Effects of Vitamin B12 and Natural Orange Extract, and Their Combination on Physical and Mental Performance (MECHEPS)

Effects of Supplementation With a New Formulation of Vitamin B12, Natural Orange Extract and a Combination of Both on Physical and Mental Performance.

The aim of this study was to determine the effects of a new formulation of vitamin B12, natural orange extract, and their combination on physical and mental performance.

Study Overview

• Status: Enrolling by invitation
• Conditions: Mental Fatigue; Flavonoids; Biomarker Feedback; Supplementation During Endurance Performance; Vitamin b12
• Intervention / Treatment: Dietary supplement: Vitamin B 12; Dietary supplement: Natural orange extract; Combination product: Vitamina B12 + natural orange extract; Dietary supplement: Microcrystalline Cellulose

Detailed Description

A triple-blind, randomized, crossover clinical trial will be conducted with three experimental phases (vitamin B12 + natural orange extract, natural orange extract, vitamin B12) and one control phase (placebo), including evaluations of the acute effect after a loading phase (three intakes, one per day including the test day).

Participants

The sample will consist of 20 well-trained male cyclists (road, MTB, or triathlon), training 3-6 sessions per week, aged between 18 and 45 years. The inclusion criteria will be:

• Healthy male (to be certified through medical examination and health analysis).
• Aged 18-45 years.
• Well-trained (3-6 training sessions per week).
• Habitual consumer of citrus fruits (orange, mandarin, lemon, or grapefruit, either whole or as juice).

Tests

• General health analysis: A general blood test (including standard biochemistry and complete blood count) will be performed during the recruitment phase.
• Medical examination: A cardiorespiratory assessment at rest will be conducted to rule out any underlying pathology.
• Specific performance-related blood analysis: Conducted at four time points (total of eight measurements): baseline during recruitment, pre- and post-test in each condition (supplement / placebo). Parameters will include: complete blood count, ferritin, CK, LDH, vitamin B12, transaminases, reticulocytes, homocysteine, C-reactive protein, IL-6, methylmalonic acid, holo-transcobalamin II, 8-OHdG (8-hydroxy-2'-deoxyguanosine), dopamine, serotonin, and Brain-Derived Neurotrophic Factor (BDNF).
• Anthropometry: Determination of body composition parameters to characterize the sample.
• Wingate test: Conducted using the standard load protocol, consisting of five maximal 30-second sprints with 3-minute recovery intervals between repetitions.
• Mental fatigue test using light-based reaction tasks: The time required to complete a light reaction sequence will be measured before and after the fatigue protocol.
• Genetic test: A saliva sample will be collected to analyze genetic profiles related to food intolerances and vitamin absorption. The objective is to identify potential explanations for interindividual differences in absorption capacity.

• Study Type: Interventional
• Enrollment (Estimated): 20
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Spain
  • Murcia
    • La Ñora, Murcia, Spain, 30107
      • Research Center for High Performance Sport. Universidad Católica de Murcia

Participation Criteria

Eligibility Criteria

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

Description

Inclusion Criteria:

• Healthy male (to be certified through medical examination and health analysis).
• Aged 18-45 years.
• Well-trained (3-6 training sessions per week).
• Habitual consumer of citrus fruits (orange, mandarin, lemon, or grapefruit, either fresh or juiced).

Exclusion Criteria:

• Smokers or regular alcohol drinkers.
• Persons suffering from a metabolic, cardiorespiratory or digestive pathology or anomaly.

digestive system.

• People who have suffered an injury in the last 6 months that limits the performance of the study tests. of the study tests.
• People taking supplements or medication in the 2 weeks before the start of the study.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Crossover Assignment
• Masking: Triple

Number of Arms

4

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Vitamin B12; This group will take 1.0 mg of vitamin B12 two days before the tests and 3 hours before each test. Therefore, the supplementation will last 3 days in total.	Dietary supplement: Vitamin B 12; Subjects in this group will intake 1 mg/day of vitamin B12.; Other Names: Vitamin B12
Experimental: Natural orange extract; This group will take 500 mg of natural orange extract two days before the tests and 3 hours before each test. Therefore, the supplementation will last 3 days in total.	Dietary supplement: Natural orange extract; Subjects in this groups will intake 500 mg/day of natural orange extract.
Experimental: Vitamin B12 + natural orange extract; This group will take 1.0 mg of vitamin B12 + 500 mg of natural orange extract two days before the tests and 3 hours before each test. Therefore, the supplementation will last 3 days in total.	Combination product: Vitamina B12 + natural orange extract; This group will take the combination of vitamin B12 and Vitamina B12 (1 mg/day) + natural orange extract (500 mg/day).
Placebo Comparator: Placebo; This group will take 500 mg of microcrystalline cellulose two days before the tests and 3 hours before each test. Therefore, the supplementation will last 3 days in total.	Dietary supplement: Microcrystalline Cellulose; Microcrystalline cellulose was used, this molecule was selected to avoid affecting the glycemia, as it can occur with dextrose or maltodextrin.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Wingate test	This test consists of 5 efforts of 30 seconds at maximum intensity with a recovery of 3 minutes on a bike. In this test the peak and average power of each effort and the total average will be evaluated. In addition, the time to maximum peak power and the fatigue index.	Throughout study completion, an average of 4 weeks

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Mental fatigue test	In this test, we will measure the speed of reaction in the resolution of a sequence of lights set by the researcher before and after the end of the performance test.	Throughout study completion, an average of 4 weeks
Hematological Parameters	A complete blood count (including red blood cell count, hemoglobin, hematocrit, and white blood cell count) will be measured from venous blood samples before and after performance tests.	Throughout study completion, an average of 4 weeks
Iron Status Marker	Serum ferritin concentration measured as an indicator of iron status, will be measured from venous blood samples before and after performance tests.	Throughout study completion, an average of 4 weeks
Vitamin B12 Status - Serum Vitamin B12	Serum vitamin B12 concentration measured from venous blood samples, will be measured from venous blood samples before and after performance tests.	Throughout study completion, an average of 4 weeks
Functional Vitamin B12 Biomarkers	Description: Plasma methylmalonic acid, holo-transcobalamin II, and homocysteine concentrations assessed as functional biomarkers of vitamin B12 status, will be measured from venous blood samples before and after performance tests.	Throughout study completion, an average of 4 weeks
Muscle Damage Markers	Description: Serum creatine kinase (CK) and lactate dehydrogenase (LDH) concentrations measured as indicators of exercise-induced muscle damage, will be measured from venous blood samples before and after performance tests.	Throughout study completion, an average of 4 weeks
Hepatic Enzymes	Description: Serum glutamic-oxaloacetic transaminase (GOT/AST) and glutamic-pyruvic transaminase (GPT/ALT) concentrations, will be measured from venous blood samples before and after performance tests.	Throughout study completion, an average of 4 weeks
Inflammatory Marker	Description: Serum C-reactive protein (CRP) concentration measured as a marker of systemic inflammation, will be measured from venous blood samples before and after performance tests.	Throughout study completion, an average of 4 weeks
Oxidative Stress Marker	Description: Plasma 8-hydroxy-2-deoxyguanosine (8-OHdG) concentration measured as a marker of oxidative DNA damage, will be measured from venous blood samples before and after performance tests.	Throughout study completion, an average of 4 weeks
Neurotransmitters	Description: Plasma serotonin, dopamine, norepinephrine, and epinephrine concentrations measured as indicators of neuroendocrine response, will be measured from venous blood samples before and after performance tests.	Throughout study completion, an average of 4 weeks
Neurotrophic Factor	Description: Serum Brain-Derived Neurotrophic Factor (BDNF) concentration, will be measured from venous blood samples before and after performance tests.	Throughout study completion, an average of 4 weeks

Collaborators and Investigators

• Sponsor: Francisco Javier Martínez Noguera
• Investigators: Principal Investigator: Francisco Javier Martinez Noguera, PhD, Universidad Católica San Antonio de Murcia

Study record dates

Study Major Dates

• Study Start (Actual): December 1, 2025
• Primary Completion (Estimated): April 1, 2026
• Study Completion (Estimated): June 1, 2026

Study Registration Dates

• First Submitted: December 18, 2025
• First Submitted That Met QC Criteria: January 7, 2026
• First Posted (Actual): January 14, 2026

Study Record Updates

• Last Update Posted (Actual): January 14, 2026
• Last Update Submitted That Met QC Criteria: January 7, 2026
• Last Verified: December 1, 2025

More Information

Terms related to this study

• Keywords: Vitamin B12; Performance; Polyphenols; Ergogenic aids; Wingate test; Natural orange extract
• Additional Relevant MeSH Terms: Behavioral Symptoms; Fatigue; Pathological Conditions, Signs and Symptoms; Behavior; Signs and Symptoms; Mental Fatigue; Heterocyclic Compounds, 1-Ring; Heterocyclic Compounds; Heterocyclic Compounds, Fused-Ring; Azoles; Polycyclic Compounds; Heterocyclic Compounds, 4 or More Rings; Pyrroles; Macrocyclic Compounds; Tetrapyrroles; Corrinoids; Vitamin B 12; microcrystalline cellulose
• Other Study ID Numbers: CE092504

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