RESUMEN
The implementation of REGULATION (EC) No 1924/2006 has led to the formation of a list of health claims that can be used in food supplements (EU 432/2012). However, such supplements are often composed of plant preparations with claims omitted from this list. The peculiarity of plants is related to their long history of use, that could allow claims based on traditionally recognized health effects. In addition, the scientific literature has been enriched over the years through clinical studies that have assessed the bioavailability and efficacy of bioactive components, and investigated their mechanisms of action. Based on existing recognized models which aim to classify research according to the level of scientific evidence, Synadiet developed a three-grade model (A, B or C) for assessing plants health claims. In this paper, the applicability of the model is illustrated through an example for which a Grade B health claim attesting the possible contribution of red clover isoflavones to the improvement of blood lipid levels in postmenopausal women has been attributed. The model appears able to be easily extrapolated to claims pertaining to other plants. If adopted by consensus at European level, this model could initiate the implementation of a positive list of health claims on plant preparations.
Asunto(s)
Suplementos Dietéticos/normas , Análisis de los Alimentos/métodos , Etiquetado de Alimentos/normas , Preparaciones de Plantas/normas , Plantas Comestibles , Adulto , Anciano , Suplementos Dietéticos/análisis , Femenino , Etiquetado de Alimentos/legislación & jurisprudencia , Humanos , Isoflavonas/análisis , Isoflavonas/normas , Legislación Alimentaria , Lípidos/sangre , Masculino , Persona de Mediana Edad , Valor Nutritivo , Preparaciones de Plantas/análisis , Posmenopausia/sangre , Trifolium/químicaRESUMEN
The side effects of kwao keur dietary supplements (obtained from the tuberous root of Pueraria mirifica) have recently been reported by the Ministry of Health, Labour and Welfare, Japan. To control the quality of kwao keur products, its ingredients need to be maintained by characteristic marker compounds, such as miroestrol, deoxymiroestrol, and kwakhurin (KWA). In this study, we described the facile synthesis of KWA, a marker compound of P. mirifica. Our revised synthetic method produced KWA with shorter steps and higher yield than the reported method. Furthermore, the absolute purity of KWA was determined by quantitative NMR analysis for standardization as a reagent, and its purity was 92.62 ± 0.12%.
Asunto(s)
Isoflavonas/síntesis química , Espectroscopía de Resonancia Magnética/normas , Pueraria/química , Suplementos Dietéticos/normas , Diseño de Fármacos , Isoflavonas/química , Isoflavonas/normas , Raíces de Plantas/química , Raíces de Plantas/metabolismo , Pueraria/metabolismo , Estándares de ReferenciaRESUMEN
Kwakhurin (Kwa) is a plant secondary metabolite solely present in Pueraria candollei var. mirifica (P. candollei), which has long been used as a Thai traditional herb for estrogen replacement therapy. Recently, health hazards have arisen in Japan regarding P. candollei-derived products containing potent estrogenic compounds. Therefore, the development of standardization methods for P. candollei materials is an urgent problem requiring resolution. The enzyme-linked immunosorbent assay (ELISA) is an effective analytical technique because it enables the development of sensitive and specific assays of the target compound through antigen-antibody reaction. Here, we produced a monoclonal antibody against Kwa (MAb 11F) by immunizing Kwa-bovine serum albumin (BSA) conjugates prepared using an N,N'-carbonyldiimidazole (CDI) mediated method. Stability and cross-reactivity tests of MAb 11F revealed that the MAb 11F is stable for at least 4â¯months at 4⯰C and is highly specific to Kwa. The detectable concentration range of an indirect competitive ELISA (icELISA) using MAb 11F exhibited values of 1.53-48.8â¯ng/mL with the limit of detection (LOD) of 1.13â¯ng/mL. Validation analyses revealed that the developed icELISA is precise, accurate, and reliable enough to be applied to P. candollei-derived samples and products for their standardization.
Asunto(s)
Ensayo de Inmunoadsorción Enzimática , Isoflavonas/química , Isoflavonas/normas , Preparaciones de Plantas/normas , Pueraria/química , Animales , Anticuerpos Monoclonales , Masculino , Ratones Endogámicos BALB C , Estructura Molecular , Fitoquímicos/química , Fitoquímicos/normas , Raíces de Plantas/químicaRESUMEN
To aid in the identification and quantification of biologically and agriculturally significant natural products, tandem mass spectrometry can provide accurate structural information with high selectivity and sensitivity. In this study, diagnostic fragmentation patterns of isoflavonoids were examined by liquid chromatography-ion trap-time of flight-mass spectrometry (LC-IT-TOF-MS). The fragmentation scheme for [M+H-2CO]+ ions derived from isoflavones and [M+H-B-ring-CO]+ ions derived from 5-hydroxyisoflavones, were investigated using different isotopically labeled isoflavones, specifically [1',2',3',4',5',6',2,3,4-13C9] and [2',3',5',6',2-D5] isoflavones. Specific isotopically labeled isoflavones were prepared through the biosynthetic incorporation of pharmacologically applied 13C- and D-labelled L-phenylalanine precursors in soybean plants following the application of insect elicitors. Using this approach, we empirically demonstrate that the [M+H-2CO]+ ion is generated by an intramolecular proton rearrangement during fragmentation. Furthermore, [M+H-B-ring-CO]+ ion is demonstrated to contain a C2H moiety derived from C-ring of 5-hydroxyisoflavones. A mechanistic understanding of characteristic isoflavone fragmentation patterns contributes to the efficacy and confidence in identifying related isoflavones by LC-MSn.
Asunto(s)
Glycine max/metabolismo , Isoflavonas/química , Isótopos/metabolismo , Espectrometría de Masa por Ionización de Electrospray/métodos , Espectrometría de Masas en Tándem/métodos , Animales , Cromatografía Liquida/métodos , Insectos/fisiología , Isoflavonas/análisis , Isoflavonas/normas , Fenilalanina/química , Protones , Estándares de Referencia , Glycine max/parasitologíaRESUMEN
An interlaboratory study was conducted to evaluate a method for determining total soy isoflavones in dietary supplements, dietary supplement ingredients, and soy foods. Isoflavones were extracted using aqueous acetonitrile containing a small amount of dimethylsulfoxide (DMSO) and all 12 of the naturally occuring isoflavones in soy were determined by high-performance liquid chromatography (HPLC) with UV detection using apigenin as an internal standard. Fifteen samples (6 pairs of blind duplicates plus 3 additional samples) of soy isoflavone ingredients, soy isoflavone dietary supplements, soy flour, and soy protein products were successfully analyzed by 13 collaborating laboratories in 6 countries. For repeatability, the relative standard deviations (RSDr) ranged from 1.07 for samples containing over 400 mglg total isoflavones to 3.31 for samples containing 0.87 mg/g total isoflavones, and for reproducibility the RSDR values ranged from 2.29 for samples containing over 400 mg/g total isoflavones to 9.36 for samples containing 0.87 mg/g total isoflavones. HorRat values ranged from 1.00 to 1.62 for all samples containing at least 0.8 mg/g total isoflavones. One sample, containing very low total isoflavones (< 0.05 mg/g), gave RSDR values of 175 and a HorRat value of 17.6. This sample was deemed to be below the usable range of the method. The method provides accurate and precise results for analysis of soy isoflavones in dietary supplements and soy foods.
Asunto(s)
Suplementos Dietéticos/análisis , Análisis de los Alimentos/métodos , Isoflavonas/análisis , Alimentos de Soja/análisis , Cromatografía Líquida de Alta Presión/métodos , Cromatografía Líquida de Alta Presión/normas , Conducta Cooperativa , Análisis de los Alimentos/normas , Glucósidos/análisis , Isoflavonas/química , Isoflavonas/normas , Estándares de Referencia , Alimentos de Soja/normas , Espectrofotometría UltravioletaRESUMEN
OBJECTIVE: To establish the optimized preparation procedure and study the method to determine the content for soybean isoflavone(SIF) Dropping Pills. METHOD: The preparation conditions, such as the proportion between SIF and PEGs, the temperature of mixture of SIF and PEGs, dropping distance, etc., were studied with Uniform Design and One-way ANOVA. SIF was identified by TLC and the content of SIF was determined by UV spectrometry at 262 nm detection wavelength. RESULT: Three batches of the prepared products meet the standards of the Chinese pharmacopoeia on dropping pills. SIF can be identified by TLC. Using UV spectrometry, the linear range of SIF was 0. 407 2 to 4. 072 g x mL(-1) and the correlation coefficient was 0. 999 8. In high, middle and low concentration, average recovery were 96. 54%, 97.27% and 97.21%, respectively (RSD were 1.3%, 0.78% and 0.71%). CONCLUSION: The preparation procedure is feasible, simple and suitable, the method established in this paper can be adopted for the quality control of SIF dropping pills, and the determination method is simple, relatively fast and accurate.
Asunto(s)
Medicamentos Herbarios Chinos/aislamiento & purificación , Glycine max/química , Isoflavonas/aislamiento & purificación , Plantas Medicinales/química , Cromatografía en Capa Delgada , Medicamentos Herbarios Chinos/análisis , Isoflavonas/análisis , Isoflavonas/normas , Tamaño de la Partícula , Polietilenglicoles/química , Control de Calidad , Espectrofotometría UltravioletaRESUMEN
OBJECTIVE: To establish the quantitative methods for calycosin glycoside and formononetin in Radix Astragali, and the samples from different sources were analyzed, in order to supply the basis for the quality control of Radix Astragali. METHOD: The content of calycosin glycoside and formononetin in 59 samples of Radix Astragali from eight with different provinces was analyzed by HPLC-DAD. RESULT: The contents of calycosin glycoside and formononetin in Radix Astragali from different sources, with different cultivating method or in different ages differed markedly, and the results showed that the quality of samples from Shannxi, Innermongolia and Shanxi were better than other sources, and the semi-wild samples were better than other cultiving samples, moreover the shorter age, the better quality. CONCLUSION: This simple, accurate and reproducible method could use to determine the contents of isoflavanoids in Radix Astragali.
Asunto(s)
Astragalus propinquus/química , Glucósidos/análisis , Isoflavonas/análisis , Plantas Medicinales/química , Astragalus propinquus/crecimiento & desarrollo , China , Cromatografía Líquida de Alta Presión/métodos , Ecosistema , Glucósidos/normas , Isoflavonas/normas , Raíces de Plantas/química , Control de Calidad , Reproducibilidad de los ResultadosRESUMEN
Isoflavones are biologically active compounds occurring naturally in a variety of plants, with relatively high levels found in soybeans. Twelve laboratories participated in a collaborative study to determine the aglycon isoflavone content of 8 test samples of soy and foods containing soy. The analytical method for the determination of isoflavones incorporates a mild saponification step that reduces the number of analytes measured and permits quantitation versus commercially available, stable reference standards. Test samples were extracted at 65 degrees C with methanol-water (80 + 20), saponified with dilute sodium hydroxide solution, and analyzed by reversed-phase liquid chromatography with UV detection at 260 nm. Isoflavone results were reported as microg/aglycon/g or microg aglycon equivalents/g. The 8 test samples included 2 blind duplicates and 4 single test samples with total isoflavone concentrations ranging from approximately 50 to 3000 microg/g. Test samples of soy ingredients and products made with soy were distributed to collaborators with appropriate reference standards. Collaborators were asked to analyze test samples in duplicate on 2 separate days. The data were analyzed for individual isoflavone components, subtotals of daidzin-daidzein, glycitin-glycitein, and genistin-genistein, and total isoflavones. The relative standard deviation (RSD) for repeatability was 1.8-7.1%, and the RSD for reproducibility was 3.2-16.1% for total isoflavone values of 47-3099 microg/g.
Asunto(s)
Cromatografía Liquida/métodos , Análisis de los Alimentos/métodos , Glycine max/química , Isoflavonas/análisis , Cromatografía Liquida/normas , Cromatografía Liquida/estadística & datos numéricos , Análisis de los Alimentos/normas , Análisis de los Alimentos/estadística & datos numéricos , Genisteína/análisis , Genisteína/química , Glucósidos/análisis , Glucósidos/química , Isoflavonas/química , Isoflavonas/normas , Laboratorios , Estructura Molecular , Estándares de ReferenciaRESUMEN
A HPLC-MS procedure for the rapid, sensitive and specific measurement of the isoflavones, daidzein, dihydrodaidzein, O-desmethylangolensin and genistein, in human plasma has been developed. Synthetic radiolabeled genistein conjugates were used for evaluation of optimum conditions for solid phase extraction. Biochanin A was added to plasma as a recovery marker for isoflavones and phenolphthalein glucuronide and 4-methylumbelliferone sulfate were added to ensure completeness of hydrolysis with beta-glucuronidase/sulfatase. Isoflavones in plasma extracts were separated using an isocratic HPLC method and analyzed by negative ion multiple reaction ion monitoring-mass spectrometry using a heated nebulizer-atmospheric pressure chemical ionization interface. Using plasma samples from four subjects consuming two servings a day of an isolated soy protein beverage for 14 days, the mean plasma genistein and daidzein concentrations were 556 and 345 nM, respectively. Within assay and between assay coefficients of variation for measurement of daidzein and genistein in five aliquots of the same plasma sample were 8.51% and 7.76%, and 5.98% and 6.12%, respectively.
Asunto(s)
Cromatografía Líquida de Alta Presión , Isoflavonas/sangre , Espectrometría de Masas , Espectrometría de Masa de Ion Secundario , Bebidas/análisis , Cromatografía Líquida de Alta Presión/normas , Genisteína , Humanos , Isoflavonas/síntesis química , Isoflavonas/aislamiento & purificación , Isoflavonas/normas , Espectrometría de Masas/normas , Reproducibilidad de los Resultados , Proteínas de Soja/análisis , Glycine max/química , Espectrometría de Masa de Ion Secundario/normasRESUMEN
Cancer chemoprevention refers to the reduction of cancer incidence by administration of agents or drugs that inhibit, reverse or retard the cancer process. Genistein has demonstrated a wide variety of biological activities that make it a good candidate for a chemopreventive agent. Many agents, such as genistein, are currently being tested with the goal of developing safe and effective chemopreventive drugs for human use. Genistein was investigated as a potential chemopreventive agent in an azoxymethane-induced colon carcinogenesis model. Genistein was tested for its ability to inhibit aberrant colon crypts in the colon of F344 rats that had been treated with azoxymethane. Genistein was administered in the diet from 1 wk before the carcinogen to 4 wk after the first carcinogen dose for a total of 5 wk. At both doses, 75 and 150 mg/kg, the mean number of foci per colon was significantly reduced. Further development of this agent includes demonstration of the preventive efficacy in an in vivo tumorigenesis model, followed by preclinical pharmacology and toxicology testing. Phase 1, 2 and 3 clinical chemoprevention trials would be then performed to determine pharmacokinetics, safe doses, and effectiveness for New Drug Approval.
Asunto(s)
Antineoplásicos/uso terapéutico , Isoflavonas/uso terapéutico , Neoplasias Experimentales/prevención & control , Animales , Antineoplásicos/farmacología , Antineoplásicos/normas , Colon/efectos de los fármacos , Colon/patología , Colon/ultraestructura , Neoplasias del Colon/tratamiento farmacológico , Neoplasias del Colon/epidemiología , Neoplasias del Colon/prevención & control , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Genisteína , Incidencia , Isoflavonas/farmacología , Isoflavonas/normas , Masculino , Microvellosidades/efectos de los fármacos , Microvellosidades/ultraestructura , Neoplasias Experimentales/tratamiento farmacológico , Neoplasias Experimentales/epidemiología , Ratas , Ratas Endogámicas F344 , Factores de TiempoRESUMEN
There is much evidence suggesting that compounds present in soybeans can prevent cancer in many different organ systems. The evidence for specific soybean-derived compounds having a suppressive effect on carcinogenesis in animal model systems is limited, however. There is evidence that the following isolated soybean derived products suppress carcinogenesis in vivo: a protease inhibitor, the Bowman-Birk inhibitor, inositol hexaphosphate (phytic acid) and the sterol beta-sitosterol. Other compounds that may be able to suppress carcinogenesis in animals are the soybean isoflavones. Soybean compounds reported to have other types of anticarcinogenic activity include soybean trypsin inhibitor, saponins and genistein. There is much evidence to suggest that diets containing large amounts of soybean products are associated with overall low cancer mortality rates, particularly for cancers of the colon, breast and prostate. It is believed that supplementation of human diets with certain soybean products shown to suppress carcinogenesis in animals could markedly reduce human cancer mortality rates.