Ignite Creation Date:
2024-05-06 @ 6:58 PM
Last Modification Date:
2024-10-26 @ 2:57 PM
Study NCT ID:
NCT05840848
Status:
COMPLETED
Last Update Posted:
2023-05-06
First Post:
2023-03-28
Brief Title:
Effect of Iron and Zinc Supplementation on B-carotene Bioavailability in Healthy Males
Sponsor:
University of Hohenheim
Organization:
University of Hohenheim
Study Overview
Official Title:
Effect of Simultaneously Consuming Iron and Zinc Supplements With That of B-carotene on the B-carotene Bioavailability in Healthy Males
Status:
COMPLETED
Status Verified Date:
2023-05
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
In vitro studies found supplemental levels of iron and zinc to inhibit the micellization and cellular uptake of β-carotene Here we investigated this in vivo in a double-blind 3-arm crossover human trial
Healthy males n6 ingested with breakfast a single dose of 15 mg β-carotene in combination with either a placebo 25 mg iron or 30 mg zinc capsule Blood samples were collected at baseline and hourly for 10 hours The triacylglycerol-rich fraction TRF was analysed for concentrations of β-carotene and plasma for β-carotene retinol triacylglycerols LDL- and HDL-cholesterol
Healthy males n6 ingested with breakfast a single dose of 15 mg β-carotene in combination with either a placebo 25 mg iron or 30 mg zinc capsule Blood samples were collected at baseline and hourly for 10 hours The triacylglycerol-rich fraction TRF was analysed for concentrations of β-carotene and plasma for β-carotene retinol triacylglycerols LDL- and HDL-cholesterol
Detailed Description:
The study followed a double-blind crossover design with three study arms separated by one week washout periods In short the participants were asked to follow a diet low in carotenoids by avoiding all orange yellow red and green fruits and vegetables for four days This was followed by three days of a strictly carotenoid-free diet which only allowed foods from a specified list Supplemental Table A2 On each study day β-carotene was administered in the morning after a 10 hour overnight fast Supplemental Figure A1 All participants orally ingested in random order a single dose of 15 mg β-carotene BIOVEA with either a placebo empty capsule 25 mg iron FeSO4 Woerwag Pharma GmbH Co KG Boeblingen Germany or 30 mg zinc ZnSO4 Woerwag Pharma capsule A standardised dinner was provided on the evening before the trial and standardised meals were provided during the entire intervention day Supplemental Table A3 On the first study day the amount of food weight or volume consumed by each participant for each meal was recorded and the same amounts provided during the following two arms to ensure similar food consumption especially of fat Water was available unrestricted for consumption throughout the day Blood samples were drawn from an indwelling venous cannula and collected at 0 hours directly before β-carotene supplementation and then every hour for 10 hours
For the determination of plasma concentrations of β-carotene LDL- and HDL-cholesterol and triacylglycerols TAG blood was collected in tubes containing EDTA Sarstedt AG Co Nuebrecht Germany and immediately centrifuged 3000 g 10 min 4 C From the obtained plasma samples three aliquots were stored at -80 C until further analysis and the rest ultracentrifuged to obtain the triacylglycerol-rich fraction TRF For the analyses of liver and kidney function markers plasma and serum were obtained from blood sampled at the 0- and 4-hour time points
The TRF was prepared according to 10 Briefly plasma 35 mL was transferred to an ultracentrifuge tube and carefully overlaid with 8 mL 13 sodium chloride and then ultracentrifuged Beckman Coulter OptimaTM L-80 XP Ultracentrifuge using a swinging bucket rotor SW41Ti at 150 000 x g for 1 hour at 4 C Afterwards the TRF was isolated by transferring the upper 6 mL which was then overlaid with nitrogen gas to minimize oxidation and stored at -80 C until extraction
The plasma samples were randomly extracted and analysed by HPLC according to 15 Briefly 40 µL plasma was extracted with an ethanoln-butanol mixture 5050 containing apo-80-carotenal-methyloxime 12µL100 mL Fluka Analytical Merck Group KGaA Darmstadt Germany as internal standard After centrifugation the clear supernatant was analysed by HPLC
The TRF was extracted and analysed by HPLC 15 For the extraction 100 µl apo-80-carotenal-methyloxime 12 µL100 mL and 2 mL ethanol for deproteination were added to 3 mL of the TRF and vortexed for 30 sec The solution was extracted twice with 2 mL hexane The hexane layers were removed combined and evaporated in a centrifugal vacuum concentrator Christ RVC 2-25 CD plus and the dried sample re-dissolved in 100 µL acetonitrile and immediately analysed by HPLC
Both the plasma and TRF samples were analysed using a Shimadzu HPLC LC-10AD equipped with a UV-Vis detector SPD 20A set at 450 nm Carotenoids were separated using a ReproSil 80 ODS-2 column 3 µm 250 x 46 mm Dr Maisch GmbH Ammerbuch Germany and an eluent in recirculation mode 82 acetonitrile 15 14-dioxin and 3 methanol volvol containing 100 mM ammonium acetate and 10 mM triethylamine at a flow rate of 15 mLmin 15 A β-carotene standard 970 purity Sigma-Aldrich was used to construct a standard curve
Plasma TAG HDL- and LDL-cholesterol were analysed by a clinical laboratory Laborärzte Sindelfingen Sindelfingen Germany
For the determination of plasma concentrations of β-carotene LDL- and HDL-cholesterol and triacylglycerols TAG blood was collected in tubes containing EDTA Sarstedt AG Co Nuebrecht Germany and immediately centrifuged 3000 g 10 min 4 C From the obtained plasma samples three aliquots were stored at -80 C until further analysis and the rest ultracentrifuged to obtain the triacylglycerol-rich fraction TRF For the analyses of liver and kidney function markers plasma and serum were obtained from blood sampled at the 0- and 4-hour time points
The TRF was prepared according to 10 Briefly plasma 35 mL was transferred to an ultracentrifuge tube and carefully overlaid with 8 mL 13 sodium chloride and then ultracentrifuged Beckman Coulter OptimaTM L-80 XP Ultracentrifuge using a swinging bucket rotor SW41Ti at 150 000 x g for 1 hour at 4 C Afterwards the TRF was isolated by transferring the upper 6 mL which was then overlaid with nitrogen gas to minimize oxidation and stored at -80 C until extraction
The plasma samples were randomly extracted and analysed by HPLC according to 15 Briefly 40 µL plasma was extracted with an ethanoln-butanol mixture 5050 containing apo-80-carotenal-methyloxime 12µL100 mL Fluka Analytical Merck Group KGaA Darmstadt Germany as internal standard After centrifugation the clear supernatant was analysed by HPLC
The TRF was extracted and analysed by HPLC 15 For the extraction 100 µl apo-80-carotenal-methyloxime 12 µL100 mL and 2 mL ethanol for deproteination were added to 3 mL of the TRF and vortexed for 30 sec The solution was extracted twice with 2 mL hexane The hexane layers were removed combined and evaporated in a centrifugal vacuum concentrator Christ RVC 2-25 CD plus and the dried sample re-dissolved in 100 µL acetonitrile and immediately analysed by HPLC
Both the plasma and TRF samples were analysed using a Shimadzu HPLC LC-10AD equipped with a UV-Vis detector SPD 20A set at 450 nm Carotenoids were separated using a ReproSil 80 ODS-2 column 3 µm 250 x 46 mm Dr Maisch GmbH Ammerbuch Germany and an eluent in recirculation mode 82 acetonitrile 15 14-dioxin and 3 methanol volvol containing 100 mM ammonium acetate and 10 mM triethylamine at a flow rate of 15 mLmin 15 A β-carotene standard 970 purity Sigma-Aldrich was used to construct a standard curve
Plasma TAG HDL- and LDL-cholesterol were analysed by a clinical laboratory Laborärzte Sindelfingen Sindelfingen Germany
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None