Effect of Betaine and Choline on Metabolic Health

ABC (Additional Betaine and Choline) Study: Effect of Betaine and Choline Intake on Biomarkers of Metabolic Health in Humans

The purpose of this research is to determine whether extra betaine and choline influence metabolic health in adults with overweight and obesity.

Study Overview

• Status: Completed
• Conditions: Obesity and Obesity-related Medical Conditions
• Intervention / Treatment: Dietary supplement: Cellulose; Dietary supplement: Trimethylglycine; Other: Eggs

Detailed Description

Betaine (N,N,N-trimethylglycine or also glycine betaine) is a derivative of choline that functions as an organic osmolyte and participates in one-carbon metabolism as a methyl donor. Betaine is naturally found in beets, wheat and spinach and sold as a food supplement without prescription. Choline is recognized as an essential nutrient that is found in various foods including eggs, nuts and beef, with the major form of choline in food found as phosphatidylcholine. In addition to being oxidized to the methyl donor betaine, choline is a precursor of several compounds involved in neurotransmitter synthesis, lipid metabolism and transport as well as the structural integrity and signaling of cell membranes. Previous studies have reported alterations in one-carbon metabolites in response to a single meal containing different forms of choline, with interindividual variability dependent on genetics and gut microbiota composition. This study will extend to longer-term impact of different forms of choline (betaine as oxidized choline and choline provided from food) with a focus on overweight and obesity, which comprise a predominant portion of the population in North America. The objective of this study is to determine the effect of betaine supplementation with or without food-form choline (eggs) on metabolic health in adults with overweight and obesity. A randomized crossover study design will be employed, which men and women of age 18-70 years with BMI 25-35 kg/m2 will participate in a 14-week study consisting of three 4-week dietary periods: 1) daily consumption of 3 grams of betaine supplement with no eggs; 2) daily consumption of 3 grams of betaine supplement with 3 whole eggs; and 3) daily consumption of 3 grams of cellulose supplement with no eggs, in a random order, each dietary period separated by a 1-week washout break. Blood, urine and fecal samples as well as anthropometric measurements will be collected at baseline, then at weeks 4, 9 and 14. The collected biological samples will be used to measure glucose and lipid markers, one-carbon metabolites and profiling of gut microbiota and genotype to determine interindividual differences in metabolism.

• Study Type: Interventional
• Enrollment (Actual): 34
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Canada
  • Ontario
    • Guelph, Ontario, Canada, N1G 2W1
      • Human Nutraceutical Research Unit

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: Yes

Description

Inclusion Criteria:

• Male or female participant of any race or ethnicity between 18-70 years of age (inclusive) at the time of informed consent
• BMI between 25-35 kg/m2
• Non-smoker
• Willing to consume 3 eggs per day for one dietary period of 4 weeks
• Willing to avoid eggs during the rest of the study including washout period breaks except for eggs that are provided
• Willing to fast before the baseline and dietary period lab visits
• Willing to provide a 24-hr prior day food record at the dietary period lab visits
• Willing to provide or collect biological specimen samples at the baseline and dietary period lab visits
• Willing to follow the study protocol including maintaining usual lifestyle during the entire study

Exclusion Criteria:

• Male or female < 18 years or > 70 years of age at the time of consent
• BMI < 25 kg/m2; or BMI > 35 kg/m2
• Pregnant or planning to become pregnant during the course of the study; currently breastfeeding or postpartum < 6 months
• Use of hormone therapy including birth control pills
• Follows a vegan diet
• Current smoker (any form of nicotine, e-cigarettes, etc)
• Use of recreational drugs (may affect metabolic pathway for choline)
• Presence of chronic illnesses, e.g., heart disease, diabetes, cancer, celiac disease, inflammatory bowel disease (Crohn's disease and ulcerative colitis), gastrointestinal liver or kidney diseases or alcoholism
• Diagnosis of trimethylaminuria (genetic disorder affecting choline use in body)
• Use of antibiotics in the last 2 months
• Use of prebiotics, probiotics or dietary fiber (i.e., Metamucil) supplements in the last 2 months
• Have allergies to eggs or any anaphylactic food allergies
• Have a schedule or lifestyle patterns incompatible with the study protocol

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Randomized
• Interventional Model: Crossover Assignment
• Masking: Quadruple

Number of Arms

3

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Daily 3 whole eggs with daily supplement containing 3 g trimethylglycine	Dietary supplement: Trimethylglycine; Betaine anhydrous from sugar beets; Other: Eggs; 3 whole eggs
Experimental: Experimental: No eggs with daily supplement containing 3 g trimethylglycine	Dietary supplement: Trimethylglycine; Betaine anhydrous from sugar beets
Placebo Comparator: Placebo Comparator: No eggs with daily supplement containing 3 g cellulose	Dietary supplement: Cellulose; Cellulose

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Lipid panel	Triglyceride, cholesterol and lipoprotein concentrations	Weeks 0, 4, 9 and 14
Glucose concentrations	Concentrations of glucose as mol units per volume	Weeks 0, 4, 9 and 14
Insulin concentrations	Concentrations of insulin as international units per volume	Weeks 0, 4, 9 and 14
Concentrations of glucose metabolism panel	Concentrations of C-peptide, GLP-1 and GIP as mol units per volume	Weeks 0, 4, 9 and 14
Body mass	Mass on a scale	Weeks 0, 4, 9 and 14
One-carbon metabolism panel	Choline metabolite concentrations	Weeks 0, 4, 9 and 14

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Gene expression	Global gene expression in isolated immune cells	Weeks 0, 4, 9 and 14
Inflammatory response in immune cells	Expression of cytokines in immune cells as assessed in response to lipopolysaccharide challenge	Weeks 0, 4, 9 and 14
Composition of the gut microbiota	Gut microbiota profiles as assessed by sequencing technologies	Weeks 0, 4, 9 and 14
Characterization of single nucleotide polymoprhisms	Allelic discrimination of variants in enzymes that influence choline metabolites	Week 0
Liver health panel	ALT, AST, GGT, ALP and bilirubin concentrations	Weeks 0, 4, 9 and 14
Blood cell counts	Complete counts of white blood cell, red blood cell, hemoglobin, hematocrit and platelet	Weeks 0, 4, 9 and 14
Blood pressure	Systolic and diastolic blood pressure	Weeks 0, 4, 9 and 14
Dietary intake	24-hour prior day food record	Weeks 0, 4, 9 and 14

Collaborators and Investigators

• Sponsor: University of Guelph

Publications and helpful links

General Publications

• Cho CE, Taesuwan S, Malysheva OV, Bender E, Tulchinsky NF, Yan J, Sutter JL, Caudill MA. Trimethylamine-N-oxide (TMAO) response to animal source foods varies among healthy young men and is influenced by their gut microbiota composition: A randomized controlled trial. Mol Nutr Food Res. 2017 Jan;61(1). doi: 10.1002/mnfr.201600324. Epub 2016 Aug 3.
• Cho CE, Aardema NDJ, Bunnell ML, Larson DP, Aguilar SS, Bergeson JR, Malysheva OV, Caudill MA, Lefevre M. Effect of Choline Forms and Gut Microbiota Composition on Trimethylamine-N-Oxide Response in Healthy Men. Nutrients. 2020 Jul 25;12(8):2220. doi: 10.3390/nu12082220.
• Zeisel SH, da Costa KA. Choline: an essential nutrient for public health. Nutr Rev. 2009 Nov;67(11):615-23. doi: 10.1111/j.1753-4887.2009.00246.x.
• Wallace TC, Blusztajn JK, Caudill MA, Klatt KC, Natker E, Zeisel SH, Zelman KM. Choline: The Underconsumed and Underappreciated Essential Nutrient. Nutr Today. 2018 Nov-Dec;53(6):240-253. doi: 10.1097/NT.0000000000000302. Epub 2018 Nov 13.
• Lever M, Slow S. The clinical significance of betaine, an osmolyte with a key role in methyl group metabolism. Clin Biochem. 2010 Jun;43(9):732-44. doi: 10.1016/j.clinbiochem.2010.03.009. Epub 2010 Mar 25.
• Craig SA. Betaine in human nutrition. Am J Clin Nutr. 2004 Sep;80(3):539-49. doi: 10.1093/ajcn/80.3.539.

Study record dates

Study Major Dates

• Study Start (Actual): January 31, 2025
• Primary Completion (Actual): August 19, 2025
• Study Completion (Actual): August 19, 2025

Study Registration Dates

• First Submitted: December 18, 2024
• First Submitted That Met QC Criteria: January 2, 2025
• First Posted (Actual): January 6, 2025

Study Record Updates

• Last Update Posted (Actual): May 12, 2026
• Last Update Submitted That Met QC Criteria: May 9, 2026
• Last Verified: May 1, 2026

More Information

Terms related to this study

• Keywords: Obesity; Overweight; Metabolism; Choline; Betaine
• Additional Relevant MeSH Terms: Nutrition Disorders; Overnutrition; Body Weight; Pathological Conditions, Signs and Symptoms; Nutritional and Metabolic Diseases; Signs and Symptoms; Overweight; Obesity; Organic Chemicals; Food; Diet, Food, and Nutrition; Physiological Phenomena; Food and Beverages; Carbohydrates; Amines; Polymers; Macromolecular Substances; Biomedical and Dental Materials; Manufactured Materials; Technology, Industry, and Agriculture; Polysaccharides; Quaternary Ammonium Compounds; Onium Compounds; Glucans; Trimethyl Ammonium Compounds; Biopolymers; Betaine; Cellulose; Eggs
• Other Study ID Numbers: 24-03-006

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No