Rapid screening of 28 alkaloids in food poisoning samples by liquid chromatography-tandem mass spectrometry / 中国热带医学

@#Abstract: Objective　To investigate a poisoning incident caused by eating eight treasure congee, and establish liquid chromatography (LC)-mass spectrometry (MS)/MS screening method of 28 alkaloids to provide references for disposal of similar poisoning incidents. Methods　LC-MS/MS was used for screening 28 alkaloids in the urine, eight treasure congee and food raw material, and the detected alkaloids were quantified. Samples were extracted with 0.4% formic acid aqueous solution and separated by a Acquity UPLC BEH C18 column (1.7 μm, 100 × 2.1 mm). Acetonitrile-0.2% formic acid aqueous solution was used as the mobile phase and gradient elution was adopted. The ionization mode was electrospray positive ionization mode, and the detection method was multi-reaction monitoring (MRM). Analytes were quantified with the external standard method. Results　In the concentration range of 0-100 ng/mL, the linear correlation coefficient r were greater than 0.999 for 28 alkaloids. The recovery of 28 alkaloids in urine sample ranged from 63.0% to 105.0%, and the relative standard deviations (RSDs) were between 5.8% and 8.6%. The recovery of 28 alkaloids in eight treasure congee sample ranged from 72.0% to 109.0%, and the RSDs were between 6.3% and 9.7%. The recovery of 28 alkaloids in semen sesami nigrum sample ranged from 60.0% to 95.0%, and the RSDs were between 4.8% and 8.2%. Hyoscyamine (2 380.0 ng/mL), scopliamine (3.6 ng/mL) and rac-anisodamine (4.7 ng/mL) were detected in the patient's urine. Hyoscyamine (63.3 μg/g), scopliamine (5.7 μg/g) and rac-anisodamine (2.1 μg/g) were detected in eight treasure congee. Hyoscyamine (901.0 μg/g), scopliamine (80.0 μg/g) and rac-anisodamine (30.1 μg/g) were detected in the seed of Datura stramonium L. The ratio of scopliamine and hyoscyamine in the seed of D. stramonium was 1∶11, which complies with the characteristics of D. stramonium L. In urine sample, the proportion of scopliamine and rac-anisodamine was 0.15% and 0.20%, and hyoscyamine accounted for 99.65%. Conclusion　Seed morphology, the content range and proportion of three alkaloids are all in accord with the characteristics of D. stramonium. Combined with the clinical symptoms of atropine poisoning, it can be deduced that this incident is a family food poisoning caused by accidental consumption of seed of D. stramonium L. The method can provide technical support for the clinical diagnosis and treatment of alkaloid poisoning patients, and also provide a basis for emergency detection and disposal of alkaloid poisoning events.

Ethnobotanical and Floristic Investigations of Datura Species in Mali

Aims: This work aimed to investigate the floristic composition and the different medicinal uses of Datura species in addition to people perceptions about them. Place and Duration of Study: The floristic study has been conducted in three sites located in the region of Sikasso (Mali) during October, 2019. The ethnobotanical one was carried out in five zones located in different regions of Mali in order to register the maximum of relevant information about Datura’s therapeutic virtues. Methodology: A floristic and ethnobotanical investigation on Datura species was carried out based on a stratified probabilistic sampling plan. The free consent and the availability of the respondents was an important criterion during the ethnobotanical survey. Besides, the characteristic parameters of floristic diversity and relative frequencies were calculated based on literature formula. Results: The floristic survey allowed to register three species of Datura: D. stramonium (65.38%), D. innoxia (19.23%) et de D. ferox (15.38%). Around these species, 148 plants belonging to 39 species divided in 32 genus and 18 families. Herbaceous represented 94.87% of these inventoried species. With high coefficients of similarity (Cs>50%), we noted an identical floristic diversity inside the three sites. Moreover, Tousseguela had the highest specific diversity index (1.21). The ethnobotanical survey targeted 118 people in five 5 localities with about 77% of men. If 50% of them found Datura to be useful, 50% considered it to be useful with distrust or dangerous. The data showed that the local populations use Datura in the treatment of many diseases; the most cited were dermatosis (16.98%) and swelling (14.15%). The leaves (93.83%) and seeds (14.81%) were the most exploited organs mainly in the form of decoction (61.18%) and by oral route, massage (15.66%) and bath (13.25%). Conclusion: These results reveal the floristic richness of Datura genus and its associated species.

Diversity and Anti-Dermatophyte Activities of Endophytic Fungi from Datura metel / 中国实验方剂学杂志

Objective:To investigate the taxonomic structure and diversity of endophytic fungi from <italic>Datura metel </italic>and screen the strains with anti-dermatophyte activities， so as to provide resources for the development of new lead compounds against dermatomycosis. Method:Endophytic fungi were isolated from the roots， stems and leaves of <italic>D. metel</italic> after tissue block incubation and then identified by morphological analysis and rDNA-internal transcribed spacer（ITS） sequencing. Their anti-dermatophyte activities were detected by agar diffusion assay. Result:A total of 292 strains of endophytic fungi were isolated from <italic>D. metel</italic>， belonging to 34 genera， with<italic> Fusarium</italic> （72.97%） in roots， <italic>Fusarium </italic>（37.25%） and <italic>Plectosphaerella </italic>（28.43%） in stems， and <italic>Colletotrichum </italic>（39.66%） in leaves as the dominant species. The isolation rate （89.23%）， colonization rate （84.62%）， and diversity index （1.82） of endophytic fungi in leaves were significantly higher than those in roots （70.48%， 70.48% and 1.23） and stems （69.39%，68.03% and 1.64）. The determination of anti-dermatophyte activities of 35 endophytic fungal fermented filtrates showed that the strains exhibiting inhibitory activities against <italic>Microsporum canis</italic>， <italic>Trichosporon mucoides</italic>， <italic>Trichophyton rubrum</italic> and <italic>Candida albicans </italic>accounted for 97.14%， 71.43%， 45.71%， and 25.71%， respectively. Among them， six strains （17.14%）， namely <italic>Fusarium </italic>sp. R1， <italic>Penicillium </italic>sp. R5， <italic>Aspergillus </italic>sp. R7， <italic>Metarhizium </italic>sp.<italic> </italic>S18， <italic>Diaporthe </italic>sp. S19， and <italic>Glomerella </italic>sp.<italic> </italic>L57， all inhibited the four types of cutaneous fungal pathogens. Conclusion:The endophytic fungi in <italic>D. metel</italic> are diverse， and the proportion of endophytic fungi possessing anti-dermatophyte activities is high， allowing them to serve as potential resources for the development of new anti-dermatophyte agents.

Evaluation of in vitro Antioxidant Activities of Ethanol Extracts of Datura innoxia Mill. Leaves and Seeds Harvested in Mali

Aims: The purpose of this study was to evaluate the in vitro antioxidant activities of Datura innoxia Mill. leaves and seeds harvested in Mali. Place and Duration of Study: Collection of plant materials were done at Kolondieba (Mali) in june 2016. Evaluation of polyphenols and flavonoids contents, determination of antioxidant activities were done at Laboratory of Plant and Food Biochemistry and Biotechnology of University of Sciences, Techniques and Technologies of Bamako (Mali) between august 2016 and March 2017. Methodology: The leaves and seeds were collected in Kolondieba, Mali. The phytochemical screening based on the standard methods of tube reactions has been performed with ethanol extracts. The quantitative estimation of total polyphenols was made by the Folin-Ciocalteu method and that of flavonoids by the use of aluminum trichloride. The in vitro antioxidant activities of the ethanol extracts was determined by three methods: the 1,1- diphenyl - 2- picrylhydrazyl (DPPH) test, the ferric reducing ability power (FRAP) test and the Phosphomolybdate test (PPM). Results: The phytochemical screening revealed that leaves and seeds of this plant contains alkaloids, polyphenols, flavonoids, coumarins, tannins, triterpenes and saponin. Phenolic contents of ethanol extracts are 30.97 ± 0.33 mg equivalent gallic acid / g in leaves and 14.02 ± 0.15 mg equivalent gallic acid / g in seeds; those of flavonoids are 15.13 ± 0.2 mg equivalent of quercetin / g in the leaves and 4.93 ± 0.41 mg equivalent of quercetin / g in the seeds. The three tests showed that the leaves have a higher level of antiradical activity in vitro than seeds. Conclusion: The results of this work showed that Datura innoxia Mill. has a good antioxidant activity which would justify its use as a potential source of natural antioxidants.

Epidemiological and clinical profile of various plant poisons in a tertiary care center in South Tamilnadu, India

Background: Plant poisoning is a common scenario in a toxicology unit . Some of the plants contain active components used as medicines. However, some of the plant constituents are poisonous with wide variety of clinical manifestations. Authors aimed to study the pattern of various types of plant poisons and clinical presentations and complications of various plant poisons admitted in our toxicological unit.Methods: A total of 87 patients admitted with history of plant poison ingestion, in toxicological unit of Tirunelveli medical college were studied . It was a retrospective observational study and was done over a period of one year.Results:  Out of 87 cases studied, 92%were due to consumption with suicidal intention. Plant poisoning ranks second only to pesticide poisoning . among the plant poisons encountered 66.6% were due to Oleander poison, followed by Datura (8%) and Glory lily(5.7%). Common parts of plant consumed by patients include seeds and plant oils . Vomiting and abdominal pain were the commonest presentations. Encephalopathy, dyselectrolytemia and arrhythmias were the notable complications in our study.Conclusions: Early admission and prompt first aid and monitoring at tertiary care center are key to the reduced mortality. The study throws light on various manifestations, toxic parts and complications of plant poisons.

Cloning and enzymatic function characterization of a novel tropinone reductase I (DaTRI 2) in Datura arborea / 药学学报

Tropinone reductase I (TRI) is a key branch point enzyme in the midstream of tropane alkaloids (TAs) biosynthesis pathway and represents an important target for TAs metabolic engineering, which can lead to metabolic flux of substrate tropinone to TAs. A novel TRI gene was isolated from Datura arborea, a woody resource plant, and designated as DaTRI2 (GenBank accession number is MH705164). The full-length cDNA of DaTRI2 with 1 135 bp exhibits a high sequence homology (96.8%) with DaTRI, and is predicted to encode a protein of 347 amino acids. Deduced DaTRI2 protein contain a conserved TGXXXGXG motif involved in NADPH binding, the catalytic N-S-Y-K tetrad motif and eleven amino acid residues important for binding to its substrate tropinone. The phylogenetic analysis revealed that DaTRI2 and other TRIs from Solanaceous plants belong to the same cluster and DaTRI2 exhibited closest phylogenetic proximity to TRIs from Datura. DaTRI2 was expressed in E. coli and the purified recombinant protein can catalyze both tropinone reduction and tropine oxidation with an optimum pH value of 8.0 and 9.6, respectively. When tropinone was used as the substrate, the Km and Vmax values of DaTRI2 at pH 6.4 were 210.05 μmol·L-1 and 69.6 nkat·mg-1 protein respectively, while the Km and Vmax values for tropine as the substrate were 188.03 μmol·L-1 and 114 nkat·mg-1 protein respectively, at pH 9.6. DaTRI2 transcript was most abundant in the young leaf, followed by the root. Cloning of DaTRI2 gene and biochemical analysis of recombinant DaTRI2 facilitate further research on the molecular mechanism on TAs biosynthesis in woody plants and provide a more potent candidate for TAs metabolic engineering.

Determination of Some Minerals and Trace Elements in Medicinal Plants - Acalypha indica (L.), Datura metel (L.) and Tylophora indica Used in the Treatment of Ashtma

In the current era, medicinal plants are the major resources of indigenous medicines in the traditional medicine system and are playing an important role. Over 80% of the world population mainly depend only on traditional/folk medicinal plants for plant-based medicines to control many of the diseases and their extracts for health care. In the present study, the medicinal plants commonly used by the local people and traditional practitioners of Kappathgudda area, Gadag District, North Karnataka region for treatment of Asthma viz. Acalypha indica L., Datura metel L. and Tylophora indica belongs to Euphorbiaceous, Solanaceae and Asclepiadaceae families were selected for elemental analysis due to its wider application in the traditional medicinal system. Analytical Atomic Absorption Spectrophotometer technique was adopted for the determination of various elements and indicated that Fe, Ca, K, Mg, Ti, Mn, V, Zn, Cu and Cd were present in all samples. The obtained results also revealed that the mineral and trace elemental contents were well within the permissible range and hence the medicinal plants are safe to be utilized in the drug formulations.

Chemical constituents from roots of Datura metel / 中国中药杂志

The chemical constituents from the n-butanol fraction of the 70% ethanol extract of Datura metel roots were separated by silica gel and ODS chromatogram columns as well as preparative HPLC. On the basis of spectral data analysis, their structures were elucidated. Twenty-one compounds were obtained and their structures were identified as citroside A (1), coniferin (2), paeoniflorin (3), (6R,7E,9R)-9-hydroxy-4,7-megastigmadien-3-one 9-O-[α-L-arabin-opyranosyl-(l→6)-β-D-glucopyranoside] (4), (1R,7R,10R,11R)-12-hydroxyl anhuienosol (5), kaurane acid glycoside A (6), ent-2-oxo-15,16-dihydroxypimar-8(14)-en-16-O-β-glucopyranoside (7), ginsenoside Rg₁(8), ginsenoside Re (9), notoginsenosides R₁(10), N-butyl-O-β-D-fructofuranoside (11), salidroside (12), hexyl β-sophoroside (13), 2,6-dimethoxy-4-hydroxyphenol 1-glucoside (14), benzyl-O-β-D-xylopyranoxyl(1→6)-β-D-glucopyranoside (15), (Z)-3-hexenyl-O-α-L-arabinopyranosyl-(1→6)-β-D-glucopyranoside (16), N-[2-(3,4-dihydro-xyphenyl)-2-hydroxyethyl]-3-(4-methoxyphenyl) prop-2-enamide (17), cannabisin D (18), cannabisin E (19), melongenamide B (20), paprazine (21). Compounds 2-17 and 20-21 were isolated from the Solanaceae family for the first time.

Datura fruit poisoning

@#Datura plants contain anticholinergic properties. Consumers may present with a spectrum of anticholinergic symptoms, including hallucination, agitation, tachycardia, delirium, hyperthermia, and dilated pupils. Prompt identification of the symptoms with appropriate treatment can be life-saving. Some patients might not be able to provide history and therefore recognition of toxidromes is imperative. Awareness should be built among the public who may be exposed to such fruits or plants.

Phenylpropanoids from roots of Daturae Flos / 中草药

Objective To study the phenylpropanoids from the roots of Datura metel. Methods The separations and purifications were taken by silica gel and ODS chromatogram columns as well as preparative HPLC, and the structural identification based on physicochemical property, 1H-NMR and 13C-NMR as well as HR-MS data. Results Nineteen compounds obtained from the butanol fraction of the 70% ethanol extract of D. metel roots, which were identified as icariside E5 (1), alangisesquin A (2), Glycopentoside F (3), conicaoside (4), (7S,8R) dehydrodiconiferyl alcohol 9’-O-β-glucopyranoside (5), iariciresinol-4’-O-β-D-glucopyranoside (6), 7R,8R-threo-4,7,9-trihy-droxy-3,3’-dimethoxy-8-O-4’-neolignan-9’-O-β-D-glucopyranoside (7), vitrifol A (8), leptolepisol D (9), thero-2,3-bis-(4-hydroxy-3-methoxypheyl)-3-methoxy-propanol (10), hyuganoside IIIb (11), officinalioside (12), iariciresinol-9-O- β-D-glucopyranoside (13), stroside A (14), erythro-buddlenol B (15), sargentodoside D (16), (+)-(7S,8S)-4-hydroxy-3,3’,5’- trimethoxy-8’,9’-dinor-8,4’-oxyneolignan-7,9-diol-7’-oic acid (17), 5’-methoxy lariciresinol (18), and dehydrodiconiferyl alcohol 4-O-β-D-glucopyranoside (19). Conclusion Compounds 2-19 are isolated from Solanaceae family for the first time and compound 1 is firstly isolated from genus Datura L.