RESUMEN
Mogroside V is one of the characteristic and effective components of luohanguo extract, a food additive used as a sweetener in Japan as per Japan's Standards and Specifications for Food Additives (JSFA; 9th ed.). JSFA stipulates that the quantitative determination for mogroside V content in luohanguo extract applies HPLC using analytical standard mogroside V. However, no mogroside V reagents with proven purities are commercially available. Therefore the current JSFA determination method is not particularly suited for daily quality control operations involving luohanguo extract. In this study, we applied an alternative quantitative method using a single reference with relative molar sensitivity (RMS). It was possible to calculate the accurate RMS by an offline combination of 1H-quantitative NMR spectroscopy (1H-qNMR) and an HPLC/variable-wavelength detector (VWD). Using the RMS of mogroside V to a commercial certified reference material grade caffeine, the mogroside V contents in luohanguo extracts could be determined using HPLC/VWD without analytical standard mogroside V. There was no significant difference between the mogroside V contents in luohanguo extracts determined using the method employing single-reference caffeine with the RMS and using the JSFA method. The absolute calibration curve for the latter was prepared using an analytical standard mogroside V whose purity was determined by 1H-qNMR. These results demonstrate that our proposed method using a single reference with RMS is suitable for quantitative determination of mogroside V in luohanguo extract and can be used as an alternative method to the current assay method in JSFA.
Asunto(s)
Cafeína/análisis , Cucurbitaceae/química , Aditivos Alimentarios/análisis , Extractos Vegetales/análisis , Triterpenos/análisis , Cafeína/normas , Cromatografía Líquida de Alta Presión/normas , Aditivos Alimentarios/normas , Japón , Espectroscopía de Resonancia Magnética/normas , Extractos Vegetales/normas , Control de Calidad , Triterpenos/normasRESUMEN
INTRODUCTION: Triterpene acids from the dried sclerotia of Poria cocos (Schw.) Wolf (poria) were recently found to possess anti-cancer activities. Identification of more triterpene acid analogues in poria is worthwhile for high throughput screening in anti-cancer drug discovery. OBJECTIVE: To establish an efficient dereplication strategy for identifying triterpene acid analogues in poria based on ultra-performance liquid chromatography with electrospray ionisation quadrupole time-of-flight tandem mass spectrometry (UPLC-ESI-QTOF-MS/MS). METHODOLOGY: The structural characteristics and mass spectrometric data profiles of known triterpene acids previously reported in poria were used to establish a predicted-analogue database. Then, the quasi-molecular ions of components in a poria extract were automatically compared with those in the predicted-analogue database to highlight compounds of potential interest. Tentative structural identification of the compounds of potential interest and discrimination of isomers were achieved by assessing ion fragmentation patterns and chromatographic behaviour prediction based on structure-retention relationship. RESULTS: A total of 62 triterpene acids were unequivocally or tentatively characterised from poria, among which 17 triterpene acids were tentatively identified for the first time in poria. CONCLUSION: This study provided more structure information of triterpene acids in poria for future high throughput screening of anti-cancer candidates. It is suggested that this semi-automated approach in which MS data are automatically compared to a predictive database may also be applicable for efficient screening of other herbal medicines for structural analogues of proven bioactives.
Asunto(s)
Ácidos/química , Cromatografía Líquida de Alta Presión/métodos , Espectrometría de Masa por Ionización de Electrospray/métodos , Espectrometría de Masas en Tándem/métodos , Triterpenos/análisis , Wolfiporia/química , Minería de Datos , Estándares de Referencia , Relación Estructura-Actividad , Triterpenos/química , Triterpenos/normasRESUMEN
CONTEXT: Dillenia indica Linn. (Dilleniaceae) is traditionally used to treat skin inflammation. OBJECTIVE: This study evaluated the healing effect of Dillenia indica fruit extracts on induced psoriasis-like wounds in Wistar rats. MATERIALS AND METHODS: Extracts were standardized to betulinic acid, including an aqueous ethanolic extract (AEE), ethyl acetate extract (EAE) and petroleum ether extract. Effects against lipid peroxidation were assessed in vitro. Wounds were created at rat tails (n = 12). Topical treatments were applied once daily for 7 days (1 mL of AEE or EAE at 5 or 50 mg/mL). Maximal dose was defined by the extract solubility. A 10-fold lower dose was also tested. Positive and negative controls were treated with clobetasol (0.5 mg/mL) or excipient. Half of each group was euthanized for histology. The remaining animals were observed for 20 days for wound measurements. RESULTS: Yields of AEE and EAE were 4.3 and 0.7%, respectively. Betulinic acid concentrations in AEE and EAE were 4.6 and 107.6 mg/g. Extracts neutralized lipid peroxidation in vitro at 0.02 µg/mL, accelerating healing at 50 mg/mL. Complete healing in mice treated with AEE occurred 16 days after wound induction. This time was 14 and 12 days in mice treated with EAE and clobetasol. Compared to orthokeratosis, parakeratosis was reduced by AEE (25%), EAE (45%) and clobetasol (55%). EAE caused superior protection against biomolecules oxidation of skin compared to AEE. DISCUSSION AND CONCLUSION: EAE exhibited activity closer to that of clobetasol. Betulinic acid may be an active constituent, which should be assessed in future studies.
Asunto(s)
Antiinflamatorios/farmacología , Antioxidantes/farmacología , Fármacos Dermatológicos/farmacología , Dilleniaceae/química , Frutas/química , Extractos Vegetales/farmacología , Psoriasis/tratamiento farmacológico , Piel/efectos de los fármacos , Triterpenos/farmacología , Rayos Ultravioleta , Cicatrización de Heridas/efectos de los fármacos , Animales , Antiinflamatorios/aislamiento & purificación , Antiinflamatorios/normas , Antioxidantes/aislamiento & purificación , Antioxidantes/normas , Biomarcadores/metabolismo , Clobetasol/farmacología , Fármacos Dermatológicos/aislamiento & purificación , Fármacos Dermatológicos/normas , Modelos Animales de Enfermedad , Peroxidación de Lípido/efectos de los fármacos , Masculino , Malondialdehído/metabolismo , Estrés Oxidativo/efectos de los fármacos , Triterpenos Pentacíclicos , Fitoterapia , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/normas , Plantas Medicinales , Carbonilación Proteica/efectos de los fármacos , Psoriasis/etiología , Psoriasis/metabolismo , Psoriasis/patología , Ratas Wistar , Piel/metabolismo , Piel/patología , Solventes/química , Factores de Tiempo , Triterpenos/aislamiento & purificación , Triterpenos/normas , Ácido BetulínicoRESUMEN
In addition to their widely recognized use as dietary supplement ingredients, plant-derived compounds are increasingly used as natural sweeteners. The search for nonnutritive sweeteners has been stimulated over the last 20-30 years by concern over demonstrated or suspected relationships between consumption of sucrose and high-fructose corn syrups and a variety of health-related conditions. In the USA, there is increased use of plant extracts known to contain highly sweet terpenoids. Purified extracts of Stevia rebaudiana (Bertoni) containing the diterpene glycosides stevioside and rebaudioside A are popular as sweeteners and are also used as dietary supplements, and soft drinks and nutritional and energy shakes incorporating extracts of Siraitia grosvenorii (Swingle) fruits containing sweet triterpene glycosides such as mogroside V are also on the market. Here, we review recent studies on these two important sources of noncaloric natural sweeteners, including analytical methods used to identify and quantify specific constituents and structural features relating to their sweetness. We also review the generally recognized as safe status of specific components and their status with respect to review by the Joint FAO/WHO Expert Committee on Food Additives.
Asunto(s)
Cucurbitaceae/química , Extractos Vegetales/análisis , Stevia/química , Edulcorantes/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Diterpenos de Tipo Kaurano/aislamiento & purificación , Diterpenos de Tipo Kaurano/normas , Glucósidos/aislamiento & purificación , Glucósidos/normas , Humanos , Extractos Vegetales/normas , Hojas de la Planta/química , Edulcorantes/normas , Triterpenos/aislamiento & purificación , Triterpenos/normasRESUMEN
BACKGROUND: Mitochondrial dysfunction and reactive oxygen species (ROS) generation cause dopaminergic neurodegeneration in Parkinson's disease. The neuroprotective approach is a promising strategy to slow disease progression in Parkinson's disease. A standardized extract of Centella asiatica ECa233 has been previously reported to have pharmacological effects in the central nervous system. PURPOSE: This study aimed to determine the neuroprotective effect and mechanisms of ECa233 in rotenone-induced parkinsonism rats. METHODS: Rats were orally given either vehicle or ECa233 (10, 30 and 100â¯mg/kg) for 20 consecutive days. Rotenone (2.5â¯mg/kg i.p.) was given to parkinsonism (PD) and ECa-treated rats from day 15 to 20. Locomotor activity was recorded on day 1, 14, 17 and 20. Tyrosine-hydroxylase (TH) immunohistological staining was used to determine dopaminergic neurons in the substantia nigra and striatum. Furthermore, mitochondrial complex I activity, malondialdehyde (MDA) levels, superoxide dismutase (SOD) and catalase protein expression were measured in brain tissue. RESULTS: Rats receiving ECa233 30â¯mg/kg showed a significant increase in distances (pâ¯<â¯0.01) together with a higher number and intensity of dopaminergic neurons in the substantia nigra and striatum (pâ¯<â¯0.001) compared to PD rats. ECa233 (30â¯mg/kg) protected against mitochondrial complex I inhibition, decreased MDA levels (pâ¯<â¯0.05) and increased SOD (pâ¯<â¯0.01) and catalase (pâ¯<â¯0.05) expression. CONCLUSION: ECa233 can protect against rotenone-induced motor deficits and dopaminergic neuronal death. These effects are mediated through the protection of mitochondrial complex I activity, the effects of antioxidants and the enhancement of antioxidant enzyme expression.
Asunto(s)
Antiparkinsonianos/farmacología , Fármacos Neuroprotectores/farmacología , Trastornos Parkinsonianos/tratamiento farmacológico , Triterpenos/farmacología , Animales , Antioxidantes/metabolismo , Catalasa/metabolismo , Centella , Cuerpo Estriado/efectos de los fármacos , Cuerpo Estriado/metabolismo , Neuronas Dopaminérgicas/efectos de los fármacos , Masculino , Malondialdehído/metabolismo , Trastornos Parkinsonianos/inducido químicamente , Trastornos Parkinsonianos/metabolismo , Extractos Vegetales , Ratas Wistar , Rotenona/toxicidad , Sustancia Negra/efectos de los fármacos , Sustancia Negra/metabolismo , Superóxido Dismutasa/metabolismo , Triterpenos/normas , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
Ganoderic acids (GAs) were bioactive secondary metabolites produced by a traditional mushroom Ganoderma lucidum. We describe a simple and efficient method for the separation and quantitative determination of four GAs, namely Ganoderic acid T (GA-T), Ganoderic acid Mk (GA-Mk), Ganoderic acid Me (GA-Me) and Ganoderic acid S (GA-S) from dried triterpene-enriched extracts of G. lucidum mycelia powder by capillary zone electrophoresis (CZE). Under the optimum conditions, the four GAs reached the baseline separation in 9 min with Glycyrrhetinic acid (GTA) as internal standard. The four GAs and internal standard (GTA) were detected at a wavelength 245 nm. All calibration curves showed good linearity (r(2)>0.9958) within test ranges. Limit of detection (LOD) and limit of quantification (LOQ) were less than 0.6 and 1.8 microg/mL, respectively. The relative standard deviation (R.S.D.) values of precision and recoveries were less than 5% and recoveries ranged from 91.4% to 103.6%. This was the first report on simultaneous determination of the four GAs and the results provided a firm basis for the trace analysis of GAs in dried fermentation mycelia powder of G. lucidum with high accuracy.
Asunto(s)
Fermentación , Lanosterol/análogos & derivados , Micelio , Reishi/química , Triterpenos/análisis , Antineoplásicos/análisis , Antineoplásicos/normas , Calibración/normas , Medicamentos Herbarios Chinos/análisis , Medicamentos Herbarios Chinos/normas , Electroforesis Capilar/métodos , Lanosterol/análisis , Lanosterol/normas , Polvos , Triterpenos/normasRESUMEN
A method has been developed for the qualitative and quantitative analysis of pharmacologically active astragalosides isolated from several species of the genus Astragalus by high performance liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry. Seven astragalosides in Radix Astragali and their commercial pharmaceutical preparations were analyzed using the developed method. The extracted ion current chromatograms were obtained from the total ion current chromatogram using the m/z of [M+Na]+ ions produced by target compounds for peak determination. The limits of detection and limits of quantification were in the range of 0.10-0.22 ng and 0.22-0.52 ng in full scan mode, respectively. All calibration curves showed good linear regression (r2 > or = 0.9965) within the test range. The overall intra- and inter-day precision was less than 2.86% for peak area and the accuracy was higher than 92.9% on using ginsenoside I as internal standard. The assay was successfully utilized to analyze the major biologically active astragalosides in six samples of Astragalus membranaceus (Fisch.) Bge var. mongholicus (Bge.) Hsiao. and eight commercial preparations. The overall results demonstrate that this method is simple, selective, and suitable for the quality control of Chinese medicine and their preparation in the low nanogram range.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/análisis , Saponinas/análisis , Espectrometría de Masa por Ionización de Electrospray/métodos , Triterpenos/análisis , Planta del Astrágalo/química , Astragalus propinquus , Cromatografía Líquida de Alta Presión/normas , Cromatografía Líquida de Alta Presión/estadística & datos numéricos , Medicamentos Herbarios Chinos/normas , Estructura Molecular , Estándares de Referencia , Saponinas/normas , Sensibilidad y Especificidad , Espectrometría de Masa por Ionización de Electrospray/normas , Espectrometría de Masa por Ionización de Electrospray/estadística & datos numéricos , Triterpenos/normasRESUMEN
Three 3beta,8beta-epoxymexicanolides, including xyloccensin K, 6-acetoxycedrodorin and a new one named xyloccensin W, were isolated from the fruit of a Chinese mangrove Xylocarpus granatum. Their structures were determined by spectroscopic analyses. The first complete assignment of (1)H and (13)C NMR data for xyloccensin W was achieved by means of 2D NMR techniques, including (1)H-(1)H COSY, HSQC, HMBC and NOESY spectra. In addition, the confusion of (1)H and (13)C NMR data previously reported for xyloccensin K was clarified.