Identification of New Biomarkers of Banana and Tomato Intake

Biomarkers Clinical Trial

— BioBanaTom  

Official title:

Identification of New Biomarkers of Banana and Tomato Intake

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT03581955
• Other study ID #: 2016-A00153-48
• Status: Completed
• Phase: N/A
• Start date: March 26, 2016
• Est. completion date: June 16, 2016

Study information

• Verified date: July 2018
• Source: Institut National de la Recherche Agronomique
• Contact: n/a
• Is FDA regulated: No
• Study type: Interventional

Clinical Trial Summary

The intake of fruits and vegetables has been associated to a lower risk of developing metabolic diseases and cancer. The intake of tomato has been proposed to decrease the risk of prostate cancer while the high content of pro-vitamine A carotenes in banana have shown to alleviate Vitamin A deficiency in different countries. Interestingly in spite of their popularity, there are no biomarkers of banana intake reported in the literature while lycopene is the most frequently used metabolite to indicate tomato consumption however, its limited specificity and between-subjects variation sets doubt of its accuracy. Therefore, the identification of novel biomarkers for both banana and tomato is of great value. Untargeted metabolomics, allows a holistic analysis of the food metabolome allowing a deeper inquiry in the metabolism of different compounds and the recognition of patterns and individual differences that may lead to new hypothesis and further research. Therefore, the aim of the present study is to identify biomarkers of acute intake of banana and tomato using an untargeted approach on urine serum of 12 volunteers that participated in a crossover, randomized, controlled study. Volunteers consumed three different test foods: 1) 240g of banana, 2) 300g of tomato and 3) Fresubin 2kcal as control. Serum and urine samples were collected in kinetics over 24h and processed to be analyzed using LC-QTof analysis. The metabolomics profiles are compared using univariate (ANOVA) and multivariate statistical methods (PCA, PLSDA). The identification of discriminant compounds was performed by tandem mass fragmentation with a high-resolution LTQ-Orbitrab Mass spectrometer and by an extensive inquiry of different online databases.

Description:

The rise of metabolomics along with different platforms such as liquid chromatography mass spectrometers (LC-MS) have allowed the assessment of thousands of metabolites simultaneously in biological samples and the recognition of patterns that may constitute a fingerprint of the intake of different foods. Recent studies demonstrated the great potential of metabolomics to discover new biomarkers of intake in intervention and cohort studies.The diversity of compounds found in food metabolomics represents a major challenge and so in an international effort to improve dietary biomarkers identification and validation, the Food Biomarkers Alliance (FOODBALL) has been created. In this project, 22 institutions from 11 different countries will collaborate in three main tasks: 1) Discovery of new dietary biomarker using a metabolomic approach, 2) systemic validation of existing and newly discovered biomarker to achieve a good coverage of food intake in different European populations and 3) exploring biological effects using biomarkers of intake (http://foodmetabolome.org/). With the latter, the necessity of building a chemical library that allows the use of standards for further identification arises. Along with FOODBALL, The Food Compound Exchange (FoodComEx) aims to improve the availability of analytical standards of biological compounds to achieve a better and easier biomarker identification (http://foodcomex.org/).

As part of INRA collaboration to FOODBALL and FoodComEx, the present project attempts to identify biomarkers of banana and tomato intake, through the exploration of the serum and urine metabolome of 12 subjects who consumed these foods following a randomized, controlled, crossover design. The present study was comprised of 3 different intervention periods and a minimum of 3 days washout between interventions. The intervention periods were comprised of 2 run in days, 1 intervention day and 1 post intervention day. In the first day of the run in period, subjects were instructed to avoid the intake of banana or tomato or any of their products; the day prior to the intervention volunteers were asked to avoid the intake of phytochemical rich foods and beverages such as wine, coffee, chocolate, tea, and other plant based foods including banana and tomato.In the morning of the intervention, day subjects arrived in fasting state to the research center at 7.30 am. Volunteers were randomly assigned to one of the three interventions, Fresubin ® 2kcal fiber, 240g of banana plus control drink, or 300g of tomato plus control drink plus 12g of refined sunflower oil. Throughout the intervention, subjects had free access to water, maximum 250ml of water per hour until 6 hours after the intake of the test food.

A trained phlebotomist placed a catheter on the subject's arm before the intake of the test foods to collect the baseline sample. Then four other samples were collected postprandially after 1h, 2h, 4h, and 6h. A total of 7 urine samples were collected. The first void of urine was collected by the subjects at home upon the morning of Day 3 and the rest of the samples after the intake of the test foods as follows: 0-1h, 1h-2h, 2h-4h, 4h-6h. The urine samples corresponding to 6h-12h and 12-24h interval were collected by volunteers at home until the morning after the intake of the food.

After the 6h collection of blood, the peripheral catheter was removed and subjects had lunch composed of white bread and cooked pasta, then subjects were allowed to go home. Before leaving the Investigation center, participants were instructed to prepare a standardized dinner based on pan fried chicken with butter and boiled rice with salt. Volunteers were not allowed to eat or drink anything except water and the standardized dinner.

On the morning of the post intervention day, subjects arrived in fasting state to the research center to give the 24h blood sample and deliver the 06-12h and 12-24h urine collection. Afterward, subjects were served breakfast at the research center and resumed their normal diet until the next run in days of the next intervention period.

Urine samples and serum samples were aliquoted and stored at -80° C until analysis.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 12
• Est. completion date: June 16, 2016
• Est. primary completion date: May 19, 2016
• Accepts healthy volunteers: Accepts Healthy Volunteers
• Gender: All
• Age group: 18 Years to 40 Years

Eligibility

Inclusion Criteria:

• Healthy males and females

• Aged 18- 40 years

• BMI >18.5 and < 30 kg/m2

• Willing/able to consume all test foods (tomato, banana, Fresubin drink) and the standardized meals (rice and chicken)

Exclusion Criteria:

• Smokers

• Diagnosed health condition (chronic or infectious disease)

• Taking nutritional supplements (e.g. vitamins, minerals) several times a week.

• Taking medication (oral contraceptive pill is allowed).

• Pregnant, lactating.

• Antibiotics treatment within 3 months prior to intervention.

• Vegetarians, as standardized meals will contain meat.

• Not willing to follow nutritional restrictions, including drinking alcohol during study days

• Not willing/able to give informed consent or to sign informed consent.

• Not affiliated to National Health Insurance.

• Being in exclusion on the National Volunteers Data file or refusing to be registered on the National Volunteers Data file.

• Currently participating or who having got 4500€ in this year to have participated in another clinical trial.

Related Conditions & MeSH terms

• Biomarkers
• Food Intake

Intervention

Other:
• Banana Cavendish
240g of fruit plus 150ml of control drink (Fresubin ® 2kcal fiber neutral flavor)
• Tomato
300g of raw tomato ("coeur de boeuf") with refined sunflower oil (12g) and 150ml of control drink (Fresubin ® 2kcal fiber neutral flavor).
• Fresubin ® 2kcal fiber neutral flavor
250 ml Fresubin ® 2kcal fiber neutral flavor

Locations

Country	Name	City	State
France	INRA	Clermont-Ferrand	Rhône-Alpes-Auvergne

Sponsors (2)

Lead Sponsor	Collaborator
Claudine MANACH	National Research Agency, France

Country where clinical trial is conducted

France, 

References & Publications (10)

Andersen MB, Kristensen M, Manach C, Pujos-Guillot E, Poulsen SK, Larsen TM, Astrup A, Dragsted L. Discovery and validation of urinary exposure markers for different plant foods by untargeted metabolomics. Anal Bioanal Chem. 2014 Mar;406(7):1829-44. doi: 10.1007/s00216-013-7498-5. Epub 2014 Jan 4. — View Citation

Giovannucci E. Tomatoes, tomato-based products, lycopene, and cancer: review of the epidemiologic literature. J Natl Cancer Inst. 1999 Feb 17;91(4):317-31. Review. — View Citation

Kesse-Guyot E, Castetbon K, Touvier M, Hercberg S, Galan P. Relative validity and reproducibility of a food frequency questionnaire designed for French adults. Ann Nutr Metab. 2010;57(3-4):153-62. doi: 10.1159/000321680. Epub 2010 Nov 16. — View Citation

Manach C, Hubert J, Llorach R, Scalbert A. The complex links between dietary phytochemicals and human health deciphered by metabolomics. Mol Nutr Food Res. 2009 Oct;53(10):1303-15. doi: 10.1002/mnfr.200800516. Review. — View Citation

Manach C., Brennan L, Drasgted L.O. Metabolomics to evaluate food intake and utilization in nutritional epidemiology. In: Metabolomics as a Tool in Nutritional Research, Woodhead publishing 2015. pp.167-196

Peralta I, Spooner DM. Genetic Improvement of Solanaceous Crops Volume 2: Tomato. CRC Press; 2006. https://books.google.com/books?hl=en&lr=&id=1m7RBQAAQBAJ&pgis=1. Accessed December 18, 2015

Pereira A, Maraschin M. Banana (Musa spp) from peel to pulp: ethnopharmacology, source of bioactive compounds and its relevance for human health. J Ethnopharmacol. 2015 Feb 3;160:149-63. doi: 10.1016/j.jep.2014.11.008. Epub 2014 Nov 13. Review. — View Citation

Pujos-Guillot E, Hubert J, Martin JF, Lyan B, Quintana M, Claude S, Chabanas B, Rothwell JA, Bennetau-Pelissero C, Scalbert A, Comte B, Hercberg S, Morand C, Galan P, Manach C. Mass spectrometry-based metabolomics for the discovery of biomarkers of fruit and vegetable intake: citrus fruit as a case study. J Proteome Res. 2013 Apr 5;12(4):1645-59. doi: 10.1021/pr300997c. Epub 2013 Mar 5. — View Citation

Re R, Bramley PM, Rice-Evans C. Effects of food processing on flavonoids and lycopene status in a Mediterranean tomato variety. Free Radic Res. 2002 Jul;36(7):803-10. — View Citation

Scalbert A, Brennan L, Manach C, Andres-Lacueva C, Dragsted LO, Draper J, Rappaport SM, van der Hooft JJ, Wishart DS. The food metabolome: a window over dietary exposure. Am J Clin Nutr. 2014 Jun;99(6):1286-308. doi: 10.3945/ajcn.113.076133. Epub 2014 Apr 23. Review. — View Citation

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Changes in metabolite profiles present in blood serum and urine before the dietary intervention (t=0) and in kinetics over 24 hours.	Metabolite profiles analyzed using a non-targeted metabolomics approach with a UPLC-MS platform. Blood serum samples collected at time 0, 1 hour, 2 hours, 4 hours, 6 hours and 24 hours. Urine fractions collected at 0-1 hours, 1-2 hours, 2-4 hours, 4-6 hours, 6-12 hours, 12-24 hours. Identification of biomarkers of acute intake of the foods of interest through the comparison of metabolomes after either single dose of tomato, banana or control drink.	0-24 hours
Secondary	Collection of pools of urine and serum samples after acute intake of tomato or banana to be used as analytical standards or for the identification of specific metabolites of banana or tomato components.	The collection of pools of biofluids that result from this study may be shared with the scientific community through the FoodComEx library in order to be used as an analytical standard or for the identification of metabolites that are specific of banana or tomato intake.	0-24 hours