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1.
In this paper, ultra performance liquid chromatography (UPLC)/quadrupole-time-of-flight mass spectrometry (QTOF) with automated data analysis software (Metabolynx?) were applied for fast analysis of hyperoside metabolites in rat after intravenous administration. MS(E) was used for simultaneous acquisition of precursor ion information and fragment ion data at high and low collision energy in one analytical run, which facilitated the fast structural characterization of 12 metabolites in rat plasma, urine and bile. The results indicated that methylation, sulfation and glucuronidation were the major metabolic pathways of hyperoside in vivo, and among them, 3'-O-methyl-hyperoside was confirmed by matching its fragmentation patterns with standard compound. The present study provided important information about the metabolism of hyperoside which will be helpful for fully understanding the mechanism of this compound's action. Furthermore, this work demonstrated the potential of the UPLC/QTOFMS approach using Metabolynx for fast and automated identification of metabolites of natural product.  相似文献   

2.
为了考查白及有效部位在肠道的可吸收成分及其代谢特征。基于在体肠灌流模型,采用超高效液相色谱-四极杆-飞行时间质谱仪(UPLC-Q-TOF/MS)对收集到的健康SD大鼠循环肠灌流液、血清、胆汁进行分析检测,并结合对照品、质谱碎片信息和Masslynx V4.1工作站中的Single Mass Analysis功能,初步推测吸收和代谢产物的结构式。在大鼠血清和胆汁中,初步鉴定出1,4-二[4-(葡萄糖氧)苄基]-2-异丁基苹果酸、4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯、α-异丁基苹果酸酯原型产物。在大鼠循环肠灌流液、血清和胆汁中,共鉴定出4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯的脱糖后硫酸化代谢产物和二氢菲5的葡萄糖醛酸化代谢产物,其代谢产物主要生水解和葡萄糖醛酸化反应。该方法初步探究了白及有效部位在大鼠循环肠灌流液中可吸收成分和代谢特征,为阐释白及药材的药效物质基础提供实验依据。  相似文献   

3.
In this paper, microdialysis combining with ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was applied to simultaneously identify components in blood and kidney dialysis after oral administration of Abelmoschus manihot extract. Microdialysis probe was implanted in the jugular vein and the kidney medulla, respectively; microdialysis samples were collected continuously, transferred to microtubes and analyzed by UPLC-Q-TOF/MS. The components in microdialysis samples were separated by an UPLC HSS T3 column and eluted with acetonitrile and water (containing 0.1% formic acid) at a flow rate of 0.4 mL/min. The results showed that unbound constituents in blood circulation of the rat include hyperoside, isoquercitrin, quercetin monoglucuronide, quercetin-3'-O-glucoside, quercetin, myricetin, and hibifolin while unbound constituents in kidney are hyperoside, isoquercitrin, quercetin monoglucuronide, which might be the potential active components in vivo. The developed method was simple and reliable, and could be adopted to rapidly screen and identify potential active components contributing to pharmacological effects of TCM and to better clarify its action mechanism.  相似文献   

4.
A protocol for the metabolic profiling of rat liver was developed based on ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) to explore metabolic state directly. Methanol/water (4:1, v:v) was selected as the optimal extraction solvent. The established method was validated with a linearity over the 10-5000 ng/mL for internal standards (IS) and got an average correlation coefficient of 0.9986. The intra-day and inter-day RSD for most endogenous compounds were below 15%. And the absolute recovery of IS was from 84.8% to 109.1%. Liver tissues from diabetic and control rats were enrolled in the subsequent study to show the usefulness of the method. A clear classification between the control and model animals was achieved, some significant metabolites were successfully filtered. These metabolites reflected the abnormal metabolism of diabetic rats. This initial application indicated that the method is suitable and reliable for liver tissue metabolic profiling. It is expected this protocol could also be extended to metabonomic studies of other tissues.  相似文献   

5.
In the present study, the metabolic profile of PAC-1, a potential anticancer drug, was investigated using liquid chromatography-mass spectrometric (LC/MS) techniques. Two different types of mass spectrometers--a quadrupole time-of-flight (Q-TOF) mass spectrometer and an ion trap (IT) mass spectrometer--were employed to acquire structural information on PAC-1 metabolites. A gradient liquid chromatographic system composed of 0.2% formic acid in methanol and 0.2% formic acid in water was used for metabolite separation on an Agilent TC-C(18) column. A total of 16 metabolites were detected. The corresponding product ion spectra were acquired and interpreted, and structures were proposed. Accurate mass measurement using LC-Q-TOF was used to determine the elemental composition of metabolites thereby confirming the proposed structures of these metabolites. Phase I metabolic changes were predominantly observed, including debenzylation, dihydrodiol formation, hydroxylation, and dihydroxylation. The detected phase II metabolites included PAC-1 and hydroxylated PAC-1 glucuronide conjugates. Based on metabolite analysis, several PAC-1 metabolic pathways in rat were proposed.  相似文献   

6.
7.
Sildenafil metabolism in liver microsomes obtained from different species was studied in vitro and compared using liquid chromatography-mass spectrometry (LC/MS) and multivariate statistical analysis. Sildenafil (1, 5, and 25 μM) was incubated with rat, mouse, dog, monkey, and human liver microsomes along with NADPH, and the reaction mixtures were analyzed by LC/MS to obtain species-specific metabolic profiles of sildenafil. A total of 12 metabolites were detected and their peak area ratio values were used as variables for multivariate analyses to evaluate the interspecies differences in sildenafil metabolism. Principal components analysis of the metabolic profiles showed that the mouse samples were generally clustered closer to the human samples on the principal component score plot. Similarity index (SI) indicated that sildenafil metabolism in mice, compared to the other animals, was highly analogous (SI=0.764 at 25 μM) to that in humans. These results suggest that LC/MS-based multivariate analytical approaches are useful for the evaluation of interspecies differences in the metabolism of xenobiotics.  相似文献   

8.
Shen Q  Li X  Qiu Y  Su M  Liu Y  Li H  Wang X  Zou X  Yan C  Yu L  Li S  Wan C  He L  Jia W 《Journal of proteome research》2008,7(5):2151-2157
Epidemiology and studies in animal models have revealed that prenatal malnutrition is highly correlated with abnormal fetal neurodevelopment. We present here a combined metabonomic and metallomic profiling technique to associate the metabolic and trace-elemental composition variations of rat amniotic fluid (AF) in malnourished pregnant rats with the retardation of fetal rat neurodevelopment. The AF samples from three groups of pregnant Sprague-Dawley rats, which were fed either a normal diet, a low-protein diet, or "a famine diet", were subjected to GC/MS and ICP/MS combined with multivariate data analysis (MVDA). PCA scores plot of both GC/MS and ICP/MS data showed similar and unique metabolic signatures of AF in response to the different diets. Rats in the famine group released increased amounts of glycine, inositol, putrescine, and rubidium and decreased amounts of methionine, dopa, tryptophan, glutamine, zinc, cobalt, and selenium in the AF. These discriminable variations in the AF may indicate the abnormality of a number of metabolic pathways in fetal rats including the folate cycle and methionine pathway, the monoamine pathway, and tri-iodothyronine (T3) metabolism. The abnormalities may be the result of metabolites or elemental differences or a combination of both. This study demonstrates the potential of combining profiling of small-molecule metabolites and trace elements to broaden the understanding of biological variations associated with fetal neurodevelopment induced by environmental perturbation.  相似文献   

9.
10.
2型糖尿病(type 2 diabetes mellitus,T2DM)是一种因胰岛素分泌不足或胰岛素抵抗而引起的慢性代谢疾病,T2DM患病人数的快速增长使治疗和预防T2DM成为世界上亟待解决的医学问题。随着微生物组学技术的进步,肠道菌群及其代谢产物与T2DM的研究亦逐渐深入,肠道菌群可能成为治疗和预防T2DM的靶点。肠道菌群及其代谢产物作用于T2DM的潜在机制,主要是参与体内炎症反应、增加肠道短链脂肪酸产量、调节肠道胆汁酸的代谢、调节支链氨基酸的代谢等。目前,治疗T2DM的药物可能会产生一些副作用,而基于肠道菌群干预T2DM的措施相对安全无害。例如,可通过严格控制的特定结构饮食长期摄入或增加益生菌的长期摄取控制血糖,或通过口服可影响肠道菌群生态结构的降糖药物(二甲双胍、阿卡波糖)有效地调控血糖水平。综述基于肠道菌群及其代谢产物诱发T2DM的潜在机制,研讨基于肠道菌群干预T2DM的措施,从肠道菌群的新视角探索治疗T2DM的新方法,为彻底治疗T2DM提供一种新可能。  相似文献   

11.
目的

探究赛前集训控体重对高水平摔跤运动员肠道菌群及代谢物的影响。

方法

招募某省摔跤队运动员11名,在赛前集训期前后测量身体成分、收集粪便样本,采用16S rRNA基因测序技术检测肠道微生物的分布和丰度,利用非靶向代谢组学分析微生物的差异代谢产物及其所富集的功能。

结果

摔跤运动员赛前集训后体重显著降低(P<0.05),但肌肉含量无显著差异。微生物组结果显示,控体重前后男女运动员肠道菌群alpha多样性和beta多样性均无显著差异。男性运动员Intestinibacter丰度显著增高(P<0.05),且与体重、四肢骨骼肌指数变化量呈正相关。代谢组学分析结果显示,男性运动员菌群差异代谢物富集于硫代谢和主要胆汁酸代谢相关通路,女性运动员菌群差异代谢物富集于神经活性配体—受体交互相关信号通路;Intestinibacter丰度变化与胆汁酸7alpha-hydroxy-3-oxo-4-cholestenoate变化呈正相关。

结论

赛前集训控体重未影响摔跤运动员肠道菌群生物多样性和整体物种构成,但可以改变男性摔跤运动员肠道菌群中产丁酸盐菌属的相对丰度,整体表现为运动员肠道菌群对控体重过程较为适应。

  相似文献   

12.
In this work, the authors present a novel, robotic, automated protocol for assessing a metabolic stability protocol assembled on a Hamilton platform and a new strategy for pooling samples (cassette analysis). To increase the high throughput of the liquid chromatography (LC) step, fast chromatography and automated liquid chromatography tandem mass spectrometry (LC/MS/MS) analytical methods were also developed, and a rapid data analysis system was generated that converts peak areas obtained by LC/MS/MS in intrinsic clearance values. All of the steps of the microsomal stability assay were carefully studied and optimized. Standard errors and confidence intervals of the measured clearances were also automatically generated in the process to allow an immediate evaluation of the significance of observed values. Methods based on pooling analysis of 2 and 4 different analytes were compared with a standard method without pooling. A simple statistical treatment was used to show their equivalence. The different protocols developed were analyzed in terms of the best compromise between accuracy and high-throughput capabilities.  相似文献   

13.
This study was conducted to compare the in vivo metabolites of salvianolic acid B (Sal B) between normal rats and antibiotic-treated rats and to clarify the role of intestinal bacteria on the absorption, metabolism and excretion of Sal B. A valid method using LC-MS(n) analysis was established for identification of rat biliary and fecal metabolites. And isolation of normal rat urinary metabolites by repeated column chromatography was applied in this study. Four biliary metabolites and five fecal metabolites in normal rats were identified on the basis of their MS(n) fragmentation patterns. Meanwhile, two normal rat urinary metabolites were firstly identified on the basis of their NMR and MS data. In contrast, no metabolites were detected in antibiotic-treated rat urine and bile, while the prototype of Sal B was found in antibiotic-treated rat feces. The differences of in vivo metabolites between normal rats and antibiotic-treated rats were proposed for the first time. Furthermore, it was indicated that the intestinal bacteria showed an important role on the absorption, metabolism and excretion of Sal B. This investigation provided scientific evidence to infer the active principles responsible for the pharmacological effects of Sal B.  相似文献   

14.
Zhang X  Choi FF  Zhou Y  Leung FP  Tan S  Lin S  Xu H  Jia W  Sung JJ  Cai Z  Bian Z 《The FEBS journal》2012,279(13):2322-2338
The incidence of inflammatory bowel disease, a relapsing intestinal condition whose precise etiology is still unclear, has continually increased over recent years. Metabolic profiling is an effective method with high sample throughput that can detect and identify potential biomarkers, and thus may be useful in investigating the pathogenesis of inflammatory bowel disease. In this study, using a metabonomics approach, a pilot study based on ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS) was performed to characterize the metabolic profile of plasma and urine samples of rats with experimental colitis induced by 2,4,6-trinitrobenzene sulfonic acid. Acquired metabolic profile data were processed by multivariate data analysis for differentiation and screening of potential biomarkers. Five metabolites were identified in urine: two tryptophan metabolites [4-(2-aminophenyl)-2,4-dioxobutanoic acid and 4,6-cihydroxyquinoline], two gut microbial metabolites (phenyl-acetylglycine and p-cresol glucuronide), and the bile acid 12α-hydroxy-3-oxocholadienic acid. Seven metabolites were identified in plasma: three members of the bile acid/alcohol group (cholic acid, 12α-hydroxy-3-oxocholadienic acid and cholestane-3,7,12,24,25-pentol) and four lysophosphatidylcholines [LysoPC(20:4), LysoPC(16:0), LysoPC(18:1) and LysoPC(18:0)]. These metabolites are associated with damage of the intestinal barrier function, microbiota homeostasis, immune modulation and the inflammatory response, and play important roles in the pathogenesis of inflammatory bowel disease. Our results positively support application of the metabonomic approach in study of the pathophysiological mechanism of inflammatory bowel disease.  相似文献   

15.
Genetic test systems involving microorganisms and liver enzyme preparations may be insufficient to detect compounds that require breakdown by enzymes provided by the microbial flora of the intestinal tract. A method is described for providing such activation and for simultaneously testing the potential genetic activity of breakdown products in an indicator organism. Parabiotic chambers containing Saccharomyces cerevisiae genetic test organisms in one chamber were separated by a membrane filter from rat cecal organisms and test chemical contained in the other chamber. The genetic activities of cycasin breakdown products for mutation, gene conversion, and mitotic crossing-over in samples incubated aerobically are reported. Samples containing cycasin alone had a small but clearly increased frequency of genetic damage. Samples containing rat cecal organisms without cycasin showed no increase in genetic activity. Anaerobic incubation resulted in no increase in genetic activity in any of the samples.  相似文献   

16.
Genetic test systems involving microorganisms and liver enzyme preparations may be insufficient to detect compounds that require breakdown by enzymes provided by the microbial flora of the intestinal tract. A method is described for providing such activation and for simultaneously testing the potential genetic activity of breakdown products in an indicator organism. Parabiotic chambers containing Saccharomyces cerevisiae genetic test organisms in one chamber were separated by a membrane filter from rat cecal organisms and test chemical contained in the other chamber. The genetic activities of cycasin breakdown products for mutation, gene conversion, and mitotic crossing-over in samples incubated aerobically are reported. Samples containing cycasin alone had a small but clearly increased frequency of genetic damage. Samples containing rat cecal organisms without cycasin showed no increase in genetic activity. Anaerobic incubation resulted in no increase in genetic activity in any of the samples.  相似文献   

17.
Daidzein (4',7-dihydroxyisoflavone), a soy phytoestrogen, is a weakly estrogenic compound that may have potential health benefits. Biotransformation of daidzein by the human gut microflora after ingestion converts it to either the highly estrogenic metabolite equol or to nonestrogenic metabolites. We investigated the metabolism of daidzein by colonic microflora of rats. Fecal samples, obtained before and after rats were exposed to daidzein at 250 or 1000 parts per million, were incubated in brain-heart infusion (BHI) broth with daidzein under anaerobic conditions. Samples were removed from the cultures daily and analyzed by high-performance liquid chromatography (HPLC) and mass spectrometry. The fecal bacteria of all rats, regardless of prior daidzein exposure, metabolized the added daidzein to dihydrodaidzein. Both compounds disappeared rapidly from BHI cultures incubated for more than 24 h, but no other daidzein metabolites were detected. Only daidzein and dihydrodaidzein were found in a direct analysis of the feces of rats that had consumed daidzein in their diets. Unlike the fecal bacteria of humans and monkeys, the rat flora rapidly metabolized daidzein to aliphatic compounds that could not be detected by HPLC or mass spectral analysis.  相似文献   

18.
Direct-infusion mass spectrometry (MS) was applied to study the metabolic effects of the symbiosis between the endophytic fungus Neotyphodium lolii and its host perennial ryegrass (Lolium perenne) in three different tissues (immature leaf, blade, and sheath). Unbiased direct-infusion MS using a linear ion trap mass spectrometer allowed metabolic effects to be determined free of any preconceptions and in a high-throughput fashion. Not only the full MS(1) mass spectra (range 150-1,000 mass-to-charge ratio) were obtained but also MS(2) and MS(3) product ion spectra were collected on the most intense MS(1) ions as described previously (Koulman et al., 2007b). We developed a novel computational methodology to take advantage of the MS(2) product ion spectra collected. Several heterogeneous MS(1) bins (different MS(2) spectra from the same nominal MS(1)) were identified with this method. Exploratory data analysis approaches were also developed to investigate how the metabolome differs in perennial ryegrass infected with N. lolii in comparison to uninfected perennial ryegrass. As well as some known fungal metabolites like peramine and mannitol, several novel metabolites involved in the symbiosis, including putative cyclic oligopeptides, were identified. Correlation network analysis revealed a group of structurally related oligosaccharides, which differed significantly in concentration in perennial ryegrass sheaths due to endophyte infection. This study demonstrates the potential of the combination of unbiased metabolite profiling using ion trap MS and advanced data-mining strategies for discovering unexpected perturbations of the metabolome, and generating new scientific questions for more detailed investigations in the future.  相似文献   

19.
人类肠道菌群能够产生多种代谢产物或与人体相互作用产生肠道菌群-宿主共代谢物,显著影响人体各大系统的生理功能。当人体健康状态以及肠道菌群发生变化时,肠道代谢物的种类和含量也会相应受到影响,因此肠道菌群代谢产物具有作为疾病诊断指针的巨大潜力。本文总结了常见的几类肠道微生物代谢产物,包括糖类、胆碱代谢物、脂质、氨基酸与肽类、维生素、胆汁酸、短链脂肪酸、酚、苯甲酰基和苯基衍生物等,及其在不同疾病状态下的作用机理,以期更好地理解肠道菌群、代谢产物和疾病之间的相关性,为疾病的预防、诊断和治疗提供新的靶点。  相似文献   

20.
We have developed a metabolic profiling scheme based on direct-infusion Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS). The scheme consists of: (1) reproducible data collection under optimized FT-ICR/MS analytical conditions; (2) automatic mass-error correction and multivariate analyses for metabolome characterization using a newly developed metabolomics tool (DMASS software); (3) identification of marker metabolite candidates by searching a species-metabolite relationship database, KNApSAcK; and (4) structural analyses by an MS/MS method. The scheme was applied to metabolic phenotyping of Arabidopsis (Arabidopsis thaliana) seedlings treated with different herbicidal chemical classes for pathway-specific inhibitions. Arabidopsis extracts were directly infused into an electrospray ionization source on an FT-ICR/MS system. Acquired metabolomics data were comprised of mass-to-charge ratio values with ion intensity information subjected to principal component analysis, and metabolic phenotypes from the herbicide treatments were clearly differentiated from those of the herbicide-free treatment. From each herbicide treatment, candidate metabolites representing such metabolic phenotypes were found through the KNApSAcK database search. The database search and MS/MS analyses suggested dose-dependent accumulation patterns of specific metabolites including several flavonoid glycosides. The metabolic phenotyping scheme on the basis of FT-ICR/MS coupled with the DMASS program is discussed as a general tool for high throughput metabolic phenotyping studies.  相似文献   

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