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.

A comparative metabolomics investigation of flavonoid variation in faba bean flowers.

INTRODUCTION: Faba bean (Vicia faba L.) flowers are edible and used as garnishes because of their aroma, sweet flavor and attractive colors. Anthocyanins are the common plant pigments that give flowers their vivid colors, whereas non-anthocyanin flavonoids can serve as co-pigments that can modify the color intensity of flowers. OBJECTIVES: To explore the polyphenol diversity and differences in standard and wing petals of faba bean flowers; and identify glycosylated flavonoids that contribute to flower color. METHODS: Flower standard and wing petals from 30 faba bean genotypes (eight color groups with a total of 60 samples) were used for polyphenol extraction. Samples were analyzed using a targeted method and a semi-untargeted analysis using liquid chromatography-high resolution mass spectrometry (LC-HRMS) combined with photodiode array (PDA) detection. Compound Discoverer software was used for polyphenol identification and multivariate analysis. RESULTS: The semi-untargeted analysis guided by the PDA detected 90 flavonoid metabolites present in faba bean flower petals. Ten anthocyanins largely influenced the flower colors, but other flavonoids (63 flavonols and 12 flavones) found with variable levels in different flower color groups appeared to also influence color, especially in mixed colors. CONCLUSION: Analysis of the different colored faba bean flowers confirmed that the color variation between the flowers was mainly controlled by anthocyanins in brown, red and purple-red flowers. Of the other flavonoids, multiglycosylated kaempferols were abundant in white and brown flowers, monoglycosylated kaempferols were common in red and purple-red flowers, and quercetin and apigenin glycosides were abundant co-pigments in purple-red flowers.

Mass Spectrometry-Based Untargeted Metabolomics Reveals the Importance of Glycosylated Flavones in Patterned Lentil Seed Coats.

Lentil seed coats are rich in antioxidant polyphenols that are important for plant defense and have potential as valorized byproducts. Although biochemical differences among lentil seed coat colors have been previously studied, differences among seed coat patterns remain largely unexplored. This study used mass spectrometry-based untargeted metabolomics to investigate polyphenol differences among lentil seed coat patterns to search for biochemical pathways potentially responsible for seed coat pattern differences. Comparing patterned with non-patterned green lentil seed coats, 28 significantly upregulated metabolites were found in patterned seed coats; 19 of them were identified as flavones. Flavones were virtually absent in non-patterned seed coats, thereby strongly suggesting a blockage in their flavone biosynthetic pathway. Although the black pattern is not readily discernible on black seed coats, many of the same flavones found in green marbled seed coats were also found in black seed coats, indicating that black seed coats likely have a marbled pattern.

Water-soluble phenolic compounds and their putative antioxidant activities in the seed coats from different lentil (Lens culinaris) genotypes.

The seed coat is a major byproduct of lentil processing with potential as a sustainable source of antioxidant polyphenols. Profiles of water-soluble phenolic compounds and antioxidant activities of seven genotypes of lentil which includes both normal-tannin and low-tannin seed coats were investigated. Antioxidant activities were assessed using four antioxidant assays, and phenolic compounds were quantified using liquid chromatography mass spectrometry (LC-MS). Total phenolic content (TPC) varied significantly among genotypes and ranged between 1519 ± 140 and 6502 ± 154 µg/g. Thirty phenolic compounds were identified with kaempferol tetraglycoside, catechin-3-glucoside and procyanidins being the dominant compounds in normal-tannin seed coats. Kaempferol tetraglycoside predominated (80-90%) in low-tannin seed coats. Antioxidant activities strongly correlated with TPC (r > 0.93) with a 6-9 times higher activity in normal-tannin than that of low-tannin lentils. Without flavan-3-ols and procyanidins, low-tannin seed coat may not exert strong antioxidant potential, whereas normal-tannin seed coat contains water-extractable natural phenolic compounds with promising antioxidant potential.

Selection of Enzymatic Treatments for Upcycling Lentil Hulls into Ingredients Rich in Oligosaccharides and Free Phenolics.

In this study, the comprehensive chemical characterization of red lentil hulls obtained from the industrial production of football and split lentils was described. The lentil hulls were rich in dietary fiber (78.43 g/100 g dry weight with an insoluble to soluble fiber ratio of 4:1) and polyphenols (49.3 mg GAE/g dry weight, of which 55% was bound phenolics), which revealed the suitability of this lentil by-product as a source of bioactive compounds with recognized antioxidant and prebiotic properties. The release of oligosaccharides and phenolic compounds was accomplished by enzymatic hydrolysis, microwave treatment and a combination of both technologies. The key role played by the selection of a suitable enzymatic preparation was highlighted to maximize the yield of bioactive compounds and the functional properties of the lentil hull hydrolysates. Out of seven commercial preparations, the one with the most potential for use in a commercial context was Pectinex® Ultra Tropical, which produced the highest yields of oligosaccharides (14 g/100 g lentil hull weight) and free phenolics (45.5 mg GAE/100 g lentil hull weight) and delivered a four-fold increase in terms of the original antioxidant activity. Finally, this enzyme was selected to analyze the effect of a microwave-assisted extraction pretreatment on the yield of enzymatic hydrolysis and the content of free phenolic compounds and oligosaccharides. The integrated microwave and enzymatic hydrolysis method, although it increased the solubilization yield of the lentil hulls (from 25% to 34%), it slightly decreased the content of oligosaccharides and proanthocyanidins and reduced the antioxidant activity. Therefore, the enzymatic hydrolysis treatment alone was more suitable for producing a lentil hull hydrolysate enriched in potential prebiotics and antioxidant compounds.

Lentil and Fava Bean With Contrasting Germination Kinetics: A Focus on Digestion of Proteins and Bioactivity of Resistant Peptides.

Germination offers advantages to improve legume protein digestibility as it disintegrates seed structure and hydrolyzes proteins and anti-nutrients. Seed permeability (related to polyphenol content of seed coats) is an important factor affecting the duration of seed germination and its impact on protein digestibility and bioactivity. The objective was to compare the effect of seed germination on protease activity, structure, and proteolysis of four selected legumes with contrasting seed coat polyphenol profiles (gray zero-tannin lentil [GZL], beluga lentil [BL], and dehulled red lentil [DL]; and zero tannin/low vicine-convicine fava bean [ZF]). Protein hydrolysis was characterized during germination and digestion with respect to proteins, peptides, and free amino acids (FAAs). In vitro antihypertensive and antioxidant activities of digests were investigated, and the peptidomic characterization [high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS)] and identification of bioactive fragments in intestinal digests were performed. Regardless of the seed type, germination increased protease activity and reduced the levels of phytic acid, trypsin inhibitors, and tannins (only in BL). A significant proteolysis of the 7S and 11S globulins and a concomitant increase of peptides and FAAs were observed in all sprouted legumes. Digestion kinetics in sprouts revealed a faster generation of FAAs and peptides than in dry seeds, with changes being more evident for DL, associated with a faster imbibition, germination, and sprout growth. In contrast, BL sprouts showed the lowest protein digestibility, likely due to a lower protease activity, seed structure disintegration, and higher anti-nutrient levels in comparison to GZL, DL, and ZF. Moreover, the digestion of sprouts resulted in a higher number of resistant peptides in DL and ZF that matched with previously reported bioactive sequences, suggesting a promising health potential of legume sprouts that was confirmed in vitro. The results suggested that the germination process improved protein digestibility and the health-promoting potential of lentil and fava bean proteins although these changes were more evident in DL due to its rapid imbibition, faster germination, and sprout development. This study will provide important information for either plant breeders to develop legume varieties with permeable seed coats or food producers that could use dehulled seeds for efficient production of sprouts as sustainable food sources of plant proteins with improved nutritional and healthy properties.

An Untargeted Metabolomics Approach for Correlating Pulse Crop Seed Coat Polyphenol Profiles with Antioxidant Capacity and Iron Chelation Ability.

Pulse crop seed coats are a sustainable source of antioxidant polyphenols, but are typically treated as low-value products, partly because some polyphenols reduce iron bioavailability in humans. This study correlates antioxidant/iron chelation capabilities of diverse seed coat types from five major pulse crops (common bean, lentil, pea, chickpea and faba bean) with polyphenol composition using mass spectrometry. Untargeted metabolomics was used to identify key differences and a hierarchical analysis revealed that common beans had the most diverse polyphenol profiles among these pulse crops. The highest antioxidant capacities were found in seed coats of black bean and all tannin lentils, followed by maple pea, however, tannin lentils showed much lower iron chelation among these seed coats. Thus, tannin lentils are more desirable sources as natural antioxidants in food applications, whereas black bean and maple pea are more suitable sources for industrial applications. Regardless of pulse crop, proanthocyanidins were primary contributors to antioxidant capacity, and to a lesser extent, anthocyanins and flavan-3-ols, whereas glycosylated flavonols contributed minimally. Higher iron chelation was primarily attributed to proanthocyanidin composition, and also myricetin 3-O-glucoside in black bean. Seed coats having proanthocyanidins that are primarily prodelphinidins show higher iron chelation compared with those containing procyanidins and/or propelargonidins.

Determination of an Acceptable Portion Size of Daal for a Bangladeshi Community-Based Iron Intervention in Adolescent Girls: A Feasibility Study.

Widely consumed daal (lentils) in Bangladesh are an ideal vehicle for iron (Fe) fortification; however, an acceptable portion size in meals needs to be determined to carry out a community feeding study in at-risk adolescent girls. A non-randomized crossover trial was conducted with n = 100 Bangladeshi girls (12.9 ± 2.0 years of age). Two recipes (thin and thick) and three portion sizes (25 g, 37.5 g, 50 g of raw lentil) of daal were served with 250 g of cooked white rice in a counter-balanced manner over 12 weeks. Each meal was fed to participants 5 days/week for two weeks. Ratings of hunger, satiety, and palatability were measured before and after each meal using Visual Analog Scales (VAS). The thick preparation in the 37.5 g portion (~200 g cooked) elicited higher VAS ratings of hunger, satiety, and palatability compared to all other meals. The 50 g portion of the thin preparation had VAS ratings similar to those of the 37.5 g thick preparation. Consuming the 37.5 g portion of fortified daal would provide 6.9 mg Fe/day to girls in a community-based effectiveness study. This would meet ~86% and ~46% of the Recommended Dietary Allowance (RDA) for Fe for girls aged 9-13 and 14-18 years, respectively.

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.

Do Faba Bean Genotypes Carrying Different Zero-Tannin Genes (zt1 and zt2) Differ in Phenolic Profiles?

Faba bean is a cool season grain legume that produces seeds with a high protein content. Seed coat tannins limit its use in food and feed. A low-tannin phenotype is controlled by either of two unlinked recessive genes zt1 and zt2. Liquid chromatography-mass spectrometry was used to characterize phenolic profiles of seed coat and flower tissue of three faba bean genotypes: CDC Snowdrop (zt1 gene), Disco/2 (zt2 gene), and ILB 938/2 (tannin-containing). For both tissues, clear differences in phenolic profiles of ILB 938/2 were observed in comparison to both low-tannin lines. Although seed coat phenolic profiles of zt1 and zt2 genotypes were similar, distinct differences were evident in flower tissue, suggesting that the gene action results in some different end products of the phenolic biosynthetic pathway. These distinctive compounds could be used as biochemical markers to distinguish between low-tannin phenotypes.

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.

Polyphenol profile comparisons of seed coats of five pulse crops using a semi-quantitative liquid chromatography-mass spectrometric method.

INTRODUCTION: Pulse crops are nutritious and therefore widely grown. Pulse seed coats are typically discarded, despite their high content of polyphenols that are known for their antioxidant properties and health benefits. A better understanding of polyphenol diversity and biochemical pathways will ultimately provide insight into how polyphenols are linked to health benefits, which will help to better utilise these seed coats. OBJECTIVES: To explore polyphenol profiles among seed coats of diverse genotypes of five pulse crops using a targeted liquid chromatography mass spectrometry (LC-MS) method. METHODS: Four genotypes of each of common bean, chickpea, pea, lentil and faba bean seed coats were selected for analysis. Following extraction, polyphenols were quantified using LC-MS. RESULTS: An LC-MS method was developed to quantify 98 polyphenols from 13 different classes in 30 min. The low-tannin seed coats had the lowest concentrations of all polyphenols. Chickpea and pea seed coats had the most similar polyphenolic profiles. The black common bean showed the most diverse seed coat polyphenol profile, including several anthocyanins not detected in any of the other seed coats. CONCLUSION: The LC-MS method reported herein was used to show polyphenol diversity within several polyphenol classes among the pulse crop seed coats. Detected in all seed coats, flavonols and hydroxybenzoic acids appear well-conserved in the edible Fabaceae. The presence of anthocyanins, flavan-3-ols and proanthocyanins in the coloured seed coats suggests that unique divergent branches were introduced in the flavonoid biosynthetic pathway, possibly in response to environmental stressors.

Iron Fortification and Bioavailability of Chickpea (Cicer arietinum L.) Seeds and Flour.

Iron (Fe) deficiency is one of the most common nutritional disorders, and is mainly due to insufficient intake of bioavailable Fe. Chickpea (Cicer arietinum L.) was examined as a potential vehicle for Fe fortification. Fortificants (FeSO4·7H2O (ferrous sulfate hepta-hydrate), FeSO4·H2O (ferrous sulfate mono-hydrate) and NaFeEDTA (ethylenediaminetetraacetic acid iron (iii) sodium salt)) were applied by a spraying and drying method. At 2000 µg g-1 iron fortificant, the fortified split desi seeds (dal), desi flour and kabuli flour supplied 18-19 mg, 16-20 mg and 11-19 mg Fe per 100 g, respectively. The overall consumer acceptability using a nine-point hedonic scale for sensory evaluation demonstrated that NaFeEDTA-fortified cooked chickpea (soup and chapatti) scored the highest among the three fortificants. Lightness (L*), redness (a*) and yellowness (b*) of Fe-fortified products changed over time. However, no organoleptic changes occurred. Fe bioavailability was increased by 5.8-10.5, 15.3-25.0 and 4.8-9.0 ng ferritin mg-1 protein for cooked split desi seeds (soup), desi chapatti and kabuli chapatti, respectively, when prepared using Fe-fortified chickpea. Desi chapatti showed significantly higher Fe bioavailability than the other two. The increase in Fe concentration and bioavailability in fortified chickpea products demonstrated that these products could provide a significant proportion of the recommended daily Fe requirement.

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.

Polyphenolic Composition of Lentil Roots in Response to Infection by Aphanomyces euteiches.

Polyphenols comprise the largest group of plant secondary metabolites and have critical roles in plant physiology and response to the biotic and abiotic environment. Changes in the content of polyphenols in the root extracts and root tissues of wild (Lens ervoides) and cultivated (Lens culinaris) lentil genotypes were examined in response to infection by Aphanomyces euteiches using liquid chromatography mass spectrometry (LC-MS). Genotype, infection and their interaction determined the composition of polyphenols in lentil roots. The levels of several polyphenols were lower in the root extract of the low-tannin genotype L. culinaris ZT-4 compared to L. ervoides L01-827A. Kaempferol derivatives including kaempferol dirutinoside and kaempferol 3-robinoside 7-rhamnoside were more concentrated in the healthy root tissues of L. ervoides L01-827A than in L. culinaris genotypes. Infection increased the concentration of kaempferol, apigenin, and naringenin in the root tissues of all genotypes, but had no effect on some polyphenols in the low-tannin genotype L. culinaris ZT-4. The concentrations of apigenin, naringenin, apigenin 4-glucoside, naringenin7-rutinoside, diosmetin, and hesperetin 7-rutinoside were higher in the infected root tissues of L. ervoides L01-827A compared with the L. culinaris genotypes. Organic acids including coumaric acid, vanillic acid, 4-aminosalicylic acid, 4-hydroxybenzoic acid, and 3,4-dihydroxybenzoic acid effectively suppressed the in-vitro hyphal growth of A. euteiches. Some of these bioactive polyphenols were more concentrated in roots of L. ervoides L01-827A but were low to undetectable in ZT-4. This study shows that genotypic differences exist in the composition of root polyphenols in lentil, and is related to the response to infection caused by A. euteiches. Polyphenols, particularly the organic acid content could be useful for selection and breeding of lentil genotypes that are resistant to Aphanomyces root rot (ARR) disease.

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.

Flowering and Growth Responses of Cultivated Lentil and Wild Lens Germplasm toward the Differences in Red to Far-Red Ratio and Photosynthetically Active Radiation.

Understanding environmental responses of pulse crop species and their wild relatives will play an important role in developing genetic strategies for crop improvement in response to changes in climate. This study examined how cultivated lentil and wild Lens germplasm responded to different light environments, specifically differences in red/far-red ratio (R/FR) and photosynthetically active radiation (PAR). Three genotypes of each the seven Lens species were grown in environmentally controlled growth chambers equipped to provide light treatments consisting of different R/FR ratios and PAR values. Our results showed that overall, days to flower of Lens genotypes were mainly influenced by the R/FR induced light quality change but not by the PAR related light intensity change. The cultivated lentil (L. culinaris) showed consistent, accelerated flowering in response to the low R/FR light environment together with three wild lentil genotypes (L. orientalis IG 72611, L. tomentosus IG 72830, and L. ervoides IG 72815) while most wild lentil genotypes had reduced responses and flowering time was not significantly affected. The longest shoot length, longest internode length, and largest leaflet area were observed under the low R/FR low PAR environment for both cultivated and wild lentils. The distinctly different responses between flowering time and elongation under low R/FR conditions among wild Lens genotypes suggests discrete pathways controlling flowering and elongation, which are both components of shade avoidance responses. The yield and above-ground biomass of Lens genotypes were the highest under high R/FR high PAR conditions, intermediate under low R/FR low PAR conditions, and lowest under high R/FR low PAR light conditions. Three L. lamottei genotypes (IG 110809, IG 110810, and IG 110813) and one L. ervoides genotype (IG 72646) were less sensitive in their time to flower responses while maintaining similar yield, biomass, and harvest index across all three light environments; these are indications of better adaptability toward changes in light environment.