Iron Bites: Formulation And Evaluation of Nutraceutical Cookies Enriched with Mucuna Pruriens and Natural Ingredients for Management of Iron Deficiency Anaemia

Figure 1: Morphological comparison of normal biconcave red blood cells and sickle-shaped (anaemic) red blood cells

.Fatigue, weakened immunity, stunted growth and development, and diminished physical performance are all possible outcomes of anemia. The primary reason is iron deficiency, which can be brought on by inadequate food, blood loss, or increased iron needs during pregnancy. Red blood cell production is impacted by vitamin B₁₂ and folic acid deficiencies, which can also result in various forms of anemia.^[1]

1.2 Nutraceutical Approach to Anaemia Management

Nutraceuticals are food-based solutions that can replace traditional iron supplements and offer health advantages beyond basic nutrition. Nutraceutical meals are often more accepted and appropriate for long-term usage than iron pills, which can induce adverse effects such nausea, constipation, and black feces. Bioactive substances including antioxidants, flavonoids, and polyphenols found in many plant-based products promote healthy red blood cells and blood production.Because they are easy to eat, have a longer shelf life, and give exact nutritional content, nutraceutical cookies are a novel and practical method of supplementing with iron. For kids and teenage females, who frequently struggle with consistent tablet consumption, they could be particularly helpful. ^[2]

1.3 Mucuna pruriens — Pharmacognostical and Nutritional Profile

Figure 2: Mucuna pruriens seeds — the primary iron-rich botanical ingredient (Family: Fabaceae; Common name: Velvet bean / Cowhage)

Mucuna pruriens, a tropical climbing legume of the Fabaceae family, is sometimes referred to as cowhage or velvet bean. Due to its nutritional and therapeutic value, it is extensively farmed in Central and South America, Africa, and India. The plant yields seed-containing pods that are high in minerals, proteins, carbs, and a number of bioactive substances. Numerous pharmacological actions of Mucuna pruriens, such as antidiabetic, antibacterial, antioxidant, neuroprotective, and anti-anaemic effects, have been documented in studies.With a high protein and carbohydrate content and minerals including iron, calcium, phosphorus, and magnesium, the seeds are highly nutritious. Important phytochemicals that support their medicinal qualities like flavonoids, tannins, saponins, alkaloids, and polyphenols. The seeds have the potential to be used as a natural nutraceutical element for the treatment of anemia because research has shown that they may increase haematological parameters such red blood cell count, packed cell volume, and platelet levels in anaemic situations.^[3]

1.4 Complementary Iron-Rich Ingredients

1.4.1 Halim Seeds (Lepidium sativum)

Figure 3: Halim seeds (Lepidium sativum L.) — Family: Brassicaceae; Common names: Garden cress, Chandrashoor — the highest iron contributor (100 mg/100 g)

Originally from Egypt and Southwest Asia, garden cress is an annual edible plant that is commonly grown in India. With about 100 mg of iron per 100 g, its seeds are regarded as one of the greatest plant sources of iron. The seeds are extremely nutritious since they are abundant in iron, folic acid, proteins, omega-3 fatty acids, calcium, phosphorus, potassium, magnesium, zinc, and dietary polysaccharides. Garden cress seeds have historically been used to treat rheumatic illnesses, digestive issues, respiratory issues, and nursing mothers' milk. The seeds are frequently utilized in nutraceutical and functional food formulations targeted at enhancing general health and avoiding iron deficiency anemia because of their great nutritional and therapeutic value.

1.4.2 Black Sesame (Sesamum indicum)

Figure 4: Black sesame seeds (Sesamum indicum L.) — Family: Pedaliaceae; Iron content: 14.6 mg/100 g

The Pedaliaceae family includes the nutrient-rich annual plant black sesame. Iron, calcium, magnesium, vitamin E, proteins, and healthful lipids may all be found in its seeds. Often referred to as the "crown of grains," black sesame is highly nutritious.Black sesame is frequently used in nutraceutical and fortified food items, such as cookies designed to control anemia, because of its high iron concentration and antioxidant components.

1.4.3 Dates (Phoenix dactylifera)

Figure 5: Dates (Phoenix dactylifera L.) — Family: Arecaceae; Common names: Dates, Khajoor — rich in iron, dietary fibre, and antioxidants [10]

Dates are extremely nutrient-dense fruits that are abundant in vitamins, minerals, natural sugars, iron, and dietary fiber. They include antioxidant chemicals and essential minerals including calcium, potassium, magnesium, and vitamins A, B complex, and C.
Because dates contain iron and folate, regular eating may boost anemic people's hemoglobin levels. They are a useful component of nutraceutical and iron-fortified food items since their natural sugars also provide them instant energy.

1.4.4 Almond (Prunus dulcis)

Figure 6: Almond (Prunus dulcis) — Family: Rosaceae — rich in protein, vitamin E, calcium, magnesium, iron, and unsaturated fatty acid

Healthy fatty acids, proteins, vitamin E, riboflavin, niacin, calcium, magnesium, phosphorus, potassium, iron, and zinc are all found in almonds, which are nutrient-rich nuts. They are prized for having strong antioxidant and nutritional qualities.Iron and other vital minerals that promote blood formation and general health are found in almonds. They are a valuable component of nutraceutical and fortified food items because their antioxidant components also aid in preventing oxidative damage to body cells, particularly hematopoietic cells.

1.5 Study Rationale

Particularly in adolescents and teenage females who are severely afflicted by anemia, conventional oral iron supplements are frequently linked to gastrointestinal side effects, an unpleasant taste, and poor patient compliance. A delicious, practical, and shelf-stable substitute for iron supplementation is offered by nutraceutical cookies enhanced with iron-rich plant ingredients.
A rich natural supply of iron, antioxidants, and other bioactive substances that promote blood formation and general health are provided by the combination of garden cress, black sesame, jaggery, mucuna pruriens, and almond.

2. Materials and Methods

2.1 Ingredients and Materials

All ingredients used in the formulation were obtained from authenticated local suppliers. Dr. V. N. Chavhan, Head of Botany, Arts, Commerce and Science College, Maregaon, botanically verified the Mucuna pruriens seeds' identification as Mucuna pruriens (L.) DC., a member of the Fabaceae family.
Mucuna pruriens seeds, dates, black sesame, almonds, garden cress seeds, oats, jaggery, ghee, and baking powder were the primary components utilized to make the cookies. During the formulation procedure, wheat flour and vanilla essence were also included as processing aids.

Figure 7: All ingredients used in the preparation of nutraceutical cookies, arranged in individual bowls prior to formulation

2.2 Formulation Design

In order to maintain the ingredient composition ratios, four cookie formulations (F1–F4) were created by methodically varying the amounts of important ingredients. Based on better sensory and physical performance, Formulation F4 was determined to be the optimal batch.

Table 1: Formulation Composition of Nutraceutical Cookie Batches F1–F4

2.3 Method of Preparation

Step 1: Gathering and preparing ingredients: Almonds and Mucuna pruriens seeds were roasted, cleaned, and milled into fine flour. Black sesame and halim seeds were dry-roasted individually. A homogenous sweet paste was created by blending dates, halim seeds, black sesame, and jaggery. A coarse powder was made from oats. Wheat flour was stored with baking powder and vanilla essence.
Step 2: Making the dough: Mucuna pruriens flour, wheat flour, oat powder, and almond powder were mixed together evenly. To create a uniformly soft dough, a tiny amount of milk was progressively added. The sweetened foundation was made by blending the jaggery, black sesame, halim seed, and dates paste with the dry flour combination. As a flavoring, vanilla essence was added.

Step 3: Shaping and baking: Using a mold or by hand, the produced dough was formed into homogeneous disc-shaped cookies. The cookies were placed on oiled baking pans and baked for 15 to 20 minutes at 160 to 180 degrees Celsius until they were firm to the touch and golden brown.
Step 4: Cooling and storage: To preserve crispness and stop moisture absorption, cookies were taken out of the oven, let to cool at room temperature for half an hour, and then kept in airtight polypropylene containers.

Figure 9: Cookies during baking at 160–180°C in preheated oven — uniform golden-brown colouration observed across all formulations

3. Evaluation

3.1 Sensory Evaluation

A trained hedonic panel evaluated all four formulations' sensory qualities, including appearance, color, scent, taste, texture, and general acceptance. With a score of 9.5/10, Sample D (which corresponds to Formulation F4) showed the best overall acceptance of all the formulations tested; most panelists assessed appearance, color, scent, taste, and texture as "Excellent" or "Very Good." Natural additives, including dates, vanilla essence, and jaggery, improved palatability without degrading sensory quality. As a result, formulation F4 was chosen as the best formulation for all subsequent studies.

Figure 10a: Sensory Evaluation Sheet — Nutritional Iron Cookies (Hedonic Scale) showing panel responses for six attributes across six panellists (Part 1)

Figure 10b: Sensory Evaluation Sheet — Nutritional Iron Cookies (Hedonic Scale) — Part 2, continued panel responses

3.2 Physical Analysis

The physical characteristics of the prepared cookies were assessed for consistency and baking quality; Figures 11–13 show the average weight, thickness, and diameter that were calculated; Table 2 shows the spread ratio for three duplicate samples, which is calculated as diameter divided by thickness; results between 1.48 and 1.50 for all three samples confirm homogeneous dough spreading and consistent baking behavior, which are indicators of adequate fat content and suitable baking powder leavening; weight uniformity (22.29–22.35 g) with minimal variation.

Figure 11: Average weight determination of formulated cookies (W1 = 22.29 g; W2 = 22.30 g; W3 = 22.35 g) using calibrated Wensar electronic analytical balance

Figure 12a: Thickness measurement of formulated cookie (20–20.3 mm) using calibrated Vernier callipers  |  Figure 12b: Diameter measurement (30–30.2 mm)

The spread ratio — calculated as diameter / thickness — for three replicate samples is presented in Table 2.

Table 2: Physical Evaluation — Average Weight, Thickness, Diameter, and Spread Ratio of Formulated Cookies (Batch F4)

Spread ratio values of 1.48–1.50 across all three samples confirm uniform dough spreading and consistent baking behaviour.

3.3 Evaluation Parameters

3.3.1 Moisture Content

Figure 13: Moisture content determination — oven-drying of cookie sample in petri dish at 105°C

As seen in Figure 13, the typical oven-drying method was used to estimate the moisture content at 105°C for three to four hours using a hot air oven.
Moisture (%) = [(31.88 − 31.81) / (31.88 − 29.88)] × 100 = [0.07 / 2.00] × 100 = 3.5%. W1 (empty petri dish) = 29.88 g; W2 (dish + sample) = 31.88 g; W3 (dish + dried sample) = 31.81 g. Good shelf stability and sufficient baking are indicated by the low moisture content of 3.5%, which is well within acceptable bounds for bakery goods (usually <10%).

3.3.2 Fat Content

Figure 14: Fat content determination by Soxhlet extraction using petroleum ether as extraction solvent

Fat content was determined by Soxhlet extraction. Fat (%) = (0.24 / 2.0) × 100 = 12%. For nutraceutical cookies, a fat level of 12% is suitable as it offers energy density and palatability while being within permissible dietary bounds.

3.3.3 Ash Content

Figure 15: Total ash determination by incineration in muffle furnace (550–600°C) — crucibles post-ashing

Ash total: W1 (crucible) = 21.31 g; W2 (crucible + sample) = 23.59 g; W3 (crucible + ash) = 21.60 g. Sample weight: 2.28 g; ash weight: 0.29 g. (0.29 / 2.28) × 100 = 12.7% is the total ash percentage. Acid-insoluble ash (%) = (0.65 / 1.7) × 100 = 38.2%. Ash that dissolves in water (%) = (0.03 / 1.7) x 100 = 1.7%. The total ash value of 12.7% reflects the mineral-rich composition attributable to halim seeds, black sesame, and jaggery.

3.3.4 Iron Detection (Qualitative)

Figure 16: Iron qualitative detection — blood-red colour development upon addition of potassium thiocyanate to HCl-acidified cookie solution, confirming presence of ferric iron

When HCl and potassium thiocyanate solution were added to the cookie extract, the potassium thiocyanate test produced a distinctive blood-red color, indicating the presence of ferric iron (Fe³⁺). This qualitative finding is in line with the high estimated iron content of 14.68 mg per batch, which is mostly attributed to jaggery (1.00 mg) and halim seeds (10.00 mg).

3.3.5 Protein Content

Figure 17: Protein content determination by Folin–Ciocalteu (Lowry) method — blue-purple colour development in test tubes indicating protein presence

The Lowry method confirmed the presence of protein in cookie samples, with blue-purple colour development in the test tubes indicating protein–Folin-Ciocalteu reagent reaction. Mucuna pruriens (6.67 g) and almonds (2.12 g) made up the majority of the estimated 13.66 g of total protein per batch.

3.3.6 Carbohydrate Determination Tests

Figure 18: Carbohydrate qualitative screening tests — left to right: Molisch (violet ring), Benedict (colour change), Fehling, Barfoed's, iodine, and hydrolysis tests

Cookie extract solutions were subjected to an extensive battery of qualitative carbohydrate testing. Table 3 displays the results. The presence of carbohydrates, reducing sugars, starch, monosaccharides, and polysaccharides was confirmed by all tests, which yielded distinctive positive findings that were compatible with the cookies' composition of oats, dates, jaggery, and Mucuna pruriens.

Table 3: Qualitative Carbohydrate Determination Tests — Observations and Inferences

3.3.7 pH Determination

Figure 19: pH determination of cookie aqueous extract using calibrated digital pH meter — reading 6.85 (near neutral)

The pH of the optimized cookies (F4) aqueous extract was found to be 6.85, which is close to neutral. This suggests that the formulation is safe for oral ingestion and does not provide a danger of mucosal irritation. A food product should have a near-neutral pH, which shows that the contributions of acidic and alkaline ingredients are balanced.

3.4 Phytochemical Screening of Mucuna pruriens

To find the bioactive phytoconstituents that provide Mucuna pruriens extract its hematopoietic and antioxidant qualities, a qualitative phytochemical screening was carried out using conventional chemical assays. Table 4 and Figure 20 show the results.

Table 4: Phytochemical Screening of Mucuna pruriens Extract

Figure 20: Qualitative phytochemical screening of Mucuna pruriens extract — test tubes (a–f) showing colour reactions for phenols, alkaloids, glycosides, saponins, tannins, and carbohydrates respectively

Phenols (dark green ferric chloride reaction), alkaloids (orange Wagner precipitate), glycosides (brownish green Keller–Kilani reaction), saponins (stable foam formation), tannins (green ferric chloride reaction), and carbohydrates (violet Molisch ring) were all detected by phytochemical screening. The nutraceutical cookie formulation benefits from the antioxidant, anti-inflammatory, and hematopoietic qualities of these bioactive phytochemicals, especially alkaloids, tannins, saponins, and flavonoids.

3.5 Nutritional Composition Analysis

3.5.1 Iron Content

Based on AOAC food composition data, the optimized F4 batch's total estimated iron content was 14.68 mg per batch. The largest individual iron component was found in halim seeds (10.00 mg), which were followed by dates (0.29 mg), almonds (0.37 mg), oats (0.22 mg), black sesame (0.73 mg), jaggery (1.00 mg), and Mucuna pruriens (1.97 mg). Table 5 shows the specific iron contribution for each element.

Table 5: Estimated Iron Content Contribution per Ingredient in Optimised Cookie Batch F4

3.5.2 Protein Content

Each batch's estimated total protein content was 13.66 g. Due to its high protein content (23.0 g/100 g), mucuna pruriens supplied the biggest percentage (6.67 g), followed by oats (0.85 g), halim seeds (2.33 g), and almonds (2.12 g). Table 6 displays the full breakdown of proteins.

Table 6: Estimated Protein Content Contribution per Ingredient in Optimised Cookie Batch F4

3.5.3 Carbohydrate Content and Energy Value

Dates (21.75 g, due to 75.0 g carbohydrate/100 g) and Mucuna pruriens (17.69 g) were the main contributions to the estimated total carbohydrate content per batch, which was 59.25 g. The full analysis of carbohydrates is shown in Table 7. The Atwater method yielded a total energy value of 399.64 kcal per batch:

Energy = (Protein × 4) + (Carbohydrate × 4) + (Fat × 9) = (13.66 × 4) + (59.25 × 4) + (12 × 9) = 54.64 + 237.00 + 108.00 = 399.64 kcal

Table 7: Estimated Carbohydrate Content Contribution per Ingredient in Optimised Cookie Batch F4

3.6 Consolidated Nutritional Composition

Table 8 provides a summary of the optimized formulation F4 nutraceutical cookies' overall nutritional makeup.

Table 8: Consolidated Nutritional Composition of Optimised Nutraceutical Cookies (Batch F4)