Consumption of Iron-Fortified Lentils Is Protective against Declining Iron Status among Adolescent Girls in Bangladesh: Evidence from a Community-Based Double-Blind, Cluster-Randomized Controlled Trial.

BACKGROUND: In many low-income countries, iron deficiency (ID) and its anemia (IDA) pose significant health challenges, particularly among females and girls. Finding sustainable and effective solutions to address this issue is critical. OBJECTIVES: This study aimed to evaluate the efficacy of incorporating iron-fortified lentils (IFLs) into the diets of rural Bangladeshi adolescent girls on their body iron (Fe) status. METHODS: A community-based, double-blind, cluster-randomized controlled trial involved n = 1195 girls aged 10-17 y. A total of 48 adolescent clubs (n = ∼27 girls each) were randomized into 3 groups: 1) 200 g cooked IFLs, 2) 200 g cooked noniron-fortified lentils (NIFLs), and 3) a control group with no lentils (usual dietary intake). The intervention, administered 5 days a week for 85 feeding days, provided ∼8.625 mg Fe from each serving of IFLs and 2.625 mg from NIFLs. Blood samples collected at baseline, midpoint (42 feeding days), and endpoint (85 feeding days) assessed key Fe and inflammation biomarkers. Statistical analyses were filtered for inflammation. RESULTS: Although all groups experienced a decline in Fe status over time, the IFL group exhibited a significantly reduced decline in serum ferritin (sFer -7.2 µg/L), and total body iron (TBI -0.48 mg/kg) level compared with NIFL (sFer -14.3 µg/L and TBI -1.36 mg/kg) and usual intake group (sFer -12.8 µg/L and TBI -1.33 mg/kg). Additionally, those in the IFL group had a 57% reduced risk of developing clinical ID (sFer <15 µg/L) compared with the usual intake group. CONCLUSIONS: Our findings suggest that incorporating IFLs into the diet can help mitigate a decline in sFer, indicating a positive impact on the body Fe status of adolescent girls. This research underscores the potential role of fortified foods in addressing ID and IDA in vulnerable populations, emphasizing the significance of food-based interventions in public health. TRIAL REGISTRATION NUMBER: This trial was registered at the clinicaltrials.gov on May 24, 2018 (https://clinicaltrials.gov/study/NCT03516734?locStr=Bangladesh&country=Bangladesh&distance=50&cond=Anemia&intr=Iron%20fortified%20lentils&rank=1) as NCT03516734.

Dual-Fortified Lentil Products-A Sustainable New Approach to Provide Additional Bioavailable Iron and Zinc in Humans.

BACKGROUND: Iron (Fe) and zinc (Zn) deficiencies are global health problems affecting 20% and 33% of the world's population, respectively. Lentil (Lens culinaris Medik.), part of the staple food supply in many countries, can be a potential vehicle for Fe and Zn fortification. OBJECTIVE: We developed a dual-fortification protocol to fortify 3 milled lentil product types (LPTs) [red-football (RF), red-split (RS), and yellow-split (YS)], with NaFeEDTA and ZnSO4.H2O to increase the bioavailable content of Fe and Zn. METHODS: Appropriate Fe and Zn doses were determined to fortify lentils based on RDAs. Relative Fe bioavailability (RFeB%) and phytic acid (PA) content were assessed using an in vitro Caco-2 cell bioassay and PA analysis, respectively. One-factor ANOVA determined the differences in colorimetric score; concentrations of Fe, Zn, and PA; and RFeB% among samples. The least significant difference was calculated with significance level set at P < 0.05. RESULTS: Fe and Zn concentration and RFeB% increased and PA concentration decreased significantly in dual-fortified lentils. Dual-fortified lentil samples had higher RFeB% compared with Fe-fortified (single) samples in all 3 LPTs, whereas RFeB% decreased in Zn-fortified (single) RF and YS samples by 43.4% and 36%, respectively. The RF, RS, and YS samples, fortified with 16 mg Fe and 8 mg Zn/100 g of lentils, provided 27 mg Fe and 14 mg Zn, 28 mg Fe and 13.4 mg Zn, and 29.9 mg Fe and 12.1 mg Zn, respectively. RFeB% of RF, RS, and YS lentil samples increased by 91-307%, 114-522%, and 122-520%, respectively. Again, PA concentrations of RF, RS, and YS lentils were reduced by 0.63-0.53, 0.83-0.71, and 0.96-0.79 mg/g, respectively. CONCLUSIONS: Dual-fortified lentil consumption can cost-effectively provide a significant part of the daily bioavailable Fe and Zn requirements of people with these 2 globally important micronutrient deficiencies.

Sensory Acceptability of Dual-Fortified Milled Red and Yellow Lentil (Lens culinaris Medik.) Dal in Bangladesh.

This study evaluated the sensory properties of uncooked and cooked milled lentils that were fortified with varying concentrations of Fe and Zn in the form of NaFeEDTA and ZnSO4.H2O, respectively. Our study was carried out among 196 lentil consumers residing in rural Bangladesh who experience with growing, processing, and marketing lentils. A nine-point hedonic scale was used to rate the appearance, odor, taste, texture and overall acceptability of three uncooked and two cooked lentil (dal) samples made from each of the three milled lentil product types (LPTs), red football, red split and yellow split. Preferences for sensory properties were found to be significantly different among all uncooked lentil samples, but not significantly different for cooked samples, with a few exceptions. This means that the fortification process minimally affects dual-fortified lentil sample (fortified with 16 mg of Fe and 8 mg of Zn per 100 g of lentil), which was compared to another cooked sample (unfortified control), in terms of consumers liking for all four attributes (appearance, odor, taste, and texture).

Iron- and Zinc-Fortified Lentil (Lens culinaris Medik.) Demonstrate Enhanced and Stable Iron Bioavailability After Storage.

Lentil (Lens culinaris Medik.) is a quick-cooking, rapidly expanding protein-rich crop with high iron (Fe) and zinc (Zn), but low bioavailability due to the presence of phytate, similar to other grains. Lentils dual fortified with Fe and Zn can significantly improve the bioavailable Fe and Zn content. Three milled lentil product types (LPTs) were fortified with Fe using NaFeEDTA [ethylenediaminetetraacetic acid iron (III) sodium salt] (Fe fortified) or Zn from ZnSO4·H2O (Zn fortified), or both (dual fortified). Fe, Zn, phytic acid (PA) concentration, and relative Fe bioavailability (RFeB%) were assessed for samples from two fortified batches (initial and for 1 year stored). Fe, Zn, and RFeB% increased significantly in two batches of samples from the three LPTs, and decreased by 5-15% after 1 year of storage. PA concentration decreased from 8 to 15% after fortification of all samples from two batches of the three LPTs but showed different patterns of influence after storage. Dual-fortified lentil fortified with 24 mg Fe and 12 mg Zn 100 g-1 lentil had the highest amount of Fe and Zn, and the lowest PA concentration, and RFeB% was increased from 91.3 to 519.5%. Significant (p ≤ 0.01) Pearson correlations were observed between Fe concentration vs. PA:Fe molar ratio (MR), Fe concentration vs. RFeB%, RFeB% vs. PA:Fe MR, and Zn concentration vs. PA:Zn MR in all samples from two batches of the three LPTs. In conclusion, dual-fortified lentil can contribute significant bioavailable Fe and Zn to populations at risk of Fe and Zn deficiency.

Iron-fortified lentils to improve iron (Fe) status among adolescent girls in Bangladesh - study protocol for a double-blind community-based randomized controlled trial.

BACKGROUND: Lentils are generally considered to be a nutrient-dense food, and a good source of iron (Fe). This study aims to establish novel evidence of the effectiveness of the consumption of Fe-fortified lentils in improving the body Fe status and thus cognitive performance in non-pregnant adolescent girls in rural Bangladesh, compared to consumption of ordinary lentils. METHODS: We have designed a double-blind (both trial participants and outcome assessors), community-based, cluster-randomized controlled trial among 1260 Bangladeshi adolescent girls between the ages of 10-17 years who are non-smoking, not married, not pregnant, not breastfeeding, and generally healthy at the time of enrollment. The intervention will include three arms who will receive: (1) Fe-fortified lentils; (2) unfortified lentils; or (3) usual intake. Participants will be served a thick preparation of cooked Fe-fortified lentils (37.5 g raw lentils, approximately 200 g cooked lentils) 5 days per week for 85 feeding days (around 4 months) using a locally acceptable recipe. Lentils were fortified with Fe in the laboratory at the Department of Plant Sciences at the University of Saskatchewan in Canada. A subsample of participants (n = 360) will be randomly invited to be included in cognitive testing. DISCUSSION: Data on socio-demographic characteristics, household food security status, adolescent food habits and cognitive testing will be collected at baseline and endline (4 months). Venous blood samples will be collected at baseline, midline (2 months) and endline to measure adolescents' Fe status. Computerized cognitive testing will include five common measures of attentional (three of attention) and mnemonic functioning (two of memory) carried out using DMDX software. The results of this study will be used to garner support for and to substantiate large-scale production and market expansion of Fe-fortified lentils, and will contribute to knowledge about how to enhance Fe status in adolescents worldwide in resource-poor settings, using staple food crops. TRIAL REGISTRATION: ClinicalTrials.gov NCT03516734 . Registered on 24 May 2018.

Relative Bioavailability of Iron in Bangladeshi Traditional Meals Prepared with Iron-Fortified Lentil Dal.

Due to low Fe bioavailability and low consumption per meal, lentil must be fortified to contribute significant bioavailable Fe in the Bangladeshi diet. Moreover, since red lentil is dehulled prior to consumption, an opportunity exists at this point to fortify lentil with Fe. Thus, in the present study, lentil was Fe-fortified (using a fortificant Fe concentration of 2800 µg g-1) and used in 30 traditional Bangladeshi meals with broad differences in concentrations of iron, phytic acid (PA), and relative Fe bioavailability (RFeB%). Fortification with NaFeEDTA increased the iron concentration in lentil from 60 to 439 µg g-1 and resulted in a 79% increase in the amount of available Fe as estimated by Caco-2 cell ferritin formation. Phytic acid levels were reduced from 6.2 to 4.6 mg g-1 when fortified lentil was added, thereby reducing the PA:Fe molar ratio from 8.8 to 0.9. This effect was presumably due to dephytinization of fortified lentil during the fortification process. A significant (p ≤ 0.01) Pearson correlation was observed between Fe concentration and RFeB% and between RFeB% and PA:Fe molar ratio in meals with fortified lentil, but not for the meal with unfortified lentil. In conclusion, fortified lentil can contribute significant bioavailable Fe to populations at risk of Fe deficiency.

Sensory Acceptability of Iron-Fortified Red Lentil (Lens culinaris Medik.) Dal.

Panelists in Saskatoon, Canada (n = 45) and Dhaka, Bangladesh (n = 98) participated in sensory evaluations of the sensory properties of both cooked and uncooked dehulled red lentil dal fortified with FeSO4 ·7H2 O, NaFeEDTA or FeSO4 ·H2 O at fortificant Fe concentrations of 800, 1,600 (both cooked and uncooked), or 2,800 ppm. Appearance, odor, and overall acceptability of cooked and uncooked samples were rated using a 9-point hedonic scale (1 = dislike extremely to 9 = like extremely). Taste and texture were rated for the cooked samples prepared as typical south Asian lentil meals. Significant differences in sensory quality were observed among all uncooked and cooked samples at both locations. Overall, scores for all sensory attributes and acceptability of uncooked lentil decreased with increasing concentration of Fe in the fortificant; however, Fe fortification (particularly with NaFeEDTA) had small effects on acceptability. Panelists from Saskatoon provided a wider range of scores than those from Bangladesh for all attributes of cooked lentil. Overall, sensory evaluation of Fe fortification using NaFeEDTA minimally affected consumer perception of color, taste, texture, odor, and overall acceptability of cooked lentil. Reliability estimates (Cronbach's alpha [CA]) indicated that consumer scores were generally consistent for all attributes of all lentil samples (mean CA > 0.80). NaFeEDTA was found to be the most suitable Fe fortificant for lentil based on consumer acceptability. Consumption of 45 to 50 g of NaFeEDTA-fortified lentil (fortificant Fe concentration of 1,600 ppm) per day meets the estimated average requirements (EARs) of Fe for humans (10.8 to 29.4 mg). PRACTICAL APPLICATION: Iron fortification of dehulled lentil dal may change organoleptic attributes that can influence consumer acceptability. Sensory evaluation by consumers helps to determine the effect on appearance, odor, taste, texture, and overall acceptability of fortified lentils. In this study, consumer acceptability was evaluated with panelists who consume lentil regularly. Panelists provided significantly different scores for 5 sensory attributes for 10 uncooked and 3 cooked lentil samples. Panelists reliably preferred NaFeEDTA as the most suitable Fe fortificant for dehulled lentils for 5 attributes. Overall, lentil dal fortified with NaFeEDTA can offer a simple and low-cost solution to human health problems associated with iron-related malnutrition.

Iron Fortification of Lentil (Lens culinaris Medik.) to Address Iron Deficiency.

Iron (Fe) deficiency is a major human health concern in areas of the world in which diets are often Fe deficient. In the current study, we aimed to identify appropriate methods and optimal dosage for Fe fortification of lentil (Lens culinaris Medik.) dal with FeSO4·7H2O (ferrous sulphate hepta-hydrate), NaFeEDTA (ethylenediaminetetraacetic acid iron (III) sodium salt) and FeSO4·H2O (ferrous sulphate mono-hydrate). We used a colorimetric method to determine the appearance of the dal fortified with fortificants at different Fe concentrations and under different storage conditions. Relative Fe bioavailability was assessed using an in vitro cell culture bioassay. We found that NaFeEDTA was the most suitable fortificant for red lentil dal, and at 1600 ppm, NaFeEDTA provides 13-14 mg of additional Fe per 100 g of dal. Lentil dal sprayed with fortificant solutions, followed by shaking and drying at 75 °C, performed best with respect to drying time and color change. Total Fe and phytic acid concentrations differed significantly between cooked unfortified and fortified lentil, ranging from 68.7 to 238.5 ppm and 7.2 to 8.0 mg g-1, respectively. The relative Fe bioavailability of cooked fortified lentil was increased by 32.2-36.6% compared to unfortified cooked lentil. We conclude that fortification of lentil dal is effective and could provide significant health benefits to dal-consuming populations vulnerable to Fe deficiency.

Marker-Trait Association Analysis of Iron and Zinc Concentration in Lentil (Lens culinaris Medik.) Seeds.

Lentil ( Medik.) seeds are relatively rich in iron (Fe) and zinc (Zn), making lentil a potential crop to aid in the global battle against human micronutrient deficiency. Understanding the genetic basis for uptake of seed Fe and Zn is required to increase sustainable concentrations of these minerals in seeds. The objectives of this study were to characterize genetic variation in seed Fe and Zn concentration and to identify molecular markers associated with these traits across diverse lentil accessions. A set of 138 cultivated lentil accessions from 34 countries were evaluated in four environments (2 sites × 2 yr) in Saskatchewan, Canada. The collection was genotyped using 1150 single-nucleotide polymorphism (SNP) markers that are distributed across the lentil genome. The germplasm tested exhibited a wide range of variation for seed Fe and Zn concentration. The marker-trait association analysis detected two SNP markers tightly linked to seed Fe and one linked to seed Zn concentration (-log10 ≥ 4.36). Additional markers were detected at -log10 ≥ 3.06. A number of putative candidate genes underlying detected loci encode Fe- and Zn-related functions. This study provides insight into the genetics of seed Fe and Zn concentration of lentil and opportunities for marker-assisted selection to improve micronutrient concentration as part of micronutrient biofortification programs.