Ignite Creation Date:
2024-05-06 @ 4:59 PM
Last Modification Date:
2024-10-26 @ 2:20 PM
Study NCT ID:
NCT05154500
Status:
COMPLETED
Last Update Posted:
2023-01-25
First Post:
2021-11-19
Brief Title:
Iron and Zinc Bioavailability From Biofortified Potatoes
Sponsor:
Instituto de Investigacion Nutricional Peru
Organization:
Instituto de Investigacion Nutricional Peru
Study Overview
Official Title:
Evaluating Iron and Zinc Bioavailability From Biofortified Potatoes to Reduce Malnutrition in the Andean Highlands
Status:
COMPLETED
Status Verified Date:
2023-01
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:
Two of the most common nutritional deficiencies are anemia and zinc deficiency One strategy for combating nutritional deficiencies is biofortification The primary objective of the proposed research is to determine in humans the bioavailability of iron and zinc from biofortified potatoes as compared to a non-fortified local variety The secondary objective is to use the data obtained from the human studies to model the potential impact of the introduction of biofortified potatoes to the alleviation of iron and zinc deficiencies in the Andean Highlands and low and middle-income countries elsewhere
Iron bioavailability studies The investigators will compare bioavailability of iron from a non-fortified potato variety with a biofortified potato that has significantly higher iron content though the use of a randomized cross-over intervention study Female volunteers will consume both biofortified potato extrinsically labelled with 58FeSO4 and a nonfortified potato labelled with 57FeSO4 Thirty women with marginal iron status plasma ferritin 25 ngml will be selected from an initial screening of 180 women Every woman will receive 2 different types of test meals in a series of 20 servings for 10 days each Blood samples will be collected during screening and on days 1 15 26 and 40 and the amount of 58Fe and 57Fe incorporated into hemoglobin and serum ferritin quantified
Zinc bioavailability study The investigators will compare bioavailability of zinc from a non-fortified potato variety with that of a biofortified potato that has significantly higher zinc content p0001 with a crossover study Forty volunteers will be randomly assigned to receive first either the biofortified or the non-fortified potato and receive the second meal 30 days later Zinc absorption from two meals will be measured using the double stable isotope technique Every volunteer will be given an iv infusion of 70zinc and two test meals of 250 g cooked potato labelled extrinsically with 67zinc separated by 3-4 hr A spot urine sample will be collected 96 hours after each set of test meal and the ratio of the two isotopes measured to calculate absorption of zinc from the test meal
Modelling and Impact assessment Data on bioavailability will be used in a Disability Adjusted Life Year model to assess the potential impact of biofortified potatoes to alleviate iron and zinc deficiencies in the Andean Highlands and low and middle-income countries elsewhere
Iron bioavailability studies The investigators will compare bioavailability of iron from a non-fortified potato variety with a biofortified potato that has significantly higher iron content though the use of a randomized cross-over intervention study Female volunteers will consume both biofortified potato extrinsically labelled with 58FeSO4 and a nonfortified potato labelled with 57FeSO4 Thirty women with marginal iron status plasma ferritin 25 ngml will be selected from an initial screening of 180 women Every woman will receive 2 different types of test meals in a series of 20 servings for 10 days each Blood samples will be collected during screening and on days 1 15 26 and 40 and the amount of 58Fe and 57Fe incorporated into hemoglobin and serum ferritin quantified
Zinc bioavailability study The investigators will compare bioavailability of zinc from a non-fortified potato variety with that of a biofortified potato that has significantly higher zinc content p0001 with a crossover study Forty volunteers will be randomly assigned to receive first either the biofortified or the non-fortified potato and receive the second meal 30 days later Zinc absorption from two meals will be measured using the double stable isotope technique Every volunteer will be given an iv infusion of 70zinc and two test meals of 250 g cooked potato labelled extrinsically with 67zinc separated by 3-4 hr A spot urine sample will be collected 96 hours after each set of test meal and the ratio of the two isotopes measured to calculate absorption of zinc from the test meal
Modelling and Impact assessment Data on bioavailability will be used in a Disability Adjusted Life Year model to assess the potential impact of biofortified potatoes to alleviate iron and zinc deficiencies in the Andean Highlands and low and middle-income countries elsewhere
Detailed Description:
Type of Study and population Double blind randomized trial of naturally biofortified potatoes to be carried out in Huancavelica Perú It will be recruited about 100 women and 80 men aged 18-45 years living in the city of Huancavelica
ETHICS Applications was submitted and approved by Research Ethics Committee REC of the University of East Anglia Faculty of Medicine and Health FMH in Norwich UK and a REC of the Instituto de Investigacion Nutricional IIN in Peru
Every participant will be fully informed of all the procedures and will be given the opportunity to ask questions and receive any further details participants require
The procedures blood samples urine samples anthropometry dietary recall will be explained to the individuals who agree to take part in the initial screening and participants will be invited to sign the first consent form
SPECIFIC METHODS FOR IRON STUDY Phase 1 Sample size The study requires an enrollment of a total of 30 women to allow for 33 who do not adhere to the protocol to reach a final sample size of 20 women The investigators estimate that the investigators may need to screen about 100 women
Nutritional Status As part of the screening all the potential candidates will be weighed and measured using standardized equipment
Measurement of physical activity All the participants will be asked to use an actigraph a tri-axial accelerometer which measures physical activity movement in three dimensions
Blood sample An appropriately trained member of the study team will collect a venous blood sample approximately 6ml in a test tube with heparin to measure ferritin hemoglobin and CRP screening and 3 more times baseline 16 and 40 days after first day of potato intake
Urine sample Participants will be asked to provide a urine sample 30ml to carry out a pregnancy test using a test strip
Sociodemographic and Food security survey The investigators will include a socioeconomic survey and the ELCSA food security questionnaire both will be applied to all enrolled participants
Study Procedures According to the results of the screening and revision of the criteria for inclusion and exclusion and following information and verification that participants understand the iron study consent form subjects will be invited to participate Consented participants will be randomly assigned to consume one or other of the two types of potato biofortified-A or not-B The first potato variety will be given during the first 2 weeks day 1-12 and the second sequence between days 15-26 in both cases excluding weekend days Saturday and Sunday The potatoes will be cooked in water and offered mashed
Blood samples will be taken on 5 occasions 1 at screening to verify compliance with the exclusion and inclusion criteria then on day 1 before participants eat the first type of potato day 15 before eating the second type of potato day 26 after eating all both potato types and on day 40 two weeks after eating all the potato
Consumption of the potato Each participant will be served 500 g of potato divided into two portions of 250 g each each participant will take depending on their allocation 03 mg of 57FeSO4 Peruanita potato or 03 mg of 58FeSO4 biofortified potato diluted in 50 ml of distilled water which should be drunk while participants are eating the potato - halfway through the meal In addition each participant will receive 200 ml of distilled water to drink The isotopes will be provided by ETH Swiss Federal Institute of Technology in Zurich In addition the isotopes that the investigators will use are stable isotopes that is participants exist naturally in the environment and as such do not pose any risk
Collection of biological samples for the measurement of iron using stable isotopes Blood samples will be collected throughout the intervention period as previously described and will be sent to ETH Zurich following standard operating procedures Participants will be informed and will provide consent for the transfer and use of their samples outside Peru for the purpose of this research study Samples will be anonymized and will not be transported with personal identifiable data
In terms of processing samples will be mixed with NO3 and H2O2 and digested in a microwave followed by separation of the iron from the blood matrix by way of exchange-anion chromatography and a solvent diluent or extraction from the solvent using distilled
The isotopes will be analyzed using a plasma coupled inductive multicollector mass spectrometer Neptune Thermo Finnigan The amount of the labelled iron isotopes 57Fe and 58Fe will be measured 14 days after consumption of both types of potato has been completed The calculation of absorption will be done considering the subjects estimated blood volume based on weight height and the subjects measured hemoglobin concentration considering the average of the initial and final blood sample The calculation is based on the principle of isotopic dilution considering as the factor for the iron isotope marker rather than the monoisotopic marker8 For the calculation of the fraction absorbed the investigators will assume 80 incorporation of the absorbed iron
SPECIFIC METHODS FOR ZINC STUDY Phase 2 Sample size The study requires a samples size of 40 women This allows for a 40 loss to reach a final total of 24 women
Procedures According to the results of screening and after checking that participants comply with all the inclusion criteria and none of the exclusion criteria participants will be invited to participate in the zinc part of the study For this participant will be given the second consent form to read and those who accept and sign the form will be invited to participate in the study
The people who accept will be randomly allocated to eat one or other of the types of potato biofortified-A or no-B All the participants will be given both types of potato but in different order in this way each participant will be their own control The first variety of potato will be given on day 1 after acceptance of participating in the study and after accepting and signing the consent form for this part consumption protocol and second variety will be eaten 29 days after the order of potato varieties will be randomly assigned
The investigators will take blood samples on three occasions enrolment and days 1 and 29 the days when participants eat the potato after fasting for 8 hours food and 6 hours liquids
Study Measurements Anthropometry As part of the screening all the potential candidates will be weighed and measured using standardized equipment According to their weight and BMI the investigators will decide whether participants are eligible
Consented participants will be asked to provide a venous blood sample approximately 10ml in a test tube with heparin Plasma and packed red cells will be obtained from the blood samples and stored at 18oC
Urine sample Consented participants will provide a urine sample 30ml as part of the screening phase to carry out a pregnancy test On days 1 3 4 5 6 7 29 32 33 34 35 and 36 the investigators will collect urine samples to analyze zinc content On each day the investigators need 100ml of urine collected in the morning between 7-12 oclock in the morning
Consumption of the potato The subjects will be given a total of 1000 g of potato in two 500 g portions The first portion will be given while participants are fasted at least 8 hours for food and 6 hours for drink and in two separate servings each one 250 g When participants have eaten the first 250 g serving the subject will receive 025 mg of 67Zn diluted in 50 ml of bottled water and then participants will receive the second serving of potato Two hours later participants will receive an intravenous infusion of 70Zn 200 ug diluted in 50 ml of bottled water After a further 2 hours participants will receive the second two servings of potato and as before after the first serving participants will receive 025 mg of 67Zn diluted in 50 ml of bottled water During this period and for 3 hours after the second portion of potato the subject will not consume any food or liquid except for water which participants will be provided with Each subject will receive 200 ml of water with each meal and 200 ml of water between meals Currently using isotopes is the best method for measuring the absorption of zinc In addition the isotopes that will be used are stable isotopes that is participants exist naturally in the environment and pose no risk No food or drink can be consumed for at least 3 hours after eating the final portion of potato
Blood samples for measurement of zinc The investigators will take a total of three blood samples in total Absorption using stable isotopes Day 29 The blood samples that have been collected will be sent to ETH Zurich ETH will undertake the analysis of CRP for Zinc The CRP will be measured using an ELISA kit The plasma zinc concentration will be measured by induction plasma coupled mass spectrometry
Urine samples for measurement of zinc using stable isotopes Samples of urine will be concentrated using freeze drying and then mineralized by digestion in a microwave oven MLS-Turbowave MLS GmbH using a mixture of HNO3 and zinc isolated from the master matrix by ion exchange chromatography The investigators will use ultrapure acids for the preparation of the samples
Determination of zinc absorption Blood and zinc isotopic composition will be analyzed in duplicate at the ETH Human Nutrition Laboratory Urine samples will be mineralized by microwave digestion MLS-ETHOS plus MLS GmbH using a mixture of HNO3 and H2O2 and zinc isolated from the sample matrix by ion-exchange chromatography Wegmuller et al 2014 All acids used for the preparation of samples will be ultrapure 70Zn66Zn and 67Zn66Zn isotopic ratios will be measured to determine 70Zn and 67Zn enrichment in the urine samples by using a high-resolution double-focusing magnetic sector field multi-collector inductively coupled plasma mass spectrometer Finnigan Neptune Thermo Electron12 Fractional absorption of the 67Zn dose from the test meals will be calculated by using the oral to iv tracers ratio method applied to spot urine samples according to principles described by Friel et al
7-day food record Using the method of registering all foods eaten over a 7-day period the investigators can obtain information on all foods eaten in a week including Saturday and Sunday To achieve this the investigators will ask the subjects to make a record of all foods consumed details of food preparation and all drinks including water that participants have eaten or drunk Participants should note the meal at which the food and drink were consumed and the time taken to eat it breakfast lunch supper and between meals snacks and the amount eaten at each time of the day To do this the investigators will give the subjects a tablet in which to register the consumptionmeals and as well take a photo of all the food eaten
METHODS SPECIFIC FOR IMPACT ASSESSMENT The data on dietary intake serum ferritin and iron bioavailability will be used for two modelling approaches Firstly the investigators will apply the Disability Adjusted Life Year DALY method which is an appropriate method to estimate nutritional and health impacts of biofortified crops This will build upon models that have been developed jointly by CIP and IIN that has made assumptions concerning iron and zinc bioavailability from potatoes Secondly the investigators will use data on habitual iron intake serum ferritin concentrations and calculated physiological requirements for iron to adapt a model estimate dietary iron bioavailability in Europe the Andean highland region and more generally to low- and middle-income countries
ETHICS Applications was submitted and approved by Research Ethics Committee REC of the University of East Anglia Faculty of Medicine and Health FMH in Norwich UK and a REC of the Instituto de Investigacion Nutricional IIN in Peru
Every participant will be fully informed of all the procedures and will be given the opportunity to ask questions and receive any further details participants require
The procedures blood samples urine samples anthropometry dietary recall will be explained to the individuals who agree to take part in the initial screening and participants will be invited to sign the first consent form
SPECIFIC METHODS FOR IRON STUDY Phase 1 Sample size The study requires an enrollment of a total of 30 women to allow for 33 who do not adhere to the protocol to reach a final sample size of 20 women The investigators estimate that the investigators may need to screen about 100 women
Nutritional Status As part of the screening all the potential candidates will be weighed and measured using standardized equipment
Measurement of physical activity All the participants will be asked to use an actigraph a tri-axial accelerometer which measures physical activity movement in three dimensions
Blood sample An appropriately trained member of the study team will collect a venous blood sample approximately 6ml in a test tube with heparin to measure ferritin hemoglobin and CRP screening and 3 more times baseline 16 and 40 days after first day of potato intake
Urine sample Participants will be asked to provide a urine sample 30ml to carry out a pregnancy test using a test strip
Sociodemographic and Food security survey The investigators will include a socioeconomic survey and the ELCSA food security questionnaire both will be applied to all enrolled participants
Study Procedures According to the results of the screening and revision of the criteria for inclusion and exclusion and following information and verification that participants understand the iron study consent form subjects will be invited to participate Consented participants will be randomly assigned to consume one or other of the two types of potato biofortified-A or not-B The first potato variety will be given during the first 2 weeks day 1-12 and the second sequence between days 15-26 in both cases excluding weekend days Saturday and Sunday The potatoes will be cooked in water and offered mashed
Blood samples will be taken on 5 occasions 1 at screening to verify compliance with the exclusion and inclusion criteria then on day 1 before participants eat the first type of potato day 15 before eating the second type of potato day 26 after eating all both potato types and on day 40 two weeks after eating all the potato
Consumption of the potato Each participant will be served 500 g of potato divided into two portions of 250 g each each participant will take depending on their allocation 03 mg of 57FeSO4 Peruanita potato or 03 mg of 58FeSO4 biofortified potato diluted in 50 ml of distilled water which should be drunk while participants are eating the potato - halfway through the meal In addition each participant will receive 200 ml of distilled water to drink The isotopes will be provided by ETH Swiss Federal Institute of Technology in Zurich In addition the isotopes that the investigators will use are stable isotopes that is participants exist naturally in the environment and as such do not pose any risk
Collection of biological samples for the measurement of iron using stable isotopes Blood samples will be collected throughout the intervention period as previously described and will be sent to ETH Zurich following standard operating procedures Participants will be informed and will provide consent for the transfer and use of their samples outside Peru for the purpose of this research study Samples will be anonymized and will not be transported with personal identifiable data
In terms of processing samples will be mixed with NO3 and H2O2 and digested in a microwave followed by separation of the iron from the blood matrix by way of exchange-anion chromatography and a solvent diluent or extraction from the solvent using distilled
The isotopes will be analyzed using a plasma coupled inductive multicollector mass spectrometer Neptune Thermo Finnigan The amount of the labelled iron isotopes 57Fe and 58Fe will be measured 14 days after consumption of both types of potato has been completed The calculation of absorption will be done considering the subjects estimated blood volume based on weight height and the subjects measured hemoglobin concentration considering the average of the initial and final blood sample The calculation is based on the principle of isotopic dilution considering as the factor for the iron isotope marker rather than the monoisotopic marker8 For the calculation of the fraction absorbed the investigators will assume 80 incorporation of the absorbed iron
SPECIFIC METHODS FOR ZINC STUDY Phase 2 Sample size The study requires a samples size of 40 women This allows for a 40 loss to reach a final total of 24 women
Procedures According to the results of screening and after checking that participants comply with all the inclusion criteria and none of the exclusion criteria participants will be invited to participate in the zinc part of the study For this participant will be given the second consent form to read and those who accept and sign the form will be invited to participate in the study
The people who accept will be randomly allocated to eat one or other of the types of potato biofortified-A or no-B All the participants will be given both types of potato but in different order in this way each participant will be their own control The first variety of potato will be given on day 1 after acceptance of participating in the study and after accepting and signing the consent form for this part consumption protocol and second variety will be eaten 29 days after the order of potato varieties will be randomly assigned
The investigators will take blood samples on three occasions enrolment and days 1 and 29 the days when participants eat the potato after fasting for 8 hours food and 6 hours liquids
Study Measurements Anthropometry As part of the screening all the potential candidates will be weighed and measured using standardized equipment According to their weight and BMI the investigators will decide whether participants are eligible
Consented participants will be asked to provide a venous blood sample approximately 10ml in a test tube with heparin Plasma and packed red cells will be obtained from the blood samples and stored at 18oC
Urine sample Consented participants will provide a urine sample 30ml as part of the screening phase to carry out a pregnancy test On days 1 3 4 5 6 7 29 32 33 34 35 and 36 the investigators will collect urine samples to analyze zinc content On each day the investigators need 100ml of urine collected in the morning between 7-12 oclock in the morning
Consumption of the potato The subjects will be given a total of 1000 g of potato in two 500 g portions The first portion will be given while participants are fasted at least 8 hours for food and 6 hours for drink and in two separate servings each one 250 g When participants have eaten the first 250 g serving the subject will receive 025 mg of 67Zn diluted in 50 ml of bottled water and then participants will receive the second serving of potato Two hours later participants will receive an intravenous infusion of 70Zn 200 ug diluted in 50 ml of bottled water After a further 2 hours participants will receive the second two servings of potato and as before after the first serving participants will receive 025 mg of 67Zn diluted in 50 ml of bottled water During this period and for 3 hours after the second portion of potato the subject will not consume any food or liquid except for water which participants will be provided with Each subject will receive 200 ml of water with each meal and 200 ml of water between meals Currently using isotopes is the best method for measuring the absorption of zinc In addition the isotopes that will be used are stable isotopes that is participants exist naturally in the environment and pose no risk No food or drink can be consumed for at least 3 hours after eating the final portion of potato
Blood samples for measurement of zinc The investigators will take a total of three blood samples in total Absorption using stable isotopes Day 29 The blood samples that have been collected will be sent to ETH Zurich ETH will undertake the analysis of CRP for Zinc The CRP will be measured using an ELISA kit The plasma zinc concentration will be measured by induction plasma coupled mass spectrometry
Urine samples for measurement of zinc using stable isotopes Samples of urine will be concentrated using freeze drying and then mineralized by digestion in a microwave oven MLS-Turbowave MLS GmbH using a mixture of HNO3 and zinc isolated from the master matrix by ion exchange chromatography The investigators will use ultrapure acids for the preparation of the samples
Determination of zinc absorption Blood and zinc isotopic composition will be analyzed in duplicate at the ETH Human Nutrition Laboratory Urine samples will be mineralized by microwave digestion MLS-ETHOS plus MLS GmbH using a mixture of HNO3 and H2O2 and zinc isolated from the sample matrix by ion-exchange chromatography Wegmuller et al 2014 All acids used for the preparation of samples will be ultrapure 70Zn66Zn and 67Zn66Zn isotopic ratios will be measured to determine 70Zn and 67Zn enrichment in the urine samples by using a high-resolution double-focusing magnetic sector field multi-collector inductively coupled plasma mass spectrometer Finnigan Neptune Thermo Electron12 Fractional absorption of the 67Zn dose from the test meals will be calculated by using the oral to iv tracers ratio method applied to spot urine samples according to principles described by Friel et al
7-day food record Using the method of registering all foods eaten over a 7-day period the investigators can obtain information on all foods eaten in a week including Saturday and Sunday To achieve this the investigators will ask the subjects to make a record of all foods consumed details of food preparation and all drinks including water that participants have eaten or drunk Participants should note the meal at which the food and drink were consumed and the time taken to eat it breakfast lunch supper and between meals snacks and the amount eaten at each time of the day To do this the investigators will give the subjects a tablet in which to register the consumptionmeals and as well take a photo of all the food eaten
METHODS SPECIFIC FOR IMPACT ASSESSMENT The data on dietary intake serum ferritin and iron bioavailability will be used for two modelling approaches Firstly the investigators will apply the Disability Adjusted Life Year DALY method which is an appropriate method to estimate nutritional and health impacts of biofortified crops This will build upon models that have been developed jointly by CIP and IIN that has made assumptions concerning iron and zinc bioavailability from potatoes Secondly the investigators will use data on habitual iron intake serum ferritin concentrations and calculated physiological requirements for iron to adapt a model estimate dietary iron bioavailability in Europe the Andean highland region and more generally to low- and middle-income countries
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