Quality markers based on phytochemical analysis and anti-inflammatory screening: An integrated strategy for the quality control of Dalbergia odorifera by UHPLC-Q-Orbitrap HRMS.

BACKGROUND: Quality control, key for the clinical application of traditional Chinese medicines (TCMs), should be connected to the authentication and efficacy of TCMs. The heartwood of Dalbergia odorifera has been widely used to treat inflammation-related diseases. However, in the Chinese pharmacopeia, only the total volatile oil, which does not sufficiently reflect the clinical efficacy, is used as a quality control indicator. PURPOSE: Establishing a "phytochemical-specificity-effectiveness-Q-marker" analytical strategy to improve the quality control of D. odorifera. METHODS: Combined with biosynthetic pathway analysis, phytochemical compositions identified by UHPLC-Q-Orbitrap HRMS were used to build substantial phytochemical groups and further discover specific Q-markers. Then, lipopolysaccharide-stimulated RAW 264.7 cells were used to screen effective anti-inflammatory ingredients. Finally, a UHPLC-HRMS method was developed and validated to quantify the selected Q-markers in D. odorifera samples. RESULTS: Along the constructed biosynthetic pathways, 93 phytochemical components were identified in D. odorifera, including 7 chalcones, 13 flavanones, 21 isoflavones, 21 isoflavanones, 3 flavonols, 19 neoflavones, etc. Among them, 31 compounds representing these 6 categories were further evaluated for their anti-inflammatory activities. It revealed that the extract of D. odorifera and nine flavonoids in the noncytotoxic range could alleviated lipopolysaccharide-stimulated inflammation in RAW 264.7 cells by decreasing the production of proinflammatory mediators such as nitric oxide and interleukin-6. Notably, neoflavones, as species-specific components, exhibited superior anti-inflammatory activities among the representative compounds. Finally, 12 Q-markers (butin, liquiritigenin, eriodictyol, melanettin, naringenin, butein, genistein, 4'-hydroxy-4-methoxydalbergione, isoliquiritigenin, 2,4-dihydroxy-5-methoxybenzophenone, medicarpin, and pinocembrin), which reflect specificity and effectiveness, were successfully quantified in 10 batches of samples from different origins. The origins and consistency of D. odorifera could be efficiently discriminated by hierarchical cluster analysis (HCA). CONCLUSION: The analysis strategy that combines phytochemical analysis with anti-inflammatory screening clarified the therapeutic material basis and discovered Q-markers, which possibly offers a more comprehensive quality assessment of D. odorifera.

A novel concept of Q-markers: Molecular connectivity index.

BACKGROUND: Drugs derived from botany have been playing essential role in both clinical treatment and pharmaceutical industry, unfortunately our worry is still that its quality and therapeutic efficacy are inconsistent. Recently many scientists launched a new project on quality (Q)-marker of medicinal herbs, this study was thus designed to generate a novel concept of quality (Q)-markers: molecular connectivity index (MCI), and to test and verify the new concept of molecular connectivity index (MCI). METHODS: The first-order term (1χ) was selected to calculate and study quality (Q)-marker for TCM. Houttuynia cordata Thunb. (HCT) was adopted as a model to verify the hypothesis. Volatile oils of HCT were determined using gas chromatography-mass (GC-MS). SIMCA 13.0 and SPSS 21.0 were used to deal with the data. RESULTS: The minimum of the MCI values was 1.273, belonging to the peak 15, but the maximum (12.822) belonged to the peak 34, and the average value of fifty volatile oils was 5.798. The results demonstrated that MCI was the principle component, and monoterpenoid and sesquiterpenoid were also the principle components in oils. Fig. 2a shows peak 5, 24, 34 were the significant ingredients, while Fig. 2b shows peak 2, 5, 24 were the significant components. CONCLUSION: The data demonstrated that MCI was associated with the structure of molecules and the therapeutic efficacy, MCI could directly exhibit the relationship between ingredients and effectiveness of Traditional Chinese Medicine (TCM). So MCI could be a potential and promising parameter for quality (Q)-marker. Therefore, MCI may be developed as a novel potential concept to control the quality of TCM.

Reflecting on a decade of metabolite screening and monitoring.

The last 10 years have witnessed robust debate within the bioanalytical community and regulatory authorities on the topic of metabolite monitoring and safety assessment. Of particular interest to regulated bioanalytical laboratories was the acceptance by the US FDA and other major regulatory bodies of a tiered approach to bioanalytical assay validation. The tiered approach defines a sliding scale of regulatory rigor for the evaluation of significant human metabolites that encompasses a range of assessments from semi-quantitative assays to fully validated assays, all of which can be used in support of regulatory submissions. This article describes the utilization of a tiered approach at Bristol-Myers Squibb and the decision trees guiding the selection of the appropriate level of assay qualification. Case studies illustrate how decisions are made, how different scientific situations influence the assay choice, and what criteria may be set to continue or discontinue metabolite monitoring in later drug development.

Target identification and mechanism of action in chemical biology and drug discovery.

Target-identification and mechanism-of-action studies have important roles in small-molecule probe and drug discovery. Biological and technological advances have resulted in the increasing use of cell-based assays to discover new biologically active small molecules. Such studies allow small-molecule action to be tested in a more disease-relevant setting at the outset, but they require follow-up studies to determine the precise protein target or targets responsible for the observed phenotype. Target identification can be approached by direct biochemical methods, genetic interactions or computational inference. In many cases, however, combinations of approaches may be required to fully characterize on-target and off-target effects and to understand mechanisms of small-molecule action.

Atividade hepatoprotetora dos extratos etanólico e hexânico das folhas de Momordica charantia / Hepatoprotective activity of hexane and ethanol extracts from Momordica charantia L. leaves

Momordica charantia L. (Cucurbitaceae) é empregada na medicina popular pelas diversas propriedades biológicas e farmacológicas. Neste trabalho, objetivou-se avaliar a atividade hepatoprotetora dos extratos hexânico (EH) e etanólico (EE) das folhas de M. charantia no modelo de lesão hepática aguda induzida pelo etanol. Em ambos os extratos foram detectados esteróides; EH apresentou atividade antioxidante mais marcante que EE. O pré-tratamento de camundongos (Swiss, machos) por via oral por três dias consecutivos com 25, 50 ou 100 mg Kg-1 de EH preveniu o aumento tanto de AST quanto de ALT (p<0,05) enquanto o pré-tratamento com EE preveniu apenas o aumento de ALT (p<0,05). Ambos os extratos reduziram a atividade das enzimas hepáticas, contudo EH apresentou efeito mais relevante, caracterizando potencial hepatoprotetor, possivelmente associado ao seu potencial antioxidante.

Momordica charantia L. (Cucurbitaceae) has been used in folk medicine based on its several biological and pharmacological properties. The aim of this work was to evaluate the hepatoprotective activity of hexane (HE) and ethanol (EE) extracts from M. charantia leaves on the acute liver injury model induced by ethanol. Steroids were detected in both extracts, and EH had higher antioxidant activity than EE. Oral pretreatment of male Swiss mice for three consecutive days with 25, 50 or 100 mg kg-1 HE prevented the increase in AST and ALT (p<0.05), whereas pretreatment with EE only prevented ALT increase (p<0.05). Both extracts reduced the activity of hepatic enzymes; however, HE had a more relevant effect, characterizing hepatoprotective potential, likely associated with its antioxidant potential.

Deficiencia de cobre en un colectivo y factores asociados / Copper deficiency in a collective and associated factors

INTRODUCCIÓN: El cobre ejerce un papel esencial como nutriente en el organismo, se necesita un aporte diario mínimo a partir de alimentos o suplementos para el buen funcionamiento de la actividad celular. Está presente en todos los tejidos y participa activamente en la síntesis de la hemoglobina e influye sobre la absorción del hierro. Su deficiencia provoca anemia, diarrea, debilidad generalizada, alteraciones respiratorias y lesión cutánea, teniendo un papel esencial como protector frente el estrés oxidativo, un importante papel en el sistema inmunológico y una gran influencia sobre la expresión de numerosos genes. OBJETIVO: Valorar el estatus de cobre en un grupo de personas adultas sanas procedente de un colectivo de personas sanas adultas, estableciéndose tanto los niveles de ingesta como los niveles plasmáticos y eritrocitarios de este mineral, y estudiando una posible relación con otros parámetros. METODOLOGÍA: El estudio se realizó en una muestra de 90 sujetos de edades comprendidas entre 21 y 59 años (31 hombres y 53 mujeres), todos de la provincia de Granada. El criterio de inclusión se basó en la aceptación de los sujetos a participar en el estudio y no presentar ningún tipo de patología que pudiera afectar su situación clínico-nutricional. El Cu se analizó mediante Espectrofotometría de Absorción Atómica (AAS) en muestras de eritrocito y plasma mineralizadas por vía húmeda. Se aplicó un cuestionario de frecuencia de consumo, y mediante programa informático Nutriber (Mataix, yGarcia Diz 2006)1 se obtuvo el % de RDA. Se contó con la aceptación del Comité ético y el consentimiento informado(AU)

RESULTADOS: La ingesta media recomendada de cobre en la población adulta española es de 1,1mg/día. Los resultados obtenidos muestran ingestas del cobre <2/3 de IR en un 58,2% de la población estudiada, de los cuales el 53,6% son hombres y el 60,8% son mujeres. En plasma, se observaron valores deficientes en cobre en el 25% de la población total, siendo el 29% hombres y el 22,6 % mujeres. En eritrocito, el 21,4% de la población general presentaron deficiencia de cobre, siendo el 28,8% hombres y el 18,9% mujeres. Se encontró una correlación significativa positiva entre la ingesta de macronutrientes y los niveles de cobre en ingesta, así como entre la ingesta de cobre y la de hierro, cinc, magnesio, calcio y selenio. Igualmente, se observa una correlación positiva significativa entre los niveles de cobre en plasma y los niveles de cinc (p<0,02). CONCLUSIÓN: Según los resultados obtenidos, podemos concluir que son necesarios estudios recientes y detallados que aborden la valoración del estatus de cobre de una manera más integral, desde una evaluación pormenorizada de la ingesta hasta la determinación de biomarcadores específicos de cobre, teniendo en cuenta su participación en numerosas reacciones biológicas celulares que pueden alterarse por la deficiencia(AU)

INTRODUCTION: The copper has a crucial role as a nutrient in the body, you need a minimum daily intake from food or supplements for the proper functioning of cellular activity. Is present in all tissues and is actively involved in the synthesis of hemoglobin and influences iron absorption. Deficiency causes anemia, diarrhea, general weakness, respiratory and skin lesion, having an essential role as a protector against oxidative stress, an important role in the immune system and a great influence on the expression of numerous genes. OBJECTIVE: To assess copper status in a group of healthy adults from a group of healthy adults, establishing both the levels of intake and plasma and erythrocyte levels of this mineral, and isexploring a possible relationship with other parameters. METHOD: The study was conducted in a sample of 90 subjects aged between 21 and 59 years (31 men and 53 women), all of the province of Granada. The inclusion criteria was based on the acceptance of individuals to participate in the study and did not show any pathology that could affect clinical and nutritional status. Cu was analyzed by Atomic Absorption Spectrometry (AAS) in erythrocytes and plasma samples mineralized by wet. We applied a food frequency questionnaire, and through software Nutriben (Mataix, and Garcia Diz 2006)1 was obtained by the% of RDA. It was accepted by the Ethics Committee and informed consent(AU)

RESULTS: The average recommended intake of copper in the Spanish adult population is 1.1 mg /day. The results show copper intakes <2 / 3 of IR in 58'2% of the population studied, of which 53'6% are men and women 60'8%. In plasma, were found deficient in copper values in 25% of the total population, being 29% male and 22.6% women. In erythrocytes, 21.4% of the general population showed copper deficiency, with 28.8% men and 18.9% women. There was a significant positive correlation between macronutrient intake and levels of copper intake, as well as between dietary copper and iron, zinc, magnesium, calcium and selenium. Similarly, there is a significant positive correlation between plasma copper levels and zinc levels (p <0.02). CONCLUSION: According to the results, we conclude that studies are needed to address recent detailed assessment of copper status in a more comprehensive, since a thorough assessment of intake to the identification of specific biomarkers of copper, taking into account participation in numerous cellular biological reactions that may be altered by the deficiency(AU)