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1.
PLoS Negl Trop Dis ; 14(4): e0008287, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32352979

RESUMO

Clonorchiasis, caused by chronic infection with Clonorchis sinensis (C. sinensis), is an important food-borne parasitic disease that seriously afflicts more than 35 million people globally, resulting in a socioeconomic burden in endemic regions. C. sinensis adults long-term inhabit the microaerobic and limited-glucose environment of the bile ducts. Energy metabolism plays a key role in facilitating the adaptation of adult flukes to crowded habitat and hostile environment. To understand energy source for adult flukes, we compared the component and content of free amino acids between C. sinensis-infected and uninfected bile. The results showed that the concentrations of free amino acids, including aspartic acid, serine, glycine, alanine, histidine, asparagine, threonine, lysine, hydroxylysine, and urea, were significantly higher in C. sinensis-infected bile than those in uninfected bile. Furthermore, exogenous amino acids could be utilized by adult flukes via the gluconeogenesis pathway regardless of the absence or presence of exogenous glucose, and the rate-limiting enzymes, such as C. sinensis glucose-6-phosphatase, fructose-1,6-bisphosphatase, phosphoenolpyruvate carboxykinase, and pyruvate carboxylase, exhibited high expression levels by quantitative real-time PCR analysis. Interestingly, no matter whether exogenous glucose was present, inhibition of gluconeogenesis reduced the glucose and glycogen levels as well as the viability and survival time of adult flukes. These results suggest that gluconeogenesis might play a vital role in energy metabolism of C. sinensis and exogenous amino acids probably serve as an important energy source that benefits the continued survival of adult flukes in the host. Our study will be a cornerstone for illuminating the biological characteristics of C. sinensis and the host-parasite interactions.


Assuntos
Aminoácidos/metabolismo , Bile/parasitologia , Clonorchis sinensis/crescimento & desenvolvimento , Clonorchis sinensis/metabolismo , Animais , Bile/química , Gatos , Clonorquíase/parasitologia , Clonorchis sinensis/enzimologia , Clonorchis sinensis/genética , Modelos Animais de Doenças , Metabolismo Energético , Perfilação da Expressão Gênica , Gluconeogênese , Redes e Vias Metabólicas/genética , Ratos
2.
Zhongguo Zhong Yao Za Zhi ; 45(5): 1064-1069, 2020 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-32237447

RESUMO

The pig bile powder, bovine bile powder, snake bile, sheep bile, goose bile powder, and bear bile powder were contained by the Chinese Pharmacopoeia. The bile power medicine has a long history in traditional Chinese medicine and definite effect. However, the medicine of bile powder(bile) are similar in morphology. Besides, many medicine lack specific microscopic identification characteristics and chemical characteristics. There is a risk of adulteration, especially when the fake medicine were mixed in authentic medicine, it is difficult to detection. The key to control the quality and ensures the clinical efficacy is the good or bad, true or false of the bile power medicine. The STR typing technology is a method that according to differential typing of PCR amplified lengths to compare and identify individual organisms. Based on the principle of STR typing, the easily, rapid DNA fingerprinting method to identify the bile power and adulteration was established.The original animal or bile powder of pigs, cattle, sheep, chickens, ducks, geese, snakes, bears, fish were collected, the 12 S-L1091/12 S-H1478 and 16 S-L3428/16 S-H3667 was obtained by sifted, the DNA fingerprinting of the bile power and adulteration was obtained by STR typing. Every species has different STR fingerprints, so different species can be identified. Besides, the fingerprints have both the authentic and fake's information, the adulteration of authentic and fake can be identified. Therefore, the method to identify the bile power and adulteration was achieved through the combination of two primers. The DNA fingerprinting method established in this study can also be used for other animal medicine.


Assuntos
Bile/química , Impressões Digitais de DNA , Materia Medica/análise , Animais , Bovinos , Galinhas , Medicina Tradicional Chinesa , Ovinos , Suínos , Ursidae
3.
Artigo em Inglês | MEDLINE | ID: mdl-31911204

RESUMO

This study was conducted to develop a highly selective, sensitive, and validated method for quantifying metronidazole in human plasma and bile fluid. Metronidazole and metronidazole-d4 (internal standard) were extracted from 100 µL of plasma and bile fluid by liquid-liquid extraction. Liquid chromatography with a Hydrosphere C18 column (50 × 2.0 mm) was performed using 10 mM ammonium formate (pH 4.0) and acetonitrile (20:80, v/v) as the mobile phase. Triple quadrupole mass spectrometry was operated with an electrospray ionization interface in multiple reaction monitoring and positive ion modes. The calibration curves were linear for bile and plasma samples over the range of 50-20,000 ng/mL (r2 > 0.999). The intra- and inter-day coefficients of variation (CVs) for plasma ranged from 2.50% to 7.85% and 3.11% to 16.9%, respectively; for bile, the intra-and inter-run precision (CVs) ranged from 2.76% to 13.2% and 3.16% to 11.5%, respectively. The mean extraction recovery for metronidazole ranged from 76.5% to 82.1% in plasma and from 78.8% to 87.8% in bile, respectively. Our proposed analytical method was successfully applied to determine metronidazole concentrations in bile as well as in plasma at multiple time points in a patient with acute cholangitis.


Assuntos
Bile/química , Cromatografia Líquida de Alta Pressão/métodos , Metronidazol/análise , Espectrometria de Massas em Tandem/métodos , Anti-Infecciosos/análise , Anti-Infecciosos/sangue , Anti-Infecciosos/química , Anti-Infecciosos/farmacocinética , Humanos , Limite de Detecção , Modelos Lineares , Extração Líquido-Líquido , Metronidazol/sangue , Metronidazol/química , Metronidazol/farmacocinética , Reprodutibilidade dos Testes
4.
J Forensic Sci ; 65(2): 570-579, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31566759

RESUMO

Only limited data exist concerning the utility of complementary specimens in heroin-related deaths. As such, this report employed a validated LC-MS-MS method to quantify 6-monoacetylmorphine (6-MAM), 6-acetylcodeine (6-AC), and their metabolites morphine and codeine in blood with (BN) and without preservative (B) and the additional unpreserved specimens of vitreous humor, urine, stomach contents, and bile from 20 postmortem cases in which heroin was the primary cause of death. The median concentration of 6-MAM in BN was 0.011 mg/L, B was 0.008 mg/L, urine was 0.186 mg/L, vitreous humor was 0.022 mg/L, stomach contents was 0.147 mg/L, and bile was 0.012 mg/L. Only one case was found to be positive for 6-AC in B (case 6, 0.002 mg/L), and the median concentration of 6-AC was 0.002 mg/L in BN, 0.012 mg/L in urine, 0.003 mg/L in vitreous humor, 0.057 mg/L in stomach contents, and 0.004 mg/L in bile. These findings present new information on the distribution of these analytes in complementary matrices and support their inclusion for accurately determining the role of heroin in opioid-related deaths.


Assuntos
Codeína/análogos & derivados , Codeína/análise , Dependência de Heroína/diagnóstico , Derivados da Morfina/análise , Morfina/análise , Detecção do Abuso de Substâncias/métodos , Adulto , Idoso , Bile/química , Biomarcadores/análise , Cromatografia Líquida , Feminino , Toxicologia Forense/métodos , Conteúdo Gastrointestinal/química , Dependência de Heroína/mortalidade , Humanos , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Corpo Vítreo/química , Adulto Jovem
5.
Forensic Sci Int ; 307: 110108, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31877542

RESUMO

Quetiapine is an atypical antipsychotic drug, frequently found in post-mortem samples. The quantitative determination of active metabolites may help in the interpretation of the potential toxic effects of the parent drug and its role in death. A fully validated LC-MS/MS method was developed for the identification and quantification of quetiapine and two main metabolites (N-desalkylquetiapine and 7-hydroxyquetiapine) in blood, biological fluids and tissues. Then, the distribution of analytes in different matrices was evaluated. LODs of 0.9, 0.3 and 0.3ng/mL were calculated for quetiapine, N-desalkylquetiapine and 7-hydroxyquetiapine respectively; while a LOQ at the concentration of 10.0ng/mL was defined for the three analytes. 13 post-mortem positive real cases have been included in the experiment. The results revealed that quetiapine and N-desalkylquetiapine might undergo a significant post-mortem redistribution, while 7-hydroxyquetiapine is less affected by this factor. N-desalkylquetiapine could be found in blood in relatively high concentrations in comparison to those of quetiapine; therefore, it should be always advisable to measure both the analytes. The analysis of tissues could provide additional data on potential intoxication with quetiapine.


Assuntos
Antipsicóticos/farmacocinética , Mudanças Depois da Morte , Fumarato de Quetiapina/farmacocinética , Tecido Adiposo/química , Adulto , Idoso , Bile/química , Química Encefálica , Cromatografia Líquida , Dibenzotiazepinas/farmacocinética , Feminino , Toxicologia Forense , Humanos , Rim/química , Limite de Detecção , Fígado/química , Pulmão/química , Masculino , Pessoa de Meia-Idade , Músculo Esquelético/química , Baço/química , Espectrometria de Massas em Tandem , Distribuição Tecidual , Adulto Jovem
6.
Food Funct ; 10(12): 8250-8262, 2019 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-31720652

RESUMO

Carotenoid esters are present in considerable amounts in most fruits, such as in citrus. Although the bioavailability of carotenoid esters is similar or even higher compared to that of free carotenoids, these molecules are generally detected only in the free form in human plasma, suggesting that hydrolysis of carotenoid esters occurs in vivo. However, the available in vitro digestion methods were not able to achieve satisfactory carotenoid ester hydrolysis so far. As bile salts play an essential role in the hydrolytic action of lipolytic enzymes from pancreatin, we evaluated the effect of increasing the bile extract/food ratio from 0.045 to 0.12 (g g-1) on the hydrolysis of ß-cryptoxanthin esters from mandarin pulp during in vitro digestion. Additionally, considering the positive effect of lipids on carotenoid bioavailability, the impact of soybean oil addition on carotenoid ester hydrolysis was studied. Finally, bioaccessibility and recovery of 33 carotenoids were assessed by LC-DAD-MS. The hydrolysis extent of ß-cryptoxanthin esters enhanced from 29% to 55% by increasing the bile extract/food ratio, but reduced respectively to 28% and 11% by the addition of 1% and 10% oil (p < 0.05). The bioaccessibility of overall carotenoids improved from 19% to 35% by increasing the bile extract/food ratio, along with that of (all-E)-ß-carotene (from 19 to 31%) and total (all-E)-ß-cryptoxanthin (17% to 49%). Soybean oil addition reduced carotenoid micellarization, regardless of the concentration (p < 0.05). Irrespective of the bile extract amount and oil addition, the bioaccessibility of carotenoids was inversely related to its hydrophobicity, with respect to the following ranking: free xanthophylls > carotenes ≥ xanthophyll esters. Altogether, these results indicate that increasing the bile extract amount is a simple and inexpensive option to improve carotenoid ester hydrolysis in in vitro digestion protocols. Additionally, the constant amounts of bile (and possibly enzymes) of static methods, such as INFOGEST, should be further optimized for experiments involving lipid addition in which carotenoid bioaccessibility is evaluated.


Assuntos
Bile/química , Carotenoides/química , Ésteres/química , Digestão , Hidrólise , Xantofilas/química
7.
Am J Forensic Med Pathol ; 40(4): 329-335, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31688050

RESUMO

Forensic pathologists are requested to select matrices alternative to blood in cases of toxicological interest in which blood is not available for different reasons. We evaluated morphine concentrations in blood, bile, and liver samples in 52 cases of heroin overdoses, relating them to each other, to understand the information that could be derived from their analysis. Gas chromatography/mass spectrometry analysis was performed for all the samples positive on screening for opiates. Shapiro-Wilk test, nonparametric Mann-Whitney test, linear regression analysis, and Bland-Altman test were used for analysis. Linear regression demonstrated that there was not a statistically significant association in morphine concentrations between blood and bile and blood and liver. Mean liver/blood ratio was 2.76, varying from 0.131 to 13.379, and bile/blood ratio was 28.79, varying from 0.28 to 559.16. According to these results, bile analysis is a "screening test"; biliary or hepatic concentration of morphine cannot provide information on hematic concentration at the time of death, having no forensic value taken individually.


Assuntos
Bile/química , Overdose de Drogas , Heroína/envenenamento , Fígado/química , Morfina/análise , Adolescente , Adulto , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Dependência de Heroína , Humanos , Modelos Lineares , Masculino , Detecção do Abuso de Substâncias , Adulto Jovem
8.
Analyst ; 144(24): 7236-7241, 2019 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-31674603

RESUMO

A whole-sample-covering near-infrared (NIR) spectroscopy scheme has been adopted for the simple drop-and-dry measurement of raw bile juice for the identification of gallbladder (GB) diseases of stone, polyp, and cancer. For reproducible measurement, a non-NIR absorbing polytetrafluoroethylene (PTFE) providing a hydrophobic surface was chosen as a substrate to form bile juice droplets of a consistent shape. To ensure representative spectroscopic sampling, NIR radiation illuminated the whole area of the dried sample for spectral acquisition. The NIR band shapes and relative band intensities of GB cancer differed moderately from those of GB stone and GB polyp. The composition of GB cancer samples was presumed to be dissimilar from other sample compositions. Differentiation between GB polyp and GB stone, however, was less facile; nevertheless, in the case of GB polyp samples, the obtained NIR features were informative in the identification of various pathological conditions such as adenomyomatosis (abnormal growth of epidermal tissue) and hepatitis B. To elucidate the NIR features of bile juice samples, separate NIR spectra of major bile constituents such as conjugated bile salts, lecithin, cholesterol, and albumin were analyzed. The demonstrated NIR spectroscopy scheme requiring no sample pretreatment or separation of bile juice could be useful for fast bile juice-based screening of GB diseases, especially the identification of early GB cancer.


Assuntos
Bile/química , Doenças da Vesícula Biliar/diagnóstico , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Estudos de Viabilidade , Humanos , Interações Hidrofóbicas e Hidrofílicas , Politetrafluoretileno/química , Análise de Componente Principal , Espectroscopia de Luz Próxima ao Infravermelho/instrumentação
9.
Artigo em Inglês | MEDLINE | ID: mdl-31627161

RESUMO

Xiao Chai Hu Tang (XCHT) is sold as traditional medicine or dietary supplement in worldwide. To understand metabolism profile of traditional medicine is key point in their logical pharmacological research and clinical application. Based on our previous research of the chemical and absorption signature of XCHT in vitro, we proposed a novel strategy to identify the bioactive components of XCHT in vivo. This strategy have two steps: firstly, based on the parents' database in vitro, built-in and editable biotransformations for phase I and phase II metabolism reactions with MassHunter Metabolite ID software (building metabolites database). Secondly, mouse plasma, bile and urine samples were analyzed by UHPLC-ESI-Q-TOF/MS technique, and the absorbed parents and metabolites were compared and identified with the XCHT's digital library using MassHunter Metabolite ID software. In total, 27 parent compounds and 26 metabolites of XCHT were identified in vivo, 2'-O-xylosyl saikosaponin b2 or b1 was reported for the first time. Saponins and their related metabolites were predominantly excreted into the bile, but flavonoids were excreted by both hepatic as well as renal excretion. Flavonoids, saponins, gingerol and their related metabolites were the absorbed components in cardiovascular system and bioactive components of XCHT. Phase I reactions (hydrolysis, hydroxylation and oxidation) and phase II reactions (glucuronidation) were identified and involved in the mouse metabolism of XCHT.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Bile/química , Bile/metabolismo , Catecóis/análise , Catecóis/química , Catecóis/metabolismo , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/química , Álcoois Graxos/análise , Álcoois Graxos/química , Álcoois Graxos/metabolismo , Flavonoides/análise , Flavonoides/química , Flavonoides/metabolismo , Masculino , Camundongos , Saponinas/análise , Saponinas/química , Saponinas/metabolismo
10.
Oncol Rep ; 42(6): 2622-2634, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31661142

RESUMO

Cholangiocarcinoma (CCA) represents a type of epithelial cancer with a late diagnosis and poor outcome. However, the molecular mechanisms responsible for the development of CCA have not yet been fully identified. Thus, in this study, we aimed to elucidate some of these mechanisms. For this purpose, isobaric tags for relative and absolute quantification (iTRAQ) was performed to analyze the secretory proteins from the 2 CCA cell lines, TFK1 and HuCCT1, as well as from a normal biliary epithelial cell line, human intrahepatic biliary epithelial cells (HiBECs). Differentially expressed proteins (DEPs) were identified and biological process analysis was performed according to the Gene Ontology (GO) functional classification annotation and KEGG metabolic pathway map analysis. tumor protein D52 (TPD52) and DnaJ heat shock protein family (Hsp40) member B1 (DNAJB1) were validated using RT­qPCR, western blot analysis and immunohistochemistry. In total, 778 proteins were identified as DEPs. Following validation, TPD52 and DNAJB1 were used for further analysis. The expression levels of TPD52 and DNAJB1 were elevated in CCA cell lines, tissues and bile samples, suggesting that these proteins may contribute to tumor pathogenesis. In addition, the expression levels of TPD52 and DNAJB1 were found to be closely associated with the clinical parameters and prognosis of patients with CCA. On the whole, the findings of this study indicate that TPD52 and DNAJB1 may serve as novel bile biomarkers for CCA.


Assuntos
Neoplasias dos Ductos Biliares/diagnóstico , Bile/química , Biomarcadores Tumorais/análise , Colangiocarcinoma/diagnóstico , Proteínas de Choque Térmico HSP40/análise , Proteínas de Neoplasias/análise , Neoplasias dos Ductos Biliares/mortalidade , Neoplasias dos Ductos Biliares/patologia , Ductos Biliares Intra-Hepáticos/citologia , Ductos Biliares Intra-Hepáticos/patologia , Biomarcadores Tumorais/metabolismo , Linhagem Celular Tumoral , Colangiocarcinoma/mortalidade , Colangiocarcinoma/patologia , Cromatografia Líquida de Alta Pressão/métodos , Biologia Computacional , Células Epiteliais/patologia , Feminino , Perfilação da Expressão Gênica , Proteínas de Choque Térmico HSP40/metabolismo , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Proteínas de Neoplasias/metabolismo , Prognóstico , Proteômica/métodos , Reação em Cadeia da Polimerase em Tempo Real , Espectrometria de Massas em Tandem/métodos , Análise Serial de Tecidos , Regulação para Cima
11.
Molecules ; 24(19)2019 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-31554153

RESUMO

Alpinetin, a bioactive flavonoid, has been known to have a diverse therapeutic effect, with namely anti-inflammatory, anticancer and antioxidant effects with low systemic toxicity. This study aimed to obtain metabolic profiles of alpinetin in orally administrated rats. The metabolites of alpinetin were systematically analyzed and identified by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The chromatographic separation was achieved on a High Strength Silica (HSS) T3 (1.8 µm, 2.1 × 100 mm) column with the mobile phase consisting of water containing 0.1% formic acid and acetonitrile with 0.1% formic acid via gradient elution. An extracted ion chromatogram strategy based on multiple prototype/metabolite intermediate templates and 71 typical metabolic reactions was proposed to comprehensively profile the metabolites of alpinetin. With the metabolite profiling strategy, altogether 15 compounds were recognized from urine, plasma, bile and feces of rats after intragastric administration of alpinetin for the first time. The prototype, glucuronide conjugates and phenolic acids metabolites were the probable predominant form of alpinetin in rats. This work showed a comprehensive study of the probable metabolic pathways of alpinetin in vivo, which could provide meaningful information for future pharmacological studies.


Assuntos
Flavanonas/administração & dosagem , Metaboloma/efeitos dos fármacos , Metabolômica , Administração Oral , Animais , Bile/química , Líquidos Corporais/química , Cromatografia Líquida de Alta Pressão , Fezes/química , Masculino , Redes e Vias Metabólicas , Metabolômica/métodos , Estrutura Molecular , Plasma/química , Ratos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
12.
Forensic Sci Int ; 303: 109959, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31546164

RESUMO

The organ distribution of 3-fluorophenmetrazine (3-FPM), pyrazolam, diclazepam as well as its main metabolites delorazepam, lormetazepam and lorazepam, was investigated. A solid phase extraction (SPE) and a QuEChERS (acronym for quick, easy, cheap, effective, rugged and safe) - approach were used for the extraction of the analytes from human tissues, body fluids and stomach contents. The detection was performed on a liquid chromatography-tandem mass spectrometry system (LCMS/MS). The analytes of interest were detected in all body fluids and tissues. Results showed femoral blood concentrations of 10 µg/L for 3-FPM, 28 µg/L for pyrazolam, 1 µg/L for diclazepam, 100 µg/L for delorazepam, 6 µg/L for lormetazepam, and 22 µg/L for lorazepam. Tissues (muscle, kidney and liver) and bile exhibited higher concentrations of the mentioned analytes than in blood. Additional positive findings in femoral blood were for 2-fluoroamphetamine (2-FA, approx. 89 µg/L), 2-flourometamphetamine (2-FMA, hint), methiopropamine (approx. 2.2 µg/L), amphetamine (approx. 21 µg/L) and caffeine (positive). Delorazepam showed the highest ratio of heart (C) and femoral blood (P) concentration (C/P ratio = 2.5), supported by the concentrations detected in psoas muscle (430 µg/kg) and stomach content (approx. 210 µg/L, absolute 84 µg). The C/P ratio indicates that delorazepam displays susceptibility for post-mortem redistribution (PMR), supported by the findings in muscle tissue. 3-FPM, pyrazolam, diclazepam, lorazepam and lormetazepam did apparently not exhibit any PMR. The cause of death, in conjunction with autopsy findings was concluded as a positional asphyxia promoted by poly-drug intoxication by arising from designer benzodiazepines and the presence of synthetic stimulants.


Assuntos
Benzodiazepinas/farmacocinética , Drogas Desenhadas/farmacocinética , Diazepam/análogos & derivados , Fenmetrazina/análogos & derivados , Mudanças Depois da Morte , Adulto , Benzodiazepinas/análise , Bile/química , Líquidos Corporais/química , Química Encefálica , Drogas Desenhadas/análise , Diazepam/análise , Diazepam/farmacocinética , Toxicologia Forense , Conteúdo Gastrointestinal/química , Humanos , Rim/química , Fígado/química , Lorazepam/análogos & derivados , Lorazepam/análise , Lorazepam/farmacocinética , Pulmão/química , Masculino , Nordazepam/análogos & derivados , Nordazepam/análise , Nordazepam/farmacocinética , Líquido Pericárdico/química , Fenmetrazina/análise , Fenmetrazina/farmacocinética , Músculos Psoas/química , Espectrometria de Massas em Tandem
13.
J Sep Sci ; 42(21): 3382-3389, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31503388

RESUMO

"Dogel ebs" was known as Sophora flavescens Ait., a classical traditional Chinese Mongolian herbal medicine, which had the effects on damp-heat dysentery, scrofula, and syndrome of accumulated dampness toxicity. Although the chemical constituents have been clarified by our previous studies, the metabolic transformation of "Dogel ebs" in vivo was still unclear. To explore the mechanism of "Dogel ebs," the metabolites in plasma, bile, and urine samples were investigated. A fast positive and negative ion switching technology was used for the simultaneous determination of flavonoids and alkaloids in "Dogel ebs" in a single run. And a target-group-change coupled with mass defect filtering strategy was utilized to analyze the collected data. 89 parent compounds and 82 metabolites were characterized by high-performance liquid chromatography with quadrupole exactive Orbitrap mass spectrometry. Both phase I and phase II metabolites were observed and the metabolic pathways involved in oxidation, demethylation, acetylation, and glucuronidation. 69 metabolites of "Dogel ebs," including three hydroxyls bonding xanthohumol, formononetin-7-O-glucuronide, 2'-hydroxyl-isoxanthohumol decarboxylation metabolite, oxysophocarpine dehydrogen, 9α-hydroxysophoramine-O-glucuronide, etc. were reported for the first time.


Assuntos
Bile/química , Medicamentos de Ervas Chinesas/análise , Animais , Bile/metabolismo , Medicamentos de Ervas Chinesas/metabolismo , Masculino , Medicina Tradicional Chinesa , Conformação Molecular , Peso Molecular , Ratos , Ratos Wistar
14.
Environ Toxicol Chem ; 38(12): 2740-2749, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31514227

RESUMO

Prior to the Deepwater Horizon oil spill, we lacked a comprehensive baseline of oil contamination in the Gulf of Mexico's sediments, water column, and biota. Gaps in prespill knowledge limit our ability to determine the aftereffects of the Deepwater Horizon blowout or prepare to mitigate similar impacts during future oil spill disasters. We examined spatiotemporal differences in exposure to and metabolism of polycyclic aromatic hydrocarbons (PAHs) in 2 hake species (Urophycis spp.) to establish a current baseline for these ecologically important, abundant, and at-risk demersal fishes. Gulf hake (Urophycis cirrata) and southern hake (Urophycis floridana) were collected throughout the Gulf of Mexico during extensive longline surveys from 2012 to 2015. Analyses of biliary PAH metabolites and liver PAH concentrations provided evidence of exposures to di- and tricyclic compounds, with the highest concentrations measured in the northern Gulf of Mexico. Species-specific differences were not detected, but temporal trends observed in biliary PAHs suggest a decrease in acute exposures, whereas increasing liver PAHs suggest chronic exposures marked by greater assimilation than metabolism rates. To our knowledge, the present study provides the first multitissue contaminant analyses, as well as the most exhaustive biometric analyses, for both gulf and southern hakes. Though sources of exposure are complex because of multiple natural and anthropogenic PAH inputs, these results will facilitate the development of much needed health metrics for Gulf of Mexico benthos. Environ Toxicol Chem 2019;38:2740-2749. © 2019 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.


Assuntos
Bile/metabolismo , Fígado/metabolismo , Poluição por Petróleo/análise , Hidrocarbonetos Policíclicos Aromáticos/análise , Poluentes Químicos da Água/análise , Animais , Bile/química , Monitoramento Ambiental , Gadiformes/crescimento & desenvolvimento , Gadiformes/metabolismo , Golfo do México , Fígado/química , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Poluentes Químicos da Água/metabolismo
15.
Forensic Sci Int ; 304: 109915, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31416646

RESUMO

Fatal intoxications due to accidental or voluntary intake of synthetic opioids represent an actual emerging issue. We report a case where we have analyzed furanyl fentanyl and its metabolite 4-anilino-N-phenetyl-piperidine (4-ANPP) in blood, urine, gastric content, bile and cerebrospinal fluid. In this case, a 53-year-old man was found dead at home with a needle still inserted in a vein; a plastic bag containing a white powder (later identified as a furanyl fentanyl-based product) was discovered in the room. Biological samples were collected during autopsy and extracted/purified onto a SPE cartridge before instrumental analysis. Qualitative and quantitative analyses were performed by LC-MS/MS on peripheral and cardiac blood, urine, cerebrospinal fluid (CSF), bile and gastric content. Furanyl fentanyl was identified and quantified in all the biological fluids collected. Interestingly, gastric content revealed an unexpected high amount of furanyl fentanyl; yet, cardiac blood and femoral blood provided significantly different concentrations (11.8 and 2.7 ng/g respectively). The concentration of furanyl fentanyl in CSF was similar to that measured in femoral blood (2.6 ng/mL), thus confirming that CSF could be a good alternative biological fluid whenever a postmortem redistribution is suspected. Concentrations of 93.5, 50.4, 171.7, 41.9, 10.2 ng/mL(g) were measured for 4-ANPP in cardiac blood, femoral blood, urine, bile and cerebrospinal fluid, respectively. The outcomes from the presented case report suggest that the two substances have been not only injected intravenously, but probably also ingested by the man. Fentanyl derivative and its precursor seemed to undergo an extensive postmortem redistribution.


Assuntos
Analgésicos Opioides/análise , Analgésicos Opioides/farmacocinética , Fentanila/análogos & derivados , Furanos/análise , Furanos/farmacocinética , Mudanças Depois da Morte , Bile/química , Cromatografia Líquida , Fentanila/análise , Fentanila/farmacocinética , Toxicologia Forense/métodos , Conteúdo Gastrointestinal/química , Humanos , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Transtornos Relacionados ao Uso de Opioides/complicações , Medicamentos Sintéticos/análise , Medicamentos Sintéticos/farmacocinética
16.
Anal Chim Acta ; 1081: 120-130, 2019 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-31446949

RESUMO

Mass spectrometry-based approaches enable us to capture changes in the metabolome in biological systems with high sensitivity and resolution. But global MS-based profiling of the bile acids (BAs) submetabolome is still a challenging task. Particularly for unconjugated BAs, the collision-induced dissociation (CID) fragment ions showed low ion intensities which were insufficient for analysis. This study is aimed at the development of an anion attachment MS-based approach for pseudotargeted profiling of the BAs submetabolome. We demonstrated that anion attachment MS with the combination use of ammonia fluoride (NH4F) and formate could provide stable anionic adduct ([M + HCOO]-) with good MS responses for unconjugated BAs. A mechanistic study revealed that the underlying rationale is due to the NH4F-induced approximate matching of attractions between BAs and anion for the 24-carboxyl hydrogen. This 24-carboxyl hydrogen regioselectivity is useful to screen for potential unconjugated BAs from the biological matrix. The stability and regioselectivity of anion attachment allowed the establishment of SRM transitions for unconjugated BAs for the first time. To profile conjugated BAs that come from the conjugation of glycine or taurine at 24-carboxyl hydrogen, specific precursor/fragment ion transitions were used for the detection. Finally, SRM-based UPLC-MS/MS method was developed for the pseudotargeted profiling of the BAs submetabolome with good linearity (r2 > 0.995) and high sensitivity (0.20-1.37 ng mL-1 for LLOQ). With this method, a total of 83 BAs, covering 45 unconjugated BAs and 38 conjugated BAs, were successfully determined in different biosamples from experimental colitis mice. The BAs metabolism homeostasis was disrupted by colitis, characterized by the decreased BAs levels in serum and excessive BAs accumuation in the gall bladder and colon. Overall, the present anion attachment MS-based approach is sufficiently sensitive and robust to comprehensively measure various BAs.


Assuntos
Compostos de Amônio/química , Ácidos e Sais Biliares/análise , Fluoretos/química , Metabolômica/métodos , Animais , Bile/química , Ácidos e Sais Biliares/química , Cromatografia Líquida de Alta Pressão/métodos , Colite/induzido quimicamente , Colite/metabolismo , Colo/metabolismo , Vesícula Biliar/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Dodecilsulfato de Sódio , Espectrometria de Massas por Ionização por Electrospray/métodos , Sulfassalazina/farmacologia
17.
Forensic Sci Int ; 304: 109892, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31395407

RESUMO

The use of New Psychoactive Substances (NPS) has become a serious global issue with increasing number of reports of their toxicities and fatalities. Likewise, in Singapore, the number of exhibits containing NPS detected had increased 80% from 2011 to 2014. This is a case series of the first four autopsy cases of fatalities due to or related to the use of NPS in Singapore. In one case, we present the first reported case of death due directly to ADB-FUBINACA toxicity (post-mortem blood concentration of 56ng/ml). Another case was due to 25B-NBOMe toxicity (post-mortem blood concentration of 10ng/ml) while the last two cases were deaths related to 5-Fluoro ADB, where the metabolites of the drug were detected.


Assuntos
Anisóis/envenenamento , Drogas Desenhadas/envenenamento , Indazóis/envenenamento , Fenetilaminas/envenenamento , Transtornos Relacionados ao Uso de Substâncias/complicações , Adolescente , Adulto , Anisóis/análise , Bile/química , Doença da Artéria Coronariana/complicações , Conteúdo Gastrointestinal/química , Humanos , Hipóxia-Isquemia Encefálica/complicações , Indazóis/análise , Masculino , Pessoa de Meia-Idade , Fenetilaminas/análise , Pneumonia/complicações , Singapura , Adulto Jovem
18.
Adv Exp Med Biol ; 1155: 35-44, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31468384

RESUMO

Taurine content in the body is maintained by both biosynthesis from sulfur-contained amino acids in the liver and ingestion from usual foods, mainly seafoods and meat. Contrary to the rodents, the maintenance of taurine content in the body depends on the oral taurine ingestion in cats as well as humans because of the low ability of the biosynthesis. Therefore, insufficient of dietary taurine intake increases the risks of various diseases such as blind and expanded cardiomyopathy in the cats. One of the most established physiological roles of taurine is the conjugation with bile acid in the liver. In addition, taurine has effect to increase the expression and activity of bile acid synthesis rate-limiting enzyme CYP7A1. Present study purposed to evaluate the influence of taurine deficiency on bile acids in the cats fed taurine-lacking diet. Adult cats were fed the soybean protein-based diet with 0.15% taurine or without taurine for 30 weeks. Taurine concentration in serum and liver was undetectable, and bile acids in the bile were significantly decreased in the taurine-deficient cats. Taurine-conjugated bile acids in the bile were significantly decreased, and instead, unconjugated bile acids were significantly increased in the taurine-deficient cats. Present results suggested that the taurine may play an important role in the synthesis of bile acids in the liver.


Assuntos
Ácidos e Sais Biliares/análise , Bile/química , Fígado/fisiopatologia , Taurina/deficiência , Animais , Gatos , Dieta
19.
Zhongguo Zhong Yao Za Zhi ; 44(12): 2538-2543, 2019 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-31359721

RESUMO

Bile acids( BAs),the major constituents of bile,are also known to be potential biomarkers of various diseases,especially liver disease. The systematic analysis of BAs is believed to be of great importance towards the clarification of the effective material basis for bile-type medicines,and the diagnosis and therapy of related diseases as well. As a part of systematic study on bile-type medicine ongoing in our group,this study lays emphasis on the isomer discrimination,and the improvement of analytical method of BAs. Further,this method was subsequently applied to elucidate in depth the chemical profile of BAs in yak bile. Regarding isomer discrimination for BAs,we constructed relative response-collision energy curves( RRCECs) by high performance liquid chromatographyion trap-time of flight-mass spectrometry( HPLC-IT-TOF-MS) in combination with high performance liquid chromatography-triple quadrupole-linear ion trap mass spectrometry( HPLC-Qtrap-MS). As a result,both the optimum collision energy( OCE) and CE_(50) exhibited great correlations with structural characteristics,thus enabling the isomer distinguishing,such as unconjugated BAs,glycine-conjugated BAs,and taurine-conjugated BAs. According to information provided by mass spectrometry,the comparison of OCE and CE_(50),retention time matching,combined with reference substances and database retrieval,a total of 30 bile acid derivatives were observed and identified in yak bile. The newly developed method could serve as a feasible tool for the in-depth characterization of BAs in bile and biological samples.


Assuntos
Ácidos e Sais Biliares/química , Bile/química , Animais , Bovinos , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas , Taurina
20.
Molecules ; 24(14)2019 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-31340434

RESUMO

Eupatorin is the major bioactive component of Java tea (Orthosiphon stamineus), exhibiting strong anticancer and anti-inflammatory activities. However, no research on the metabolism of eupatorin has been reported to date. In the present study, ultra-high-performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) combined with an efficient online data acquisition and a multiple data processing method were developed for metabolite identification in vivo (rat plasma, bile, urine and feces) and in vitro (rat liver microsomes and intestinal flora). A total of 51 metabolites in vivo, 60 metabolites in vitro were structurally characterized. The loss of CH2, CH2O, O, CO, oxidation, methylation, glucuronidation, sulfate conjugation, N-acetylation, hydrogenation, ketone formation, glycine conjugation, glutamine conjugation and glucose conjugation were the main metabolic pathways of eupatorin. This was the first identification of metabolites of eupatorin in vivo and in vitro and it will provide reference and valuable evidence for further development of new pharmaceuticals and pharmacological mechanisms.


Assuntos
Flavonoides/farmacocinética , Glicoconjugados/isolamento & purificação , Microssomos Hepáticos/metabolismo , Orthosiphon/química , Acetilação , Animais , Bile/química , Biotransformação , Fezes/química , Flavonoides/sangue , Flavonoides/urina , Microbioma Gastrointestinal/fisiologia , Glicoconjugados/metabolismo , Hidrogenação , Masculino , Metilação , Oxirredução , Extratos Vegetais/química , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
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