Exploring Little Millet (Panicum sumatrense) Strain DHLM 36-3 and OLM‑203 for Functional Food Development and Nutritional Enhancement

Millets, often termed miracle crops or nutri‑cereals, are resilient C4 plants capable of thriving in low rainfall regions with minimal external inputs. Among them, little millet (Panicum sumatrense) has emerged as a promising minor millet due to its rich nutritional profile and adaptability to diverse agro‑ecological conditions. It is a valuable source of B‑vitamins, calcium, iron, zinc, potassium, and essential fatty acids, making it an important dietary component for addressing micronutrient deficiencies (Reddy, 2017).

Recent studies highlight the functional and health‑promoting properties of little millet, including anti‑diabetic, anti‑obesity, and cardioprotective effects, largely attributed to its polyphenols, flavonoids, and hydroxycinnamic acid derivatives (Ambati & Sucharitha, 2019; Gujral, 2023). Germination and cooking significantly improve protein and starch digestibility, reduce anti‑nutritional factors, and enhance antioxidant activity, thereby increasing nutrient bioavailability (Gujral, 2023).

 

Specific strains such as DHLM 36‑3 and OLM‑203 have been reported to exhibit high levels of essential amino acids, soluble dietary fiber, and antioxidants, positioning them as potential functional food ingredients (Gujral, 2023). Advanced omics‑based crop improvement strategies have further enhanced the nutritional value and environmental resilience of these strains, making them suitable for food and nutritional security programs (Mishra et al., 2024). Moreover, metabolomic profiling of OLM‑203 under water stress has revealed key pathways contributing to drought tolerance, reinforcing its suitability for arid regions (Dhawale et al., 2022). Agronomic studies also demonstrate that OLM‑203 performs well under optimized row spacing, yielding superior grain productivity compared to other varieties (Barad et al., 2024).

 

In terms of disease resistance, DHLM 36‑3 has shown moderate resistance to banded blight caused by Rhizoctonia solani, making it a valuable candidate for breeding programs aimed at developing resilient cultivars (Salam et al., 2023). Collectively, these findings underscore the potential of little millet strains as functional food crops with nutritional, therapeutic, and ecological significance.

 

The present research aims to assess the nutritional and economic viability of malt‑based formulations using little millet strains DHLM 36‑3 and OLM‑203, with the goal of developing value‑added functional food products. By optimizing processing methods to enhance nutrient bioavailability and reduce anti‑nutritional factors, this work contributes to promoting millet‑based innovations that align with sustainable health, wellness, and food security.

Sample Collection

The study utilized two distinct millet samples, selected for their nutritional potential and agronomic viability. These include:

1.     Little Millet (Panicum sumatrense) – Variety: DHLM 36-3

2.     Little Millet (Panicum sumatrense) – Variety: OLM-203

“Both the strains were authenticated and procured from the Agriculture University, Raichur, Karnataka, India. The strains were selected for its nutritional traits, adaptability to local agro-climatic conditions, and relevance to millet-based formulation research, ensuring credibility in malt development and nutritional profiling.”

 

Physical Evaluation of the Samples:

Colour and shape

The colour and shape of the selected millet samples were observed from their physical and visual appearance

Thousand kernel weight

Thousand kernels weight was determined by weight of randomly selected 100 kernels by means of electronic/analyticalbalance (precision of 0.001 g) and multiplying their weight by 10.

True density

The true density was measured by toluene displacement method. One thousand grains of millet samples were weighed and put in graduated cylinder containing known amount of toluene. Rise in toluene level was noted and true density was reported by using the formula   

 

𝑇𝑟𝑢𝑒 𝑑𝑒𝑠𝑛𝑠𝑖𝑡𝑦 (𝑔/𝑚𝑙) = 𝑊 (𝑔) / 𝑉 (𝑚𝑙)

Where ‘W’ is weight of one thousand grains and ‘V’ is rise in toluene level after the addition of the grains.

 

Bulk density

The millet samples were filled in measuring cylinders up to certain level from the constant height followed by weighing. The bulk density was determined by using the formula

 

𝐵𝑢𝑙𝑘 𝑑𝑒𝑠𝑖𝑡𝑦 (𝑔/𝑚𝑙) = 𝑊𝑒𝑖𝑔ℎ𝑡 (𝑔)/ 𝑉𝑜𝑙𝑢𝑚𝑒 (𝑚𝑙)

 

Porosity

Porosity was analyzed by using the relationship of bulk density and true density as follows.

 

𝑃𝑜𝑟𝑜𝑠𝑖𝑡𝑦 = 1 − (𝑡𝑟𝑢𝑒𝑑𝑒𝑛𝑠𝑖𝑡𝑦 − 𝑏𝑢𝑙𝑘𝑑𝑒𝑛𝑠𝑖𝑡𝑦) /𝐵𝑢𝑙𝑘𝑑𝑒𝑛𝑠𝑖𝑡𝑦 × 100

 

Chemical Analysis of Samples

 

Moisture (%)

Moisture content (%) was determined using the Hot Air Oven Method, following the standardized procedure outlined by the Association of Official Analytical Chemists (AOAC, 1984). This gravimetric technique involves measuring the mass of a food or grain sample before and after drying in a controlled-temperature oven. The sample is placed in a pre-weighed moisture dish and heated at 130 ± 1°C for 1 hour in a forced-air oven, ensuring uniform evaporation of water without decomposition of other volatile constituents. After drying, the sample is cooled in a desiccator to prevent moisture reabsorption and then reweighed. The moisture content is calculated as the percentage loss in weight, representing the amount of water originally present in the sample. This method is widely accepted for cereals, flours, and other food products due to its reliability and reproducibility, and corresponds to AOAC Official Method 925.10 (AOAC, 1984).

 

Ash (%) Content Determination by Muffle Furnace Method

The ash content of powdered millet samples was determined using the dry ashing technique in a muffle furnace, following the protocol described by Pande et al. (2018). Approximately 2 grams of the powdered sample were placed in a pre-weighed silica crucible and incinerated at 500°C for 5 hours. After complete combustion, the crucible was allowed to cool in a desiccator and then reweighed. The resulting ash was white in color, indicating the absence of residual carbon, and confirming complete oxidation of organic matter. This method provides a reliable estimate of the total mineral content and is commonly employed in the standardization of plant-based food products.

 

Crude Fiber Determination (%)

Crude fiber content was estimated following the protocol outlined in Biochemical Methods by Sadasivam and Manickam (2018). The method involves subjecting the defatted millet sample to sequential acid and alkali digestion to remove soluble materials, leaving behind indigestible fibrous components such as cellulose and lignin. The residue is then filtered, dried, weighed, and incinerated in a muffle furnace to eliminate any remaining organic matter. The crude fiber percentage is calculated based on the weight difference before and after ashing, providing a reliable measure of the structural carbohydrate content in the sample.

 

Mineral Analysis by Atomic Absorption Spectrophotometry

The concentrations of essential minerals Zinc (Zn), Iron (Fe), Manganese (Mn), and Calcium (Ca) were quantitatively determined using Atomic Absorption Spectrophotometry (AAS). This technique involves aspirating the digested sample into a flame or graphite furnace, where the target metal atoms absorb ultraviolet light at element-specific wavelengths. The absorbance is directly proportional to the concentration of the element in the sample, as governed by the Beer-Lambert law. For accurate detection, air-acetylene flame was employed for Zn, Fe, and Mn, while lanthanum nitrate was added as a releasing agent for Ca to prevent interference. Calibration curves were constructed using standard solutions, and absorbance values were measured at wavelengths of 213.9 nm (Zn), 248.3 nm (Fe), 279.5 nm (Mn), and 422.7 nm (Ca). This method ensures high sensitivity and specificity for trace mineral quantification in food matrices (Paul et al. 2014; FSSAI, 2024).

 

Biochemical Analysis of Samples

 

Protein Estimation by Lowry’s Method

Protein content was estimated using the Lowry method, as described by Sadasivam and Manickam (2018). This colorimetric assay is based on two sequential reactions:

·       Biuret Reaction: Proteins react with copper ions in an alkaline medium, forming a cuprous complex.

·       Folin-Ciocalteu Reaction: The cuprous ions, along with aromatic amino acids (tyrosine and tryptophan), reduce the phosphomolybdic-phosphotungstic components of the Folin-Ciocalteu reagent, producing a blue-purple chromophore.

The intensity of the color, measured spectrophotometrically at 660 nm, is directly proportional to the protein concentration. A standard curve is constructed using bovine serum albumin (BSA), and unknown sample concentrations are interpolated accordingly. This method is highly sensitive and suitable for detecting microgram quantities of protein in plant and food matrices.

 

Carbohydrate Estimation by Anthrone Method

Total carbohydrate content was determined using the Anthrone method, as described by Sadasivam and Manickam (2018). This colorimetric assay involves the acid hydrolysis of polysaccharides into monosaccharides, followed by dehydration to form furfural derivatives. These intermediates react with Anthrone reagent in a strongly acidic medium to produce a blue-green chromogen, which is measured spectrophotometrically at 620 nm.

The procedure includes:

·       Hydrolyzing the sample with 2.5 N hydrochloric acid in a boiling water bath for 3 hours.

·       Neutralizing with solid sodium carbonate and centrifuging to collect the supernatant.

·       Reacting aliquots of the supernatant with freshly prepared Anthrone reagent (200 mg Anthrone in 100 mL concentrated H₂SO₄).

·       Heating the mixture in a boiling water bath for 10 minutes and cooling rapidly.

·       Measuring absorbance at 620 nm and calculating carbohydrate concentration using a standard glucose curve.

 

Fat Estimation by Soxhlet Extraction Method

Crude fat content was determined using the Soxhlet extraction technique, as described by Sadasivam and Manickam (2018). This method involves the continuous extraction of lipids from a dried and finely ground sample using an organic solvent typically petroleum ether or diethyl ether in a Soxhlet apparatus.

The procedure includes:

·       Drying the sample thoroughly to eliminate moisture, which can interfere with solvent efficiency.

·       Weighing a known quantity of the sample and placing it in a cellulose thimble.

·       Extracting the sample with solvent for 6–8 hours or until the solvent runs clear, indicating complete lipid removal.

·       Evaporating the solvent from the flask and drying the residue in an oven at 105 ± 1°C to constant weight.

·       Calculating the fat content based on the weight difference of the flask before and after extraction.

 

 

Nutritional Dynamics of Little Millet Strains

 

The proximate and mineral profiling of little millet (Panicum sumatrense) strains DHLM 36‑3 and OLM‑203** under different processing conditions (raw powder, germinated shadow‑dry, and germinated sun‑dry) revealed distinct nutritional dynamics that highlight their functional food potential.

 

Ash Content (%):

Mineral concentration increased notably upon germination and sun‑drying. OLM‑203 recorded the highest ash content (3.65%) in the germinated sun‑dry state, indicating enhanced mineral density due to moisture loss and nutrient mobilization. DHLM 36‑3 also showed improved ash levels, supporting its role in mineral‑rich formulations.

 

Moisture Content (%):

Moisture decreased progressively across treatments, with all sun‑dried samples falling below 7.7%. This reduction enhances shelf‑life and microbial stability, making both strains suitable for long‑term storage and malt‑based product development.

 

Crude Fiber (%):

Both strains exhibited modest increases in crude fiber after germination, maintaining structural balance. This fiber enrichment supports gastrointestinal health and satiety, while avoiding excessive toughness in processed products.

 

Carbohydrate (g):

Carbohydrate content declined slightly post germination due to enzymatic hydrolysis. Raw OLM‑203 showed the highest carbohydrate value (63 g), making it energy‑dense, while germination improved digestibility and bioactive release in both strains.

 

Protein (g):

Protein content increased by ~2 g in both DHLM 36‑3 and OLM‑203 after germination and sun‑drying. This improvement reflects enzymatic mobilization during sprouting, positioning little millet as a functional base for protein‑enriched foods.

 

Fat (g):

Fat content varied, with raw OLM‑203 showing the highest fat (3.92 g). However, lipid degradation during sun‑drying reduced fat levels, suggesting the need for optimized processing to retain energy value and flavor.

 

Calcium (mg):

Calcium behavior was non‑linear. OLM‑203 displayed a resurgence in calcium after sun‑drying (400 mg), likely due to mineral redistribution and concentration effects. This highlights its potential for bone health applications.

 

Iron (mg):

Iron bioavailability improved significantly with germination. DHLM 36‑3 increased from 7.57 mg (raw) to 8.01 mg (sun‑dry), attributed to phytate breakdown and enhanced solubility. This supports its antianemic potential.

 

Zinc (mg):

Zinc remained stable across treatments, with DHLM 36‑3 consistently high (~3.1 mg). This resilience underscores its importance for immune and metabolic functions in millet‑based diets.

 

Manganese (mg):

Manganese was undetectable in both strains, possibly due to genetic variation or soil mineral availability, warranting further trace analysis.

 

Overall, germination followed by sun‑drying enhanced protein, iron, and ash content in DHLM 36‑3 and OLM‑203, while carbohydrate and calcium retention were particularly strong in OLM‑203. These nutritional shifts validate the potential of little millet strains as functional food ingredients, offering balanced energy, improved mineral bioavailability, and resilience against nutrient loss during processing. Their profiles support the development of nutrient‑dense, sustainable malt‑based products that can serve as healthier alternatives to conventional snacks.

The physical and nutritional evaluation of Little millet strains DHLM 36‑3 and OLM‑203 revealed distinct traits that directly enhance their functional suitability in malt‑based product development. Both strains exhibited smaller grain sizes but demonstrated complementary strengths: DHLM 36‑3 showed the highest true density (1.358 g/ml), indicating compact grain architecture favorable for nutrient retention and oxidative stability, while OLM‑203 displayed lower porosity yet maintained balanced bulk and density values, ensuring predictable milling and formulation behavior (Sushmita et al., 2020). Moisture reduction during germination and sun‑drying improved storability and microbial safety, aligning with AOAC protocols for stable cereal processing. Ash content increased notably in OLM‑203 (3.65%), reflecting enhanced mineral concentration, while calcium resurgence after sun‑drying (400 mg/100 g) highlights its potential for bone health applications (Sushmita et al., 2020). Protein content improved by ~2 g in both strains post germination, supporting their role in protein‑enriched functional foods, and iron bioavailability was significantly enhanced in DHLM 36‑3 (from 7.57 to 8.01 mg), consistent with phytate degradation and enzymatic activation during sprouting (Kumar et al., 2023). Zinc levels remained stable across treatments, with DHLM 36‑3 consistently high (~3.1 mg), underscoring mineral resilience and immune support potential. These findings corroborate earlier reports that little millet varieties are superior to staple cereals in terms of fiber, fat, and iron content, and highlight their adaptability to diverse agro‑ecological conditions (Sushmita et al., 2020). Collectively, the results signify that DHLM 36‑3 is particularly suited for nutrient preservation and iron enrichment, while OLM‑203 offers advantages in carbohydrate energy density and calcium retention. Thus, both strains demonstrate strong potential as nutrient‑dense, sustainable ingredients for malt‑based functional food formulations, contributing to micronutrient recovery, dietary diversification, and healthier alternatives to ultra‑processed snacks.

 

The present study underscores the nutritional and functional potential of Little Millet (Panicum sumatrense) strains DHLM 36 3 and OLM 203 in the development of malt based nutri products. Through comprehensive physicochemical, biochemical, and sensory evaluations, the research establishes a robust framework for valorizing these underutilized grains in functional food innovation. The choice of little millet strains was guided by their local availability, resilience under low input cultivation, and potential to enhance food and nutritional security.

Processing steps soaking, germination, and sun drying were optimized to improve nutrient bioavailability and reduce anti nutritional factors. Physicochemical characterization revealed distinct grain attributes: DHLM 36 3 exhibited higher true density, favoring nutrient retention and oxidative stability, while OLM 203 demonstrated balanced carbohydrate energy density and calcium resurgence post sun drying, highlighting its suitability for bone health applications. Germination significantly enhanced protein and iron content in both strains, while zinc remained stable, ensuring mineral resilience. These metabolic shifts reflect the functional versatility of little millets and support their use in health oriented formulations.

 

Anti nutritional profiling confirmed reductions in tannins, saponins, and oxalates after germination, while enzymatic screening (amylase, protease, L asparaginase, L glutaminase) and antioxidant detection (flavonoids, phenols) emphasized their bioactive potential. Sensory evaluation of malt based formulations incorporating little millet flour demonstrated favorable consumer acceptance, validating their practical applicability in product development.

 

Overall, the findings highlight that little millet strains DHLM 36 3 and OLM 203 are nutrient dense, resilient, and functionally versatile crops, capable of delivering protein enrichment, mineral bioavailability, and antioxidant benefits. Their integration into malt based formulations offers a sustainable, health promoting alternative to nutrient poor snack foods, contributing to public health, local food security, and agro biodiversity conservation. This study advocates for mainstreaming little millets into modern diets and encourages further research into their bioactive compounds, enzymatic activity, and therapeutic potential, ultimately supporting the transition toward sustainable, indigenous, and nutritionally optimized food systems.

 

Data Availability Statement

Data are contained within the article.

Ethics Statement

All the experiments set with animals by following and adhering to Institutional Animal Ethics Committee (Ref: CBPL-IAEC-072/04/2024) before performing any animal experiments.

Conflicts of Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Funding

No funding

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