RESUMEN
Bamboo leaf extract (BLE) is a pale brown powder extracted from bamboo leaves, and it is listed in the Chinese Standard GB-2760 as a legal and safe food additive. The present study aims to identify and characterize the major flavonoids in BLE. The identification of major flavonoids was carried out using ultra performance liquid chromatography combined with electrospray ionization quadruple time-of-flight tandem mass spectrometry (HPLC/ESI-QTOF-MS/MS). A total of 31 flavonoid compounds were identified and tentatively characterized base on reference standards and MS dissociation mechanisms. HPLC/ESI-QTOF-MS can serve as an important analytical platform to identification structure of bamboo leaf flavonoids (BLF).
Asunto(s)
Flavonoides , Hojas de la Planta , Espectrometría de Masa por Ionización de Electrospray , Espectrometría de Masas en Tándem , Flavonoides/química , Flavonoides/análisis , Hojas de la Planta/química , Cromatografía Líquida de Alta Presión/métodos , Espectrometría de Masa por Ionización de Electrospray/métodos , Espectrometría de Masas en Tándem/métodos , Estructura Molecular , Sasa/química , Extractos Vegetales/química , Medicamentos Herbarios Chinos/química , Bambusa/químicaRESUMEN
Five new glycosides, namely methyl 3-methoxybenzoate-4,5-di-O-ß-D-glucopyranoside (1), (1aS,3aS,3R)-3-(4'-O-ß-D-glucopyranosyl-3'-methoxyphenyl)-5,6-dioxa-bicyclo[3.3.0]octane-1-one (2), quinolin-4(1H)-one-3-O-ß-D-glucopyranoside (3), 3-methoxy-propiophenone 4-O-(6'-ß-D-xylopyranosyl)-ß-D-glucopyranoside (4), methyl 3-methoxybenzoate 4-O-(6'-ß-D-xylopyranosyl)-ß-D-glucopyranoside (5), and one known compound, bambulignan B (6) were isolated from the culms of Phyllostachys nigra var. henonis. Their structures were determined using spectroscopic analysis. All compounds were evaluated for their DPPH radical scavenging activity. Compound 6 exhibited antioxidant activity with IC50 value of 59.5 µM (positive control, L-ascorbic acid, IC50 = 12.4 µM; 2,6-ditertbutyl-4-methyl phenol, IC50 = 11.8 µM).
RESUMEN
The utilization of natural products in food preservation represents a promising strategy for the dual benefits of controlling foodborne pathogens and enhancing the nutritional properties of foods. Among the phytonutrients, flavonoids have been shown to exert antibacterial effects by disrupting bacterial cell membrane functionality; however, the underlying molecular mechanisms remain elusive. In this study, we investigated the effect of quercetin on the cell membrane permeability of Staphylococcus aureus ATCC 27217. A combined metabolomic and transcriptomic approach was adopted to examine the regulatory mechanism of quercetin with respect to the fatty acid composition and associated genes. Kinetic analysis and molecular docking simulations were conducted to assess quercetin's inhibition of ß-ketoacyl-acyl carrier protein reductase (FabG), a potential target in the bacterial fatty acid biosynthesis pathway. Metabolomic and transcriptomic results showed that quercetin increased the ratio of unsaturated to saturated fatty acids and the levels of membrane phospholipids. The bacteria reacted to quercetin-induced stress by attempting to enhance fatty acid biosynthesis; however, quercetin directly inhibited FabG activity, thereby disrupting bacterial fatty acid biosynthesis. These findings provide new insights into the mechanism of quercetin's effects on bacterial cell membranes and suggest potential applications for quercetin in bacterial inhibition.
Asunto(s)
Antibacterianos , Ácidos Grasos , Quercetina , Staphylococcus aureus , Quercetina/farmacología , Quercetina/química , Staphylococcus aureus/efectos de los fármacos , Ácidos Grasos/metabolismo , Ácidos Grasos/biosíntesis , Antibacterianos/farmacología , Simulación del Acoplamiento Molecular , Metabolómica/métodos , Transcriptoma/efectos de los fármacos , Fitoquímicos/farmacología , Fitoquímicos/química , Perfilación de la Expresión Génica , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Metaboloma/efectos de los fármacos , Permeabilidad de la Membrana Celular/efectos de los fármacosRESUMEN
Two new sesquiterpenoids, named 9,10-dihydroxy-albaflavenone (1) and 5-hydroxy-albaflavenone (2) were isolated from Dictyophora indusiata. Their structures and absolute configurations were determined by NMR, ECD and HRESIMS. Compounds 1 and 2 showed anti-inflammatory activity by inhibiting TNF-α and NO secretion to varying degrees.
Asunto(s)
Basidiomycota , Sesquiterpenos , Basidiomycota/química , Sesquiterpenos/farmacología , Factor de Necrosis Tumoral alfaRESUMEN
As an edible and medicinal fungus, Dictyophora indusiata is well-known for its morphological elegance, distinctive taste, high nutritional value, and therapeutic properties. In this study, eighteen compounds (1-18) were isolated and identified from the ethanolic extract of D. indusiata; four (1-4) were previously undescribed. Their molecular structures and absolute configurations were determined via a comprehensive analysis of spectroscopic data (1D/2D NMR, HRESIMS, ECD, and XRD). Seven isolated compounds were examined for their anti-inflammatory activities using an in vitro model of lipopolysaccharide (LPS)-simulated BV-2 microglial cells. Compound 3 displayed the strongest inhibitory effect on tumor necrosis factor-α (TNF-α) expression, with an IC50 value of 11.9 µM. Compound 16 exhibited the highest inhibitory activity on interleukin-6 (IL-6) production, with an IC50 value of 13.53 µM. Compound 17 showed the most potent anti-inflammatory capacity by inhibiting the LPS-induced generation of nitric oxide (NO) (IC50: 10.86 µM) and interleukin-1ß (IL-1ß) (IC50: 23.9 µM) and by significantly suppressing induced nitric oxide synthase (iNOS) and phosphorylated nuclear factor-kappa B inhibitor-α (p-IκB-α) expression at concentrations of 5 µM and 20 µM, respectively (p < 0.01). The modes of interactions between the isolated compounds and the target inflammation-related proteins were investigated in a preliminary molecular docking study. These results provided insight into the chemodiversity and potential anti-inflammatory activities of metabolites with small molecular weights in the mushroom D. indusiata.
Asunto(s)
Agaricales , Agaricales/metabolismo , Lipopolisacáridos/farmacología , FN-kappa B/metabolismo , Simulación del Acoplamiento Molecular , Antiinflamatorios/química , Óxido Nítrico/metabolismoRESUMEN
In 2015, HTC and Valve launched a virtual reality headset empowered with Lighthouse, the cutting-edge space positioning technology. Although Lighthouse is superior in terms of accuracy, latency and refresh rate, its algorithms do not support base station expansion, and is flawed concerning occlusion in moving targets, that is, it is unable to calculate their poses with a small set of sensors, resulting in the loss of optical tracking data. In view of these problems, this paper proposes an improved pose estimation algorithm for cases where occlusion is involved. Our algorithm calculates the pose of a given object with a unified dataset comprising of inputs from sensors recognized by all base stations, as long as three or more sensors detect a signal in total, no matter from which base station. To verify our algorithm, HTC official base stations and autonomous developed receivers are used for prototyping. The experiment result shows that our pose calculation algorithm can achieve precise positioning when a few sensors detect the signal.
RESUMEN
Breast cancer is one of the most common cancers and is the second leading cause of cancer mortality in women worldwide. Novel therapies and chemo-therapeutic drugs are urgently needed to be developed for the treatment of breast cancer. Increasing evidence suggests that fatty acid synthase (FAS) plays an important role in breast cancer, for the expression of FAS is significantly higher in human breast cancer cells than in normal cells. Tannic acid (TA), a natural polyphenol, possesses significant biological functions, including bacteriostasis, hemostasis, and anti-oxidant. Our previous studies demonstrated that TA is a natural FAS inhibitor whose inhibitory activity is stronger than that of classical FAS inhibitors, such as C75 and cerulenin. This study further assessed the effect and therapeutic potential of TA on FAS over-expressed breast cancer cells, and as a result, TA had been proven to possess the functions of inhibiting intracellular FAS activity, down-regulating FAS expression in human breast cancer MDA-MB-231 and MCF-7 cells, and inducing cancer cell apoptosis. Since high-expressed FAS is recognized as a molecular marker for breast cancer and plays an important role in cancer prognosis, these findings suggest that TA is a potential drug candidate for treatment of breast cancer.
Asunto(s)
Antineoplásicos/farmacología , Neoplasias de la Mama/enzimología , Acido Graso Sintasa Tipo I/antagonistas & inhibidores , Taninos/farmacología , Apoptosis/efectos de los fármacos , Western Blotting , Línea Celular Tumoral , Inhibidores Enzimáticos/farmacología , Femenino , Citometría de Flujo , HumanosRESUMEN
The purpose of this work was to characterize the functions of two sesquiterpene synthase genes from moso bamboo (Phyllostachys edulis). Two novel sesquiterpene synthase genes, belonging to the Tpsa subfamily, were isolated from moso bamboo. MoTPS2 was 1641 bp in length and encoded a protein of 63 kDa, whereas MoTPS6 was 1626 bp in length, encoded protein 62.4 kDa. Both genes were expressed in Pichia pastoris for heterologous expression, and protein contents reached 0.243 µg µL(-1) for MoTPS2 and 0.088 µg µL(-1) for MoTPS6. The soluble enzymes were catalytically active, and capable of converting farnesyl pyrophosphate to two distinct sesquiterpene compounds. The MoTPS2 gene encoded a farnesol synthase which was responsible for the production of (E, E)-farnesol. MoTPS6 showed nerolidol synthase activity, catalyzing the formation of (E)-nerolidol. Functional characterization of both MoTPSs should prove beneficial for future research into large-scale fermentation of sesquiterpenes.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Ligasas/metabolismo , Proteínas de Plantas/metabolismo , Poaceae/enzimología , Secuencia de Aminoácidos , Clonación Molecular , Farnesol/metabolismo , Ligasas/genética , Datos de Secuencia Molecular , Filogenia , Pichia/genética , Proteínas de Plantas/genética , Poaceae/genética , Fosfatos de Poliisoprenilo/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alineación de Secuencia , Sesquiterpenos/metabolismoRESUMEN
Two new compounds, erythro-syringylglycerol-9-O-trans-4-hydroxycinnamate 7-O-ß-d-glucopyranoside (1) and indocalatin A (2), together with three known ones, 5,7,3'-trihydroxy-6-C-ß-d-digitoxopyranosyl-4'-O-ß-d-glucopyranosyl flavonoid (3), 5,4'-dihydroxy-3',5'-dimethoxy-7-O-[ß-d-apiose-(1â2)]-ß-d-glucopyranosyl flavonoid (4), and tricin-6-C-ß-boivinopyranosyl-8-C-ß-glucopyranoside (5), were isolated from the 95% EtOH extract of Indocalamus latifolius leaves. Their molecular structures were determined by UV, IR, HRESIMS, CD, and 1D and 2D NMR data analyses.
Asunto(s)
Glucósidos/aislamiento & purificación , Lignanos/aislamiento & purificación , Poaceae/química , Glucósidos/química , Lignanos/química , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Hojas de la Planta/química , EstereoisomerismoRESUMEN
The glutinous rice dumpling named "Zongzi" in Chinese is a type of traditional food that is popular in East Asian countries. "Zongzi" is made of glutinous rice and wrapped in the leaves of Indocalamus latifolius McClure as the packaging material. Four new compounds, latifoliusine A (2), (7S,8R) syringylglycerol-8-O-4'-sinapyl ether 4-O-ß-d-glucopyranoside (7), (7S,8S) syringylglycerol-8-O-4'-sinapyl ether 7-O-ß-d-glucopyranoside (8), and (7R,8S) syringylglycerol-8-O-4'-sinapyl ether 7-O-ß-d-glucopyranoside (10), along with six known compounds (1, 3-6 and 9) were isolated from I. latifolius McClure leaves. The structures and relative configurations of the compounds were determined by detailed spectroscopic analysis, high-resolution electrospray ionization mass spectroscopy (HRESIMS), heteronuclear single quantum correlation (HSQC), heteronuclear multiple bond correlation (HMBC), nuclear overhauser enhancement (NOE) and circular dichroism (CD). All of the isolated compounds were screened for their antibacterial activities in vitro. The results indicated that apigenin 6-C-α-l-arabinopyranosyl-8-C-ß-d-glucopyranoside (5) and apigenin 7-O,8-C-di-glucopyranoside (6) have antibacterial activities against four bacterial strains (Staphylococcus aureus, Bacillus thuringiensis, Escherichia coli and Pseudomonas solanacearum).
Asunto(s)
Apigenina/farmacología , Glicósidos/farmacología , Hojas de la Planta/química , Poaceae/química , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Apigenina/química , Apigenina/aislamiento & purificación , Bacillus thuringiensis/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Glicósidos/química , Glicósidos/aislamiento & purificación , Modelos Moleculares , Estructura Molecular , Ralstonia solanacearum/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacosRESUMEN
Resveratrol is an important antioxidant that confers several beneficial effects on human health. 4-coumarate coenzyme A ligase (4CL) and resveratrol synthase (RS) are key rate-limiting enzymes in the biosynthetic pathway of resveratrol. Using gene fusion technology, the fusion gene, 4CL::RS, was constructed by the 4CL gene from Arabidopsis thaliana and RS gene from Arachis hypogaea. DNAMAN analysis showed that the fusion gene encoded a 964-amino acid protein with an approximate weight of 104.7 kDa and a pI of 5.63. A prokaryotic expression vector containing Nco-I and EcoR-I restriction sites, pET-30a/4CL::RS, was identified by liquid culture bacterial PCR, enzyme digestion, and sequencing, and then used in the induction of expression. Subsequently, a biosynthetic pathway of resveratrol was constructed in Escherichia coli BL21(DE3) that harbored pET-30a/4CL::RS. The recombinant strains were induced to express the fusion protein at 28 °C for 8 h. After bacterial cells were disrupted by hypothermic ultrasonication, the 4CL::RS fusion protein was thoroughly separated from tags using Ni-NTA affinity chromatography, and then detected by SDS-PAGE analysis. When the recombinant strains expressed the fusion protein, the precursor, p-coumaric acid, was converted to resveratrol. In the present study, the final concentration of resveratrol derived from 1 mM p-coumaric acid was 80.524 mg/L, with a 35.28 % (mol/mol) conversion yield.
Asunto(s)
Aciltransferasas/genética , Arabidopsis/enzimología , Arachis/enzimología , Coenzima A Ligasas/genética , Proteínas Recombinantes de Fusión/metabolismo , Aciltransferasas/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Arachis/genética , Arachis/metabolismo , Vías Biosintéticas , Clonación Molecular/métodos , Coenzima A Ligasas/metabolismo , Ácidos Cumáricos/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Propionatos , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/aislamiento & purificación , Resveratrol , Estilbenos/metabolismoRESUMEN
Two new compounds, xylitol 1-O-(6'-O-p-hydroxylbenzoyl)-glucopyranoside (1) and bambulignan B (2), together with three known ones gastrodin (3), glucovanillin (4), and rel-(7S,7'R,8R,8'S)-4,4'-dihydroxy-3,3',5,5'-tetramethoxy-7,7'-epoxyligna-9,9'-diol-9(or)9'-O-ß-glucopyranoside (5), were isolated from the 95% EtOH extract of the dry leaves of Pleioblastus amarus (Keng) keng f. Their structures were determined by UV, IR, HR-ESI-MS, CD, and 1D and 2D NMR data analyses as well as GC experiments.
Asunto(s)
Medicamentos Herbarios Chinos/aislamiento & purificación , Glucósidos/aislamiento & purificación , Lignanos/aislamiento & purificación , Poaceae/química , Xilitol/análogos & derivados , Xilitol/aislamiento & purificación , Medicamentos Herbarios Chinos/química , Glucósidos/química , Lignanos/química , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Hojas de la Planta/química , Xilitol/químicaRESUMEN
Three new phenylpropanoids, namely (7'R,8'R) guaiacylglycerol 4'-O-ß-D-[6â³-O-(4-O-ß-D-glucopyranosyl)-p-hydroxyl-benzoyl]-glucopyranoside (1), (7 R,8R) guaiacylglycerol 8-O-1'-(2',6'-dimethoxy-4'-O-ß-D-glucopyranosyl)-benzene (2), (7'R,8'R) guaiacylglycerol 4'-O-ß-D-[6â³-O-3,5-dimethoxy-4-hydroxylbenzoyl]-gluco-pyranoside (3), along with one known phenylpropanoid (4) were isolated from the ethanol extract of Phyllostachys nigra var. henonis fresh culm. The structures of all compounds were determined by analysis of UV, 1D NMR, 2D NMR, HR-ESI-MS and CD data. All compounds were evaluated for their DPPH radical scavenging activity. Compound 2 (IC50 54.9 µM) and 3 (IC50 77.2 µM) exhibited moderate antioxidant activity compared with two positive control compounds L-ascorbic acid (IC50 15.5 µM) and 2,6-ditertbutyl-4-methyl phenol (IC50 19.1 µM).
RESUMEN
Antifungalmycin 702, a novel polyene macrolide antibiotic produced by Streptomyces padanus JAU4234, strongly inhibited mycelial growth of the rice blast fungus, Magnaporthe grisea, with EC50 of 37 µg/ml and EC90 of 136 µg/ml. Significant reduction in the number of conidia was observed at above 20 µg/ml. Conidia germination and appressorium formation were also suppressed and were not viable with >40 µg/ml. When treated with antifungalmycin 702, hyphae morphology became irregular. Based on microscopic examination, antifungalmycin 702 may exert its antifungal activity by changing the structure of cell membranes and the cytoskeleton and interacting with the organelles. Antifungalmycin 702 thus has potential as a new fungicide in the treatment of rice blast disease.
Asunto(s)
Fungicidas Industriales/farmacología , Macrólidos/farmacología , Magnaporthe/efectos de los fármacos , Polienos/farmacología , Streptomyces/metabolismo , Membrana Celular/efectos de los fármacos , Citoesqueleto/efectos de los fármacos , Fungicidas Industriales/aislamiento & purificación , Macrólidos/aislamiento & purificación , Magnaporthe/crecimiento & desarrollo , Viabilidad Microbiana/efectos de los fármacos , Micelio/efectos de los fármacos , Micelio/crecimiento & desarrollo , Orgánulos/efectos de los fármacos , Oryza/microbiología , Polienos/aislamiento & purificaciónRESUMEN
Flavonoids exhibit a wide range of biological activities and the fragmentation characterization and differentiation of isomeric diglycosyl flavonoids is an important area of research. In this study, ultra-high-performance liquid chromatography electrospray ionization quadrupole time-of-flight tandem mass spectrometry was used to identify two pairs of isomeric diglycosyl flavonoids: naringin/narirutin and neohesperidin/hesperidin. The two pairs of flavonoids were identified by product ion tandem mass spectra of the precursor ions [M-H]-. The results showed that characteristic product ions distinguishing flavonoids glycosylated with rhamnosyl-(alpha1 --> 2)-glucosyl from isomeric flavonoids glycosylated with rhamnosyl-(alpha1 --> 6)-glucosyl were the precursor ion [M-H]- and its characteristic product ions Y0- and [0.2X0-H]-.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Flavanonas/química , Espectrometría de Masas en Tándem/métodos , Flavanonas/análisis , Isomerismo , Metanol/química , Espectrometría de Masa por Ionización de Electrospray/métodosRESUMEN
A large amount of waste from Bambusa chungii culms is generated from the bamboo pulping industry, causing disposal problems. Nevertheless, bamboo culms are a suitable source of functional ingredients, such as antioxidant compounds. However, because of the high compactness and tightness in their material structure, obtaining phytochemicals from bamboo culms using conventional organic solvent extraction methods can be inefficient. In this research, we developed a pressurized hot water extraction (PHWE) method to recover 19 target phenolic compounds from Bambusa chungii culms. The extracted compounds were determined by ultra-high performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UPLC-QTOF-MS). The antioxidant potential of the extracts was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays. We investigated the effects of temperature, extraction time, and the material-to-liquid ratio on PHWE, and these parameters were optimized with a Box-Behnken design experiment and response surface tool. The optimal extraction condition was found at 170 °C, with a 1:30 g/mL material-to-liquid ratio and a 14 min extraction time. Following these optimal parameters, the total yield of target phenolic compounds (TYPC) reached 3.85 mg/g of raw material, and the half-maximal inhibitory concentrations (IC50) for the DPPH and ABTS tests were 94.7 mg/L and 21.88 mg/L, respectively. The FRAP value was 1.23 µmol FSE/mg of dried extract. A strong correlation between TYPC and the antioxidant activity of the extract was confirmed. The TYPC and antioxidant capacity of the optimal PHWE extract of the Bambusa chungii culms were both considerably higher than those of extracts obtained from conventional solvent extractions. These results indicated that PHWE is an excellent green technique for recovering phenolic compounds from bamboo culms, and the PHWE extracts of Bambusa chungii culms may be a good source of natural antioxidants.
RESUMEN
Seven lignans and eight phenylpropanoids, including one new lignan, 7S,8R,8'R-5,5'-dimethoxyariciresinol-4-O-ß-D-glucopyranoside (1), were isolated from the liquid juice of Phyllostachys edulis. Their structures were established by extensive spectroscopic analyses. The absolute configuration of the new compound was determined by comparing its experimental electronic circular dichroism (ECD) spectra with calculated ECD spectra. All compounds were evaluated for their anti-inflammatory activity and xanthine oxidase inhibitor activity, and the results showed that compound 9 exhibited a moderate activity in these two bioassays. In addition, all the compounds can be detected in health panda faeces by LC-MS.
Asunto(s)
Lignanos , Poaceae/química , Propionatos/farmacología , Animales , Antiinflamatorios/aislamiento & purificación , Antiinflamatorios/farmacología , Dicroismo Circular , Heces/química , Lignanos/aislamiento & purificación , Lignanos/farmacología , Espectrometría de Masas , Estructura Molecular , Fitoquímicos/aislamiento & purificación , Fitoquímicos/farmacología , Extractos Vegetales/química , Propionatos/aislamiento & purificación , Ursidae , Xantina Oxidasa/antagonistas & inhibidoresRESUMEN
By studying on absorption spectrum of red compound coming from the reaction system of Fe2+ and 1, 10-phenanthroline and the capacity of antioxidant TBHQ and bamboo leaf extract for scavenging hydroxyl free radical, some results were drawn as follows: the determining wavelength of bamboo leaf extract for scavenging hydroxyl free radical by spectrophotometric method is 509.1 nm, and IC50 (the value of antioxidant concentration at scavenging half of hydroxyl free radical)was used as the index to evaluate scavenging capacity. The determined IC50 values were TBHQ (0.040), M20 (0.378), M40 (0.323), M60 (0.334), and bamboo leaf extract could be used as natural antioxidant.
Asunto(s)
Bambusa/química , Depuradores de Radicales Libres/química , Extractos Vegetales/química , Radical Hidroxilo , Hojas de la Planta , EspectrofotometríaRESUMEN
Neem (Azadirachta indica) extract is well-known as a natural pesticide for the control of agricultural pests. Azadirachtin A and its structural analogues are considered as active compounds. However, the amounts of azadirachtins varies in neem extracts, providing a variety of insecticidal activities. In this study, a novel method of automated online solid-phase extraction coupled with liquid chromatography/quadrupole-time-of-flight mass spectrometry (SPE-LC-Q-TOF-MS) was developed and validated for simultaneous quantification of five azadirachtins (azadirachtins A, B, D, H and I) in seed and leaf extracts of A. indica. Different experimental parameters (such as SPE cartridge, injection volume and washing step) were optimized. The optimized SPE-LC-Q-TOF-MS method showed good recovery (82.0-102.8%), linearity (r2 ≥ 0.9991) and precision (0.83-4.83%). The limit of detections (LODs) for the five analytes ranged from 0.34 to 0.76 ng mL-1. The validated method was successfully applied for determination of the analytes in the neem leaves and seeds from different locations and a neem formulation. The online SPE-LC-Q-TOF-MS method was found to be a simple, precise and accurate and can be used as a powerful tool for quality control of neem extracts or its formulations.
RESUMEN
Chinese chive (jiu cai) is a popular vegetable in China and has a unique flavour and aroma. The molecular basis of the characteristic fragrance and nutritional properties of Chinese chive has not been previously identified. Sequential extractions in a series of solvents and high-performance liquid chromatography were used to isolate 40 compounds from Chinese chive. The compounds were identified based on high-resolution electrospray ionization mass spectra, 1D and 2D nuclear magnetic resonance techniques, and circular dichroism spectra. Eight novel compounds were identified-four new pyrazines, which have distinctive flavour; one new lignan; and three new flavonoids-together with 32 known compounds. Several of these compounds have potential applications as health-promoting dietary supplements, food additives, or seasonings. Additionally, the volatile organic compounds in fresh and steamed Chinese chive were compared, and the toxicological activity of extracts from fresh and steamed Chinese chive was tested in normal rat liver (IAR20) and kidney (NRK) cells. The results showed that Chinese chive is toxic to liver and kidney cells when fresh, but is safe after heating. This could explain why it is traditional to eat cooked Chinese chive. A possible metabolic rule regarding pyrazines is postulated based on this data, and a human metabolic pathway is suggested for two of the novel compounds which have the highest amount of Chinese chive extracts.