, the permeability coefficients obtained for the four main alkaloids allocryptopine, protopine, sanguinarine, and chelerythrine were 1.66, 1.99, 2.98, and 3.06 x 10(-4) cm/s, and the transport efficiencies were as high as 68, 77, 83, and 85%, respectively.

Methicillin-resistant Staphylococcus aureus (MRSA) is serious clinical urgent problems worldwide. In the present study, the antibacterial activity of Hylomecon hylomeconoides was investigated. The EtOH extract and its fraction (n-hexane, CH(2)Cl(2), EtOAc, and H(2)O) were investigated against MRSA. The most active extract (CH(2)Cl(2)) led to the isolation of 6-methoxydihydrosanguinarine (6-MS), 6-acetonylhydrosanguinarine, and dihydrosanguinarine. These compounds were very active against MRSA strains with minimum inhibitory concentrations (MICs) ranging from 1.95 to 250 microg/ml. Our study did however focus on 6-MS as it appeared to be the most active with MICs in the range of 1.9 to 3.9 microg/ml. These results encourage us to think that 6-MS can be used as a natural antibacterial agent.

A metabolic pathway of 2,3,10,11-oxygenated tetrahydroprotoberberines having the OH group on ring D was demonstrated. Metabolism of (13)C- or D(2)-labeled precursors was studied in cell cultures of Macleaya, Corydalis, and Nandina species. The structures of alkaloid metabolites obtained from feeding experiments were determined by application of combined LC-NMR, LC-MS/MS, and LC-CD techniques. (S)-Tetrahydropseudoprotoberberine (5) was stereospecifically O-methylated to the S-isomer (12) in cell cultures of three plant species. This S-isomer was further N-methylated to the (S)-alpha-N-methyl salt (15), which was oxidized to produce the pseudoprotopine-type alkaloid (10) in cell cultures of Macleaya and Corydalis species. These transformations were the same as those of 2,3,9,10-oxygenated protoberberines. The tetrahydropseudoprotoberberines (5, 6, and 12) were dehydrogenated to pseudoprotoberberines (13, 16, and 14), respectively. Both the R- and S-enantiomers of 5 were dehydrogenated in Macleaya cordata different from the case of 2,3,9,10-oxygenated protoberberines. Precursor 7, with OH groups at C-10 and C-11, was O-methylated at C-10 in M. cordata and C. ochotensis var. raddeana, which was distinct from O-methylation in N. domestica, in which 7 was O-methylated at both C-11 and C-10. Stereoselective O-demethylation [(S)-5 to (S)-18] occurred in N. domestica.

A biologically active benzophenanthridine alkaloid, 6-methoxydihydrosanguinarine (MS), was isolated from Hylomecon plants. Although enantiomers of MS can be separated by chiral HPLC, its isomers rapidly form a racemic mixture in methanol. The rate constants for the racemization of MS enantiomers were 9.20x10(-4)s(-1) and 9.95x10(-4)s(-1) for (+)-MS and (-)-MS, respectively, as determined by dynamic HPLC and chiral chromatography. This unusually rapid racemization may originate from the formation of a stable iminium ion intermediate, sanguinarine. Therefore, the variety of biological activities exhibited by MS may be attributable to a combination of (+)-MS, (-)-MS, and sanguinarine.

Phytochemical investigation of the whole plant of Dicranostigma leptopodum (Maxim) Fedde led to the isolation of a new hopane triterpene, dicranostigmone (1), and a known compound, erythrodiol-3-O-palmitate (2). The structure of the new compound (1) was elucidated by various spectroscopic methods including 2D NMR techniques (gCOSY, HMQC, HMBC, NOESY), HR-ESI-MS, and X-ray.

Bioassay-guided fractionation of the crude extract of the whole plant of Macleaya cordata R. Br. led to the isolation of four alkaloids, which were identified as sanguinarine (1), chelerythrine (2), protopine (3) and alpha-allocryptopine (4) on the basis of their physicochemical and spectrometric data. Compounds 1 and 2 demonstrated a significant antifungal activity against the six test fungi with median inhibitory concentrations (IC50) ranging from 0.47 to 6.13 microg/mL. Compound 1 was the most effective with an IC50 of 0.47 microg/mL on Rhizoctonia solani. Furthermore, compounds 1 and 2 also demonstrated strong antibacterial activity, with IC50 values ranging from 5.01 to 11.3 microg/mL, and minimum inhibitory concentrations (MIC) ranging from 8.0 to 32.0 microg/mL. This is the first report on the activity of the alkaloids from M. cordata against economically important phytopathogenic fungi and bacteria.

A statistic approach using response surface methodology (RSM) for optimization of the ultra-high performance liquid chromatography (UHPLC) gradient and ionization response of electrospray ionization mass spectrometry (ESI-MS) to analyze the main alkaloids from the plant matrices of six Meconopsi species is presented. The optimization was performed with Box-Behnken designs (BBD) and the multicriteria response variables were described using global Derringer's desirability. Four parameters of UHPLC and six major parameters of ESI-MS were investigated for their contribution to analytes separation and response, leading to a total of 27 and 54 experiments being performed for each instrument, respectively. Quantitative analysis of four main alkaloids in nine samples from six Meconopsis species was employed to evaluate the statistical significance of the parameters on UHPLC-QTOF/ESI-MS analytes response. The results indicated that the optimized UHPLC-QTOF-MS method is very sensitive with the limit of detections (LODs) ranging from 0.5 to 0.1 ng/ml. The overall intra-day and the inter-day variations were less than 2.45%. The recovery of the method was in the range of 94.3-104.8% with RSD less than 4.0%. This approach has important implication in sensitivity enhancement of the ultra-trace determination of alkaloids from complex matrixes in the fields of natural products, metabolomics and pharmacokinetics.

Two isochinoline alkaloids, glaucine and oxoglaucine were investigated for their suggested anti-inflammatory influence concerning nitric oxide and cytokine production. Mouse peritoneal macrophages were stimulated with different Toll-like receptor (TLR) ligands such as LPS for TLR4, zymosan for TLR2 and CpG for TLR9. The alkaloids inhibited TNF-alpha and IL-6 production induced by these ligands. In regard to IL-12 suppressive effect was registered in the case of CpG stimulation. Glaucine succeeded to enhance LPS and zymosan-induced IL-10 production. The reduction of pro-inflammatory cytokines and increase of anti-inflammatory IL-10 are indicative for their use in different acute and chronic inflammatory diseases.

Objective: To observe the expression of Bcl-2 and Bax proteins in rat's myocardial cells after Macleaya cordata alkaloids poisoning, and to provide certain molecular biology references for the detection of Macleaya cordata alkaloids poisoning., Methods: Experimental model of Macleaya cordata alkaloids poisoning was established, the expression levels of Bcl-2 and Bax proteins in these cells were detected by immunohistochemistry, and the results were analyzed by computer image system., Results: The expression levels of Bcl-2 and Bax proteins in myocardial cells in poisoning groups were much greater than those in the control groups (P<0.05)., Conclusion: If the clinical symptoms may not be obvious, the detection of Bcl-2 and Bax proteins level by immunohistochemistry still could be ancillary method.

Objective: To study the alkaloids of Macleaya cordata., Method: The alkaloids were isolated and purified by various column chromatography as well as recrystallization. Their structures were identified by physico-chemical properties and spectral analysis., Result: Thirteen alkaloids were isolated and identified as 6-methoxy-dihydrosanguinarine (1), norsanguinarine (2), 6-acetonyl-dihyrochelerythrine (3), 6-acetonyl-dihyrosanguinnarine (4), sanguidimerine (5), chelidimerine (6), (+/-) -bocconarborine A (7), (+/-)-bocconarborine B (8), cryptopine (9), dihydrosanguinarine (10), dihydrochelerythrine (11), protopine (12), alpha-allocryptopine (13)., Conclusion: Compounds 2, 3, 5 - 9 were firstly isolated from the genus Macleaya and compound 1 was obtained from M. cordata for the first time.

A new proaporphine alkaloid, 8, 9-dihydroprooxocryptochine (1), together with three known alkaloids, was isolated from the aerial parts of Meconopsishorridula Hook. f. & Thomson (Papaveraceae), a traditional Tibetan medicine. The structure of 1 was determined by spectroscopic methods.

A method for the analysis of alkaloids in Macleaya cordata (Willd.) R. Br. using high-performance liquid chromatography with diode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESI/MS) was developed. Using protopine (PRO), allocryptopine (ALL), sanguinarine (SA), and chelerythrine (CHE) as the model components, different columns for the separation and different mobile phases for the signal intensities of alkaloids in ESI/MS were investigated, respectively. The results showed that good separation and high signal intensities can be obtained on a high carbon loading (17%) reversed-phase C(18) column with 30 mM formic acid in mobile phase for the analysis of alkaloids. Under the optimal separation condition and UV detection (284 nm), linearity of the six alkaloids was obtained over concentration range from 0.05 to 100.00 microg/ml. The limit of detection (LOD) was 1.62, 1.87, 1.79, 1.76, 1.10, and 0.94 ng/ml for SA, CHE, PRO, ALL, dihydrosanguinarine (DHSA), and dihydrochelerythrine (DHCHE), respectively. The LODs with ESI/MS detection were lower three orders of magnitude than those obtained with UV detection. The proposed method could be used to control quality of the raw materials of the herb more comprehensively.

A new secoberbine alkaloid (-)-N-methylcorydalisol was isolated from the aerial parts of Hypecoum lactiflorum Kar. et Kir. Pazij. (Papaveraceae) of Mongolian origin and was characterised. The known alkaloids of protopine and protoberberine type protopine, allocryptopine, (-)-N-methylcanadine and (-)-N-methylstylopine were also isolated. (-)-N-methylstylopine is a new alkaloid for the genus, while (-)-N-methylcanadine is new for the species. All structures were established by physical and spectral analysis.

Macleaya cordata (Willd.) (Papaveraceae) is used as an active component in the natural feed additive Sangrovit. Sangrovit contains mixture of the intact aerial parts and the fraction of quaternary benzo[c]phenanthridine alkaloids from M. cordata (FQBA). In a 90-day pilot toxicity trial, Sangrovit and the FQBA were tested for safety. Male Wistar rats were fed for 90 days with 100, 7000 or 14000mg of Sangrovit or 600mg of FQBA in 1kg of feed. Body and organ weights, clinical chemistry and hematology markers, oxidative stress parameters, morphological structure of tongue, liver, ileum, kidney and heart samples, and total cytochrome P450 in liver were monitored. The results showed no statistically significant alterations in any parameter between control and treated animals, except for the group treated with 14000ppm Sangrovit that resulted in elevation of reduced glutathione level and superoxide dismutase activity in liver.

Sanguinarine (SAN) and chelerythrine (CHE) are known as major effective components in the quaternary benzo[c]phenanthridine isoquinoline alkaloids (QBA) fraction of Macleaya cordata (Willd) R. Br. but possess different biological activities. In this study, a method for the separation of SAN and CHE based on methyl acrylate-co-divinylbenzene (MA-co-DVB) macroporous adsorbents was established. The relationship between the polarities of the adsorbents and their adsorption-desorption behaviors towards SAN and CHE was investigated. The results showed that, among three selected commercial adsorbents and seven synthesized macroporous polymeric adsorbents with different MA content, the adsorbent No. 5 with 50% MA content provided the best separation power, and the two alkaloids were separated successfully in a gradient eluent process with 60% (v/v) ethanol aqueous and 80% ethanol aqueous contained 8% acetic acid. Dynamic adsorption and desorption tests had been performed in the column packed with the adsorbent No. 5 for optimizing the process parameters. Under the optimized conditions, the ratio of SAN and CHE transformed from 2:1 in the QBA fraction of M. cordata to 1:13 and 25:1 in the products obtained from the two-step gradient elution, and the recoveries of both SAN and CHE were nearly 90%.

Mycobacterium tuberculosis is a serious worldwide health threat, killing almost 2 million people per year. Alternative antimycobacterial drugs are urgently needed; studies have shown that medicinal plants traditionally used to treat respiratory diseases are a potential source of compounds to treat tuberculosis. This paper studied the antimycobacterial activity of 28 extracts from four different plant species that have been used in traditional Mexican medicine to treat tuberculosis. Bark and leaf crude extracts of Juglans regia L., Juglans mollis Engelm., Carya illinoensis (Wangenh) K. Koch and Bocconia frutescens showed in vitro anti-M. tuberculosis activity. Hexane bark extracts from C. illinoensis, J. mollis and J. regia were the most active with a minimal inhibitory concentration (MIC) of 31, 50 and 100 microg/mL, respectively. Ethanol bark extracts from C. illinoensis and J. mollis showed activity at 100 and 125 microg/mL, respectively. Leaf extracts had the lowest activity. Methanol and hexane leaves extracts from B. frutescens had a MIC of 125 microg/mL. None of the aqueous extracts showed antimycobacterial activity., ((c) 2008 John Wiley & Sons, Ltd.)

Purpose: This study describes the antibacterial effect of extracts and compounds isolated from the aerial part of Chelidonium majus Linn. (Papaveraceae) acting against clinical strains of methicillin-resistant Staphylococcus aureus (MRSA)., Methods: The activities were evaluated by using the macrobroth dilution method and reported as the MICs/MBCs., Results: Bioassay-guided fractionation of the most active extract from the aerial parts (EtOAc) led to the isolation of benzo[c]phenanthridine-type alkaloids 8-hydroxydihydrosanguinarine (hhS), 8-hydroxydihydrochelerythrine (hhC), which were potently active against MRSA strains., Conclusions: The selective antibacterial activity reported in this paper for 8-hydroxylated benzo[c]phenanthridine-type alkaloids isolated from C.majus opens the possibility that they could be helpful for the developing of new antibacterial agents for treating the infection of MRSA which has created nosocomial problem worldwide.

A four-component coupling process involving sequential reactions of aldehydes, primary amines, acid chlorides, and nucleophiles has been developed to prepare multifunctional substrates that may be employed in subsequent ring-forming reactions to generate a diverse array of functionalized heterocyclic scaffolds. This new approach to diversity-oriented synthesis was then applied to the first total synthesis of the isopavine alkaloid (+/-)-roelactamine.

The effects of aqueous root extracts from five species of the family Papaveraceae on the growth of cyanobacteria, algae, and other non-target aquatic organisms were investigated to evaluate their potential use as algicides or cyanocides in the aquatic environment. Dicranostigma lactucoides and Sanguinaria canadensis featured the highest toxicity while Macleaya microcarpa was found to be the least toxic to all aquatic organisms tested. The Chelidonium majus extract had the best properties as a potential algicide or cyanocide because of its significant toxicity to phytoplankton and lower toxicity to non-target aquatic organisms as compared with the other Papaveraceae family members., (2007 Wiley Periodicals, Inc.)

Crude methanolic extracts from 22 Mexican medicinal plants were screened for antitrichomonal activity against Trichomonas vaginalis, which is the etiological agent of trichomoniasis. Among the plants tested Carica papaya and Cocos nucifera showed the best antitrichomonal activity with IC(50) values of 5.6 and 5.8 microg/ml, respectively. The extracts of Bocconia frutescens, Geranium mexicanum, and Lygodium venustum showed moderate activity with IC(50) values ranging from 30.9 to 60.9 microg/ml. All the other plant extracts were inactive (IC(50)>100 microg/ml). All extracts tested were less active than metronidazole (IC(50) 0.037 microg/ml), an antiprotozoal drug used as positive control. The results of the antiprotozoal screening support the popular uses of five of the plants tested for the treatment of some urogenital tract disorders in Mexican traditional medicine. However, seeds of Carica papaya and aerial parts of Bocconia frutescens should be used in herbal medicine with care to avoid toxicity.

A method for separation and purification of total alkaloids from Macleaya cordata with macroporous resin was studied, with the content and recovery rate of total alkaloids as indexes. The results were as follows: The static adsorption capacity of AB-8 type resin was 104. 65 mg/g, the static elution ratio were 95.9% , the dynamic adsorption capacity of AB-8 type resin was 96.5 mg/g, the recovery rate was more than 91.24% and the purity was more than 90%. AB-8 type resin was the best for separating and purificating Macleaya cordata in total alkaloids. The optimum conditions is: the eluant is 90% alcohol and 2-3 times as the volume of the resin, the volume of the resin is 10.4 times of total alkaloids in sample, concentration of total alkaloids of sample is 21.57 mg/ml and current velocity of 2-3 ml/min, pH value of sample is 7-8.

The content of the seven quaternary benzo[c]phenanthridine alkaloids (QBA) sanguinarine (SA), chelerythrine (CHE), chelirubine (CHR), chelilutine (CHL), sanguilutine (SL), sanguirubine (SR) and macarpine (MA) was determined in the underground part of six plant species of the family Papaveraceae (Sanguinaria canadensis L., Dicranostigma lactucoides HOOK.f.et THOMS, Chelidonium majus L., Macleaya cordata (Willd.), Macleaya microcarpa (Maxim) and Stylophorum lasiocarpum (Oliv.)). HPLC method with reversed phase column Synergi Max-RP C-12 Phenomenex was used, mobile phase consisted of heptanesulfonic acid (0.01 mol/l) with triethanolamine (0.1 mol/l) in redistilled water, pH 2.5, acetonitrile gradient 25-60% during 25 min. Detection was performed at 280 nm. The highest content of SA and CHE was found in the roots of D. lactucoides (1.99%, resp. 3.43% of the dried roots). In rhizomes of S. canadensis was their content more then two times lower.

An isocratic high-performance liquid chromatographic method coupled with electrospray mass spectrometry was developed to determine protopine, allocryptopine, sanguinarine and chelerythrine in fruits of Macleaya cordata. The sample was extracted with hydrochloric acid aqueous solution using microwave-assisted extraction method. The extracts were separated on a C8 reversed-phase HPLC column with acetonitrile:acetate buffer as mobile phase, and full elution of all analytes was realized isocratically within 10 min. The abundance of pseudomolecule ions was recorded using selected ion recording at m/z 354.4, 370.1, 332.5, 348.5 and 338.5 for protopine, allocryptopine, sanguinarine, chelerythrine and the internal standard, jatrorrhizine, respectively. Internal standard curves were used for the quantification of protopine, allocryptopine, sanguinarine and chelerythrine, which showed a linear range of 0.745-74.5, 0.610-61.0, 0.525-105 and 0.375-75 microg/mL, respectively, with correlation coefficients of 0.9995, 0.9992, 0.9993 and 0.9989, and limits of detection of 3.73, 3.05, 1.60 and 1.11 ng/mL, respectively.

Objective: To observe pathological changes and apoptosis in rats myocardial cells after Macleaya cordata total alkaloids poisoning, and to provide some references for Macleaya cordata total alkaloids poisoning detection., Methods: An experimental model of Macleaya cordata total alkaloids poisoning was established, and the technology of TUNEL staining was used.The results were analyzed by computer image analysis competitive system., Results: Quantities of apoptosis in myocardial cells in poisoning groups were much more than those in the control groups at different tages (P<0.01). In addition the quantities of apoptosis were different after different poisoning duration., Conclusion: Although clinical symptoms was not obvious and could not be detected by poison analysis. Pathological changes induced by Macleaya cordata total alkaloids could be found through the apoptosis detection.

After consumption of poppy seeds various substances were detected in urine or blood samples using an immunoassay and a sophisticated liquid chromatographic-tandem mass spectrometric procedure. These compounds are widely considered to be putative markers of heroin (HER) abuse whereas acetylcodeine was regarded as a marker for illicit preparations ("street HER"). Besides positive urinary opiate immunoassay results during a 48 hours monitoring period, peak concentrations of morphine (MOR), codeine and their glucuronides appeared 4 to 8 hours after ingestion of poppy seeds, and concentrations of total MOR higher than 10 microg/mL were observed. Also, in serum samples taken up to 6 hours after consumption, MOR glucuronides were found. Free MOR was only detected in traces (1 to 3 ng/mL) within 2 hours of consumption. In addition, 3 of 6 onsite opiate sweat tests revealed positive results 6.5 hours after ingestion. Furthermore, it was demonstrated that neither noscapine (NOS) nor papaverine (PAP) was detectable in urine or blood samples after the consumption of poppy seeds containing up to 94 microg NOS and up to 3.3 mug PAP. NOS and PAP were rapidly metabolized, whereas desmethylpapaverine and, especially, its glucuronide were found in urine samples of poppy seed consumers even 48 hours after consumption. According to these results PAP metabolites should not be regarded as markers of illicit HER abuse. In conclusion, only acetylcodeine can be regarded as a specific marker but has the problem of a short half-life. Therefore, we suggest that NOS and PAP, but not their metabolites, might be used cautiously as additional markers of illicit HER abuse as they have not been detected after oral intake of poppy seeds in normal doses. But it must be kept in mind that in some cases poppy seeds with an unusually high content of these alkaloids could be available, and that these substances are also agents in some pharmaceuticals.

The Himalayan blue poppy, Meconopsis grandis, has sky blue-colored petals, although the anthocyanidin nucleus of the petal pigment is cyanidin. The blue color development in this blue poppy involving ferric ions was therefore studied. We analyzed the vacuolar pH, and the organic and inorganic components of the colored cells. A direct measurement by a proton-selective microelectrode revealed that the vacuolar pH value was 4.8. The concentrations of the total anthocyanins in the colored cells were around 5mM, and ca. three times more concentrated flavonols were detected. Fe was detected by atomic analysis of the colored cells, and the ratio of Fe to anthocyanins was ca. 0.8 eq. By mixing the anthocyanin, flavonol and metal ion components in a buffered aq. solution at pH 5.0, we were able to reproduce the same blue color; the visible absorption spectrum and CD were identical to those in the petals, with Fe(3+), Mg(2+) and flavonol being essential for the blue color. The blue pigment in Meconopsis should be a new type of metal complex pigment that is different from a stoichiometric supramolecular pigment such as commelinin or protocyanin.

The contents of total alkaloids in Meconopsis quintuplinervia Regel, grown in the different localtites of Qinghai Province, are detected by the method of spectrophometry. The result showed that total alkaloid in different localities were 0. 0262% to approximately 0.0788% , its mean was 0.0502%. The content of total alkaloids in the herb increased with elevation, not with latitude.

Objective: To study the chemical constituents of a Tibetan medicine Meconopsis quintuplinervia., Method: Column chromatographic techniques were applied to isolate constituents. A combination of IR, MS and NMR spectroscopy was used to identify structures of constituents., Result: Twelve compounds were isolated from the ethanolic extract and their structures were elucidated as quercetin 3-O-beta-D-glucopyranoside (I), quercetin 3-O-beta-D-galactopyranosyl-(1-->6)-glucopyranoside (II), kaempferol 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranoside (III), isorhamnetin 3-0-beta-D-galactopyranosyl-(1-->6)-beta-D-glucopyranoside (IV), caffeic acid (V), protocatechuic acid (VI), p-hydroxycinnamic (VII), 2-(3,4-dihydroxyphenyl )-ethyl-O-beta-D-glucopyranoside (VIII), p-hydroxybenzoyl-beta-D-glucopyranoside (IX), 4-O-beta-D-glucopyranosyl-(Z)-p-coumaric acid (X), 5, 7-dihydroxy-4H-4-chromenone (XI), daucosterol (XII)., Conclusion: Ten compounds were isolated from this genus for the first time except for XI and XII.