Nutritional Requirements
Conditions
Keywords
Vitamin E, Egg, Spinach
Brief summary
Malnutrition of the fat-soluble nutrient vitamin E (α-tocopherol; αT) is problematic. Since αT is rich in plant foods (e.g. spinach) that are mostly absent of accessible lipid, dietary patterns that can potentiate αT bioavailability by pairing vegetables with lipid-rich foods have been emphasized. The purpose of this study is to use deuterium-labeled spinach (containing stable isotopes of αT) to validate eggs as a dietary tool to improve αT bioavailability directly from a model plant food, and hence achieve nutrient adequacy. It is expected that compared with deuterium-labeled spinach alone, co-ingestion of eggs will dose- and time-dependently increase plasma bioavailability of spinach-derived deuterium-labeled αT without affecting time to maximal concentrations or half-lives. Further, phospholipid-rich egg yolk lipid will enhance nutrient bioavailability compared with vegetable oil. The outcome will therefore support an egg-based food pairing that can enhance the health benefits of plant-centric dietary patterns.
Detailed description
In the US, 92-96% of men and women do not meet recommended intakes for αT. Dietary recommendations strongly encourage a diet rich in fruits and vegetables to meet dietary αT requirements. However, αT bioavailability from most plant foods is quite poor, thereby emphasizing a need for effective food pairings that can enhance the absorption and promote adequate status of these health-promoting nutrients. The objective of this application is to use deuterium-labeled spinach (containing stable isotopes of αT) to validate eggs as a dietary tool to improve αT bioavailability directly from a model plant food, and hence achieve nutrient adequacy. Our hypothesis is that the bioavailability of αT from deuterium-labeled spinach will be potentiated by egg intake in a dose-dependent manner by increasing their secretion in intestinal-derived chylomicrons. Furthermore, phospholipid-rich whole eggs will enhance spinach-derived αT bioavailability compared with vegetable oil, and will be most functionally responsible for the benefits of eggs to enhance nutrient absorption. Additionally, egg whites will more greatly promote nutrient bioaccessibility compared with spinach alone. To test this, our specific aim is to assess egg-mediated improvements in αT bioavailability by conducting a cross-over pharmacokinetic study in healthy men and women. In Study Arms 1-4, participants will ingest deuterium-labeled spinach (containing 5 mg αT) with 0, 1, 2, or 3 hardboiled eggs (containing 0, 4.8, 9.6, or 14.4 g total fat, respectively). In Study Arm 5, participants will ingest spinach with two egg whites. In Study Arm 6, participants will ingest spinach with 9.6 grams of vegetable oil. In Study Arm 5, participants will ingest spinach alone followed by 1 egg 3-hours later. In Study Arm 7, participants will ingest spinach with 1 egg followed by another egg 3-hours later. In Study Arm 8, participants will ingest spinach with 1 egg followed by another egg 3-hours later. Thus, Study Arms 1-6 will test the dose-dependent and food matrix effects of eggs on αT bioavailability, and study Arms 7-8 will test the meal timing effects of eggs on αT bioavailability. Eucaloric diets will be controlled for αT intakes for 3 d prior to and during the initial 24 h of each trial to minimize heterogeneity of pharmacokinetic responses. Spinach-derived deuterium-labeled αT will be measured in plasma and isolated chylomicrons collected at timed intervals from 0-72 h post-meal ingestion, and biomarkers of antioxidant status and oxidative distress will be assessed at baseline (0 h) of each trial. Outcomes from this study are expected to demonstrate a dose- and time-dependent function of eggs to increase deuterium-labeled αT bioavailability (based on area under the curve (AUC) 0-72 h, Cmax, and % estimated absorption). The rationale for this study is that, by establishing the efficacy of eggs to potentiate plant-derived fat-soluble nutrient bioavailability, a strong framework will exist for an easily implementable health-promoting food pairing strategy to overcome malnutrition of αT.
Interventions
No eggs will be consumed on test day along with spinach consumption
One egg will be consumed on test day along with spinach consumption
Two eggs will be consumed on test day along with spinach consumption
Three eggs will be consumed on test day along with spinach consumption
One egg will be consumed on test day three hours after spinach consumption
Two eggs will be consumed on test day: one along with spinach consumption and the other one three hours after spinach consumption
Two egg whites will be consumed on test day along with spinach consumption
OTHERVegetable oil at 0 h
9.6 grams of Vegetable oil will be consumed on test day along with spinach consumption
Sponsors
Ohio State University
Study design
Allocation
NON_RANDOMIZED
Intervention model
CROSSOVER
Primary purpose
PREVENTION
Masking
NONE
Eligibility
Sex/Gender
ALL
Age
18 Years to 65 Years
Healthy volunteers
Yes
Inclusion criteria
* Body Mass Index (BMI) = 19-25 kg/m2 * Normolipidemic (total cholesterol \<240 mg/dL; triglyceride \<150 mg/dL) * Fasting glucose \<100 mg/dL * Normal hematocrit level (41%-50% for men and 36%-48% for women) * Normal hemoglobin level (13.5-17.5 g/dL for men and 12.0-15.5 g/dL for women) * No use of dietary supplements for \>1 month * No use of medications that affect lipid or glucose metabolism * Non-smoker * No history of gastrointestinal disorders
Exclusion criteria
* Egg allergy * Alcohol intake \> 2 drinks per day * Aerobic activity \>7 h/wk * Body mass change \>2 kg in the past 1 month * Women who are pregnant, lactating, or initiated or changed birth control in the past 3 month * Vegetarian
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Vitamin E Area Under the Curve (0-72 h) | 0, 3, 4.5, 6, 7.5, 9, 12, 24, 36, 48, 72 hours post-ingestion of spinach | Area under the concentration-time curve of plasma deuterium-labeled alpha-tocopherol over 72 hours post ingestion. |
| Vitamin E Cmax | 0-72 hours post-ingestion of spinach | Maximum plasma concentration of deuterium-labeled alpha-tocopherol |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Vitamin E Tmax (hr) | 0-72 hours post-ingestion of spinach | Time to reach maximum plasma concentration of deuterium-labeled alpha-tocopherol |
| Half-Life (hr) | 0-72 hours post-ingestion of spinach | The time required for the plasma concentration of vitamin E to decrease by 50%. |
| Estimated Absorption (%Dose) of Vitamin E | 0-72 hours post-ingestion of spinach | Absorption of deuterium-labeled alpha-tocopherol |
| Chylomicron Vitamin E | 0, 3, 4.5, 6, 7.5, 9, 12 hours post-ingestion of spinach | Deuterium-labeled alpha-tocopherol concentration in chylomicron |
Countries
United States
Participant flow
Participants by arm
| Arm | Count |
|---|---|
| Baseline Characteristics Characteristics of participants upon enrollment into the study | 8 |
| Total | 8 |
Baseline characteristics
| Characteristic | Baseline Characteristics | — |
|---|---|---|
| Age, Categorical <=18 years | 0 Participants | — |
| Age, Categorical >=65 years | 0 Participants | — |
| Age, Categorical Between 18 and 65 years | 8 Participants | — |
| Body Mass Index | 21.8 kg/m^2 STANDARD_DEVIATION 2 | — |
| Diastolic blood pressure | 70.8 mmHg STANDARD_DEVIATION 5.4 | — |
| Glucose | 4.81 mmol/L STANDARD_DEVIATION 0.39 | — |
| Race and Ethnicity Not Collected | — | — Participants |
| Region of Enrollment United States | 8 participants | — |
| Sex: Female, Male Female | 5 Participants | — |
| Sex: Female, Male Male | 3 Participants | — |
| Systolic blood pressure | 109.6 mmHg STANDARD_DEVIATION 8.3 | — |
| Total Cholesterol | 4.19 mmol/L STANDARD_DEVIATION 0.47 | — |
| Total Lipid | 4.75 mmol/L STANDARD_DEVIATION 0.57 | — |
| Triglyceride | 0.57 mmol/L STANDARD_DEVIATION 0.15 | — |
| Waist circumference Female | 74.1 cm STANDARD_DEVIATION 4.6 | — |
| Waist circumference Male | 76.3 cm STANDARD_DEVIATION 4.2 | — |
Adverse events
| Event type | EG000 affected / at risk | EG001 affected / at risk |
|---|---|---|
| deaths Total, all-cause mortality | 0 / 8 | 0 / 5 |
| other Total, other adverse events | 0 / 8 | 0 / 5 |
| serious Total, serious adverse events | 0 / 8 | 0 / 5 |
Outcome results
Primary
Vitamin E Area Under the Curve (0-72 h)
Area under the concentration-time curve of plasma deuterium-labeled alpha-tocopherol over 72 hours post ingestion.
Time frame: 0, 3, 4.5, 6, 7.5, 9, 12, 24, 36, 48, 72 hours post-ingestion of spinach
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| SPINACH | Vitamin E Area Under the Curve (0-72 h) | 2.90 umol/L*h | Standard Error 0.88 |
| SPINACH + 1 EGG0 h | Vitamin E Area Under the Curve (0-72 h) | 5.24 umol/L*h | Standard Error 1 |
| SPINACH + 2 EGGS0h | Vitamin E Area Under the Curve (0-72 h) | 5.18 umol/L*h | Standard Error 1.06 |
| SPINACH + 3 EGGS0 h | Vitamin E Area Under the Curve (0-72 h) | 4.39 umol/L*h | Standard Error 0.69 |
| SPINACH + WHITE | Vitamin E Area Under the Curve (0-72 h) | 5.10 umol/L*h | Standard Error 0.65 |
| SPINACH + OIL | Vitamin E Area Under the Curve (0-72 h) | 6.88 umol/L*h | Standard Error 0.74 |
| SPINACH + EGG3 h | Vitamin E Area Under the Curve (0-72 h) | 1.67 umol/L*h | Standard Error 0.45 |
| SPINACH + EGGS0 & 3 h | Vitamin E Area Under the Curve (0-72 h) | 4.88 umol/L*h | Standard Error 1.53 |
Primary
Vitamin E Cmax
Maximum plasma concentration of deuterium-labeled alpha-tocopherol
Time frame: 0-72 hours post-ingestion of spinach
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| SPINACH | Vitamin E Cmax | 0.09 umol/L | Standard Error 0.02 |
| SPINACH + 1 EGG0 h | Vitamin E Cmax | 0.18 umol/L | Standard Error 0.03 |
| SPINACH + 2 EGGS0h | Vitamin E Cmax | 0.15 umol/L | Standard Error 0.03 |
| SPINACH + 3 EGGS0 h | Vitamin E Cmax | 0.14 umol/L | Standard Error 0.02 |
| SPINACH + WHITE | Vitamin E Cmax | 0.15 umol/L | Standard Error 0.02 |
| SPINACH + OIL | Vitamin E Cmax | 0.2 umol/L | Standard Error 0.02 |
| SPINACH + EGG3 h | Vitamin E Cmax | 0.06 umol/L | Standard Error 0.01 |
| SPINACH + EGGS0 & 3 h | Vitamin E Cmax | 0.14 umol/L | Standard Error 0.04 |
Secondary
Chylomicron Vitamin E
Deuterium-labeled alpha-tocopherol concentration in chylomicron
Time frame: 0, 3, 4.5, 6, 7.5, 9, 12 hours post-ingestion of spinach
Population: Chylomicron measurements of vitamin E content were initially planned as part of the study protocol. While chylomicrons were isolated from blood samples and cryopreserved during the intervention, sample analysis was not performed because resources and personnel became unavailable. No data are available and these measures are not anticipated to be completed.
Secondary
Estimated Absorption (%Dose) of Vitamin E
Absorption of deuterium-labeled alpha-tocopherol
Time frame: 0-72 hours post-ingestion of spinach
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| SPINACH | Estimated Absorption (%Dose) of Vitamin E | 1.93 Percent | Standard Error 0.5 |
| SPINACH + 1 EGG0 h | Estimated Absorption (%Dose) of Vitamin E | 3.52 Percent | Standard Error 0.46 |
| SPINACH + 2 EGGS0h | Estimated Absorption (%Dose) of Vitamin E | 2.95 Percent | Standard Error 0.53 |
| SPINACH + 3 EGGS0 h | Estimated Absorption (%Dose) of Vitamin E | 2.96 Percent | Standard Error 0.44 |
| SPINACH + WHITE | Estimated Absorption (%Dose) of Vitamin E | 3.02 Percent | Standard Error 0.44 |
| SPINACH + OIL | Estimated Absorption (%Dose) of Vitamin E | 3.86 Percent | Standard Error 0.43 |
| SPINACH + EGG3 h | Estimated Absorption (%Dose) of Vitamin E | 1.10 Percent | Standard Error 0.22 |
| SPINACH + EGGS0 & 3 h | Estimated Absorption (%Dose) of Vitamin E | 2.70 Percent | Standard Error 0.73 |
Secondary
Half-Life (hr)
The time required for the plasma concentration of vitamin E to decrease by 50%.
Time frame: 0-72 hours post-ingestion of spinach
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| SPINACH | Half-Life (hr) | 25.36 hr | Standard Error 4.54 |
| SPINACH + 1 EGG0 h | Half-Life (hr) | 22.52 hr | Standard Error 3.26 |
| SPINACH + 2 EGGS0h | Half-Life (hr) | 26.92 hr | Standard Error 2.25 |
| SPINACH + 3 EGGS0 h | Half-Life (hr) | 24.34 hr | Standard Error 3.65 |
| SPINACH + WHITE | Half-Life (hr) | 26.79 hr | Standard Error 1.57 |
| SPINACH + OIL | Half-Life (hr) | 30.45 hr | Standard Error 2.11 |
| SPINACH + EGG3 h | Half-Life (hr) | 16.89 hr | Standard Error 2.53 |
| SPINACH + EGGS0 & 3 h | Half-Life (hr) | 21.24 hr | Standard Error 5.37 |
Secondary
Vitamin E Tmax (hr)
Time to reach maximum plasma concentration of deuterium-labeled alpha-tocopherol
Time frame: 0-72 hours post-ingestion of spinach
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| SPINACH | Vitamin E Tmax (hr) | 9.64 hr | Standard Error 0.86 |
| SPINACH + 1 EGG0 h | Vitamin E Tmax (hr) | 10.10 hr | Standard Error 0.55 |
| SPINACH + 2 EGGS0h | Vitamin E Tmax (hr) | 10.90 hr | Standard Error 0.55 |
| SPINACH + 3 EGGS0 h | Vitamin E Tmax (hr) | 9.94 hr | Standard Error 0.63 |
| SPINACH + WHITE | Vitamin E Tmax (hr) | 9.94 hr | Standard Error 0.63 |
| SPINACH + OIL | Vitamin E Tmax (hr) | 9.38 hr | Standard Error 0.79 |
| SPINACH + EGG3 h | Vitamin E Tmax (hr) | 10.20 hr | Standard Error 0.74 |
| SPINACH + EGGS0 & 3 h | Vitamin E Tmax (hr) | 9.90 hr | Standard Error 0.9 |