RESUMO
RATIONALE: Chlorogenic acid (CA) is well known for its various biological activities. Here, a clinical study was performed in patients with advanced malignant cancer to explore its therapeutic effects. We aimed to develop a method to quantify CA in human plasma and urine to assist the clinical pharmacokinetic study. METHODS: Ultra-performance liquid chromatography (UPLC) coupled with a triple quadruple mass spectrometry was used to separate and detect CA, with puerarin serving as the internal standard. RESULTS: The method presents an excellent linearity ranging from 5 to 2000 ng/mL for plasma analysis and 50 to 20 000 ng/mL for urine analysis. Intra- and inter-day precision and accuracy were both less than 15% for plasma and urine. CONCLUSIONS: These results showed that the novel UPLC method was robust and sensitive, and fulfilled the requirements for a clinical pharmacokinetic study of CA in patients with advanced cancer.
Assuntos
Ácido Clorogênico/sangue , Ácido Clorogênico/urina , Cromatografia Líquida/métodos , Espectrometria de Massas em Tandem/métodos , Calibragem , Fracionamento Químico , Ácido Clorogênico/administração & dosagem , Ácido Clorogênico/farmacocinética , Cromatografia Líquida/normas , Humanos , Injeções Intramusculares , Isoflavonas/análise , Limite de Detecção , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Manejo de Espécimes , Espectrometria de Massas em Tandem/normasRESUMO
A novel anti-tumor candidate drug, 5-chlorogenic acid (5-CQA) injection, was used for the treatment of malignant glioma in clinical trial (phase I) in China. The isolation and identification of the metabolites of 5-CQA injection in humans were investigated in the present study. Urine and feces samples obtained after intramuscular administration of 5-CQA injection to healthy adults have been analyzed by high-performance liquid chromatography coupled with high-resolution mass and multiple-stage mass spectrometry (HPLC-HRMS/MSn). No metabolite was detected in human feces; however, in human urine, a total of six metabolites were identified including isomerized 5-CQA (P1 and P2), hydrolyzed 5-CQA (M1and M2), and methylated 5-CQA (M3 and M4). Among them, M3 and M4 were the main metabolites and target analytes for human mass balance study. Additionally, the structure of M3 and M4 was characterized by high-performance liquid chromatography-solid phase extraction-nuclear magnetic resonance (HPLC-SPE-NMR), and the results demonstrated that the methoxy group of M3 and M4 was exclusively attributed to C-3' and C-4', respectively. Due to the unavailability of commercial reference, the pure products of M3 and M4 were synthesized by 5-CQA methylation and followed by isolation and purification. Moreover, the potential activity of M3 and M4 on malignant glioma was predicted using a reverse molecular docking analysis on eight malignant glioma-related pathways. The results showed that M3 and M4 had various interactions against malignant glioma-related targets. Our study provides an insight into the metabolism of 5-CQA injection in humans and supports the clinical human mass balance study. Graphical abstract á .
Assuntos
Antineoplásicos/metabolismo , Ácido Clorogênico/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Espectroscopia de Ressonância Magnética/métodos , Espectrometria de Massas/métodos , Adulto , Antineoplásicos/química , Antineoplásicos/urina , Ácido Clorogênico/química , Ácido Clorogênico/urina , Fezes/química , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estrutura MolecularRESUMO
The health-promoting effects of phenolic compounds depend on their bioaccessibility from the food matrix and their consequent bioavailability. We carried out a randomized crossover pilot clinical trial to evaluate the matrix effect (raw flesh and juice) of 'Ataulfo' mango on the bioavailability of its phenolic compounds. Twelve healthy male subjects consumed a dose of mango flesh or juice. Blood was collected for six hours after consumption, and urine for 24 h. Plasma and urine phenolics were analyzed by electrochemical detection coupled to high performance liquid chromatography (HPLC-ECD). Five compounds were identified and quantified in plasma. Six phenolic compounds, plus a microbial metabolite (pyrogallol) were quantified in urine, suggesting colonic metabolism. The maximum plasma concentration (Cmax) occurred 2-4 h after consumption; excretion rates were maximum at 8-24 h. Mango flesh contributed to greater protocatechuic acid absorption (49%), mango juice contributed to higher chlorogenic acid absorption (62%). Our data suggests that the bioavailability and antioxidant capacity of mango phenolics is preserved, and may be increased when the flesh is processed into juice.
Assuntos
Antioxidantes/administração & dosagem , Cinamatos/administração & dosagem , Manipulação de Alimentos , Sucos de Frutas e Vegetais , Frutas , Mangifera , Fenóis/administração & dosagem , Adulto , Antioxidantes/análise , Antioxidantes/metabolismo , Ácido Clorogênico/administração & dosagem , Ácido Clorogênico/sangue , Ácido Clorogênico/metabolismo , Ácido Clorogênico/urina , Cinamatos/sangue , Cinamatos/metabolismo , Cinamatos/urina , Produtos Agrícolas/química , Produtos Agrícolas/economia , Produtos Agrícolas/crescimento & desenvolvimento , Estudos Cross-Over , Frutas/química , Frutas/economia , Frutas/crescimento & desenvolvimento , Sucos de Frutas e Vegetais/análise , Microbioma Gastrointestinal , Humanos , Hidroxibenzoatos/administração & dosagem , Hidroxibenzoatos/sangue , Hidroxibenzoatos/metabolismo , Hidroxibenzoatos/urina , Absorção Intestinal , Masculino , Mangifera/química , Mangifera/crescimento & desenvolvimento , México , Valor Nutritivo , Fenóis/sangue , Fenóis/metabolismo , Fenóis/urina , Projetos Piloto , Pirogalol/sangue , Pirogalol/urina , Especificidade da Espécie , Adulto JovemRESUMO
Isochlorogenic acid A is widely present in fruits, vegetables and herbal medicines, and is characterized by anti-inflammatory, hepatoprotective and antiviral properties. However, little is known about its metabolic fate and pharmacokinetic properties. This study is thus designed to investigate the metabolic fate of isochlorogenic acid A. An analytical method based on high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC/Q-TOF MS) was established to characterize the metabolites of isochlorogenic acid A in the plasma, urine and feces of rats. A total of 32 metabolites were identified. The metabolic pathways mainly include hydrolyzation, dehydroxylation, hydrogenation and conjugation with methyl, glucuronic acid, glycine, sulfate, glutathione and cysteine. Moreover, the pharmacokinetic profiles of all the circulating metabolites were investigated. M11 resulting from hydrolyzation, dehydroxylation and hydrogenation was the dominant circulating metabolite after the intragastric administration of isochlorogenic acid A. The results obtained will be useful for further study of elucidating potential bioactive metabolites which can provide better explanation of the pharmacological and/or toxicological effects of this compound.
Assuntos
Anti-Inflamatórios/metabolismo , Anti-Inflamatórios/farmacocinética , Ácido Clorogênico/análogos & derivados , Cromatografia Líquida de Alta Pressão/métodos , Animais , Anti-Inflamatórios/sangue , Anti-Inflamatórios/urina , Antivirais/sangue , Antivirais/metabolismo , Antivirais/farmacocinética , Antivirais/urina , Ácido Clorogênico/sangue , Ácido Clorogênico/metabolismo , Ácido Clorogênico/farmacocinética , Ácido Clorogênico/urina , Fezes/química , Masculino , Espectrometria de Massas/métodos , Redes e Vias Metabólicas , Plantas Medicinais/metabolismo , Substâncias Protetoras/metabolismo , Substâncias Protetoras/farmacocinética , Ratos , Ratos Sprague-DawleyRESUMO
A feeding study was carried out in which six healthy ileostomists ingested a juice drink containing a diversity of dietary (poly)phenols derived from green tea, apples, grapes and citrus fruit. Ileal fluid and urine collected at intervals over the ensuing 24 h period were then analysed by HPLC-MS. Urinary excretions were compared with results obtained in an earlier study in which the juice drink was ingested by ten healthy control subjects with an intact colon. Some polyphenol components, such as (epi)catechins and (epi)gallocatechin(s), were excreted in urine in similar amounts in ileostomists and subjects with an intact colon, demonstrating that absorption took place principally in the small intestine. In the urine of ileostomists, there were reduced levels of other constituents, including hesperetin-7-O-rutinoside, 5-O-caffeoylquinic acid and dihydrochalcones, indicating their absorption in both the small and large intestine. Ileal fluid analysis revealed that even when absorption occurred in the small intestine, in subjects with a functioning colon a substantial proportion of the ingested components still pass from the small into the large intestine, where they may be either absorbed before or after catabolism by colonic bacteria.
Assuntos
Bebidas/análise , Intestino Grosso/efeitos dos fármacos , Intestino Delgado/efeitos dos fármacos , Polifenóis/administração & dosagem , Polifenóis/farmacocinética , Absorção , Adulto , Idoso , Disponibilidade Biológica , Catequina/administração & dosagem , Catequina/farmacocinética , Chalconas/farmacocinética , Chalconas/urina , Ácido Clorogênico/análogos & derivados , Ácido Clorogênico/farmacocinética , Ácido Clorogênico/urina , Cromatografia Líquida de Alta Pressão , Citrus/química , Dieta , Feminino , Hesperidina/farmacocinética , Hesperidina/urina , Humanos , Absorção Intestinal/efeitos dos fármacos , Intestino Grosso/metabolismo , Intestino Delgado/metabolismo , Masculino , Malus/química , Pessoa de Meia-Idade , Ácido Quínico/análogos & derivados , Ácido Quínico/farmacocinética , Ácido Quínico/urina , Chá/química , Vitis/químicaRESUMO
SCOPE: Until now, the question of how the ingested doses of chlorogenic acids (CGA) from coffee influence their absorption and metabolism remains unresolved. To assess absorption in the small intestine, we performed a dose-response study with a randomized, double-blinded, crossover design with ileostomist subjects. METHODS AND RESULTS: After a polyphenol-free diet, the volunteers consumed, on three separate occasions, coffee with different total CGA contents (high 4525 µmol; medium 2219 µmol; low 1053 µmol). CGA concentrations in plasma, ileal effluent, and urine were subsequently determined by HPLC-DAD-ESI-MS and -ESI-MS/MS. The results show that the consumption of higher CGA concentrations leads to a faster ileal excretion. This corresponds to a renal excretion of 8.0 ± 4.9% (high), 12.1 ± 6.7% (medium), and 14.6 ± 6.8% (low) of total CGA and metabolites. Glucuronidation of CGA became slightly greater with increasing dose. After enzyme treatment, the area under the curve (AUC)(0-8h) for CGA metabolites in plasma was 4412 ± 751 nM × h(0-8) (-1) (high), 2394 ± 637 nM × h(0-8) (-1) (medium), 1782 ± 731 nM × h(0-8) (-1) (low), respectively. Additionally, we were able to identify new metabolites of CGA in urine and ileal fluid. CONCLUSION: We conclude that the consumption of high CGA concentrations via coffee might influence the gastrointestinal transit time and consequently affect CGA absorption and metabolism.
Assuntos
Ácido Clorogênico/farmacocinética , Café/química , Intestino Delgado/efeitos dos fármacos , Absorção , Adulto , Disponibilidade Biológica , Ácido Clorogênico/administração & dosagem , Ácido Clorogênico/sangue , Ácido Clorogênico/urina , Cromatografia Líquida de Alta Pressão , Creatinina/urina , Estudos Cross-Over , Relação Dose-Resposta a Droga , Método Duplo-Cego , Feminino , Humanos , Ileostomia/métodos , Íleo/metabolismo , Intestino Delgado/metabolismo , Espectrometria de Massas em TandemRESUMO
Different studies have shown that milk may interact with polyphenols and affect their bioavailability in humans. The present study investigated the effect of the simultaneous consumption of coffee and milk on the urinary excretion of chlorogenic acids (CGA) and metabolites. Subjects were submitted to consumption of water, instant coffee (609 mmol of CGA) dissolved in water, and instant coffee dissolved in whole milk. Urine was collected for 24 h after consumption of each treatment for analysis of CGA and metabolites by HPLC/LC-MS. The amount of CGA and metabolites recovered after consumption of combined coffee-milk (40% ± 27%) was consistently lower in all subjects compared to that of coffee alone (68% ± 20%). Concluding, the simultaneous consumption of milk and coffee may impair the bioavailability of coffee CGA in humans.
Assuntos
Ácido Clorogênico/metabolismo , Coffea/química , Café/química , Leite/química , Adulto , Animais , Disponibilidade Biológica , Bovinos , Ácido Clorogênico/urina , Feminino , Humanos , Masculino , Proteínas do Leite/química , Ligação Proteica , Adulto JovemRESUMO
Chlorogenic acid (5-CQA) is one of the major components in some Chinese herbal injections. However, the metabolism of 5-CQA in rats after intravenous injection has not been determined. An ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF MS) method was applied to identify the metabolites in bile, urine, feces and plasma after a single intravenous administration of 10 mg x kg(-1) 5-CQA to rats. Using MSE and mass defect filter techniques, a total of 35 metabolites were detected in bile, urine, feces and plasma. The predominant metabolites in bile were glutathione conjugates of O-methyl-5-CQA, accounting for approximately 80% of the metabolites excreted in bile. The major components in urine were parent drug, O-methyl-5-CQA, hydrolyzed metabolites and glucuronide conjugates. The major components in feces were O-methyl-5-CQA and its cysteine conjugates. The major component in plasma was the parent drug. The urinary and fecal excretion pathways were equally important to 5-CQA in rats. These results demonstrate that 5-CQA undergoes extensively metabolism in rats and are highly reactive to nucleophiles such as GSH. This finding indicates that attention should be paid on the injections containing 5-CQA, which may covalently bind to proteins, leading to allergenic drug reactions.
Assuntos
Ácido Clorogênico/farmacocinética , Glutationa/metabolismo , Ligação Proteica , Animais , Bile/metabolismo , Biotransformação , Ácido Clorogênico/administração & dosagem , Ácido Clorogênico/sangue , Ácido Clorogênico/urina , Cromatografia Líquida de Alta Pressão/métodos , Cisteína/metabolismo , Fezes/química , Glucuronídeos/metabolismo , Injeções Intravenosas , Masculino , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodosRESUMO
A systematic investigation of the human metabolism of hydroxycinnamic acid conjugates was carried out. A set of 24 potential human metabolites of coffee polyphenols has been chemically prepared, and used as analytical standards for unequivocal identifications. These included glucuronide conjugates and sulfate esters of caffeic, ferulic, isoferulic, m-coumaric and p-coumaric acids as well as their dihydro derivatives. A particular focus has been made on caffeic and 3,4-dihydroxyphenylpropionic acid derivatives, especially the sulfate conjugates, for which regioselective preparation was particularly challenging, and have so far never been identified as human metabolites. Ten out of the 24 synthesized conjugates have been identified in human plasma and/or urine after coffee consumption. A number of these conjugates were synthesized, characterized and detected as hydroxycinnamic acid metabolites for the first time. This was the case of dihydroisoferulic acid 3'-O-glucuronide, caffeic acid 3'-sulfate, as well as the sulfate and glucuronide derivatives of 3,4-dihydroxyphenylpropionic acid.
Assuntos
Líquidos Corporais/metabolismo , Ácidos Cafeicos/sangue , Ácidos Cafeicos/urina , Café/metabolismo , Ácidos Cumáricos/sangue , Ácidos Cumáricos/urina , Comportamento de Ingestão de Líquido , Glucuronídeos/sangue , Glucuronídeos/urina , Ésteres do Ácido Sulfúrico/sangue , Ésteres do Ácido Sulfúrico/urina , Ácidos Cafeicos/química , Ácido Clorogênico/sangue , Ácido Clorogênico/urina , Cromatografia Líquida de Alta Pressão , Ácidos Cumáricos/química , Glucuronídeos/química , Humanos , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Ésteres do Ácido Sulfúrico/químicaRESUMO
Chlorogenic acid possessed a weak caffeine-like psychostimulant property when assessed for its effect on spontaneous locomotor activity in mice. In the evaluation of the effects for the major metabolites of chlorogenic acid which were detected upon incubation with rat feces and/or excreted in urine after oral administration to rats, caffeic and m-coumaric acids were found to be the principal active metabolites, while the others contributed little to this caffeine-like psychostimulant activity.
Assuntos
Ácido Clorogênico/metabolismo , Ácido Clorogênico/farmacologia , Atividade Motora/efeitos dos fármacos , Animais , Ácidos Cafeicos/análise , Ácidos Cafeicos/farmacologia , Estimulantes do Sistema Nervoso Central/metabolismo , Estimulantes do Sistema Nervoso Central/farmacologia , Ácido Clorogênico/urina , Cromatografia Líquida de Alta Pressão , Ácidos Cumáricos/análise , Ácidos Cumáricos/farmacologia , Fezes , Flavonoides , Masculino , Camundongos , Camundongos Endogâmicos , Fenóis , Polifenóis , RatosRESUMO
Estimation of dietary intake of polyphenols is difficult, due to limited availability of food composition data and bias inherent to dietary assessment methods. The aim of the present study was to evaluate the associations between the intake of polyphenol-rich foods and the urinary excretion of several phenolic compounds and therefore explore whether these phenolic compounds could be used as a biomarker of intake. Fifty-three participants of the SU.VI.MAX study (a randomised primary-prevention trial evaluating the effect of daily antioxidant supplementation on chronic diseases) collected a 24 h urine and a spot urine sample and filled a dietary record during a 2 d period. Thirteen polyphenols and metabolites, chlorogenic acid, caffeic acid, m-coumaric acid, gallic acid, 4-O-methylgallic acid, quercetin, isorhamnetin, kaempferol, hesperetin, naringenin, phloretin, enterolactone and enterodiol, were measured using HPLC-electrospray ionisation-MS-MS. In spot samples apple consumption was positively correlated to phloretin, grapefruit consumption to naringenin, orange to hesperetin, citrus fruit consumption to both naringenin and hesperetin, with r coefficients ranging from 0.31 to 0.57 (P < 0.05). The combination of fruits and/or fruit juices was positively correlated to gallic acid and 4-O-methylgallic acid, isorhamnetin, kaempferol, hesperetin, naringenin and phloretin (r 0.24-0.44, P < 0.05). Coffee consumption was positively correlated to caffeic and chlorogenic acids (r 0.29 and 0.63, P < 0.05 respectively). Black tea and wine consumption were positively correlated with gallic and 4-O-methylgallic acids (r 0.37-0.54, P < 0.001). The present results suggest that several polyphenols measured in a spot urine sample can be used as biomarkers of polyphenol-rich food intake.