Boron Enrichment and Retention viaTraditional Leaf-Based Cooking Methods in Idly Preparation

Boron Enrichment and Retention viaTraditional Leaf-Based Cooking Methods in Idly Preparation | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Boron Enrichment and Retention via Traditional Leaf-Based Cooking Methods in Idly Preparation Pradeep Bhatu Patil, Ms. Shivani Padala, Ms. B. Tulja, Mr. Swapnil Deshmukh, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6928254/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Boron is a vital trace element essential for bone health, cognitive function, and metabolic processes. However, boron deficiency remains a global concern due to soil depletion, poor dietary intake, limited water boron content, and general unawareness. The World Health Organization acceptable daily intake of 1–13 mg/day, yet populations in several developed and developing countries often fail to meet even the minimum requirement. This study aimed to investigate the role of traditional leaf-based steaming practices in enhancing boron retention and enrichment in idly , a steamed rice cake. Five leaf types—Palash, Jackfruit, Banana, Badam, and Banyan—were analysed for boron baseline levels, leaching, retention, and transfer efficiency (enrichment) using Colorimetry and Inductively Coupled Plasma Mass Spectrometry. Leaves were subjected to hot water treatment (60°C for 15 minutes) to simulate blanching, followed by steaming in autoclave at 121 0 C for 15 minutes with one-sided and two-sided leaf contact configurations. Results indicated significant differences in boron dynamics among leaf types. Palash and Jackfruit leaves exhibited superior boron retention and transfer capabilities, while Banana and Badam leaves showed higher leaching. Seasonal variations, leaf maturity, regional geography, gravitational effects during steaming, and leaf orientation further influenced boron dynamics. Additionally, leaves exhibited dual roles—some enriched the idly , while others neutralized boron loss during steaming. The findings highlight the potential of traditional cooking practices in addressing dietary boron deficiency naturally and sustainably. Future studies should focus on optimizing steaming conditions, understanding leaf microstructures, and expanding research across diverse regions to ensure reproducible and practical dietary interventions for boron supplementation. Food Science & Technology Nutrition & Dietetics Boron Traditional Cooking using leaves Nutritional Enhancement Trace Elements Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Ayurveda, the 5,000-year-old Indian system of holistic medicine, emphasizes the importance of a balanced diet and natural food preparation techniques to maintain physical, mental, and spiritual well-being. In Ayurveda, food is considered medicine, and every aspect of food preparation, from ingredient selection to cooking methods, is viewed as a critical factor influencing health. Leaves from trees such as Palash, Banana, Jackfruit, Badam, and Banyan have traditionally been revered not just for their culinary utility but also for their medicinal and nutritional properties. Despite its growing recognition, dietary boron deficiency remains a significant but underexplored issue, especially in Indian dietary contexts. Soil depletion has led to lower boron content in crops, whereas cooking practices could also have an impact on diminishing the natural availability of this trace element [ 1 – 4 ]. Furthermore, while Western dietary patterns have been studied extensively for boron intake, limited data exist on traditional Indian culinary practices and their contribution to boron consumption. Traditional cooking methods, particularly the use of natural leaf-based steaming techniques, have been practiced for generations but remain scientifically under-investigated for their role in micronutrient retention and enrichment [ 5 ]. The culinary use of leaves in steaming idly is deeply rooted in Indian culture, offering sustainability and potential nutritional benefits. This study investigates boron retention, leaching, and enrichment in idly prepared using traditional leaf-based steaming methods. Boron plays a crucial role in bone health, hormonal balance, and anti-inflammatory processes. Understanding its behavior during cooking can guide the development of dietary recommendations and public health policies, promoting traditional cooking methods as an accessible strategy to improve boron intake in boron-deficient populations[ 6 ]. Rationale for the Study Despite boron's critical role in bone health, immunity, and hormonal balance, it remains one of the most overlooked micronutrients in Indian diets. Its transient presence in the body—neither stored in significant amounts nor formally classified as an essential nutrient—has pushed it into the shadows of nutritional research. Yet, boron exerts a profound influence on hidden hunger , impacting the absorption and homeostasis of key micronutrients like zinc, iron, and magnesium. This silent regulator plays a pivotal role in preventing stunting (via zinc), combating anemia (via iron), and addressing hypothyroidism (via magnesium). Beyond these effects, boron's role in hormonal balance suggests potential links to obesity outcomes. Ignoring boron in dietary strategies isn't just an oversight—it's a missed opportunity to tackle some of the most persistent public health challenges. Boron deficiency remains an invisible crisis, overlooked in both research and public health strategies. Western studies highlight insufficient boron intake due to modern food processing and cooking techniques. In India, the situation is likely even graver, with soil depletion caused by heavy rainfall and declining organic carbon levels exacerbating the deficiency. Yet, a forgotten treasure lies in our traditional culinary wisdom—leaf-based steaming methods. Could our ancestors have unknowingly devised a solution to this micronutrient shortfall? This study dares to ask that question, hypothesizing that steaming food in leaves like Palash, Banana, and Jackfruit may naturally enrich it with boron. Using cutting-edge techniques like ICP-MS, we aim to uncover whether these age-old practices hold the key to addressing a modern nutritional dilemma, potentially revolutionizing dietary guidelines and public health policies. Materials and Methods The study aimed to investigate the dynamics of boron estimation, enrichment, retention, and leaching during the steaming of idly using traditional leaves as molds. The selected leaves included Palash, Jackfruit, Banana, Badam, and Banyan. The experimental design was divided into two key phases: Phase 1 (Fig. 1a), which focused on boron leaching from raw and hot-water-treated leaves, and Phase 2 (Fig. 1b), which analyzed boron retention and transfer in idly steamed with one-sided (OSLSI) and two-sided leaf (TSLSI) configurations. The study employed Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and colorimetry-based boron quantification methods to ensure high accuracy and reproducibility. Phase -I The initial phase involved the collection and preparation of leaf samples. Fresh leaves were sourced personally across three districts (Amalner District in Maharashtra, Nizamabad and Wanaparthy Districts in Telangana) with an emphasis on quality and authenticity (Fig. 2). Each type of leaf was thoroughly washed using boron-free distilled water to eliminate surface contaminants. Post-washing, the leaves were gently air-dried under controlled conditions and stored at + 4°C until further analysis (Fig. 3). To ensure uniformity across samples, circular discs of 9.5 cm in diameter were carefully cut from each leaf using a standardized mold (Fig. 3). These discs were then divided into two groups: raw leaves and hot-water-treated leaves. The treatment group was subjected to immersion in 60°C water for 15 minutes to mimic blanching or serving hot food on leaves. These conditions are commonly used in Indian traditional food preparation. Both raw and treated leaves were subsequently analyzed for boron content and leaching dynamics. Phase -II The second phase involved the preparation of idly , a traditional South Indian steamed rice cake, using the selected leaves as steaming molds. The idly batter was prepared in a standardized ratio of 3:1 (fermented rice to black gram flour) and allowed to ferment for 8 hours at room temperature to achieve the required microbial activity for optimal texture and flavor. The batter was then carefully portioned and steamed using two distinct configurations. In the one-sided steaming method, the batter was placed directly on a single leaf surface, while in the two-sided steaming method, the batter was sandwiched between two leaves to ensure maximal leaf-to-batter interaction. For baseline comparison, a control group was maintained where the idly batter was steamed without any leaves. After steaming, the idly samples and the corresponding leaves underwent a series of processing steps for boron estimation. Both the steamed leaves and idly samples were oven-dried at 70–80°C until a constant weight was achieved. The dried samples were then transferred to a muffle furnace (Model:AI-106, Manufactured by Accuma, India) and ashed at 600°C for 5–6 hours until white ash remained. The resulting ash was dissolved in 50 mM hydrochloric acid (HCl), vortexed thoroughly, and centrifuged at 8000 rpm for 15 minutes to ensure complete extraction of boron from the ash matrix. The resulting supernatants were collected and stored in boron-free tubes for analysis. To quantify boron content accurately, two analytical methods were employed: colorimetry and ICP-MS (Inductively Coupled Plasma Mass Spectrometry). The preliminary screening of boron content was performed using a colorimetric boron estimation kit (Quantichrome ™ Phosphate Assay Kit, Bioassay Systems, USA) and absorbance readings were recorded at 550 nm using a multi-mode spectrophotometric reader. However, for higher precision and trace-level detection, ICP-MS analysis was performed using the Agilent 7900 ICP-MS system. Calibration standards ranging from 0.01 µg/L to 500 µg/L were used to ensure the accuracy of boron quantification. The data were expressed in multiple formats, including mg/g of ash, µg/cm² of leaf area, and mg/100 g wet weight of idly , allowing for cross-comparison across different phases and configurations. To assess boron leaching during the hot-water treatment, leaf samples were analyzed for boron content both before and after immersion in 60°C water for 15 minutes. The boron content in the leachate (water after immersion) was also quantified to determine the extent of boron migration from the leaves to the water medium. Similarly, steamed idly samples were analyzed to quantify boron enrichment across the OSLSI and TSLSI approach of steaming configurations. The leaves placed on upper and lower surfaces of idly (for two-sided leaf contact) were also examined independently to evaluate any differences in boron retention patterns. To ensure experimental rigor, multiple quality control measures were implemented. Separate gloves and instruments were used for each leaf type to avoid cross-contamination. All instruments, including the ICP-MS system, spectrophotometric reader, and muffle furnace, were calibrated at regular intervals to maintain analytical accuracy. Each experiment was performed in triplicate, with a total of 36 replicates per treatment group to ensure reproducibility and statistical robustness. The study’s primary outcome measures included boron content in raw leaves, boron leaching in hot water, boron retention in steamed idly , and fold-change analysis comparing control and experimental groups. Additional parameters, such as percent boron retention and migration efficiency, were derived from the primary data to provide a comprehensive understanding of boron dynamics. Comparative analyses were performed across steaming configurations, leaf types, and pre- versus post-treatment conditions. Leaf sourcing was performed sustainably, and all waste materials were disposed of responsibly in compliance with environmental safety protocols. The equipment used for sample preparation and analysis included multi-mode spectrophotometric readers, Agilent 7900 ICP-MS systems, muffle furnaces, analytical balances, centrifuges, hot air ovens, and stainless-steel steamers. All laboratory procedures were carried out under sterile and controlled conditions to prevent sample contamination. In summary, this study followed a meticulously designed workflow encompassing leaf preparation, hot-water treatment, idly steaming, sample drying, ashing, boron extraction, quantification, and statistical analysis. The integration of ICP-MS and colorimetry provided a dual-layered analytical approach, ensuring both accuracy and reproducibility. By investigating boron leaching, retention, and transfer under different experimental conditions, this study provides valuable insights into the potential of traditional leaves as natural boron carriers in steamed food products. Statistical analysis played a crucial role in validating the findings and ensuring data reliability. Brown-Forsythe and Welch’s ANOVA tests were performed to identify significant differences between groups. Post-hoc pairwise comparisons were conducted using Sidak’s multiple comparisons test to evaluate mean differences across treatments. All statistical analyses were carried out using GraphPad Prism (v9.0), and p-values < 0.05 were considered statistically significant. Graphical representations in the form of bar plots were used using estimated values, extrapolated values and percent increase or fold change to visualize the results effectively. Error bars denoting standard deviation (SD) of the mean were included in each graph to highlight variability across replicates. Results This study meticulously evaluated boron dynamics in five traditional leaves (Palash, Jackfruit, Badam, Banana, and Banyan) used for steaming idly, focusing on boron leaching, retention, enrichment, and the impact of one-sided (OSLSI) versus two-sided (TSLSI) leaf steaming configurations. Quantitative assessments through ICP-MS and colorimetry provided precise insights into boron mobility, retention, and transfer across various cooking treatments. Boron Content in Raw and Hot-Water Treated Leaves (Fig. 4a) Boron Content in Raw and Hot-Water Treated Leaves (Fig. 4a) In raw leaf samples, Palash exhibited the highest boron content (1.90 mg/g ash), followed by Banana (1.17 mg/g ash), Badam (0.96 mg/g ash), and Jackfruit (0.79 mg/g ash), while Banyan recorded the lowest boron levels (0.25 mg/g ash). After hot-water treatment at 60°C for 15 minutes, a significant reduction in boron content was observed across all leaves. Palash retained 1.34 mg/g Leaf ash, reflecting a 29.5% loss which is a significant variance, suggesting structural differences influencing boron retention post-treatment. Boron Leaching in Water During Hot-Water Treatment (Fig. 4b&4c) Boron leaching into water during hot-water treatment revealed striking differences among leaves. Badam demonstrated the highest leaching (13.50 µg/g), followed by Banana (12.34 µg/g) and Jackfruit (9.80 µg/g). Palash (2.07 µg/g) and Banyan (0.79 µg/g) exhibited significantly lower leaching levels. When analyzed per cm² of leaf surface, Banana showed the highest boron leaching efficiency (0.35 µg/cm²), followed by Badam (0.29 µg/cm²) and Jackfruit (0.22 µg/cm²). Palash (0.08 µg/cm²) and Banyan (0.02 µg/cm²) had minimal leaching, suggesting that Palash leaf structure effectively retained boron even under thermal stress. These differences were statistically significant (p < 0.001), emphasizing the variability in leaf permeability and boron-binding capacity. Boron Levels of Un-steamed Leaves (Control Leaves – Fig. 4d) The analysis of boron levels in un-steamed control leaves revealed distinct variations across different leaf types, highlighting their inherent boron content before any thermal processing. Palash leaves exhibited the highest boron levels at 1.90 mg/g of leaf ash, indicating their natural richness in boron. Following this, Banana leaves recorded 1.17 mg/g, while Badam leaves displayed slightly lower levels at 0.96 mg/g. Jackfruit leaves contained 0.79 mg/g, suggesting moderate boron accumulation. In contrast, Banyan leaves showed the lowest boron levels at 0.25 mg/g, reflecting their limited boron uptake capacity. Boron Retention in Leaves Post-Idly (Single sided - SSLSI) Preparation (Fig. 4e) The analysis of boron retention in leaves post-idly preparation under Single-Sided Leaf Steamed Idly (SSLSI) conditions revealed distinct differences across Palash, Banana, Badam, and Jackfruit leaves. Palash leaves exhibited the highest retention efficiency, with boron content decreasing minimally from 1.90 mg/g ash in control leaves to 1.75 mg/g ash after steaming, reflecting a 7.89% loss. Banana leaves followed, showing a reduction from 1.17 mg/g ash to 1.05 mg/g ash (10.25% loss), while Badam leaves retained boron moderately, dropping from 0.96 mg/g ash to 0.89 mg/g ash (7.29% loss). Jackfruit leaves demonstrated the highest loss, with boron levels declining from 0.79 mg/g ash to 0.59 mg/g ash, marking a 25.3% decrease. Boron Retention in Leaves Post-Idly (double/Two sided - TSLSI) Preparation (Fig. 4f) After idly preparation, boron retention varied significantly across the upper and lower leaf surfaces. Palash leaves showed higher boron retention on the upper surface (72.81 µg/70.7 cm²) compared to the lower surface (56.28 µg/70.7 cm²) reflecting a 1.29-fold difference. In contrast, Banana leaves displayed marginal differences between upper (66.27 µg/70.7 cm²) and lower surfaces (70.65 µg/70.7cm 2 ), indicating a more uniform retention pattern. Jackfruit leaves retained 47.88 µg on the lower side and 41.07 µg on the upper side, while Banyan leaves showed the lowest retention levels with 27.41 µg on the upper and 17.54 µg on the lower surface. The significant differences were found in palash leaf (p < 0.0001) suggest that gravitational pull and leaf structure strongly influence boron retention during steaming. Boron Enrichment in Idly (OSLSI) Across Leaf Types (Fig. 5a – 5c) The boron content in one-sided leaf steamed idly (OSLSI), analyzed through colorimetry, demonstrated notable differences across the five leaf types. Jackfruit leaf idly exhibited the highest boron levels at 0.46 mg/g of idly ash, indicating relatively effective boron transfer despite the single-sided steaming configuration. Palash leaf idly followed with a boron content of 0.41 mg/g, suggesting moderate enrichment efficiency. Banyan leaf idly and Badam leaf idly showed identical boron levels at 0.40 mg/g, while Banana leaf idly registered slightly lower boron content at 0.39 mg/g. In comparison, the control idly, prepared without leaf interaction, had the lowest boron content at 0.37 mg/g. Boron Enrichment in Idly (TSLSI vs OSLSI) Across Leaf Types (Fig. 5d) In TSLSI idly samples, Jackfruit leaves demonstrated the highest boron enrichment (12.19 mg/g ash), followed by Palash (11.26 mg/g ash), Banana (7.73 mg/g ash), Banyan (7.20 mg/g ash), and Badam (6.53 mg/g ash). Compared to the control idly (5.97 mg/g ash), Jackfruit leaves exhibited a 2.04-fold increase in boron content, while Palash showed a 1.88-fold increase. In one-sided leaf steaming (OSLSI), the boron content in idly was notably lower across all leaf types. Jackfruit idly (0.46 mg/g ash) and Palash idly (0.41 mg/g ash) recorded the highest boron content, while Banana (0.39 mg/g ash) and Banyan (0.40 mg/g ash) showed marginally lower values. These results suggest that TSLSI is significantly more effective in boron enrichment compared to OSLSI, attributed to the enhanced surface contact and controlled diffusion during two-sided steaming. Boron Levels in Idly Batter vs. Steamed Idly (Fig. 5e) Boron content in raw idly batter was higher (2.16 mg/100 g wet weight) compared to control idly post-steaming (1.38 mg/100 g wet weight), indicating a 36.1% loss during steaming. This suggests gravitational leaching during steaming contributes significantly to boron loss. The decrease in boron content highlights the need for optimizing steaming conditions to minimize such losses. Methodology Comparison: ICP-MS vs. Colorimetry (Fig. 5f) Comparing ICP-MS and colorimetry for boron quantification revealed a notable discrepancy. Control idly analysed through ICP-MS showed 5.97 mg/g ash, whereas colorimetry detected only 0.37 mg/g ash, reflecting a 16.13-fold difference. Similarly, for Palash idly, ICP-MS detected 11.26 mg/g ash, while colorimetry suggested a far lower value of 0.41 mg/g ash. This disparity underscores the higher sensitivity and accuracy of ICP-MS in detecting boron content, particularly in complex matrices like idly ash. Extrapolation of ICP-MS values in 100gram of wet weight of Idly The extrapolated boron levels in idly (mg/100 g wet weight) revealed significant variation across leaf types, emphasizing the differential boron enrichment capacity of each leaf during steaming. Jackfruit leaf idly demonstrated the highest boron content at 3.50 mg/100 g wet weight, showcasing its superior efficiency in boron transfer during the steaming process. Palash leaf idly followed with a boron content of 2.67 mg/100 g wet weight, indicating notable boron enrichment. Banyan leaf idly and Badam leaf idly displayed moderate boron levels at 1.76 mg/100 g wet weight and 1.97 mg/100 g wet weight, respectively. Banana leaf idly showed comparatively lower boron content at 1.55 mg/100 g wet weight, indicating weaker boron retention and transfer efficiency. In contrast, the control idly, prepared without leaf interaction, registered the lowest boron content at 1.38 mg/100 g wet weight. Percentage increase in Boron content after steaming Idly with Two-sided Leaves with respect to control Idly (Fig. 6b) The percent increase in boron content in idly compared to control idly, as analysed via ICP-MS, highlights substantial differences in boron enrichment efficiency across different leaf types. Palash leaf idly demonstrated the highest percentage increase at 193%, underscoring its exceptional ability to transfer and retain boron during steaming. Jackfruit leaf idly followed with a significant increase of 153%, further confirming its strong boron enrichment potential. In contrast, Badam leaf idly and Banyan leaf idly showed moderate increases of 43% and 27%, respectively, reflecting limited efficiency in boron retention and transfer. Banana leaf idly displayed the lowest percentage increase at only 12%, indicating minimal contribution to boron enrichment during steaming. Fold-Change and Percent Retention Across Leaf Types (Fig. 6c and 6d) The percent increase in boron content in idly compared to idly batter, analyzed through ICP-MS, reveals critical insights into the efficiency of leaf-based steaming in preserving and enriching boron levels. The results indicate that steaming idly inherently results in boron losses of approximately 56%, primarily due to leaching and gravitational effects. However, the TSLSI idly approach demonstrated a protective effect in mitigating these losses, with Palash and Jackfruit leaves emerging as the most effective barriers. Palash leaf idly not only prevented boron losses but achieved an additional 23% increase in boron content compared to the batter. Similarly, Jackfruit leaf idly exhibited an impressive 62% increase, underscoring its exceptional boron retention and enrichment capabilities during steaming. In contrast, Banana (-7.1%), Banyan (-8.1%), and Badam (-9.1%) leaves failed to completely counteract boron losses. However, it is noteworthy that these leaves managed to recover approximately 47–48% of the boron losses, indicating a partial but limited protective effect during the steaming process. Gravitational Influence on Boron Retention in Leaves Gravitational effects were observed to influence boron leaching patterns in two-sided steaming configurations. Lower leaf surfaces consistently exhibited higher leaching compared to upper surfaces, especially in leaves like Badam (72.81 µg vs. 30.67 µg) and Palash (72.81 µg vs. 56.28 µg). In contrast, Banana leaves showed minimal variation, suggesting leaf porosity and density also mediate gravitational effects. Discussion The present study provides a comprehensive evaluation of boron leaching, retention, and enrichment across diverse leaf types subjected to thermal conditions during idly preparation. Through a combination of ICP-MS and colorimetric analysis, we explored the nuanced interplay between thermal gradients, leaf structural integrity, and boron mobility, offering critical insights into optimizing traditional food preparation practices for enhanced boron bioavailability. Temperature-Dependent Boron Dynamics: A Tale of Two Thermal Realms Boron leaching displayed a stark contrast between mild thermal exposure at 60°C and intense autoclave steaming at 121°C (steam flows unidirectionally and horizontally from one door toward the opposite side door). Palash leaves, which exhibited minimal boron leaching during hot water treatment (2.07 µg/g wet weight), demonstrated significant boron enrichment (2.67 mg/100 g wet weight) under high-pressure steaming. In contrast, Badam and Banana leaves showed higher boron losses (13.50 µg/g and 12.34 µg/g, respectively) at 60°C but failed to match the enrichment efficiency of Palash and Jackfruit leaves under high-temperature steaming. The fundamental difference lies in the temperature gradient: at 60°C, boron diffusion occurs at the leaf surface, while at 121°C, the high-pressure environment disrupts cellular barriers, enabling deeper boron mobilization into the idly matrix. This thermal sensitivity underscores the importance of cooking conditions in maximizing nutrient transfer. One-Sided vs. Two-Sided Leaf Steaming: A Structural Dilemma The comparison between OSLSI and TSLSI idly sample approaches revealed critical differences in boron enrichment. While ICP-MS detected significantly higher boron levels in TSLSI (e.g., Palash idly: 11.26 mg/g ash), colorimetric extrapolation from OSLSI fell short of capturing this enrichment accurately (6.61 mg/g ash). This discrepancy highlights not only the sensitivity of ICP-MS but also the structural advantage of two-sided steaming. The increased leaf-batter interface in TSLSI allows for superior boron transfer, particularly in structurally robust leaves such as Palash and Jackfruit. Conversely, thinner and more porous leaves like Banyan and Banana demonstrated limited benefits from double-sided steaming, likely due to gravitational boron leaching. Gravitational Forces: The Invisible Hand Governing Boron Leaching The gravitational effect emerged as a silent yet powerful factor dictating boron leaching patterns. Across all leaves, the lower surface consistently exhibited higher boron leaching compared to the upper surface, as seen in Badam (lower: 30.67 µg, upper: 83.36 µg) and Palash (lower: 56.28 µg, upper: 72.81 µg). The pronounced differences indicate gravitational pull as a key driver of nutrient losses during steaming, particularly in thinner and more porous leaves. Interestingly, steamed idly batter appeared to act as a partial sealing agent, reducing boron losses by blocking surface pores during thermal exposure. This phenomenon emphasizes the critical role of both leaf structure and batter consistency in nutrient retention. Leaf-Specific Variability: Not All Leaves Are Created Equal Leaf type played a decisive role in boron mobility during steaming. Jackfruit and Palash leaves consistently emerged as the most effective nutrient carriers, with boron levels reaching 3.50 mg/100 g wet weight and 2.67 mg/100 g wet weight, respectively. Their structural robustness and favourable thermal response likely contributed to higher boron bioavailability in the final idly product. In contrast, Banana and Badam leaves showed higher boron leaching during water treatment but failed to translate these initial boron reserves into effective idly enrichment. Banyan leaves, despite their minimal boron losses at 60°C, demonstrated limited enrichment efficiency, suggesting intrinsic structural or chemical limitations. Boron Loss from Batter to Idly: The Missing Nutrient One of the study's most intriguing findings was the boron loss observed during the transition from idly batter to steamed idly. While the batter retained higher boron levels (2.16 mg/100 g wet weight), steaming resulted in a 56.5% reduction (1.38 mg/100 g wet weight in control idly). This significant loss indicates gravitational leaching as a dominant contributor to boron depletion. The downward movement of condensed steam likely accelerates boron mobility away from the idly matrix, further compounded by the structural characteristics of the leaf used. Microstructural Insights: A Window into Boron Retention The variability observed across leaf types calls for a deeper understanding of their microstructural attributes. Thin, porous leaves like Banana and Banyan demonstrated higher gravitational leaching, while thicker leaves with robust cuticles, such as Palash and Jackfruit, displayed superior boron retention. Scanning Electron Microscopy (SEM) and advanced spectroscopy techniques could provide critical insights into the pore density, cuticular thickness, and boron-binding affinities, offering potential avenues for engineering leaf surfaces to optimize boron retention during thermal exposure. Nutritional Implications: Bridging Tradition and Micronutrient Deficiencies From a nutritional standpoint, the results highlight the immense potential of using leaves as natural boron fortifiers in traditional steamed foods. Due to lack of adequate data, there's no established Recommended Dietary Allowance (RDA) for boron. The World Health Organization (WHO) suggests an acceptable safe range of intake for adults is 1–13 mg per day. The Tolerable Upper Intake Level (UL) for boron is 20 mg per day for adults. The consumption of 100 g of idly steamed using Jackfruit or Palash leaves could contribute significantly towards meeting daily requirements. This approach not only aligns with culturally relevant dietary habits but also presents an affordable alternative to commercial boron supplements often used in sports nutrition and clinical interventions. Analytical Sensitivity: ICP-MS vs. Colorimetry The 16.13-fold difference observed between ICP-MS and colorimetry underscores the limitations of conventional colorimetric assays in capturing boron concentrations accurately, especially at lower ranges. While colorimetry serves as a useful screening tool, ICP-MS remains the gold standard for precise boron quantification in complex matrices like idly. Future studies aiming to standardize analytical protocols must account for these methodological disparities to ensure reproducibility and accuracy in boron analysis. Future Directions: Towards Precision Nutritional Culinary Practices Several avenues for future research emerge from these findings. Advanced imaging techniques, including SEM and confocal microscopy, could unravel the microstructural dynamics governing boron mobility during steaming. Further, chemical profiling of boron-binding compounds in leaves may help identify molecular interactions responsible for boron retention or leaching. Optimizing steaming parameters—time, temperature, and leaf orientation—could enhance boron transfer efficiency. Additionally, exploring region-specific leaf varieties and their seasonal variations could help create a robust database for informed leaf selection in culinary practices. This study bridges traditional culinary wisdom with contemporary nutritional science, showcasing how a simple steaming process using leaves can address boron deficiencies in a sustainable and culturally relevant manner. Palash and Jackfruit leaves emerge as superior candidates for boron enrichment, demonstrating remarkable resilience to thermal stress and gravitational losses. Meanwhile, the interplay of temperature, structural properties, and analytical precision highlights the need for a holistic approach to optimizing boron retention during food preparation. In a world grappling with micronutrient deficiencies and rising reliance on synthetic supplements, these findings present an innovative, cost-effective strategy to enhance boron intake through time-honoured culinary practices. As we unlock the molecular secrets of leaf-boron interactions, we move closer to a future where food is not only a source of sustenance but also a powerful tool for precision nutrition. Ayurveda and Modern Science: Bridging Tradition and Nutrition Ayurvedic texts highlight the unique ‘Prakriti’ (inherent nature) of different leaves, categorizing them based on their taste ( Rasa ), potency ( Veerya ), and post-digestive effect ( Vipaka ). For example, Banana leaves are considered cooling ( Sheeta Veerya ) and purifying, making them ideal for serving hot, steamed foods [ 7 ]. Palash leaves, on the other hand, are known for their detoxifying and antimicrobial properties, believed to cleanse impurities from food during cooking [ 8 , 9 ]. Similarly, Jackfruit leaves are said to promote digestive health and nutrient absorption when used in food preparation [ 10 ]. Ayurveda also emphasizes the concept of ‘Samskara’, which refers to the transformation of food properties through cooking techniques. Steaming food in natural leaves is seen as an act of infusing the food with the medicinal essence of the leaves, creating a synergistic relationship between the cooking medium and the food. While numerous minerals and bioactive compounds, such as phenolics and alkaloids, may infuse into food during preparation, this manuscript specifically emphasizes boron infusion. Traditional cooking methods utilizing leaves are explored, highlighting our hypothesis that boron levels in food could play a pivotal role in nutrient absorption and homeostasis[ 11 ]. Boron, classified as a metalloid with an atomic number of 5, is recognized as a possibly essential trace element that plays a crucial role in various biological processes. Historically, it has been underappreciated in the context of human nutrition; however, recent research has highlighted its significant impact on bone health, cognitive function, hormone regulation, and immune response[ 12 – 19 ]. Naturally occurring in a variety of plant-based foods, including fruits, vegetables, nuts, and legumes, boron is found in two primary forms: inorganic borates and organic sugar-borate esters. Although this research has focused on total boron levels, further studies are required to find out type of boron present in leaves and idly. Upon ingestion, boron is metabolized into boric acid within the gastrointestinal tract, exhibiting an absorption efficiency of approximately 85–90%, with the majority being excreted through urine[ 20 , 21 ]. Despite the growing acknowledgment of its physiological importance, the World Health Organization (WHO) has not established a specific Recommended Daily Allowance (RDA) for boron, instead proposing an acceptable safe range of 1–13 mg/day for adults[ 22 ], additionally few countries are debating on the increase in acceptable daily range is required. Initially regarded as a non-essential trace element, boron has since been reclassified by scientists as potentially essential due to emerging evidence of its critical physiological roles. However, since boron is not stored in the body and is rapidly excreted, its full spectrum of importance remains an area of ongoing scientific exploration. Ayurveda has long recognized leaves as natural bioenhancers, preserving nutrients and infusing food with therapeutic vitality. The practice of steaming food on leaves aligns with Ayurvedic wisdom, minimizing nutrient loss and maximizing bioavailability—a principle now validated by science. Palash and Jackfruit leaves excel in boron retention and controlled release, while Banana and Badam leaves, despite their popularity, show significant boron leaching, highlighting the critical role of leaf structure in nutrient dynamics. Seasonality and geography are equally influential, with Sharad Ritu (autumn) leaves traditionally prized for peak nutrient content—a fact now supported by modern research. Beyond nutrition, leaf-based cooking reflects environmental stewardship, as leaves are biodegradable, locally sourced, and ecologically sustainable, contributing to a balanced nutrient cycle. Boron is a trace mineral or essential faciliatory nutrient? Boron is known to play a pivotal role in calcium metabolism, brain function, insulin regulation, and hormone activity [ 23 , 24 ]. Studies suggest that boron helps regulate key metabolic pathways, including those associated with steroid hormones (e.g., oestrogen and vitamin D), immune responses, and oxidative stress management [ 25 , 26 ]. Boron's deficiency has been linked to a range of health complications, including reduced bone density, impaired cognitive function, increased urinary calcium excretion, and elevated inflammatory markers[ 27 ]. Despite its physiological importance, boron remains one of the least studied micronutrients, with dietary intake data primarily limited to Western populations [ 28 , 29 ]. Surveys estimate median boron intake in adults to range between 0.87 to 1.35 mg/day, with higher values observed in vegetarians due to their plant-rich diets [ 30 , 31 ]. However, in India, where traditional diets heavily rely on plant-based foods, there is a lack of concrete data on dietary boron intake and its bioavailability in cooked food samples. This knowledge gap is particularly concerning given the decline in soil boron levels, which directly impacts the boron content of locally grown crops. More than physical nourishment, Ayurveda views leaf-steamed food as energetically therapeutic, with Prana —a vital life force—transferring into food during steaming. Scientifically, leaves serve as efficient carriers of boron, a micronutrient essential for human health. While boron has often been described as a "cosmic particle" due to its unique role in both plant and human physiology, a more fitting term would be the "Chaitanya particle" , signifying its life-giving and facilitating properties. In plants, boron functions as a flushing agent, mobilizing and transporting other micronutrients, while in humans, it regulates hormones, strengthens bones, and supports cognitive function. Interestingly, recent research highlights boron as an "interkingdom chemical communication tool." Studies using pig models reveal that boron acts as a mediator, allowing animals to maintain health even in the presence of pathogenic bacteria—a sort of negotiating agent between different biological kingdoms. In this context, greater boron loss during cooking might not always represent a failure; instead, it symbolizes boron's dynamic role as a facilitator and communicator across biological systems. The more boron mobilizes, the more it serves its interkingdom purpose—a silent battle won for maintaining biological harmony. Establishing a Recommended Dietary Allowance (RDA) for boron remains a significant challenge due to the limited availability of comprehensive and consistent human data. However, the development of provisional intake recommendations, supported by emerging evidence and region-specific dietary data, is a necessary step toward formulating a sustainable, evidence-based RDA. Accordingly, a preliminary draft proposing such intake ranges has been prepared and is included as supplementary material to facilitate further discussion and research-based validation. This synergy between Ayurvedic wisdom and scientific validation underscores the potential of leaf-based steaming in addressing boron deficiency—a rising global concern. Policymakers must prioritize education, awareness campaigns, and incentives for adopting these traditional culinary practices. Future research integrating Ayurvedic principles with advanced analytical tools will further optimize these sustainable and culturally resonant interventions, transforming them into scalable solutions for global health and nutrition security. Conclusion This study highlights the potential of traditional leaf-based culinary practices in addressing boron deficiency, a critical yet often overlooked micronutrient gap. Leaves like Palash and Jackfruit demonstrated superior boron retention during idly preparation, influenced by factors such as temperature, leaf microstructure, and steaming duration. These findings suggest that natural leaves can serve as sustainable and cost-effective alternatives to synthetic boron supplements. Policymakers should incorporate these insights into public health strategies, including awareness campaigns targeting schools, health workers, and culinary professionals. Government programs, such as mid-day meal schemes, could adopt these practices to ensure nutrient-rich meals. Standardization protocols and certification systems are essential to maintain hygiene and consistency in leaf-based cooking methods. Continued research is necessary to optimize boron transfer efficiency and validate regional variations in leaf properties for broader implementation. Abbreviations OSLSI One–sided leaf steaming approach on Idly making TSLSI Two–sided leaf steaming approach on Idly making ICP MS–inductive coupled plasma–mass spectrophotometry Declarations Acknowledgments: The authors thank the Indian Council of Medical Research-National Institute of Nutrition (ICMR-NIN) and Osmania University College of Technology (OUCT) for their support and resources. Gratitude is extended to our mentors and collaborators for their guidance and expertise. We acknowledge the Director of ICMR-NIN for the intramural grant (No. ST/1/Intra-Pro-Allot/2023-24) and thank Mr. Saida Naik, Mr. Subash Tatikayala, and Mr. Mohan Kumar for their assistance during experimentation. I also thank Director ICMR-NIV, Pune for allowing me to continue working on this manuscript while I am working at ICMR-NIV. References Singh MV (2008) Micronutrient deficiencies in crops and soils in India. Micronutrient deficiencies in global crop production. Springer, pp 93–125 Rego TJ et al (2007) Widespread deficiencies of sulfur, boron, and zinc in Indian semi-arid tropical soils: On-farm crop responses. 30(10):1569–1583 Rerkasem B, Jamjod S (2004) Boron deficiency in wheat: a review. Field Crops Res 89(2):173–186 Ahmed S, Hossain MB (1997) The problem of boron deficiency in crop production in Bangladesh , in Boron in Soils and Plants: Proceedings of the International Symposium on Boron in Soils and Plants held at Chiang Mai, Thailand, 7–11 September, 1997 , R.W. Bell and B. Rerkasem, Editors. Springer Netherlands: Dordrecht. pp. 1–5 Mustafa M et al (2012) Looking back to the past: revival of traditional food packaging . in 2nd Regional Conference on Local Knowledge (KEARIFAN TEMPATAN). Jerejak Island Pizzorno L (2015) Nothing boring about boron 14(4):35 Patil S, PA C (2017) Paradiya parpati kalpa: A mercurial preparation of ayurveda for medicinal purpose. Kumar A et al (2023) A SYSTEMIC REVIEW ON PALASH (BUTEA MONOSPERMA). 9(2): pp. 871–899 Saroj P, Shah N Butea monosperma (Palash)–Its Ethnobotanical Knowledge, Phytochemical Studies, Pharmacological Aspects and Future Prospects. Sew Y-S et al (2020) Effects of Fermented Jackfruit Leaf and Pulp Beverages on Gut Microbiota and Faecal Short Chain Fatty Acids Content in Sprague-Dawley Rats. 29(3):22542–22550 Patil P, Jaleel A, Qadri S (2023) The interplay of 'twin nutrients' (vitamin D and boron) in an allegedly understated anthropometric status (stunting, anaemia, malnutrition) that refuses to improve - missing links, missed opportunities and actionable lessons from the field of veterinary medicine. Research Square Pietrzkowski Z et al (2014) Short-term efficacy of calcium fructoborate on subjects with knee discomfort: a comparative, double-blind, placebo-controlled clinical study. Clin Interv Aging 9:895–899 Nielsen FH, Eckhert CD (2020) Boron Adv Nutr 11(2):461–462 Nielsen FH (2014) Update on human health effects of boron. J Trace Elem Med Biol 28(4):383–387 Naghii MR et al (2011) Comparative effects of daily and weekly boron supplementation on plasma steroid hormones and proinflammatory cytokines. J Trace Elem Med Biol 25(1):54–58 Uluisik I, Karakaya HC, Koc A (2018) The importance of boron in biological systems. J Trace Elem Med Biol 45:156–162 Khaliq H, Juming Z, Ke-Mei P (2018) The Physiological Role of Boron on Health. Biol Trace Elem Res 186(1):31–51 Hunt CD (2012) Dietary boron: progress in establishing essential roles in human physiology. J Trace Elem Med Biol 26(2–3):157–160 Kobylewski SE et al (2017) Activation of the EIF2α/ATF4 and ATF6 Pathways in DU-145 Cells by Boric Acid at the Concentration Reported in Men at the US Mean Boron Intake. Biol Trace Elem Res 176(2):278–293 Moseman RF (1994) J.E.h.p. Chem Dispos boron Anim Hum 102(suppl 7):113–117 Kot F (2015) S.J.B.s.p. Boron Environ 1:33 NIH. Boron - Fact Sheet for Health Professionals (2022) ; Available from: https://ods.od.nih.gov/factsheets/Boron-HealthProfessional/#en2 Khaliq H, Juming Z (2018) J.B.t.e.r. Ke-Mei. physiological role boron health 186:31–51 Dubey P, Thakur V, Chattopadhyay MJN (2020) Role minerals trace Elem diabetes insulin Resist 12(6):1864 Estevez-Fregoso E et al (2023) Effects of boron-containing compounds on liposoluble hormone functions. 11(2):84 Estevez-Fregoso E et al (2023) Eff Boron-Containing Compd Liposoluble Hormone Funct 11(2):84 Nielsen F, Biology (2014) Update Hum health Eff boron 28(4):383–387 Meacham S et al (2010) Boron in human health: evidence for dietary recommendations and public policies. 3(1) Nielsen F (2012) H.J.P.k.i.n., Manganese, molybdenum, boron, chromium, and other trace elements. : pp. 586–607 Russell RB, Cousins JL, Dunn RJ, Ferland JT, Hambidge G, Lynch KM, Penland S, Ross JG, Stoeker AC, Suttie BJ, Turnlund JW, West JR, Zlotkin KP (2001) S., Dietary Reference Intakes for Vitamin A, Vitamin K, Arsenic, Boron, Chromium, Copper, Iodine, Iron, Manganese, Molybdenum, Nickel, Silicon, Vanadium, and Zinc. , ed. I. o.M.U.P.o . Micronutrients. Washington (DC): National Academies Press (US) Buck GMC, Dourson R, Foster ML, Goyer P, Chairman RA, Howe P, Co-Rapporteur); Luoto R, Nielsen FH, Price CJ, Woods WG (1998) In: Organization WH (ed) Environmental Health Criteria 204 - Boron. World Health Organization, Editor Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6928254","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":473446168,"identity":"239c022a-d1c3-4ff5-a784-5d40d351c834","order_by":0,"name":"Pradeep Bhatu Patil","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4UlEQVRIiWNgGAWjYJACCTB5mIHxAQPDAWI0MMO1MBuQqOUAA5sEUVoMjvcfvF25454833HeY9U8NXfk+BmYHz66gU/LmcPMlmfPFBvOPMyXdpvn2DNjyQY2Y+McPFokZySzSTa2JTBuOMxjdpuH7XDihgM8bNJ4tcx/DNZiD9JSzPOPCC38EsxgLYkgLcy8bcRo4Uk2tmw8k5AM9Euy5Ny+w8aSzQT8wsZ+8OHNxh0Jtn3nzx788ObbYTl+9uaHj/FpAQPGBhDJw8DEA6KZCSlH1sL4gxjVo2AUjIJRMOIAAEmvTfjwl+h9AAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0001-7638-5558","institution":"ICMR-NIN","correspondingAuthor":true,"prefix":"","firstName":"Pradeep","middleName":"Bhatu","lastName":"Patil","suffix":""},{"id":473446169,"identity":"f677b1bb-9ead-4fbc-be3e-f4fd19f94916","order_by":1,"name":"Ms. Shivani Padala","email":"","orcid":"","institution":"PG dessertation Student","correspondingAuthor":false,"prefix":"Ms.","firstName":"Shivani","middleName":"","lastName":"Padala","suffix":""},{"id":473446170,"identity":"697f190c-1e5c-47f0-8f0c-64920612f495","order_by":2,"name":"Ms. B. 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In Ayurveda, food is considered medicine, and every aspect of food preparation, from ingredient selection to cooking methods, is viewed as a critical factor influencing health. Leaves from trees such as Palash, Banana, Jackfruit, Badam, and Banyan have traditionally been revered not just for their culinary utility but also for their medicinal and nutritional properties.\u003c/p\u003e \u003cp\u003eDespite its growing recognition, dietary boron deficiency remains a significant but underexplored issue, especially in Indian dietary contexts. Soil depletion has led to lower boron content in crops, whereas cooking practices could also have an impact on diminishing the natural availability of this trace element [\u003cspan additionalcitationids=\"CR2 CR3\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Furthermore, while Western dietary patterns have been studied extensively for boron intake, limited data exist on traditional Indian culinary practices and their contribution to boron consumption. Traditional cooking methods, particularly the use of natural leaf-based steaming techniques, have been practiced for generations but remain scientifically under-investigated for their role in micronutrient retention and enrichment [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe culinary use of leaves in steaming idly is deeply rooted in Indian culture, offering sustainability and potential nutritional benefits. This study investigates boron retention, leaching, and enrichment in idly prepared using traditional leaf-based steaming methods. Boron plays a crucial role in bone health, hormonal balance, and anti-inflammatory processes. Understanding its behavior during cooking can guide the development of dietary recommendations and public health policies, promoting traditional cooking methods as an accessible strategy to improve boron intake in boron-deficient populations[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eRationale for the Study\u003c/h3\u003e\n\u003cp\u003eDespite boron's critical role in bone health, immunity, and hormonal balance, it remains one of the most overlooked micronutrients in Indian diets. Its transient presence in the body\u0026mdash;neither stored in significant amounts nor formally classified as an essential nutrient\u0026mdash;has pushed it into the shadows of nutritional research. Yet, boron exerts a profound influence on \u003cem\u003ehidden hunger\u003c/em\u003e, impacting the absorption and homeostasis of key micronutrients like zinc, iron, and magnesium. This silent regulator plays a pivotal role in preventing \u003cem\u003estunting\u003c/em\u003e (via zinc), combating \u003cem\u003eanemia\u003c/em\u003e (via iron), and addressing \u003cem\u003ehypothyroidism\u003c/em\u003e (via magnesium). Beyond these effects, boron's role in hormonal balance suggests potential links to \u003cem\u003eobesity\u003c/em\u003e outcomes. Ignoring boron in dietary strategies isn't just an oversight\u0026mdash;it's a missed opportunity to tackle some of the most persistent public health challenges. Boron deficiency remains an invisible crisis, overlooked in both research and public health strategies. Western studies highlight insufficient boron intake due to modern food processing and cooking techniques. In India, the situation is likely even graver, with soil depletion caused by heavy rainfall and declining organic carbon levels exacerbating the deficiency. Yet, a forgotten treasure lies in our traditional culinary wisdom\u0026mdash;leaf-based steaming methods. Could our ancestors have unknowingly devised a solution to this micronutrient shortfall? This study dares to ask that question, hypothesizing that steaming food in leaves like Palash, Banana, and Jackfruit may naturally enrich it with boron. Using cutting-edge techniques like ICP-MS, we aim to uncover whether these age-old practices hold the key to addressing a modern nutritional dilemma, potentially revolutionizing dietary guidelines and public health policies.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003eThe study aimed to investigate the dynamics of boron estimation, enrichment, retention, and leaching during the steaming of \u003cem\u003eidly\u003c/em\u003e using traditional leaves as molds. The selected leaves included Palash, Jackfruit, Banana, Badam, and Banyan. The experimental design was divided into two key phases: Phase 1 (Fig.\u0026nbsp;1a), which focused on boron leaching from raw and hot-water-treated leaves, and Phase 2 (Fig.\u0026nbsp;1b), which analyzed boron retention and transfer in \u003cem\u003eidly\u003c/em\u003e steamed with one-sided (OSLSI) and two-sided leaf (TSLSI) configurations. The study employed Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and colorimetry-based boron quantification methods to ensure high accuracy and reproducibility.\u003c/p\u003e\n\u003ch3\u003ePhase -I\u003c/h3\u003e\n\u003cp\u003eThe initial phase involved the collection and preparation of leaf samples. Fresh leaves were sourced personally across three districts (Amalner District in Maharashtra, Nizamabad and Wanaparthy Districts in Telangana) with an emphasis on quality and authenticity (Fig.\u0026nbsp;2). Each type of leaf was thoroughly washed using boron-free distilled water to eliminate surface contaminants. Post-washing, the leaves were gently air-dried under controlled conditions and stored at +\u0026thinsp;4\u0026deg;C until further analysis (Fig.\u0026nbsp;3). To ensure uniformity across samples, circular discs of 9.5 cm in diameter were carefully cut from each leaf using a standardized mold (Fig.\u0026nbsp;3). These discs were then divided into two groups: raw leaves and hot-water-treated leaves. The treatment group was subjected to immersion in 60\u0026deg;C water for 15 minutes to mimic blanching or serving hot food on leaves. These conditions are commonly used in Indian traditional food preparation. Both raw and treated leaves were subsequently analyzed for boron content and leaching dynamics.\u003c/p\u003e\n\u003ch3\u003ePhase -II\u003c/h3\u003e\n\u003cp\u003eThe second phase involved the preparation of \u003cem\u003eidly\u003c/em\u003e, a traditional South Indian steamed rice cake, using the selected leaves as steaming molds. The \u003cem\u003eidly\u003c/em\u003e batter was prepared in a standardized ratio of 3:1 (fermented rice to black gram flour) and allowed to ferment for 8 hours at room temperature to achieve the required microbial activity for optimal texture and flavor. The batter was then carefully portioned and steamed using two distinct configurations. In the one-sided steaming method, the batter was placed directly on a single leaf surface, while in the two-sided steaming method, the batter was sandwiched between two leaves to ensure maximal leaf-to-batter interaction. For baseline comparison, a control group was maintained where the \u003cem\u003eidly\u003c/em\u003e batter was steamed without any leaves.\u003c/p\u003e \u003cp\u003eAfter steaming, the \u003cem\u003eidly\u003c/em\u003e samples and the corresponding leaves underwent a series of processing steps for boron estimation. Both the steamed leaves and \u003cem\u003eidly\u003c/em\u003e samples were oven-dried at 70\u0026ndash;80\u0026deg;C until a constant weight was achieved. The dried samples were then transferred to a muffle furnace (Model:AI-106, Manufactured by Accuma, India) and ashed at 600\u0026deg;C for 5\u0026ndash;6 hours until white ash remained. The resulting ash was dissolved in 50 mM hydrochloric acid (HCl), vortexed thoroughly, and centrifuged at 8000 rpm for 15 minutes to ensure complete extraction of boron from the ash matrix. The resulting supernatants were collected and stored in boron-free tubes for analysis.\u003c/p\u003e \u003cp\u003eTo quantify boron content accurately, two analytical methods were employed: colorimetry and ICP-MS (Inductively Coupled Plasma Mass Spectrometry). The preliminary screening of boron content was performed using a colorimetric boron estimation kit (Quantichrome\u003csup\u003e\u0026trade;\u003c/sup\u003e Phosphate Assay Kit, Bioassay Systems, USA) and absorbance readings were recorded at 550 nm using a multi-mode spectrophotometric reader. However, for higher precision and trace-level detection, ICP-MS analysis was performed using the Agilent 7900 ICP-MS system. Calibration standards ranging from 0.01 \u0026micro;g/L to 500 \u0026micro;g/L were used to ensure the accuracy of boron quantification. The data were expressed in multiple formats, including mg/g of ash, \u0026micro;g/cm\u0026sup2; of leaf area, and mg/100 g wet weight of \u003cem\u003eidly\u003c/em\u003e, allowing for cross-comparison across different phases and configurations.\u003c/p\u003e \u003cp\u003eTo assess boron leaching during the hot-water treatment, leaf samples were analyzed for boron content both before and after immersion in 60\u0026deg;C water for 15 minutes. The boron content in the leachate (water after immersion) was also quantified to determine the extent of boron migration from the leaves to the water medium. Similarly, steamed \u003cem\u003eidly\u003c/em\u003e samples were analyzed to quantify boron enrichment across the OSLSI and TSLSI approach of steaming configurations. The leaves placed on upper and lower surfaces of idly (for two-sided leaf contact) were also examined independently to evaluate any differences in boron retention patterns.\u003c/p\u003e \u003cp\u003eTo ensure experimental rigor, multiple quality control measures were implemented. Separate gloves and instruments were used for each leaf type to avoid cross-contamination. All instruments, including the ICP-MS system, spectrophotometric reader, and muffle furnace, were calibrated at regular intervals to maintain analytical accuracy. Each experiment was performed in triplicate, with a total of 36 replicates per treatment group to ensure reproducibility and statistical robustness.\u003c/p\u003e \u003cp\u003eThe study\u0026rsquo;s primary outcome measures included boron content in raw leaves, boron leaching in hot water, boron retention in steamed \u003cem\u003eidly\u003c/em\u003e, and fold-change analysis comparing control and experimental groups. Additional parameters, such as percent boron retention and migration efficiency, were derived from the primary data to provide a comprehensive understanding of boron dynamics. Comparative analyses were performed across steaming configurations, leaf types, and pre- versus post-treatment conditions.\u003c/p\u003e \u003cp\u003eLeaf sourcing was performed sustainably, and all waste materials were disposed of responsibly in compliance with environmental safety protocols. The equipment used for sample preparation and analysis included multi-mode spectrophotometric readers, Agilent 7900 ICP-MS systems, muffle furnaces, analytical balances, centrifuges, hot air ovens, and stainless-steel steamers. All laboratory procedures were carried out under sterile and controlled conditions to prevent sample contamination.\u003c/p\u003e \u003cp\u003eIn summary, this study followed a meticulously designed workflow encompassing leaf preparation, hot-water treatment, \u003cem\u003eidly\u003c/em\u003e steaming, sample drying, ashing, boron extraction, quantification, and statistical analysis. The integration of ICP-MS and colorimetry provided a dual-layered analytical approach, ensuring both accuracy and reproducibility. By investigating boron leaching, retention, and transfer under different experimental conditions, this study provides valuable insights into the potential of traditional leaves as natural boron carriers in steamed food products.\u003c/p\u003e \u003cp\u003eStatistical analysis played a crucial role in validating the findings and ensuring data reliability. Brown-Forsythe and Welch\u0026rsquo;s ANOVA tests were performed to identify significant differences between groups. Post-hoc pairwise comparisons were conducted using Sidak\u0026rsquo;s multiple comparisons test to evaluate mean differences across treatments. All statistical analyses were carried out using GraphPad Prism (v9.0), and p-values\u0026thinsp;\u0026lt;\u0026thinsp;0.05 were considered statistically significant. Graphical representations in the form of bar plots were used using estimated values, extrapolated values and percent increase or fold change to visualize the results effectively. Error bars denoting standard deviation (SD) of the mean were included in each graph to highlight variability across replicates.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eThis study meticulously evaluated boron dynamics in five traditional leaves (Palash, Jackfruit, Badam, Banana, and Banyan) used for steaming idly, focusing on boron leaching, retention, enrichment, and the impact of one-sided (OSLSI) versus two-sided (TSLSI) leaf steaming configurations. Quantitative assessments through ICP-MS and colorimetry provided precise insights into boron mobility, retention, and transfer across various cooking treatments.\u003c/p\u003e\n\u003ch3\u003eBoron Content in Raw and Hot-Water Treated Leaves (Fig. 4a)\u003c/h3\u003e\n\u003cdiv class=\"Heading\"\u003eBoron Content in Raw and Hot-Water Treated Leaves (Fig.\u0026nbsp;4a)\u003c/div\u003e \u003cp\u003eIn raw leaf samples, Palash exhibited the highest boron content (1.90 mg/g ash), followed by Banana (1.17 mg/g ash), Badam (0.96 mg/g ash), and Jackfruit (0.79 mg/g ash), while Banyan recorded the lowest boron levels (0.25 mg/g ash). After hot-water treatment at 60\u0026deg;C for 15 minutes, a significant reduction in boron content was observed across all leaves. Palash retained 1.34 mg/g Leaf ash, reflecting a 29.5% loss which is a significant variance, suggesting structural differences influencing boron retention post-treatment.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eBoron Leaching in Water During Hot-Water Treatment (Fig.\u0026nbsp;4b\u0026amp;4c)\u003c/h2\u003e \u003cp\u003eBoron leaching into water during hot-water treatment revealed striking differences among leaves. Badam demonstrated the highest leaching (13.50 \u0026micro;g/g), followed by Banana (12.34 \u0026micro;g/g) and Jackfruit (9.80 \u0026micro;g/g). Palash (2.07 \u0026micro;g/g) and Banyan (0.79 \u0026micro;g/g) exhibited significantly lower leaching levels. When analyzed per cm\u0026sup2; of leaf surface, Banana showed the highest boron leaching efficiency (0.35 \u0026micro;g/cm\u0026sup2;), followed by Badam (0.29 \u0026micro;g/cm\u0026sup2;) and Jackfruit (0.22 \u0026micro;g/cm\u0026sup2;). Palash (0.08 \u0026micro;g/cm\u0026sup2;) and Banyan (0.02 \u0026micro;g/cm\u0026sup2;) had minimal leaching, suggesting that Palash leaf structure effectively retained boron even under thermal stress. These differences were statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), emphasizing the variability in leaf permeability and boron-binding capacity.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eBoron Levels of Un-steamed Leaves (Control Leaves – Fig. 4d)\u003c/h3\u003e\n\u003cp\u003eThe analysis of boron levels in un-steamed control leaves revealed distinct variations across different leaf types, highlighting their inherent boron content before any thermal processing. Palash leaves exhibited the highest boron levels at 1.90 mg/g of leaf ash, indicating their natural richness in boron. Following this, Banana leaves recorded 1.17 mg/g, while Badam leaves displayed slightly lower levels at 0.96 mg/g. Jackfruit leaves contained 0.79 mg/g, suggesting moderate boron accumulation. In contrast, Banyan leaves showed the lowest boron levels at 0.25 mg/g, reflecting their limited boron uptake capacity.\u003c/p\u003e\n\u003ch3\u003eBoron Retention in Leaves Post-Idly (Single sided - SSLSI) Preparation (Fig. 4e)\u003c/h3\u003e\n\u003cp\u003eThe analysis of boron retention in leaves post-idly preparation under Single-Sided Leaf Steamed Idly (SSLSI) conditions revealed distinct differences across Palash, Banana, Badam, and Jackfruit leaves. Palash leaves exhibited the highest retention efficiency, with boron content decreasing minimally from 1.90 mg/g ash in control leaves to 1.75 mg/g ash after steaming, reflecting a 7.89% loss. Banana leaves followed, showing a reduction from 1.17 mg/g ash to 1.05 mg/g ash (10.25% loss), while Badam leaves retained boron moderately, dropping from 0.96 mg/g ash to 0.89 mg/g ash (7.29% loss). Jackfruit leaves demonstrated the highest loss, with boron levels declining from 0.79 mg/g ash to 0.59 mg/g ash, marking a 25.3% decrease.\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eBoron Retention in Leaves Post-Idly (double/Two sided - TSLSI) Preparation (Fig.\u0026nbsp;4f)\u003c/h2\u003e \u003cp\u003eAfter idly preparation, boron retention varied significantly across the upper and lower leaf surfaces. Palash leaves showed higher boron retention on the upper surface (72.81 \u0026micro;g/70.7 cm\u0026sup2;) compared to the lower surface (56.28 \u0026micro;g/70.7 cm\u0026sup2;) reflecting a 1.29-fold difference. In contrast, Banana leaves displayed marginal differences between upper (66.27 \u0026micro;g/70.7 cm\u0026sup2;) and lower surfaces (70.65 \u0026micro;g/70.7cm\u003csup\u003e2\u003c/sup\u003e), indicating a more uniform retention pattern. Jackfruit leaves retained 47.88 \u0026micro;g on the lower side and 41.07 \u0026micro;g on the upper side, while Banyan leaves showed the lowest retention levels with 27.41 \u0026micro;g on the upper and 17.54 \u0026micro;g on the lower surface. The significant differences were found in palash leaf (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) suggest that gravitational pull and leaf structure strongly influence boron retention during steaming.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eBoron Enrichment in Idly (OSLSI) Across Leaf Types (Fig.\u0026nbsp;5a \u0026ndash; 5c)\u003c/h2\u003e \u003cp\u003eThe boron content in one-sided leaf steamed idly (OSLSI), analyzed through colorimetry, demonstrated notable differences across the five leaf types. Jackfruit leaf idly exhibited the highest boron levels at 0.46 mg/g of idly ash, indicating relatively effective boron transfer despite the single-sided steaming configuration. Palash leaf idly followed with a boron content of 0.41 mg/g, suggesting moderate enrichment efficiency. Banyan leaf idly and Badam leaf idly showed identical boron levels at 0.40 mg/g, while Banana leaf idly registered slightly lower boron content at 0.39 mg/g. In comparison, the control idly, prepared without leaf interaction, had the lowest boron content at 0.37 mg/g.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eBoron Enrichment in Idly (TSLSI vs OSLSI) Across Leaf Types (Fig.\u0026nbsp;5d)\u003c/h2\u003e \u003cp\u003eIn TSLSI idly samples, Jackfruit leaves demonstrated the highest boron enrichment (12.19 mg/g ash), followed by Palash (11.26 mg/g ash), Banana (7.73 mg/g ash), Banyan (7.20 mg/g ash), and Badam (6.53 mg/g ash). Compared to the control idly (5.97 mg/g ash), Jackfruit leaves exhibited a 2.04-fold increase in boron content, while Palash showed a 1.88-fold increase.\u003c/p\u003e \u003cp\u003eIn one-sided leaf steaming (OSLSI), the boron content in idly was notably lower across all leaf types. Jackfruit idly (0.46 mg/g ash) and Palash idly (0.41 mg/g ash) recorded the highest boron content, while Banana (0.39 mg/g ash) and Banyan (0.40 mg/g ash) showed marginally lower values. These results suggest that TSLSI is significantly more effective in boron enrichment compared to OSLSI, attributed to the enhanced surface contact and controlled diffusion during two-sided steaming.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eBoron Levels in Idly Batter vs. Steamed Idly (Fig.\u0026nbsp;5e)\u003c/h2\u003e \u003cp\u003eBoron content in raw idly batter was higher (2.16 mg/100 g wet weight) compared to control idly post-steaming (1.38 mg/100 g wet weight), indicating a 36.1% loss during steaming. This suggests gravitational leaching during steaming contributes significantly to boron loss. The decrease in boron content highlights the need for optimizing steaming conditions to minimize such losses.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eMethodology Comparison: ICP-MS vs. Colorimetry (Fig.\u0026nbsp;5f)\u003c/h2\u003e \u003cp\u003eComparing ICP-MS and colorimetry for boron quantification revealed a notable discrepancy. Control idly analysed through ICP-MS showed 5.97 mg/g ash, whereas colorimetry detected only 0.37 mg/g ash, reflecting a 16.13-fold difference. Similarly, for Palash idly, ICP-MS detected 11.26 mg/g ash, while colorimetry suggested a far lower value of 0.41 mg/g ash. This disparity underscores the higher sensitivity and accuracy of ICP-MS in detecting boron content, particularly in complex matrices like idly ash.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eExtrapolation of ICP-MS values in 100gram of wet weight of Idly\u003c/h2\u003e \u003cp\u003eThe extrapolated boron levels in idly (mg/100 g wet weight) revealed significant variation across leaf types, emphasizing the differential boron enrichment capacity of each leaf during steaming. Jackfruit leaf idly demonstrated the highest boron content at 3.50 mg/100 g wet weight, showcasing its superior efficiency in boron transfer during the steaming process. Palash leaf idly followed with a boron content of 2.67 mg/100 g wet weight, indicating notable boron enrichment.\u003c/p\u003e \u003cp\u003eBanyan leaf idly and Badam leaf idly displayed moderate boron levels at 1.76 mg/100 g wet weight and 1.97 mg/100 g wet weight, respectively. Banana leaf idly showed comparatively lower boron content at 1.55 mg/100 g wet weight, indicating weaker boron retention and transfer efficiency. In contrast, the control idly, prepared without leaf interaction, registered the lowest boron content at 1.38 mg/100 g wet weight.\u003c/p\u003e \u003cp\u003e \u003cb\u003ePercentage increase in Boron content after steaming Idly with Two-sided Leaves with respect to control Idly (Fig.\u0026nbsp;6b)\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThe percent increase in boron content in idly compared to control idly, as analysed via ICP-MS, highlights substantial differences in boron enrichment efficiency across different leaf types. Palash leaf idly demonstrated the highest percentage increase at 193%, underscoring its exceptional ability to transfer and retain boron during steaming. Jackfruit leaf idly followed with a significant increase of 153%, further confirming its strong boron enrichment potential.\u003c/p\u003e \u003cp\u003eIn contrast, Badam leaf idly and Banyan leaf idly showed moderate increases of 43% and 27%, respectively, reflecting limited efficiency in boron retention and transfer. Banana leaf idly displayed the lowest percentage increase at only 12%, indicating minimal contribution to boron enrichment during steaming.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eFold-Change and Percent Retention Across Leaf Types (Fig.\u0026nbsp;6c and 6d)\u003c/h2\u003e \u003cp\u003eThe percent increase in boron content in idly compared to idly batter, analyzed through ICP-MS, reveals critical insights into the efficiency of leaf-based steaming in preserving and enriching boron levels. The results indicate that steaming idly inherently results in boron losses of approximately 56%, primarily due to leaching and gravitational effects. However, the TSLSI idly approach demonstrated a protective effect in mitigating these losses, with Palash and Jackfruit leaves emerging as the most effective barriers.\u003c/p\u003e \u003cp\u003ePalash leaf idly not only prevented boron losses but achieved an additional 23% increase in boron content compared to the batter. Similarly, Jackfruit leaf idly exhibited an impressive 62% increase, underscoring its exceptional boron retention and enrichment capabilities during steaming.\u003c/p\u003e \u003cp\u003eIn contrast, Banana (-7.1%), Banyan (-8.1%), and Badam (-9.1%) leaves failed to completely counteract boron losses. However, it is noteworthy that these leaves managed to recover approximately 47\u0026ndash;48% of the boron losses, indicating a partial but limited protective effect during the steaming process.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eGravitational Influence on Boron Retention in Leaves\u003c/h2\u003e \u003cp\u003eGravitational effects were observed to influence boron leaching patterns in two-sided steaming configurations. Lower leaf surfaces consistently exhibited higher leaching compared to upper surfaces, especially in leaves like Badam (72.81 \u0026micro;g vs. 30.67 \u0026micro;g) and Palash (72.81 \u0026micro;g vs. 56.28 \u0026micro;g). In contrast, Banana leaves showed minimal variation, suggesting leaf porosity and density also mediate gravitational effects.\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe present study provides a comprehensive evaluation of boron leaching, retention, and enrichment across diverse leaf types subjected to thermal conditions during idly preparation. Through a combination of ICP-MS and colorimetric analysis, we explored the nuanced interplay between thermal gradients, leaf structural integrity, and boron mobility, offering critical insights into optimizing traditional food preparation practices for enhanced boron bioavailability.\u003c/p\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eTemperature-Dependent Boron Dynamics: A Tale of Two Thermal Realms\u003c/h2\u003e \u003cp\u003eBoron leaching displayed a stark contrast between mild thermal exposure at 60\u0026deg;C and intense autoclave steaming at 121\u0026deg;C (steam flows unidirectionally and horizontally from one door toward the opposite side door). Palash leaves, which exhibited minimal boron leaching during hot water treatment (2.07 \u0026micro;g/g wet weight), demonstrated significant boron enrichment (2.67 mg/100 g wet weight) under high-pressure steaming. In contrast, Badam and Banana leaves showed higher boron losses (13.50 \u0026micro;g/g and 12.34 \u0026micro;g/g, respectively) at 60\u0026deg;C but failed to match the enrichment efficiency of Palash and Jackfruit leaves under high-temperature steaming. The fundamental difference lies in the temperature gradient: at 60\u0026deg;C, boron diffusion occurs at the leaf surface, while at 121\u0026deg;C, the high-pressure environment disrupts cellular barriers, enabling deeper boron mobilization into the idly matrix. This thermal sensitivity underscores the importance of cooking conditions in maximizing nutrient transfer.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003eOne-Sided vs. Two-Sided Leaf Steaming: A Structural Dilemma\u003c/h2\u003e \u003cp\u003eThe comparison between OSLSI and TSLSI idly sample approaches revealed critical differences in boron enrichment. While ICP-MS detected significantly higher boron levels in TSLSI (e.g., Palash idly: 11.26 mg/g ash), colorimetric extrapolation from OSLSI fell short of capturing this enrichment accurately (6.61 mg/g ash). This discrepancy highlights not only the sensitivity of ICP-MS but also the structural advantage of two-sided steaming. The increased leaf-batter interface in TSLSI allows for superior boron transfer, particularly in structurally robust leaves such as Palash and Jackfruit. Conversely, thinner and more porous leaves like Banyan and Banana demonstrated limited benefits from double-sided steaming, likely due to gravitational boron leaching.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003eGravitational Forces: The Invisible Hand Governing Boron Leaching\u003c/h2\u003e \u003cp\u003eThe gravitational effect emerged as a silent yet powerful factor dictating boron leaching patterns. Across all leaves, the lower surface consistently exhibited higher boron leaching compared to the upper surface, as seen in Badam (lower: 30.67 \u0026micro;g, upper: 83.36 \u0026micro;g) and Palash (lower: 56.28 \u0026micro;g, upper: 72.81 \u0026micro;g). The pronounced differences indicate gravitational pull as a key driver of nutrient losses during steaming, particularly in thinner and more porous leaves. Interestingly, steamed idly batter appeared to act as a partial sealing agent, reducing boron losses by blocking surface pores during thermal exposure. This phenomenon emphasizes the critical role of both leaf structure and batter consistency in nutrient retention.\u003c/p\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003eLeaf-Specific Variability: Not All Leaves Are Created Equal\u003c/h2\u003e \u003cp\u003eLeaf type played a decisive role in boron mobility during steaming. Jackfruit and Palash leaves consistently emerged as the most effective nutrient carriers, with boron levels reaching 3.50 mg/100 g wet weight and 2.67 mg/100 g wet weight, respectively. Their structural robustness and favourable thermal response likely contributed to higher boron bioavailability in the final idly product. In contrast, Banana and Badam leaves showed higher boron leaching during water treatment but failed to translate these initial boron reserves into effective idly enrichment. Banyan leaves, despite their minimal boron losses at 60\u0026deg;C, demonstrated limited enrichment efficiency, suggesting intrinsic structural or chemical limitations.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003eBoron Loss from Batter to Idly: The Missing Nutrient\u003c/h2\u003e \u003cp\u003eOne of the study's most intriguing findings was the boron loss observed during the transition from idly batter to steamed idly. While the batter retained higher boron levels (2.16 mg/100 g wet weight), steaming resulted in a 56.5% reduction (1.38 mg/100 g wet weight in control idly). This significant loss indicates gravitational leaching as a dominant contributor to boron depletion. The downward movement of condensed steam likely accelerates boron mobility away from the idly matrix, further compounded by the structural characteristics of the leaf used.\u003c/p\u003e \u003cdiv id=\"Sec25\" class=\"Section3\"\u003e \u003ch2\u003eMicrostructural Insights: A Window into Boron Retention\u003c/h2\u003e \u003cp\u003eThe variability observed across leaf types calls for a deeper understanding of their microstructural attributes. Thin, porous leaves like Banana and Banyan demonstrated higher gravitational leaching, while thicker leaves with robust cuticles, such as Palash and Jackfruit, displayed superior boron retention. Scanning Electron Microscopy (SEM) and advanced spectroscopy techniques could provide critical insights into the pore density, cuticular thickness, and boron-binding affinities, offering potential avenues for engineering leaf surfaces to optimize boron retention during thermal exposure.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec26\" class=\"Section3\"\u003e \u003ch2\u003eNutritional Implications: Bridging Tradition and Micronutrient Deficiencies\u003c/h2\u003e \u003cp\u003eFrom a nutritional standpoint, the results highlight the immense potential of using leaves as natural boron fortifiers in traditional steamed foods. Due to lack of adequate data, there's no established Recommended Dietary Allowance (RDA) for boron. The World Health Organization (WHO) suggests an acceptable safe range of intake for adults is 1\u0026ndash;13 mg per day. The Tolerable Upper Intake Level (UL) for boron is 20 mg per day for adults. The consumption of 100 g of idly steamed using Jackfruit or Palash leaves could contribute significantly towards meeting daily requirements. This approach not only aligns with culturally relevant dietary habits but also presents an affordable alternative to commercial boron supplements often used in sports nutrition and clinical interventions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec27\" class=\"Section3\"\u003e \u003ch2\u003eAnalytical Sensitivity: ICP-MS vs. Colorimetry\u003c/h2\u003e \u003cp\u003eThe 16.13-fold difference observed between ICP-MS and colorimetry underscores the limitations of conventional colorimetric assays in capturing boron concentrations accurately, especially at lower ranges. While colorimetry serves as a useful screening tool, ICP-MS remains the gold standard for precise boron quantification in complex matrices like idly. Future studies aiming to standardize analytical protocols must account for these methodological disparities to ensure reproducibility and accuracy in boron analysis.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec28\" class=\"Section2\"\u003e \u003ch2\u003eFuture Directions: Towards Precision Nutritional Culinary Practices\u003c/h2\u003e \u003cp\u003eSeveral avenues for future research emerge from these findings. Advanced imaging techniques, including SEM and confocal microscopy, could unravel the microstructural dynamics governing boron mobility during steaming. Further, chemical profiling of boron-binding compounds in leaves may help identify molecular interactions responsible for boron retention or leaching. Optimizing steaming parameters\u0026mdash;time, temperature, and leaf orientation\u0026mdash;could enhance boron transfer efficiency. Additionally, exploring region-specific leaf varieties and their seasonal variations could help create a robust database for informed leaf selection in culinary practices. This study bridges traditional culinary wisdom with contemporary nutritional science, showcasing how a simple steaming process using leaves can address boron deficiencies in a sustainable and culturally relevant manner. Palash and Jackfruit leaves emerge as superior candidates for boron enrichment, demonstrating remarkable resilience to thermal stress and gravitational losses. Meanwhile, the interplay of temperature, structural properties, and analytical precision highlights the need for a holistic approach to optimizing boron retention during food preparation. In a world grappling with micronutrient deficiencies and rising reliance on synthetic supplements, these findings present an innovative, cost-effective strategy to enhance boron intake through time-honoured culinary practices. As we unlock the molecular secrets of leaf-boron interactions, we move closer to a future where food is not only a source of sustenance but also a powerful tool for precision nutrition.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec29\" class=\"Section2\"\u003e \u003ch2\u003eAyurveda and Modern Science: Bridging Tradition and Nutrition\u003c/h2\u003e \u003cp\u003eAyurvedic texts highlight the unique \u0026lsquo;Prakriti\u0026rsquo; (inherent nature) of different leaves, categorizing them based on their taste (\u003cem\u003eRasa\u003c/em\u003e), potency (\u003cem\u003eVeerya\u003c/em\u003e), and post-digestive effect (\u003cem\u003eVipaka\u003c/em\u003e). For example, Banana leaves are considered cooling (\u003cem\u003eSheeta Veerya\u003c/em\u003e) and purifying, making them ideal for serving hot, steamed foods [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Palash leaves, on the other hand, are known for their detoxifying and antimicrobial properties, believed to cleanse impurities from food during cooking [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Similarly, Jackfruit leaves are said to promote digestive health and nutrient absorption when used in food preparation [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Ayurveda also emphasizes the concept of \u0026lsquo;Samskara\u0026rsquo;, which refers to the transformation of food properties through cooking techniques. Steaming food in natural leaves is seen as an act of infusing the food with the medicinal essence of the leaves, creating a synergistic relationship between the cooking medium and the food. While numerous minerals and bioactive compounds, such as phenolics and alkaloids, may infuse into food during preparation, this manuscript specifically emphasizes boron infusion. Traditional cooking methods utilizing leaves are explored, highlighting our hypothesis that boron levels in food could play a pivotal role in nutrient absorption and homeostasis[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eBoron, classified as a metalloid with an atomic number of 5, is recognized as a possibly essential trace element that plays a crucial role in various biological processes. Historically, it has been underappreciated in the context of human nutrition; however, recent research has highlighted its significant impact on bone health, cognitive function, hormone regulation, and immune response[\u003cspan additionalcitationids=\"CR13 CR14 CR15 CR16 CR17 CR18\" citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Naturally occurring in a variety of plant-based foods, including fruits, vegetables, nuts, and legumes, boron is found in two primary forms: inorganic borates and organic sugar-borate esters. Although this research has focused on total boron levels, further studies are required to find out type of boron present in leaves and idly. Upon ingestion, boron is metabolized into boric acid within the gastrointestinal tract, exhibiting an absorption efficiency of approximately 85\u0026ndash;90%, with the majority being excreted through urine[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Despite the growing acknowledgment of its physiological importance, the World Health Organization (WHO) has not established a specific Recommended Daily Allowance (RDA) for boron, instead proposing an acceptable safe range of 1\u0026ndash;13 mg/day for adults[\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], additionally few countries are debating on the increase in acceptable daily range is required. Initially regarded as a non-essential trace element, boron has since been reclassified by scientists as \u003cem\u003epotentially essential\u003c/em\u003e due to emerging evidence of its critical physiological roles. However, since boron is not stored in the body and is rapidly excreted, its full spectrum of importance remains an area of ongoing scientific exploration.\u003c/p\u003e \u003cp\u003eAyurveda has long recognized leaves as natural bioenhancers, preserving nutrients and infusing food with therapeutic vitality. The practice of steaming food on leaves aligns with Ayurvedic wisdom, minimizing nutrient loss and maximizing bioavailability\u0026mdash;a principle now validated by science. Palash and Jackfruit leaves excel in boron retention and controlled release, while Banana and Badam leaves, despite their popularity, show significant boron leaching, highlighting the critical role of leaf structure in nutrient dynamics.\u003c/p\u003e \u003cp\u003eSeasonality and geography are equally influential, with \u003cem\u003eSharad Ritu\u003c/em\u003e (autumn) leaves traditionally prized for peak nutrient content\u0026mdash;a fact now supported by modern research. Beyond nutrition, leaf-based cooking reflects environmental stewardship, as leaves are biodegradable, locally sourced, and ecologically sustainable, contributing to a balanced nutrient cycle.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eBoron is a trace mineral or essential faciliatory nutrient?\u003c/h3\u003e\n\u003cp\u003eBoron is known to play a pivotal role in calcium metabolism, brain function, insulin regulation, and hormone activity [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Studies suggest that boron helps regulate key metabolic pathways, including those associated with steroid hormones (e.g., oestrogen and vitamin D), immune responses, and oxidative stress management [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Boron's deficiency has been linked to a range of health complications, including reduced bone density, impaired cognitive function, increased urinary calcium excretion, and elevated inflammatory markers[\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eDespite its physiological importance, boron remains one of the least studied micronutrients, with dietary intake data primarily limited to Western populations [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Surveys estimate median boron intake in adults to range between 0.87 to 1.35 mg/day, with higher values observed in vegetarians due to their plant-rich diets [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. However, in India, where traditional diets heavily rely on plant-based foods, there is a lack of concrete data on dietary boron intake and its bioavailability in cooked food samples. This knowledge gap is particularly concerning given the decline in soil boron levels, which directly impacts the boron content of locally grown crops.\u003c/p\u003e \u003cp\u003eMore than physical nourishment, Ayurveda views leaf-steamed food as energetically therapeutic, with \u003cem\u003ePrana\u003c/em\u003e\u0026mdash;a vital life force\u0026mdash;transferring into food during steaming. Scientifically, leaves serve as efficient carriers of boron, a micronutrient essential for human health. While boron has often been described as a \u003cem\u003e\"cosmic particle\"\u003c/em\u003e due to its unique role in both plant and human physiology, a more fitting term would be the \u003cem\u003e\"Chaitanya particle\"\u003c/em\u003e, signifying its life-giving and facilitating properties. In plants, boron functions as a flushing agent, mobilizing and transporting other micronutrients, while in humans, it regulates hormones, strengthens bones, and supports cognitive function.\u003c/p\u003e \u003cp\u003eInterestingly, recent research highlights boron as an \u003cem\u003e\"interkingdom chemical communication tool.\"\u003c/em\u003e Studies using pig models reveal that boron acts as a mediator, allowing animals to maintain health even in the presence of pathogenic bacteria\u0026mdash;a sort of negotiating agent between different biological kingdoms. In this context, greater boron loss during cooking might not always represent a failure; instead, it symbolizes boron's dynamic role as a facilitator and communicator across biological systems. The more boron mobilizes, the more it serves its interkingdom purpose\u0026mdash;a silent battle won for maintaining biological harmony. Establishing a Recommended Dietary Allowance (RDA) for boron remains a significant challenge due to the limited availability of comprehensive and consistent human data. However, the development of provisional intake recommendations, supported by emerging evidence and region-specific dietary data, is a necessary step toward formulating a sustainable, evidence-based RDA. Accordingly, a preliminary draft proposing such intake ranges has been prepared and is included as supplementary material to facilitate further discussion and research-based validation.\u003c/p\u003e \u003cp\u003eThis synergy between Ayurvedic wisdom and scientific validation underscores the potential of leaf-based steaming in addressing boron deficiency\u0026mdash;a rising global concern. Policymakers must prioritize education, awareness campaigns, and incentives for adopting these traditional culinary practices. Future research integrating Ayurvedic principles with advanced analytical tools will further optimize these sustainable and culturally resonant interventions, transforming them into scalable solutions for global health and nutrition security.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study highlights the potential of traditional leaf-based culinary practices in addressing boron deficiency, a critical yet often overlooked micronutrient gap. Leaves like Palash and Jackfruit demonstrated superior boron retention during idly preparation, influenced by factors such as temperature, leaf microstructure, and steaming duration. These findings suggest that natural leaves can serve as sustainable and cost-effective alternatives to synthetic boron supplements. Policymakers should incorporate these insights into public health strategies, including awareness campaigns targeting schools, health workers, and culinary professionals. Government programs, such as mid-day meal schemes, could adopt these practices to ensure nutrient-rich meals. Standardization protocols and certification systems are essential to maintain hygiene and consistency in leaf-based cooking methods. Continued research is necessary to optimize boron transfer efficiency and validate regional variations in leaf properties for broader implementation.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eOSLSI\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eOne\u0026ndash;sided leaf steaming approach on Idly making\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eTSLSI\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eTwo\u0026ndash;sided leaf steaming approach on Idly making\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eICP\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eMS\u0026ndash;inductive coupled plasma\u0026ndash;mass spectrophotometry\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAcknowledgments:\u003c/h2\u003e \u003cp\u003eThe authors thank the Indian Council of Medical Research-National Institute of Nutrition (ICMR-NIN) and Osmania University College of Technology (OUCT) for their support and resources. Gratitude is extended to our mentors and collaborators for their guidance and expertise. We acknowledge the Director of ICMR-NIN for the intramural grant (No. ST/1/Intra-Pro-Allot/2023-24) and thank Mr. Saida Naik, Mr. Subash Tatikayala, and Mr. Mohan Kumar for their assistance during experimentation. I also thank Director ICMR-NIV, Pune for allowing me to continue working on this manuscript while I am working at ICMR-NIV.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eSingh MV (2008) Micronutrient deficiencies in crops and soils in India. Micronutrient deficiencies in global crop production. 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I.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003eo.M.U.P.o\u003c/span\u003e\u003cspan address=\"http://o.M.U.P.o\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Micronutrients. Washington (DC): National Academies Press (US)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBuck GMC, Dourson R, Foster ML, Goyer P, Chairman RA, Howe P, Co-Rapporteur); Luoto R, Nielsen FH, Price CJ, Woods WG (1998) In: Organization WH (ed) Environmental Health Criteria 204 - Boron. World Health Organization, Editor\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"ICMR-National Institute of Nutrition","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Boron, Traditional Cooking using leaves, Nutritional Enhancement, Trace Elements ","lastPublishedDoi":"10.21203/rs.3.rs-6928254/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6928254/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBoron is a vital trace element essential for bone health, cognitive function, and metabolic processes. However, boron deficiency remains a global concern due to soil depletion, poor dietary intake, limited water boron content, and general unawareness. The World Health Organization acceptable daily intake of 1\u0026ndash;13 mg/day, yet populations in several developed and developing countries often fail to meet even the minimum requirement. This study aimed to investigate the role of traditional leaf-based steaming practices in enhancing boron retention and enrichment in \u003cem\u003eidly\u003c/em\u003e, a steamed rice cake.\u003c/p\u003e \u003cp\u003eFive leaf types\u0026mdash;Palash, Jackfruit, Banana, Badam, and Banyan\u0026mdash;were analysed for boron baseline levels, leaching, retention, and transfer efficiency (enrichment) using Colorimetry and Inductively Coupled Plasma Mass Spectrometry. Leaves were subjected to hot water treatment (60\u0026deg;C for 15 minutes) to simulate blanching, followed by steaming in autoclave at 121\u003csup\u003e0\u003c/sup\u003eC for 15 minutes with one-sided and two-sided leaf contact configurations. Results indicated significant differences in boron dynamics among leaf types. Palash and Jackfruit leaves exhibited superior boron retention and transfer capabilities, while Banana and Badam leaves showed higher leaching. Seasonal variations, leaf maturity, regional geography, gravitational effects during steaming, and leaf orientation further influenced boron dynamics. Additionally, leaves exhibited dual roles\u0026mdash;some enriched the \u003cem\u003eidly\u003c/em\u003e, while others neutralized boron loss during steaming. The findings highlight the potential of traditional cooking practices in addressing dietary boron deficiency naturally and sustainably. Future studies should focus on optimizing steaming conditions, understanding leaf microstructures, and expanding research across diverse regions to ensure reproducible and practical dietary interventions for boron supplementation.\u003c/p\u003e","manuscriptTitle":"Boron Enrichment and Retention viaTraditional Leaf-Based Cooking Methods in Idly Preparation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-23 05:51:58","doi":"10.21203/rs.3.rs-6928254/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"eaa69493-7897-403e-9431-f61b2178ff06","owner":[],"postedDate":"June 23rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":50278984,"name":"Food Science \u0026 Technology"},{"id":50278985,"name":"Nutrition \u0026 Dietetics"}],"tags":[],"updatedAt":"2025-06-23T05:51:58+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-23 05:51:58","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6928254","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6928254","identity":"rs-6928254","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}