Metabolomics Approach After Acute Intake of Grumixama Juice
2016. június 13. frissítette: Luciane de Lira Teixeira, University of Sao Paulo
Metabolomics Approach After Acute Intake of Grumixama Juice (Eugenia Brasiliensis Lam)
Investigate the bioavailability of the main flavonoid, their colonic transformation and untargeted metabolites by an metabolomic approach following acute intake of a grumixama juice by humans.
A tanulmány áttekintése
Állapot
Befejezve
Körülmények
Beavatkozás / kezelés
Részletes leírás
The grumixama is a sweet and little cherry native of the South and Southeast regions of the Atlantic Forest of Brazil rich in phenolic compounds, mainly anthocyanins and ellagitannins. The objective of this study is Investigate the bioavailability of the main flavonoid, their colonic transformation and untargeted metabolites by an metabolomic approach following acute intake of a grumixama juice by humans.
Fifteen healthy subjects consumed grumixama juice at single dose, and urine and plasma samples were collected at different time points over 24 h period. The metabolites were analyzed by LC-ESI-MSn, LC-Q-TOF plasma and urine were also analyzed using untargeted metabolomic approach for amino acids and organic acids by GCMS.
Tanulmány típusa
Beavatkozó
Beiratkozás (Tényleges)
15
Fázis
- Nem alkalmazható
Részvételi kritériumok
A kutatók olyan embereket keresnek, akik megfelelnek egy bizonyos leírásnak, az úgynevezett jogosultsági kritériumoknak. Néhány példa ezekre a kritériumokra a személy általános egészségi állapota vagy a korábbi kezelések.
Jogosultsági kritériumok
Tanulmányozható életkorok
18 év (Felnőtt)
Egészséges önkénteseket fogad
Igen
Tanulmányozható nemek
Női
Leírás
Inclusion Criteria:
- Not have medical historical of cardiovascular, gastrointestinal, hepatic, renal, thyroid and/or diabetes dysfunction
- Not be alcohol addicts, not use vitamins and other supplements
- Not are using any kind of medication which affect the digestion and absorption of food, not be smokers
- Not are pregnant.
Exclusion Criteria:
- Have medical historical of cardiovascular, gastrointestinal, hepatic, renal, thyroid and/or diabetes dysfunction
- Be alcohol addicts, use vitamins and other supplements
- Are using any kind of medication which affect the digestion and absorption of food, be smokers
- Are pregnant.
Tanulási terv
Ez a rész a vizsgálati terv részleteit tartalmazza, beleértve a vizsgálat megtervezését és a vizsgálat mérését.
Hogyan készül a tanulmány?
Tervezési részletek
- Kiosztás: N/A
- Beavatkozó modell: Egyetlen csoportos hozzárendelés
- Maszkolás: Nincs (Open Label)
Fegyverek és beavatkozások
Résztvevő csoport / kar |
Beavatkozás / kezelés |
|---|---|
|
Egyéb: Grumixama Juice
Juice of grumixama purple fruit (Eugenia brasiliensis Lam.), which is good source of anthocyanins and ellagitannins. It was made with filtered water and sanitized fruits, with a blender. It was administered in single dose of 0.97 mg of anthocyanins and of 4.71 mg of ellagitannins per mL of juice. Which volunteers ingested 10 mL of juice per each Kg body weight. The metabolomic approach of plasma and urine samples following acute intake of grumixama juice was done by the collection of blood samples and urine, following the intake of grumixama juice. |
In the morning of experimental day after 10 h fasting, the volunteers have been collect the first urine in the morning and it was collected the blood before the intake of grumixama juice, after they drink the juice (0 min), it was collected blood at the times: 15 min, 30 min, 60 min,120 min, 240 min and 24 h; and it was also collected urine at the period: 0 -1h, 1 - 2h, 2 - 4h, 4 - 6 h, 6 - 12 h, 12 - 14 h, the first urine in the morning in the second day (24 h).
During the period in the first 4 h of experiment the volunteers only intake the juice.
|
Mit mér a tanulmány?
Elsődleges eredményintézkedések
Eredménymérő |
Intézkedés leírása |
Időkeret |
|---|---|---|
|
Identification of metabolites in plasma and urine samples of 15 healthy volunteers after intake of grumixama juice by LCMS
Időkeret: 2 months
|
The blood and urine samples were collected in the times describe in "Arms and Interventions".
The samples were acidified and the anthocyanins and ellagitannins metabolites were extracted by SPE column, these metabolites extracted were concentrated and analyzed by LC-ESI-MSn (for phenolic acids and anthocyanins) and LC-Q-TOF (for urolithins), the mass fragments were expressed in m/z.
|
2 months
|
|
Identification of metabolites in plasma and urine samples of 15 healthy volunteers after intake of grumixama juice by GCMS.
Időkeret: 2 months
|
The samples were collected as describe in "Arms and Interventions", but for urine samples were used only the samples collected at times 0 (before the juice intake), 0-1h, 1-2h, 2-4h and at 24h after intake.
The plasma samples had their proteins precipitated with cold methanol, the metabolites were extracted, concentrated and the amino acids and organic acids were derivatized to became volatile and the samples were analyzed by GCMS.
For urine samples the procedures were similar.
The mass fragment profile of derivatized compounds were compared to a GCMS library.
|
2 months
|
Másodlagos eredményintézkedések
Eredménymérő |
Intézkedés leírása |
Időkeret |
|---|---|---|
|
Urinary creatinine values of 15 healthy volunteers for data normalization
Időkeret: 15 days
|
The creatinine was measured in the 15 volunteers by using a commercial kit, the results obtained in mg/dL, and these values were transformed in mol/L.
The values of creatinine were used to normalize the quantification data.
|
15 days
|
|
Cycle cell analysis in breast cancer cells treated with metabolites extracted from plasma and urine of 10 healthy volunteers after grumixama juice intake
Időkeret: 2 days
|
Plasma and urine preparations were obtained by SPE extraction were dried and tested against breast cancer cells (MDA-MB 231) MDA-MB 231 cells were cultured in monolayer on Dulbecco Eagle modified culture medium supplemented with 10% fetal bovine serum, it was used 5x10-4 cells per well. The cells were incubated in a humidified chamber including 5% CO2 at 37ºC into a confluence of approximately 80%. Cells were further exposed to 200 mg/mL of urine and plasma extract preparations and phenolic compound standards (PCA, VA, HA, C3G) at 0.25 and 25 mg/mL, and maintained at 37ºC, under an atmosphere of 5% CO2 for 48 hours. Negative control was supplied with growth media and DMSO, and blank wells containing growth media only. The cells were lysed, the RNA were degraded and the DNA were marked with propidium iodide (1mg/mL) and the fluorescence was measure by flow cytometer and the cell cycle analysis were done by FlowJo. The results were expressed in % of cell for each cell cycle phase. |
2 days
|
|
Multivariate data analysis of metabolites identified in plasma and urine of 15 healthy volunteers after intake of grumixama juice by GCMS
Időkeret: 2 months
|
The peak intensity of each compound identified by GCMS were normalized by using a internal standards (D4-alanine).
For urine these data also were normalized by creatinine values to normalize sample volume.
Multivariate data analysis were done observe changes in these metabolites (amino acids and organic acids) during the 24h after grumixama juice intake.
The data were analyzed by MetaboAnalyst 3.0 (http://www.metaboanalyst.ca/).
|
2 months
|
|
Pathway analysis of metabolites identified by GCMS
Időkeret: 1 month
|
The samples were collected as describe in "Arms and Interventions", but for urine samples were used only the samples collected at times 0 (before the juice intake), 0-1h, 1-2h, 2-4h and at 24h after intake.
The plasma samples had their proteins precipitated with cold methanol, the metabolites were extracted, concentrated and the amino acids and organic acids were derivatized to became volatile and the samples were analyzed by GCMS.
For urine samples the procedures were similar.
The mass fragment profile of derivatized compounds were compared to a GCMS library.
|
1 month
|
|
Identification of anthocyanins in urine samples of 10 healthy volunteers by LCMS.
Időkeret: 2 months
|
The blood and urine samples were collected in the times describe in "Arms and Interventions".
The samples were acidified and the anthocyanins metabolites were extracted by SPE column, these metabolites extracted were concentrated and analyzed by LC-ESI-MSn (for phenolic acids and anthocyanins) the mass fragments were expressed in m/z.
|
2 months
|
|
Identification of phenolic acids in plasma and urine samples of 10 healthy volunteers by LCMS
Időkeret: 2 months
|
The blood and urine samples were collected in the times describe in "Arms and Interventions".
The samples were acidified and the anthocyanins metabolites were extracted by SPE column, these metabolites extracted were concentrated and analyzed by LC-ESI-MSn (for phenolic acids and anthocyanins) the mass fragments were expressed in m/z.
|
2 months
|
|
Identification of urolithins in urine samples of 10 healthy volunteers by LCMS
Időkeret: 2 months
|
The urine samples were collected in the times describe in "Arms and Interventions".
The samples were acidified and the ellagitannins metabolites were extracted by SPE column, these metabolites extracted were concentrated and analyzed by LC-Q-TOF (for urolithins), the mass fragments were expressed in m/z.
|
2 months
|
|
Quantification of anthocyanins in urine samples of 10 healthy volunteers by HPLC
Időkeret: 2 months
|
The urine samples were collected in the times describe in "Arms and Interventions".
The samples were acidified and the anthocyanins metabolites were extracted by SPE column, these metabolites extracted were concentrated and analyzed by HPLC, the results were expressed nmol/mol of creatinine for urine samples.
It was made to each volunteer, in each point of collection sample.
|
2 months
|
|
Quantification of urolithins in urine samples of 10 healthy volunteers by HPLC
Időkeret: 2 months
|
The urine samples were collected in the times describe in "Arms and Interventions".
The samples were acidified and the ellagitannins metabolites were extracted by SPE column, these metabolites extracted were concentrated and analyzed by HPLC, the results were expressed in mmol/mol of creatinine.
It was made to each volunteer, in each point of collection sample.
|
2 months
|
|
Antiproliferative activity of metabolites extracted from plasma of 10 healthy volunteers after grumixama juice intake
Időkeret: 2 days
|
Plasma preparations were obtained by SPE extraction were dried and tested against breast cancer cells (MDA-MB 231) MDA-MB 231 cells were cultured in monolayer on Dulbecco Eagle modified culture medium supplemented with 10% fetal bovine serum, it was used 5x10-4 cells per well.
The cells were incubated in a humidified chamber including 5% CO2 at 37ºC into a confluence of approximately 80%.
Cells were further exposed to 200 mg/mL of plasma extract preparations and phenolic compound standards (PCA, VA, HA, C3G) at 0.25 and 25 mg/mL, and maintained at 37ºC, under an atmosphere of 5% CO2 for 48 hours.
Negative control was supplied with growth media and DMSO, and blank wells containing growth media only.
Cells were marked with CFSE, and fluorescence was measured by flow cytometer.
The results were expressed as % of proliferation inhibition.
|
2 days
|
|
Antiproliferative activity of metabolites extracted from urine of 10 healthy volunteers after grumixama juice intake
Időkeret: 2 days
|
Urine preparations were obtained by SPE extraction were dried and tested against breast cancer cells (MDA-MB 231) MDA-MB 231 cells were cultured in monolayer on Dulbecco Eagle modified culture medium supplemented with 10% fetal bovine serum, it was used 5x10-4 cells per well.
The cells were incubated in a humidified chamber including 5% CO2 at 37ºC into a confluence of approximately 80%.
Cells were further exposed to 200 mg/mL of urine extract preparations and phenolic compound standards (PCA, VA, HA, C3G) at 0.25 and 25 mg/mL, and maintained at 37ºC, under an atmosphere of 5% CO2 for 48 hours.
Negative control was supplied with growth media and DMSO, and blank wells containing growth media only.
Cells were marked with CFSE, and fluorescence was measured by flow cytometer.
The results were expressed as % of proliferation inhibition
|
2 days
|
|
Quantification of amino acids identified by GCMS
Időkeret: 1 month
|
Standards curves of the amino acids that changed significantly after grumixama juice intake were made by GCMS.
Through these standards curves the amino acids and organic acids were quantified and the results were expressed in mmol/μmol of creatinine for urine samples and mmol/mL of plasma.
|
1 month
|
|
Quantification of organic acids identified by GCMS
Időkeret: 1 month
|
Standards curves of organic acids that changed significantly after grumixama juice intake were made by GCMS.
Through these standards curves the organic acids were quantified and the results were expressed in mmol/μmol of creatinine for urine samples and mmol/mL of plasma.
|
1 month
|
|
Quantification of phenolic acids in plasma samples of 10 healthy volunteers by HPLC
Időkeret: 2 months
|
The blood samples were collected in the times describe in "Arms and Interventions".
The samples were acidified and the anthocyanins metabolites were extracted by SPE column, these metabolites extracted were concentrated and analyzed by HPLC, the results were expressed in μmol/mL of plasma.
It was made to each volunteer, in each point of collection sample.
|
2 months
|
|
Quantification of phenolic acids in urine samples of 10 healthy volunteers by HPLC
Időkeret: 2 months
|
The urine samples were collected in the times describe in "Arms and Interventions".
The samples were acidified and the anthocyanins metabolites were extracted by SPE column, these metabolites extracted were concentrated and analyzed by HPLC, the results were expressed in mmol/mol of creatinine for urine samples.
It was made to each volunteer, in each point of collection sample.
|
2 months
|
|
Pharmacokinetic parameters (AUC) of anthocyanins in urine of 10 healthy volunteers after intake of grumixama juice
Időkeret: 1 month
|
Using the quantification results for each collection time was calculated the area under curve (AUC) of anthocyanins in urine (expressed in μmol/h).
|
1 month
|
|
Pharmacokinetic parameters (AUC) of phenolic acids in urine of 10 healthy volunteers after intake of grumixama juice
Időkeret: 1 month
|
Using the quantification results for each collection time was calculated the area under curve (AUC) of phenolic acids in urine (expressed in mmol/h).
|
1 month
|
|
Pharmacokinetic parameters (AUC) of urolithins in urine of 10 healthy volunteers after intake of grumixama juice
Időkeret: 1 month
|
Using the quantification results for each collection time was calculated the area under curve (AUC) of urolithins in urine (expressed in mmol/h).
|
1 month
|
|
Urine volume
Időkeret: 1 day
|
The volume of each urine sample was measured in mL.
|
1 day
|
|
The total elimination of anthocyanins in urine of 10 healthy volunteers after intake of grumixama juice
Időkeret: 1 month
|
The total elimination amount of anthocyanins was calculated using the urine volume and it was expressed in nmol.
|
1 month
|
|
The total elimination of phenolic acids in urine of 10 healthy volunteers after intake of grumixama juice
Időkeret: 1 month
|
The total elimination amount of phenolic acids was calculated using the urine volume and it was expressed in µmol.
|
1 month
|
|
The total elimination of urolithins in urine of 10 healthy volunteers after intake of grumixama juice
Időkeret: 1 month
|
The total elimination amount of urolithins was calculated using the urine volume and it was expressed in µmol.
|
1 month
|
Egyéb eredményintézkedések
Eredménymérő |
Intézkedés leírása |
Időkeret |
|---|---|---|
|
Normal blood glycemia values of 15 volunteers before juice intake
Időkeret: 1 day
|
The glycemia was measured in 15 volunteers to evaluate if the glycemia values were acceptable.
The values were expressed in mg/dL.
|
1 day
|
|
Normal total cholesterol values of 15 volunteers before juice intake
Időkeret: 1 day
|
The total cholesterol was measured in 15 volunteers to evaluate if the cholesterol values were acceptable.
The values were expressed in mg/dL.
|
1 day
|
|
Normal triglycerides values of 15 volunteers before juice intake
Időkeret: 1 day
|
The triglycerides were measured in 15 volunteers to evaluate if the triglycerides values were acceptable.
The values were expressed in mg/dL.
|
1 day
|
|
Normal HDL values of 15 volunteers before juice intake
Időkeret: 1 day
|
The HDL values were measured in 15 volunteers to evaluate if the HDL and LDL values were acceptable.
The values were expressed in mg/dL.
|
1 day
|
|
Similar weight of 15 volunteers before juice intake
Időkeret: 1 day
|
The weight was measured in 15 volunteers to calculate the dose of juice to be give to the volunteers and also to calculate the BMI.
The results were expressed in Kg of body mass.
|
1 day
|
|
Similar height of 15 volunteers before juice intake
Időkeret: 1 day
|
The height was measured in 15 volunteers to calculate the body index mass (BMI).
The results were expressed in meters (m).
|
1 day
|
|
Similar BMI of 15 volunteers before juice intake
Időkeret: 1 day
|
The height was measured in 15 volunteers to calculate the body index mass (BMI).
The results were expressed in Kg/m2.
This assessment was made to check if the volunteers had a similar BMI.
|
1 day
|
|
Normal LDL values of 15 volunteers before juice intake
Időkeret: 1 day
|
The LDL values were measured in 15 volunteers to evaluate if the HDL and LDL values were acceptable.
The values were expressed in mg/dL.
|
1 day
|
|
Normal serum creatinine of 15 volunteers before juice intake
Időkeret: 1 day
|
The serum creatinine values were measured in 15 volunteers to evaluate if the values were acceptable.
The values were expressed in mg/dL.
|
1 day
|
|
Alanine aminotransferase measures in serum samples of 15 volunteers before juice intake
Időkeret: 1 day
|
The alanine aminotransferase was measured in the serum of 15 volunteers to evaluate if the values were acceptable.
The values were expressed in U/L.
|
1 day
|
|
The aspartate transaminase measures in serum samples of 15 volunteers before juice intake
Időkeret: 1 day
|
The aspartate transaminase was measured in the serum of 15 volunteers to evaluate if the values were acceptable.
The values were expressed in U/L.
|
1 day
|
Együttműködők és nyomozók
Itt találhatja meg a tanulmányban érintett személyeket és szervezeteket.
Szponzor
Együttműködők
Nyomozók
- Kutatásvezető: Luciane L. Teixeira, MS, University of Sao Paulo
- Tanulmányi szék: Neuza M. Aymoto Hassimotto, PhD, University of Sao Paulo
- Tanulmányi szék: Franco M. Lajolo, PhD, University of Sao Paulo
Publikációk és hasznos linkek
A vizsgálattal kapcsolatos információk beviteléért felelős személy önkéntesen bocsátja rendelkezésre ezeket a kiadványokat. Ezek bármiről szólhatnak, ami a tanulmányhoz kapcsolódik.
Általános kiadványok
- Flores G, Dastmalchi K, Paulino S, Whalen K, Dabo AJ, Reynertson KA, Foronjy RF, D'Armiento JM, Kennelly EJ. Anthocyanins from Eugenia brasiliensis edible fruits as potential therapeutics for COPD treatment. Food Chem. 2012 Oct 1;134(3):1256-62. doi: 10.1016/j.foodchem.2012.01.086. Epub 2012 Feb 18.
- Reynertson KA, Yang H, Jiang B, Basile MJ, Kennelly EJ. Quantitative analysis of antiradical phenolic constituents from fourteen edible Myrtaceae fruits. Food Chem. 2008 Aug 15;109(4):883-890. doi: 10.1016/j.foodchem.2008.01.021.
- Teixeira Lde L, Bertoldi FC, Lajolo FM, Hassimotto NM. Identification of Ellagitannins and Flavonoids from Eugenia brasilienses Lam. (Grumixama) by HPLC-ESI-MS/MS. J Agric Food Chem. 2015 Jun 10;63(22):5417-27. doi: 10.1021/acs.jafc.5b01195. Epub 2015 Jun 2.
Tanulmányi rekorddátumok
Ezek a dátumok nyomon követik a ClinicalTrials.gov webhelyre benyújtott vizsgálati rekordok és összefoglaló eredmények benyújtásának folyamatát. A vizsgálati feljegyzéseket és a jelentett eredményeket a Nemzeti Orvostudományi Könyvtár (NLM) felülvizsgálja, hogy megbizonyosodjon arról, hogy megfelelnek-e az adott minőség-ellenőrzési szabványoknak, mielőtt közzéteszik őket a nyilvános weboldalon.
Tanulmány főbb dátumok
Tanulmány kezdete
2014. március 1.
Elsődleges befejezés (Tényleges)
2014. augusztus 1.
A tanulmány befejezése (Tényleges)
2016. március 1.
Tanulmányi regisztráció dátumai
Először benyújtva
2016. április 2.
Először nyújtották be, amely megfelel a minőségbiztosítási kritériumoknak
2016. május 30.
Első közzététel (Becslés)
2016. június 6.
Tanulmányi rekordok frissítései
Utolsó frissítés közzétéve (Becslés)
2016. június 14.
Az utolsó frissítés elküldve, amely megfelel a minőségbiztosítási kritériumoknak
2016. június 13.
Utolsó ellenőrzés
2016. május 1.
Több információ
A tanulmányhoz kapcsolódó kifejezések
Egyéb vizsgálati azonosító számok
- 17449613.8.0000.0067
Terv az egyéni résztvevői adatokhoz (IPD)
Tervezi megosztani az egyéni résztvevői adatokat (IPD)?
NEM
Ezt az információt közvetlenül a clinicaltrials.gov webhelyről szereztük be, változtatás nélkül. Ha bármilyen kérése van vizsgálati adatainak módosítására, eltávolítására vagy frissítésére, kérjük, írjon a következő címre: register@clinicaltrials.gov. Amint a változás bevezetésre kerül a clinicaltrials.gov oldalon, ez a webhelyünkön is automatikusan frissül. .