RESUMO
In this work, a high-speed shear extraction off-line coupling high-speed countercurrent chromatography method was developed to separate maslinic acid and oleanolic acid from olive pomace. To improve extraction efficiency, the polar disparity between maslinic acid and oleanolic acid necessitated the concurrent utilization of both polar and non-polar solvents during high-speed shear extraction. Then, the high-speed shear extraction was directly feed to high-speed countercurrent chromatography for subsequently separation. A total of 250 min were needed to complete the extraction and separation process. This yielded two molecules from 3.3 g of defatted olive pomace: 7.2 mg of 93.8 % pure maslinic acid and 2.3 mg of 90.1 % pure oleanolic acid, both determined by HPLC at 210 nm. Furthermore, the compounds exhibited inhibitory activity against Escherichia coli and Staphylococcus aureus. At a concentration of 100 µg/mL, its efficacy in inhibiting hyaluronidase was comparable to that of the standard drug indomethacin. Compared with the conventional separation method, this coupled technique reduced the whole time due to the direct injection of sample extraction solution. This technique provides a useful approach for the separation of natural products with significant polarity differences.
Assuntos
Olea , Ácido Oleanólico , Ácido Oleanólico/análogos & derivados , Triterpenos , Ácido Oleanólico/análise , Olea/química , Distribuição Contracorrente , Antibacterianos/farmacologia , Triterpenos/química , Cromatografia Líquida de Alta Pressão , Extratos Vegetais/farmacologia , Extratos Vegetais/análiseRESUMO
Comprehensive and rapid analysis of secondary metabolites like saponins remains challenging. This study aimed to establish a semi-automated workflow for filtration, identification, and characterization of saikosaponins in six Bupleurum species. Radix Bupleuri, a high-sales herbal medicine, is often adulterated, restricting its quality control and applications. Two authentic Radix Bupleuri species and four major adulterants were analyzed through UHPLC-LTQ-Orbitrap-MS for targeted saikosaponin analysis. To reveal trace saikosaponins and obtain quality fragment data, a MATLAB-based process automatically enumerating "sugar chain + aglycone + side chain" combinations and deduplicating generated a predicted saikosaponin database covering all possible saikosaponins as a precursor ion list for comprehensive targeted acquisition. To focus on informative ions and reduce MS analysis workload, we utilized MATLAB to automatically filtrate the false positive ions by MS1 and MS2 spectrometry. The newly established MATLAB-assisted data acquisition approach exhibited 50 % improvement in characterization of targeted saikosaponins. Furthermore, positive and negative ionization workflows were designed for accurate saikosaponins characterization based on fragmentation rules. In total, 707 saikosaponins were characterized, including over 500 potential new compounds and previously unreported C29 aglycones. We identified 25 saikosaponins present in both authentic species but absent in adulterants as potential markers. This unprecedented comprehensive multi-origin species differentiation demonstrates the promise of MATLAB-assisted acquisition and processing to advance saponin identification and standardize the Radix Bupleuri market.
Assuntos
Bupleurum , Medicamentos de Ervas Chinesas , Ácido Oleanólico , Saponinas , Medicamentos de Ervas Chinesas/química , Bupleurum/química , Extratos Vegetais , Saponinas/análise , Ácido Oleanólico/análise , Cromatografia Líquida , Espectrometria de Massas , Íons , Cromatografia Líquida de Alta Pressão/métodosRESUMO
It is generally believed that high-quality Bupleurum scorzonerifolium roots possess specific morphological characteristics, being red, robust, and long with strong odor. However, the scientific connotation of these characteristics has not been elucidated. According to the theory of "quality evaluation through morphological identification", we studied the correlations between appearance traits(the RGB value of root surface, root length, root diameter, dry weight, and ratio of phloem to xylem) and content of main chemical components(volatile oils, total saponins, total flavonoids, total polysaccharides, and seven saikosaponins) of B. scorzonerifolium roots. Epson Scanner and ImageJ were used to scan the root samples and measure the appearance traits. Ultraviolet spectrophotometry and HPLC were employed to determine the content of chemical components. The correlation, regression, and cluster analyses were performed to study the correlations between the appearance traits and the content of chemical components. The results showed that the content of volatile oils and saikosaponins were significantly correlated with RGB value, root length, and root diameter, indicating that within a certain range, the roots being redder, longer, and thicker had higher content of volatile oils and saikosaponins. According to the appearance traits and chemical component content, the 14 samples from different producing areas were classified into four grades, and the differences in morphological traits and chemical component content were consistent among different grades. The findings in this study demonstrate that appearance traits(RGB value, root length, and root diameter) can be used to evaluate the quality of B. scorzonerifolium roots. Meanwhile, this study lays a foundation for establishing an objective quality evaluation method for B. scorzonerifolium roots.
Assuntos
Bupleurum , Óleos Voláteis , Ácido Oleanólico , Saponinas , Bupleurum/química , Saponinas/análise , Ácido Oleanólico/análise , Óleos Voláteis/análise , Raízes de Plantas/químicaRESUMO
OBJECTIVES: During the spread of pandemic diseases, immunity boosting herbal drugs are taken as a preventive medicine. Kapacurak Kutinir Curanam is a Siddha drug used for flu like viral infections, cold and fever. Developing an analytical method to estimate the content of active phytoconstituents in such antiviral immune boosting drug will be useful in the phyto pharmaceutical industry. METHODS: A precise, reliable and sensitive ordinary phase high performance thin layer chromatography (HPTLC) method has been developed and validated for identification and simultaneous estimation of six bioactive components namely like andrographolide, columbin, gallic acid, ρ-coumaric acid, piperine and oleanolic acid from any Indian traditional medicine, medicinal plant, drugs and food materials etc. The separation was achieved on silica gel 60F254 TLC plates using toluene: ethyl acetate: formic acid (7:3:0.5, v/v) as mobile phase. The gallic acid, ρ-coumaric acid, piperine markers were estimated using the densitometric scanning in absorption mode at 254 nm. The densitometric scanning was done after derivatization (vanillin-sulphuric acid reagent) at λ=520 nm for andrographolide, columbin and oleanolic acid. RESULTS: The linear regression analysis data for the calibration plots showed a correlation coefficient in the concentration range 1-5 µg per band for the bioactive markers with respect to area. The method was validated for accuracy, precision, limit of detection (LOD), and quantitation of limit (LOQ). CONCLUSIONS: Developed method was accurate, precise and fast to ensure the quality of Kapacurak Kutinir Curanam.
Assuntos
Ácido Oleanólico , Ácido Oleanólico/análise , Cromatografia em Camada Fina/métodos , Ácidos Cumáricos , Ácido GálicoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Xiaojin Pills (XJPs), which has the function of dissipating knots and dispersing swelling, removing blood stasis, and relieving pain, is a classic prescription for the treatment of mammary glands hyperplasia. It is also the first choice of Chinese patent medicine for the clinical treatment of mammary glands hyperplasia in contemporary traditional Chinese medicine clinics. Previous studies have shown that the efficacy of XJPs "taken orally after soaked with Chinese Baijiu" in tradition was significantly better than that of taking it orally with water in modern in terms of activating the blood, anti-inflammation, analgesia, anti-mammary gland hyperplasia, anti-breast cancer and its metastasis in vitro and in vivo, especially under low-dose conditions. However, the material basis for the difference in efficacy between XJP&B and XJP&W is still unclear. AIM OF THE STUDY: To analyze the material basis of the significant difference in efficacy between XJP&B and XJP&W from the perspective of serum pharmacochemistry and pharmacokinetics, and clarified the scientific connotation of XJPs "taken orally after soaked with Chinese Baijiu". MATERIALS AND METHODS: Ultra-high performance liquid chromatography-mass spectrometry combined with a multivariate statistical analysis method were used to screen the differential components in the Chinese Baijiu extract and the water extract of XJPs and the corresponding residues, so as to clarify the differential components between XJP&B and XJP&W in vitro. The migrating components in the blood after XJP&B and XJP&W were characterized by serum pharmacochemical methods, in order to clarify the differential components in rats. The pharmacokinetic parameters of the representative components absorbed into the blood were compared between XJP&B and XJP&W by the pharmacokinetics study method, in order to determine the dynamic changes of the representative components in rats. RESULTS: The identification results of different components in vitro showed that there were 34 and 12 different compounds between the Chinese Baijiu extract and water extract of XJPs, and the residues after Chinese Baijiu extraction and water extraction, respectively. The content of different components such as arachidonic acid, ursolic acid, 3-acetyl-11-keto-ß-boswellic acid, 2α-hydroxyursolic acid, and oleanolic acid was higher in the Chinese Baijiu extract, which was more than twice the content in the water extract. The results of the serum pharmacochemistry study indicated that 42 prototype components were identified in the serum of rats after XJP&B and XJP&W, including organic acids, alkaloids, steroids, and terpenoids. And XJP&B increased the absorption of the prototype components of organic acids in XJPs into the blood. The pharmacokinetic study results of representative components demonstrated that the mean plasma concentration-time profile and pharmacokinetic parameters of muscone, aconitine, and 3-acetyl-11-keto-ß-boswellic acid were significantly different between XJP&B and XJP&W. Compared with XJP&W, the Cmax and AUC0-t of muscone and aconitine in XJP&B were higher, and the T1/2 and MRT0-t of 3-acetyl-11-keto-ß-boswellic acid in XJP&B were relatively longer. CONCLUSION: This research proved that "taking XJPs orally after being soaked with Chinese Baijiu" can increase the dissolution and absorption of active ingredients in XJPs, increase the plasma concentration and content of representative ingredients, and prolong its action time, thus enhancing the biological activity of XJPs in vitro and in vivo. To a certain extent, this study revealed the material basis of the significantly better efficacy of XJP&B than XJP&W and clarified the scientific connotation of XJPs "taken orally after soaked with Chinese Baijiu", which can provide a theoretical basis for the optimization of XJPs' clinical administration method.
Assuntos
Alcaloides , Medicamentos de Ervas Chinesas , Ácido Oleanólico , Aconitina/análise , Animais , Ácidos Araquidônicos , China , Cromatografia Líquida de Alta Pressão/métodos , Cicloparafinas , Medicamentos de Ervas Chinesas/química , Hiperplasia , Medicamentos sem Prescrição , Ácido Oleanólico/análise , Ratos , Triterpenos , ÁguaRESUMO
Apples are rich sources of ursolic acid (UA) and oleanolic acid (OA) which are the major and most prominent triterpenes in the peel of an apple. Pentacyclic triterpenes are ideal nutraceuticals due to their ability to reduce the risk of many life-threatening diseases such as cancer, cardiovascular and diabetes. This study was to determine the content of UA and OA in the apple peel extract from different cultivars grown in South Africa as well as the correlation of their content level with antioxidant capacity. Quantitative analysis of UA and OA in apple peels from three cultivars; red delicious (RD), royal gala (RG) and granny smith (GS) apples was carried out using HPLC and their antioxidant capacity was analyzed using the DPPH assay. The RD showed the highest content of UA and OA (248.02 ± 0.08 µg/ml and 110.00 ± 0.08 µg/ml respectively) in the apple peel extract and also displayed a significantly high level of antioxidant capacity (97.3 ± 0.40%; p < 0.0001) compared to the RG and GS cultivars. A strong positive correlation was noted between the UA, OA and antioxidant capacities of all the cultivars. Only the RD cultivar showed a significant correlation though; UA (r = 0.9570; p = 0.0027) and OA (r = 0.8503; p = 0.0319). This study demonstrated that the RD and RG apple peels possess the highest UA and OA content which invariably increases their antioxidant activities compared to GS apple. Thus, both apple cultivars would be useful and recommended for food consumption and nutraceuticals values to improve human health.
Assuntos
Malus , Ácido Oleanólico , Antioxidantes/análise , Antioxidantes/farmacologia , Frutas/química , Humanos , Ácido Oleanólico/análise , Ácido Oleanólico/química , Ácido Oleanólico/farmacologia , Extratos Vegetais/químicaRESUMO
Cannabis sativa L. has been used for a long time to obtain food, fiber, and as a medicinal and psychoactive plant. Today, the nutraceutical potential of C.sativa is being increasingly reappraised; however, C. sativa roots remain poorly studied, despite citations in the scientific literature. In this direction, we identified and quantified the presence of valuable bioactives (namely, ß-sitosterol, stigmasterol, campesterol, friedelin, and epi-friedelanol) in the root extracts of C. sativa, a finding which might pave the way to the exploitation of the therapeutic potential of all parts of the C. sativa plant. To facilitate root harvesting and processing, aeroponic (AP) and aeroponic-elicited cultures (AEP) were established and compared to soil-cultivated plants (SP). Interestingly, considerably increased plant growth-particularly of the roots-and a significant increase (up to 20-fold in the case of ß-sitosterol) in the total content of the aforementioned roots' bioactive molecules were observed in AP and AEP. In conclusion, aeroponics, an easy, standardized, contaminant-free cultivation technique, facilitates the harvesting/processing of roots along with a greater production of their secondary bioactive metabolites, which could be utilized in the formulation of health-promoting and health-care products.
Assuntos
Cannabis/química , Cannabis/crescimento & desenvolvimento , Hidroponia , Colesterol/análogos & derivados , Colesterol/análise , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/análise , Fitosteróis/análise , Extratos Vegetais/química , Raízes de Plantas/química , Raízes de Plantas/crescimento & desenvolvimento , Sitosteroides/análise , Estigmasterol/análise , Triterpenos/análiseRESUMO
As the main active component of Panacis majoris Rhizoma, Chikusetsu saponin IVa has the activity of anti-oxidation, anti-inflammatory pain, and so on. Obtaining high purity Chikusetsu saponin IVa by simple purification steps is a prerequisite for its deep development. In this paper, the separation and purification of Chikusetsu saponin IVa were studied by molecular imprinting technique. By ultraviolet and visible spectrophotometry and computer molecular simulation, it was concluded that water-soluble 3-(2-carboxyethyl)-1-vinylimidazolium bromide ionic liquid was the best functional monomer compared with acrylic acid and acrylamide. The molecularly imprinted polymers were prepared by precipitation polymerization at 60â with Chikusetsu saponin IVa as template molecule, 3-(2-carboxyethyl)-1-vinylimidazolium bromide as functional monomer, ethylene glycol dimethacrylate as cross-linker, 2, 2'-azobisisobutyronitrile as initiator, and ethanol as porogen. The properties of molecularly imprinted polymers were studied by scanning electron microscopy, Fourier transform infrared spectroscopy, thermo-gravimetric analysis, nitrogen adsorption/desorption isotherm, and X-ray photoelectron spectroscopy. The maximum adsorption capacity was 171.33 mg/g, and the imprinting factor was 2.6. Finally, the polymers can be successfully used in the purification of Chikusetsu saponin IVa from Panacis majoris Rhizoma through a simple procedure, the content was significantly increased. The recoveries of the spiked samples for the CS-IVa ranged from 94.05 to 99.95% with relative standard deviation values lower than 2.67%. The results showed that the polymers demonstrated good adsorption capacity for Chikusetsu saponin IVa. Meanwhile, the polymers showed great stability and reusability during the application.
Assuntos
Impressão Molecular/métodos , Ácido Oleanólico/análogos & derivados , Saponinas/isolamento & purificação , Extração em Fase Sólida/métodos , Cromatografia Líquida de Alta Pressão , Ácido Oleanólico/análise , Ácido Oleanólico/química , Ácido Oleanólico/isolamento & purificação , Panax/química , Reprodutibilidade dos Testes , Rizoma/química , Saponinas/análise , Saponinas/química , Sensibilidade e EspecificidadeRESUMO
Shuganzhi Tablets (SGZT) is developed on the basis of a clinical empirical formula as a hospital preparation for the treatment of fatty liver. In this study, a rapid and highly sensitive LC-MS/MS method was established and validated for simultaneous determination of ginsenoside Re, ginsenoside Rg1, notoginsenoside R1, naringin, specnuezhenide, emodin, polydatin, hesperidin and saikosaponin A in rat plasma. Multiple reaction monitoring mode played an important role in simultaneous quantitative analysis of multiple components. The analytes were separated by the action of an ACQUITY UPLC® BEH C18 column (2.1 × 50 mm, 1.7 µm) in five minutes. The validated LC-MS/MS method was successfully applied to the pharmacokinetic analysis of hesperidin, emodin, polydatin and naringin of SGZT in rat plasma after administration. A UHPLC system couple with a quadrupole combined with time of flight mass spectrometer was used for qualitatively analyzing of the composition of SGZT and its metabolites in serum, urine, bile and feces of rats. The results showed that a total of 65 components were detected in rat biological samples, including 10 prototype components and 55 metabolites. It was speculated that the ingredients of SGZT experienced mainly the following reactions in rats: phase I reaction such as hydrolysis, oxidation, hydroxylation, carboxylation and dehydroxylation and phase â ¡ reaction such as glucuronidation and sulfation. These results provide useful information for the further study of its active ingredients.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas , Espectrometria de Massas em Tandem/métodos , Animais , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/análise , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacocinética , Ginsenosídeos/análise , Ginsenosídeos/química , Ginsenosídeos/farmacocinética , Modelos Lineares , Masculino , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/análise , Ácido Oleanólico/química , Ácido Oleanólico/farmacocinética , Ratos , Ratos Wistar , Reprodutibilidade dos Testes , Saponinas/análise , Saponinas/química , Saponinas/farmacocinética , Sensibilidade e Especificidade , Espectrometria de Massas por Ionização por Electrospray/métodos , ComprimidosRESUMO
Ardisia crenata Sims (Primulaceae) occurs in natural habitats in two varieties, bearing red or white fruits. While roots of the red-berried ardisia are valued as a medicinal product, the pharmacological activity of which is attributed to triterpene saponins, including ardisiacrispin A, data on the white-berried variety are scarce. A TLC-densitometric method was developed and validated to estimate the levels of saponins, calculated as ardisiacrispin A, in different plant parts in both varieties. Their content amounted to 22.17±4.75 and 25.72±1.46â mg/g d.w. in roots, and 2.64±0.74 and 3.43±0.70â mg/g d.w. in fruits of red-berried and white-berried ardisia, respectively. Assessment of cytotoxicity of ardisiacrispin A and A. crenata extracts on a panel of human cancer cell lines revealed a similar effect of root extracts from both varieties, with the highest potency against melanoma WM793 and colon cancer Caco2. Thus, roots of the white-berried variety may be treated as a substitute for red-berried ardisia and serve as an alternative source for the acquisition of plant material rich in bioactive saponins.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Ardisia/química , Ácido Oleanólico/análogos & derivados , Extratos Vegetais/farmacologia , Saponinas/farmacologia , Antineoplásicos Fitogênicos/análise , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Ácido Oleanólico/análise , Ácido Oleanólico/farmacologia , Extratos Vegetais/análise , Raízes de Plantas/química , Saponinas/análiseRESUMO
The content of ursolic acid and oleanolic acid was determined in different plant parts of two Glechoma species, G. hederacea and G. hirsuta. To achieve optimal extraction conditions of ursolic acid and oleanolic acid from plant material, several methods including maceration, heat reflux, Soxhlet, and ultrasonic extraction, as well as various solvents (methanol, dichloromethane, ethyl acetate), were investigated and compared.For the simultaneous quantification of pentacyclic triterpenes in extracts from Glechoma sp., an UPLC-MS/MS was developed and validated. The method exhibited good linearity, precision, and recovery, and it also was simple, specific, and fast. We developed the method for future application in the quality control of plant materials and botanical extracts containing ursolic acid and oleanolic acid. With regard to the triterpene constituents, both G. hederacea and G. hirsuta can be used equally, and the aboveground parts of both species, but the leaves especially, are abundant sources of ursolic acid (7.1â-â7.5 mg/g dry weight [DW]). Dichloromethane as an extractant provided the best extraction efficiency as well as selectivity to obtain Glechoma extracts rich in triterpenes as compared to methanol and ethyl acetate, regardless of the particular extraction technique. Dry dichloromethane extracts from aerial parts of Glechoma sp. obtained by the heat reflux method resulted in products with a high content of UA (17â-â25% w/w) are considered to be convenient and rich sources of this compound.
Assuntos
Lamiaceae , Ácido Oleanólico , Triterpenos , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Ácido Oleanólico/análise , Triterpenos Pentacíclicos , Extratos Vegetais , Espectrometria de Massas em Tandem , Triterpenos/análiseRESUMO
Chloroform extracts from leaves, inflorescences and fruits of Prunus padus were analysed for anti-inflammatory activity and accumulation of corosolic (CA), ursolic (UA) and oleanolic (OA) acids. The analytes were identified and quantified by GC-MS and UHPLC-PDA. Their total levels depend on plant material type and harvesting time, and varied from 0.25 mg/g DW in fruits, through 0.76-1.09 mg/g DW in flowers, to 1.41-4.54 mg/g DW in leaves. Significant variation in the leaf analytes contents was observed during vegetation with the peak amounts in autumn, which indicated the optimal harvesting season. The plant extracts inhibited pro-inflammatory enzymes (lipoxygenase and hyaluronidase) in a concentration-dependent manner, and their activity parameters correlated with the levels and activity of pure triterpene acids, especially CA and UA. The results of the comparison with the positive controls (heparin, indomethacin, dexamethasone) might partly support the application of P. padus in anti-inflammatory therapies, reported by traditional medicine.
Assuntos
Frutas/química , Inflorescência/química , Ácido Oleanólico/análise , Extratos Vegetais/química , Folhas de Planta/química , Prunus/química , Triterpenos/análise , Cromatografia Líquida de Alta Pressão , Cromatografia Gasosa-Espectrometria de Massas , Lipídeos/química , Extratos Vegetais/farmacologia , Ácido UrsólicoRESUMO
Differing sensitivity is the main obstacle for a direct combination of HPTLC with NMR spectroscopy. A sufficient amount of the isolated compound zone must be provided by HPTLC for subsequent offline NMR detection (HPTLC//NMR). To fill the gap, a straightforward procedure was developed using the same analytical HPTLC system for both bioprofiling and isolation of bioactive zones from multicomponent mixtures. The HPTLC-effect-directed analysis (EDA) revealed several bioactive compounds in five botanical extracts, i.e. Salvia officinalis, Thymus vulgaris and Origanum vulgare, all Lamiaceae, and peels of red and green apples (Jonagored and Granny Smith, respectively), both Rosaceae. A tricky case study was designed to show how to deal with potentially coeluting bioactive structural isomers, e.g., ursolic (UA), oleanolic (OA) and betulinic acids (all C30H48O3), which are most difficult to identify and assign. A multipotent bioactive HPTLC zone showed the same hRF value and mass signal in HPTLCHRMS, though containing the coeluting structural isomers UA and OA. After zone isolation from the HPTLC plate, first the 1H NMR spectrum allowed to distinguish distinct allylic H-18 protons, i.e. 2.20 ppm for UA and 2.85 ppm for OA, and at the same time, to quantify the two isomers by using the PUlse Length-based CONcentration methodology (HPTLC//1H qNMR-PULCON). In case of a partial overlap of the diagnostic signal with that of the matrix, results were corroborated with those obtained by using the 1H deconvoluted or 2D 1H-13C Heteronuclear Single Quantum Coherence spectra. The comparison of the quantitative results showed a good correlation (R2 = 0.9718) between the two orthogonal methods HPTLC-Vis and HPTLC//1H qNMR-PULCON. A sufficient zone isolation from the HPTLC plate (mean isolation rate of 82%) for both UA and OA (0.27 - 4.67 mM) was achieved for HPTLC//qNMR, comparing the isolated bioactive compound zone with the respective zone in the botanical extract via HPTLC-Vis densitometry. The HPTLC-EDA-Vis//1H qNMR-PULCON procedure for bioprofiling and quantification/identification/confirmation of bioactive compounds in botanical extracts is considered as straightforward, eco-friendly (only 16 mL solvent required), simple (NMR calibration used over weeks) and reliable new alternative to the status quo of bioactivity-guided fractionation.
Assuntos
Bioensaio/métodos , Espectroscopia de Ressonância Magnética , Compostos Fitoquímicos/análise , Fracionamento Químico , Cromatografia em Camada Fina , Lamiaceae/química , Ácido Oleanólico/análise , Ácido Oleanólico/química , Triterpenos Pentacíclicos , Extratos Vegetais/química , Padrões de Referência , Triterpenos/análise , Triterpenos/química , Ácido Betulínico , Ácido UrsólicoRESUMO
In Phytolaccaceae family, Phytolacca americana L. (American pokeweed) and P. esculenta Van Houtte (Chinese pokeweed) are the two representative species among the genus. Pokeweeds have triterpenoid saponins as toxic compounds in every part of the plant. The saponins phytolaccoside A, B, D, E, and G were isolated from P. americana, and esculentoside H, J, L, K, M, I, and N were isolated from P. esculenta. Along with saponins, their aglycones (phytolaccagenin, phytolaccagenic acid, esculentic acid and jaligonic acid) were also isolated from P. americana and P. esculenta. Two people who unknowingly ate misidentified pokeweed plant roots were transferred to the emergency room. Urine and gastric content after irrigation were collected from the first patient (patient 1), and blood and urine were collected from the second patient (patient 2). The samples were analyzed to identify toxic substances with liquid chromatography-tandem mass spectrometry (LC-MS/MS). In the blood sample, 1.9â¯ng/mL of esculentoside A and 1.5â¯ng/mL of esculentoside C were detected, while the concentration of esculentoside B and H were below the LLOQ. In gastric contents and ingested roots, esculentoside A, B, C, and H were identified. Esculentoside A, C, and H were identified in the urine of patient 1, and esculentoside A and C were identified in the urine sample of patient 2. The developed analytical method was validated for parameters such as linearity, limit of detection, precision, accuracy, matrix effect, recovery, and process efficiency, and they showed clear and unbiased results.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Ácido Oleanólico , Phytolacca americana/química , Intoxicação por Plantas/diagnóstico , Saponinas , Humanos , Limite de Detecção , Modelos Lineares , Ácido Oleanólico/análise , Ácido Oleanólico/intoxicação , Extratos Vegetais/química , Reprodutibilidade dos Testes , Saponinas/análise , Saponinas/intoxicação , Espectrometria de Massas em TandemRESUMO
The Brazilian red propolis (BRP) constitutes an important commercial asset for northeast Brazilian beekeepers. The role of Dalbergia ecastaphyllum (L.) Taub. (Fabaceae) as the main botanical source of this propolis has been previously confirmed. However, in addition to isoflavonoids and other phenolics, which are present in the resin of D. ecastaphyllum, samples of BRP are reported to contain substantial amounts of polyprenylated benzophenones, whose botanical source was unknown. Therefore, field surveys, phytochemical and chromatographic analyses were undertaken to confirm the botanical sources of the red propolis produced in apiaries located in Canavieiras, Bahia, Brazil. The results confirmed D. ecastaphyllum as the botanical source of liquiritigenin (1), isoliquiritigenin (2), formononetin (3), vestitol (4), neovestitol (5), medicarpin (6), and 7-O-neovestitol (7), while Symphonia globulifera L.f. (Clusiaceae) is herein reported for the first time as the botanical source of polyprenylated benzophenones, mainly guttiferone E (8) and oblongifolin B (9), as well as the triterpenoids ß-amyrin (10) and glutinol (11). The chemotaxonomic and economic significance of the occurrence of polyprenylated benzophenones in red propolis is discussed.
Assuntos
Clusiaceae/química , Dalbergia/química , Isoflavonas/química , Compostos Fitoquímicos/análise , Compostos Fitoquímicos/química , Benzofenonas/análise , Benzofenonas/química , Brasil , Chalconas/análise , Cromatografia Líquida de Alta Pressão , Desenho de Fármacos , Flavanonas/análise , Flavonoides/análise , Isoflavonas/análise , Espectroscopia de Ressonância Magnética , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/análise , Extratos Vegetais/análise , Pterocarpanos/análise , Terpenos/análise , Triterpenos/análiseRESUMO
Natural product studies explore potential and interesting new compounds to discover innovative drugs. Nigella sativa (N. sativa) (Ranunculaceae) is traditionally used to treat diabetes. Flavonoids and triterpenoid mostly show anti-diabetic activity. The current study aim to identify new compounds by a systematic study of the anti-oxidant and anti-diabetic activity of aerial parts of N. sativa concerning. Phytochemicals were isolated from the methanolic extract of aerial parts of the plant by column chromatography and identified by nuclear magnetic resonance spectroscopy and mass spectroscopy. A new triterpenoid saponin glycoside was isolated along with flavonoids. The anti-diabetic study was carried out by DPPH, ABTS, α -glucosidase, and protein tyrosine phosphatase 1B assays at doses of 12.5 to 250 µM. The isolated phytochemicals were identified as 3-O-(ß-d-xylopyranosyl-(1-3)-α-l-rhamnopyrnaosyl-(1-2)-α-l-arabinopyranosyl]-28-O-(α-l-rhamno-pyranosyl-(1-4)-ß-d-glucopyranosyl-(1-6)-ß-d-glucopyranosyl] hederagenin (1), flaccidoside III (2), catechol (3), quercetin-3-gentiobiosides (4), magnoflorine (5), nigelflavonoside B (6), nigelloside (7), quercetin sphorotrioside (8), kaempferol-3, 7-diglucoside (9), kaempferol 3-O-rutinoside (10), rutin (11), 3-O-[α-l-rhamnopyranosyl-(1â2)-α-l-arabinopyranpsylhederagenin (12), 3ß,23,28-trihydroxyolean-12-ene-3-O-α-l-arabinopyranoside(1â4)-a-rhamnopyranosyl,(1â4)-ß-d-gluco-pyranoside (13), 3-O-α-l-rhamnopyranosyl-(1â2)-α-l-arabinopyranpsyl]-28-O-ß-d-gluco-pyranosyl hederagenin (14), and α-hederin (15). These were isolated and are reported for the first time in this study. Compared 13 was identified as a new compound. Compound 2 was isolated for first time from the genus Nigella. Compound 6 was found to be the most active in the DPPH, and ABTS assays and compound 10 was found to be the most active in the α-glucosidase assay, with IC50 32.7 ± 0.1, 95.18 ± 0.9, 214.5 ± 0.0 µΜ, respectively. Compound 12, at a dose of 125 µΜ, showed anti-diabetic activity in a PTP1B assay with IC50 91.30 ± 2.5 µΜ. In conclusion, the anti-diabetic activity of N. sativa is due to its flavonoids and TTSGs. Therefore, our studies suggest that the aerial parts of N. sativa are also a valuable and alternate source of valuable phytochemicals that could be used to develop anti-oxidant and anti-diabetic medicines.
Assuntos
Antioxidantes/análise , Diabetes Mellitus/tratamento farmacológico , Nigella sativa/química , Ácido Oleanólico/análogos & derivados , Componentes Aéreos da Planta/química , Extratos Vegetais/análise , Saponinas/análise , Antioxidantes/química , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Benzotiazóis/química , Compostos de Bifenilo/química , Cromatografia Líquida de Alta Pressão , Flavonoides/análise , Flavonoides/isolamento & purificação , Flavonoides/farmacologia , Inibidores de Glicosídeo Hidrolases/farmacologia , Concentração Inibidora 50 , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Estrutura Molecular , Nigella sativa/enzimologia , Ácido Oleanólico/análise , Ácido Oleanólico/química , Ácido Oleanólico/isolamento & purificação , Ácido Oleanólico/farmacologia , Picratos/química , Componentes Aéreos da Planta/enzimologia , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Proteínas Tirosina Fosfatases/antagonistas & inibidores , Saponinas/química , Saponinas/isolamento & purificação , Saponinas/farmacologia , Ácidos Sulfônicos/química , Triterpenos/análise , Triterpenos/isolamento & purificação , Triterpenos/farmacologiaRESUMO
An ultrasound-assisted supercritical carbon dioxide (USC-CO2) procedure was developed for the extraction of ursolic acid, oleanolic acid and oridonin from Rabdosia rubescens, with yields that were 9.84-10.46 and 15.43-21.10% higher than those of the conventional SC-CO2 and heat-reflux extractions, respectively. USC-CO2 uses a shorter extraction time (1.83-2.09 times) and less organic solvent (3.39-173.25 times) to operate at a lower extraction temperature (5-16 °C). The dominant component in the extract was oridonin, which may indicate that the kinetic behavior in the extraction system is predominated by that of oridonin. Furthermore, the USC-CO2 and conventional SC-CO2 dynamic extraction kinetics of oridonin from R. rubescens were well described by the second-order rate and Fick's second law models. The extraction rate constant, energy of activation for diffusion, Biot number and thermodynamic parameters were deduced from the data obtained. These results provide valuable insights into the USC-CO2 and conventional SC-CO2 procedures.
Assuntos
Fracionamento Químico/métodos , Diterpenos do Tipo Caurano/isolamento & purificação , Isodon/química , Ácido Oleanólico/isolamento & purificação , Triterpenos/isolamento & purificação , Dióxido de Carbono/química , Cromatografia Líquida de Alta Pressão , Diterpenos do Tipo Caurano/análise , Química Verde , Cinética , Ácido Oleanólico/análise , Extratos Vegetais/química , Plantas Medicinais/química , Reprodutibilidade dos Testes , Solventes/química , Espectrometria de Massas em Tandem , Triterpenos/análise , Ultrassom , Ácido UrsólicoRESUMO
Saponins are an important group found in Chenopodium quinoa. They represent an obstacle for the use of quinoa as food for humans and animal feeds because of their bitter taste and toxic effects, which necessitates their elimination. Several saponins elimination methods have been examined to leach the saponins from the quinoa seeds; the wet technique remains the most used at both laboratory and industrial levels. Dry methods (heat treatment, extrusion, roasting, or mechanical abrasion) and genetic methods have also been evaluated. The extraction of quinoa saponins can be carried out by several methods; conventional technologies such as maceration and Soxhlet are the most utilized methods. However, recent research has focused on technologies to improve the efficiency of extraction. At least 40 saponin structures from quinoa have been isolated in the past 30 years, the derived molecular entities essentially being phytolaccagenic, oleanolic and serjanic acids, hederagenin, 3ß,23,30 trihydroxy olean-12-en-28-oic acid, 3ß-hydroxy-27-oxo-olean-12en-28-oic acid, and 3ß,23,30 trihydroxy olean-12-en-28-oic acid. These metabolites exhibit a wide range of biological activities, such as molluscicidal, antifungal, anti-inflammatory, hemolytic, and cytotoxic properties.
Assuntos
Chenopodium quinoa/química , Saponinas/química , Saponinas/isolamento & purificação , Sementes/química , Extração em Fase Sólida/métodos , Anti-Inflamatórios/análise , Chenopodium quinoa/genética , Cromatografia Líquida de Alta Pressão , Temperatura Alta , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/análise , Extratos Vegetais/farmacologia , Saponinas/análise , Saponinas/genética , Sementes/genética , Sonicação/métodos , ÁguaRESUMO
A method for sample pretreatment using liquid-liquid chromatography combined with a conventional liquid chromatography was developed for quantitative determination of trace chemical component in traditional Chinese medicine. The main effective component, wilforlide A, in the traditional Chinese medicinal herb Tripterygium wilfordii as well as in its Chinese patent medicine glycosides tablets was successfully determined after sample pretreatment by liquid-liquid chromatography. A biphasic solvent system n-hexane-ethyl acetate-ethanol-water (6:4:6:4, v/v) was screened for crude sample treatment by liquid-liquid chromatography. The collection time of eluted fractions containing target component could be well predicted using a continuous-stirred tank reactors model after determination of its retention time. Then, quantitative analysis of wilforlide A in Tripterygium wilfordii as well as in its tablets could be successfully determined by conventional reversed-phase high performance liquid chromatography with UV detector. Under the optimized conditions, the method showed good linearity (R2 = 0.9999) for wilforlide A in the range of 0.01 mg mL-1 -0.10 mg mL-1. The limit of detection and limit of quantity were 1.35 ng mL-1 and 4.50 ng mL-1, respectively. The average recovery rate, intra-day and inter-day precisions of wilforlide A were 96.43%, 0.67% and 1.14%, respectively. Compared with previous studies, the present method showed advantages of complete recovery of target component in the sample pretreatment and good repeatability.
Assuntos
Química Farmacêutica/métodos , Cromatografia Líquida , Medicamentos de Ervas Chinesas/química , Glicosídeos/análise , Medicina Tradicional Chinesa/normas , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/análise , Tripterygium/químicaRESUMO
A new pentacyclic triterpenoid, 3-oxo-urs-11,13(18)-dien-28-oic acid (1), along with twelve known triterpenoids, α-amyrin (2), 19α-hydroxy-α-amyrin (3), triptohypol E (4), uvaol (5), 2α,3α-dihydroxyurs-11-en-13ß,28-olide (6), 3ß-hydroxyurs-11-en-13ß,28-olide (7), ursolic acid (8), asiatic acid (9), oleanolic acid (10), aegiceradienol (11), obtusalin (12) and betulinic acid (13) were isolated from the leaves of Rhododendron dauricum L. Their structures were established from spectroscopic data and comparison with reported values. Among them, compounds 3, 4, 6, 7 and 11 were isolated from the Ericaceae family for the first time. Compounds 2, 5, 9, 12 and 13 were obtained from R. dauricum for the first time. Additionally, compounds 6, 10 and 11 significantly inhibited the levels of NO in LPS-stimulated RAW 264.7 cells at 3 µM.