Formulation and Evaluation of a Polyherbal Anticancer Tablet Containing Moringa Oleifera and Tulsi

Herbal Medicine

Herbal medicine, also known as phytotherapy, is the practice of using medicinal plants and plant-derived substances to prevent or treat various health conditions. It is one of the oldest forms of healthcare in human history and continues to be a vital part of traditional and modern medicine systems, especially in countries like India, China, and many parts of Africa and Latin America. Among the vast range of medicinal plants, Moringa oleifera and Ocimum sanctum—commonly known as Tulsi—stand out for their remarkable therapeutic potential and extensive use in traditional as well as modern healthcare systems. Moringa oleifera, often referred to as the “Miracle Tree,” is native to the Indian subcontinent and is widely cultivated in tropical and subtropical regions. Every part of the Moringa tree, particularly its leaves, is rich in essential nutrients including vitamins A, C, and E, calcium, potassium, and iron. Beyond its nutritional value, Moringa contains a wide range of bioactive compounds such as flavonoids (quercetin, kaempferol), phenolic acids, isothiocyanates, and glucosinolates. These phytochemicals contribute to its diverse pharmacological actions, including antioxidant, anticancer, anti-inflammatory, antidiabetic, antimicrobial, and hepatoprotective effects. Studies have shown that Moringa leaf extracts can neutralize free radicals, inhibit the growth of cancer cells, reduce inflammatory mediators, and support immune modulation, making it a potent natural agent for chronic disease management. Ocimum sanctum, commonly known as Tulsi or Holy Basil, holds sacred significance in Indian culture and is revered as the "Queen of Herbs" in Ayurveda. It is a well-known adaptogen and is valued for promoting longevity, resilience, and vitality. Tulsi is rich in pharmacologically active constituents such as eugenol, ursolic acid, rosmarinic acid, apigenin, and carvacrol, which endow it with powerful anti-inflammatory, immunomodulatory, antioxidant, antimicrobial, and antistress properties. Scientific studies have validated Tulsi’s efficacy in enhancing immune function, reducing oxidative stress, improving respiratory health, and protecting against chemical-induced toxicity. It has also demonstrated antimicrobial activity against a range of bacterial, viral, and fungal pathogens, making it useful in supporting the body’s defense mechanisms. Together, Moringa and Tulsi represent a synergy of nutrition and pharmacology, offering a comprehensive profile of therapeutic benefits that address oxidative stress, inflammation, immune dysregulation, and cellular damage—factors that are central to many chronic and lifestyle-related diseases. Their use in traditional systems is now being increasingly supported by modern scientific methodologies, including phytochemical analysis, in vitro and in vivo pharmacological studies, and clinical trials. With the integration of traditional knowledge and contemporary research, these herbs are paving the way for the development of effective, safe, and natural health-supporting formulations that align with global health priorities and sustainable wellness. Herbal medicines are generally considered to be safe, cost-effective, and environmentally friendly. However, their effectiveness depends on proper selection, standardization, formulation, and quality control of the plant materials used.

Key Advantages of Polyherbal Formulations:

• Synergistic Action

The therapeutic effect of combined herbs may be greater than the sum of their individual effects. One herb may enhance the efficacy or absorption of another, leading to improved overall outcomes.

• Multi-targeted Approach

Polyherbal formulations can act on multiple biological pathways simultaneously, making them particularly useful in the management of complex and chronic diseases such as cancer, diabetes, and inflammatory disorders.

• Enhanced Bioavailability

Certain herbs in the formulation may increase the solubility, absorption, or stability of active compounds from other herbs, thereby improving the overall bioavailability of the preparation.

• Reduced Toxicity and Side Effects

Combining herbs allows for the use of lower doses of individual components, which can reduce the likelihood of adverse effects while maintaining or enhancing therapeutic efficacy.

• Broader Therapeutic Spectrum

Polyherbal formulations typically include herbs with different pharmacological activities—such as antioxidant, immunomodulatory, antimicrobial, and adaptogenic properties—offering comprehensive treatment.

• Improved Stability and Preservation

Some plant constituents possess natural antioxidant or antimicrobial properties that help stabilize the overall formulation and prolong shelf life.

• Prevention of Resistance

In infectious or cancer therapies, the use of multiple bioactive compounds with varied mechanisms of action helps prevent the development of resistance in pathogens or tumor cells.

Herbal Tablet Overview:

• Definition and Composition:
  Herbal tablets are solid dosage forms that are designed to deliver medicinal benefits from plants in a compact and easy-to-administer form. These tablets are typically made by compressing powdered herbs or herbal extracts (such as aqueous or ethanolic extracts) into solid shapes.
• Types of Herbal Tablets:
  • Conventional tablets: Designed for rapid release of active ingredients.
  • Controlled-release tablets: Formulated to release active ingredients gradually over time, providing prolonged effects.
  • Chewable tablets: Easy to chew and often formulated for children or those who have difficulty swallowing pills.

Importance of Herbal Tablets:

• Natural Alternative to Pharmaceuticals:
  Herbal tablets offer a natural alternative to synthetic drugs. They provide an option for individuals seeking relief from various health issues with minimal risk of side effects, especially in conditions where natural healing is preferred or required.
• Therapeutic Benefits:
  • Anti-inflammatory: Herbs like turmeric and ginger are widely used in tablet formulations for their anti-inflammatory properties.
  • Antioxidant: Plants like Moringa Oleifera contain powerful antioxidants that combat oxidative stress.
  • Immune Boosting: Tulsi (Ocimum sanctum) and Echinacea are used for enhancing the immune system.
• Growing Market Demand:
  With increasing awareness and a preference for holistic health, there is a growing demand for natural remedies. Many consumers now prefer herbal treatments over conventional drugs due to concerns about the side effects of synthetic drugs.

Growing Popularity of Herbal Tablets:

• Consumer Preferences for Natural Products:

Increasing health awareness has led to a surge in the demand for herbal tablets. Consumers are seeking natural, plant-based treatments for everyday health issues like digestive problems, stress management, immune support, and skin health.

• Wide Use for Chronic Conditions:

Many herbal tablets are used to manage chronic health conditions such as arthritis, high blood pressure, diabetes, and respiratory disorders. Herbs like turmeric, garlic, and ginger are popular for their anti-inflammatory properties and antioxidant activity.

Market Expansion:

With the global interest in wellness and natural products, the market for herbal tablets continues to grow, with many companies focusing on developing herbal products that meet the needs of the wellness-conscious consumer. Organic certification and sustainable sourcing are becoming key considerations for consumers when selecting herbal products.

Drug Profile:

5. Moringa oleifera:

Moringa oleifera, also known as the drumstick tree or Moringa, is native to regions of South Asia and Africa. The plant is renowned for its nutritional value and medicinal properties, particularly for its anticancer, anti-inflammatory, and antioxidant effects.

Key Bioactive Compounds in Moringa

• Isothiocyanates
• Flavonoids
• Phenolic Acids

Moringa's Anticancer Mechanisms

Tulsi (Ocimum sanctum):

Tulsi (Ocimum sanctum), or Holy Basil, is considered a sacred plant in Hindu culture and is widely used in traditional medicine for its medicinal properties. The plant is rich in essential oils, flavonoids, and alkaloids, which have demonstrated a wide range of anticancer effects.

Key Bioactive Compounds in Tulsi

• Eugenol
• Rosmarinic Acid
• Ursolic Acid

Tulsi's Anticancer Mechanisms:

Synergistic Effects of Moringa and Tulsi in Cancer Therapy

When combined, the anticancer properties of Moringa oleifera and Tulsi (Ocimum sanctum) are enhanced, offering a multifaceted approach to cancer treatment. These plants provide complementary bioactive compounds that can work in concert to:

• Enhance Apoptotic Pathways: Both Moringa and Tulsi contain compounds that activate apoptosis through distinct pathways, increasing the overall effectiveness in killing cancer cells.

• Suppress Tumor Growth and Metastasis: By inhibiting angiogenesis, metastasis, and tumor progression, the combination of Moringa and Tulsi helps prevent the spread of cancer.

• Regulate Oxidative Stress and Inflammation: The combination of antioxidants and anti-inflammatory agents from both plants reduces the molecular pathways that fuel cancer development and progression.

• Complement Conventional Cancer Treatments: Moringa and Tulsi may act as adjuvants to conventional therapies, reducing the side effects of chemotherapy and radiation while boosting the immune system and promoting recovery.

Literature Review

1. Al-Asmari et al. (2015)

Investigated Moringa oleifera extracts against breast and colorectal cancer cell lines. Results revealed dose-dependent cytotoxicity with inhibition of proliferation and induction of apoptosis in MCF-7 and HCT-8 cells.
PLOS ONE, 10(8): e0135814.

2. Saini et al. (2016)

Reviewed the phytochemical profile of Moringa oleifera, identifying quercetin, kaempferol, and niazimicin as key anticancer compounds. These phytochemicals have antioxidant, anti-inflammatory, and antiproliferative properties.
3 Biotech, 6:203.

3. Manikandan et al. (2007)

Explored ethanolic extracts of Ocimum sanctum and reported significant apoptosis in oral squamous carcinoma cells via caspase activation and ROS modulation.
Cell Biochemistry and Function, 25(2): 189–194.

4. Mondal et al. (2009)

A comprehensive review of Tulsi’s pharmacological profile highlights anticancer effects, particularly via modulation of antioxidant defense and immune response.
International Journal of Clinical Pharmacology and Therapeutics, 47(11): 601–612.

5. Bharali et al. (2003)

Demonstrated that Moringa oleifera leaf extract can significantly reduce tumor burden in mice models bearing Dalton’s lymphoma, attributed to its antiangiogenic and cytotoxic properties.
Indian Journal of Experimental Biology, 41(11): 1239–1243.

6. Siddiqui et al. (2013)

Found that Ocimum sanctum modulates key carcinogenic pathways such as NF-κB, STAT3, and COX-2, suggesting chemopreventive potential.
Nutritional and Cancer, 65(Suppl 1): 26–34.

7. Tiloke et al. (2013)

Explored the molecular effects of Moringa oleifera in A549 lung cancer cells. The extract triggered DNA fragmentation, reduced mitochondrial membrane potential, and caused apoptosis.
Asian Pacific Journal of Cancer Prevention, 14(10): 6067–6072.

8. Goyal et al. (2011)

Outlined Moringa oleifera’s role as a chemopreventive agent, including its suppression of oxidative stress, DNA damage, and pro-inflammatory markers.
Phytotherapy Research, 25(1): 17–25.

9. Prakash & Gupta (2005)

Summarized Tulsi's traditional and modern applications, including its anticancer effects through adaptogenic and detoxification pathways.
Journal of Ethnopharmacology, 93(1): 1–15.

10. Verma et al. (2013)

Reported that Moringa’s benzyl isothiocyanate exhibits strong antiproliferative effects in prostate and pancreatic cancer cell lines via apoptotic signaling.
Cancer Letters, 341(1): 30–38.

11. Pattanayak et al. (2010)

Validated Tulsi’s role in cancer management, citing its ability to scavenge free radicals and suppress tumor promotion through epigenetic modulation. Pharmacognosy Reviews, 4(7): 95–105.

Aim

To formulate, standardize, and evaluate a polyherbal tablet comprising Moringa oleifera and Ocimum sanctum, exploring its physicochemical, phytochemical, and potential anticancer properties for integrative therapeutic use.

Objectives

1. To authenticate the selected medicinal plants
   Confirm the identity and purity of Moringa oleifera and Ocimum sanctum using macroscopic, microscopic, and powder microscopy techniques in accordance with Ayurvedic and pharmacopoeial standards.
2. To prepare and standardize the herbal powders
   Subject both plant materials to proper drying, pulverization, and sieving, ensuring a consistent particle size and stability suitable for tablet formulation.
3. To develop a polyherbal tablet formulation
   Combine Moringa and Tulsi powders in a 5:1 ratio (250 mg:50 mg) with optimized pharmaceutical excipients for tablet cohesion, disintegration, and flow properties.
4. To evaluate physicochemical characteristics of the tablet
   Analyze critical quality parameters such as tablet hardness, friability, weight variation, disintegration time, moisture content, and in vitro dissolution behavior.
5. To quantify bioactive compounds using analytical methods
   Employ UV-Visible spectroscopy or High-Performance Liquid Chromatography (HPLC) for standardization by estimating marker compounds like quercetin, eugenol, and ursolic acid.
6. To ensure stability and quality consistency
   Conduct short-term stability testing under accelerated conditions to examine any physical, chemical, or pharmacological changes in the formulation.

Plan Of Work

1. Literature Review & Plant Authentication

O Conduct a thorough review of existing literature on the pharmacological properties of Moringa oleifera and Ocimum sanctum, focusing on their anticancer, antioxidant, anti-inflammatory, and immunomodulatory effects.

O Authenticate the plant materials using botanical and pharmacognostic methods to ensure proper identification and quality.

2. Collection and Preparation of Plant Materials

O Source fresh leaves of Moringa and Tulsi from certified suppliers or cultivation areas.

O Dry the leaves at controlled temperatures to preserve bioactive compounds, then grind them into fine powders suitable for tablet formulation.

3. Formulation Development

O Develop the polyherbal tablet formulation with Moringa (250 mg) and Tulsi (50 mg) per tablet, ensuring the total active ingredient (API) content comprises 60% of the tablet weight.

O Select appropriate excipients (binders, fillers, lubricants, disintegrants) to ensure tablet cohesion, stability, and ease of manufacture.

O Conduct preliminary formulation trials to optimize the composition for efficacy and stability.

4. Tablet Evaluation (Physicochemical Testing)

O Conduct essential quality control tests, such as hardness, friability, disintegration time, and dissolution rate, to ensure the formulation meets pharmaceutical standards for tablet quality and performance.

5. Quantification of Bioactive Compounds

O Use HPLC or UV-Vis spectroscopy to quantify active compounds (e.g., quercetin, eugenol, ursolic acid) in the final tablet formulation.

O Standardize the formulation based on the active compound content, ensuring the desired therapeutic potency.

6. Stability and Shelf-life Testing

O Test the stability of the tablets under different environmental conditions, such as varying temperature and humidity.

O Monitor physical and chemical changes over time, including degradation of active compounds, and assess the shelf life of the formulation.

Material And Equipment

1. Raw Plant Materials

2.Excipients

3.Testing Equipment

1. Pre-Manufacturing Preparations

Step 1: Cleaning and Sanitization

Equipment Used:

• • Cleaning agents (70% ethanol, disinfectants)
  • Stainless steel weighing balance
  • Blender, Granulator, Sieves, Tray dryer, Tablet press
  • Hardness Tester, Friability Tester, Disintegration Tester
  • Stainless steel trays, spatulas, gloves, and masks

Procedure:

1. Equipment Cleaning:

o Disassemble all processing equipment (if applicable).

o Clean each part using an appropriate cleaning solution.

o Use hot water and mild detergent for primary cleaning, followed by 70% ethanol for disinfection.

2. Sanitization of Work Area:
   • • Wipe down work surfaces, tables, and floors with disinfectant.
     • Ensure the tablet compression area is free from dust and contaminants.
     • Mop the production area with approved disinfectant solution.
3. Operator Hygiene:

o Operators must wear sterile gloves, masks, lab coats, and hairnets before entering the manufacturing area.

 o Ensure hands are sanitized before handling materials.

Step 2: Raw Material Weighing

Equipment Used:

• Digital Weighing Balance (Calibrated)
• Stainless Steel Trays
• Glass Beakers & Measuring Cylinders

Procedure:

1. Tare the balance before weighing to ensure accuracy.
2. Weigh each ingredient as per formulation:
3. Cross-check each measurement against the formulation sheet.
4. Transfer weighed materials into labeled stainless steel trays for easy identification.

2. Manufacturing Process

Step 3: Dry Blending

Equipment Used:

• • High-Speed Mixer / Blender
  • Mortar & Pestle (for small batches)
  • Stainless steel scoops and spatulas

Procedure:

1. 1. Transfer the API (Moringa Oleifera leaf powder), binder, filler, and disintegrant into the blender.
   2. Mix at low speed (30–40 rpm) for 10–15 minutes to ensure uniform distribution.
   3. If using a mortar and pestle, blend in circular motions until homogenous.
   4. Visually inspect the uniformity of the blend.

Step 4: Wet Granulation

Equipment Used:

• • Granulator
  • Measuring Cylinder, Stirring Rod

Procedure:

1. 1. Prepare the binder solution using purified water or alcohol.
   2. Slowly add the binding solution to the dry mix while continuously stirring.
   3. Mix until wet granules form with a slightly damp, non-sticky texture.

Instructions:

• • Ensure the granules are not overly wet, as excessive moisture can lead to poor drying.
  • Stop mixing when the granules form a soft clump that easily breaks upon pressure.  

Step 5: Wet Sieving (Granule Sizing Before Drying)

Purpose:

• • To break down large lumps formed during wet granulation.
  • To ensure uniform granule size, improving drying efficiency.
  • To eliminate ungranulated fine powder, ensuring batch consistency.

Equipment Used:

• • 16-Mesh Stainless Steel Sieve (For wet granule sizing)
  • Wet Granulator / Hand Sieving Setup
  • Stainless Steel Trays
  • Spatula / Wooden Spoon

Procedure:

Prepare Sieving Equipment:

• Place a 16-mesh sieve over a stainless-steel collection tray.

• Ensure the sieve is clean and dry before use.

• Transfer Wet Granules:

• Carefully transfer freshly prepared wet granules onto the sieve.

• Do not overload the sieve, as excessive material may reduce efficiency.

• Sieving Process:

• Using a spatula or wooden spoon, gently press the granules through the mesh.

• Move the material circularly to break down lumps.

• Continue sieving until most granules pass through, leaving only large ungranulated lumps.

Reprocessing Oversized Granules:

• If there are large unprocessed lumps, return them to the granulation stage.

• Adjust binder or liquid content if excessive large lumps form.

• Collect and Transfer:

• Transfer sieved wet granules into stainless steel trays for drying.

• Label each batch correctly for tracking. 

Step 6: Drying of Granules

Equipment Used:

• Tray Dryer / Hot Air Oven

• Stainless Steel Drying Trays

Procedure:

1. 1. Spread granules evenly on trays and dry at 40–50°C for 30–60 minutes.
   2. Ensure moisture content is below 5% before proceeding.

Instructions:

• • Use a Moisture Analyzer to check the residual moisture content.

Step 7: Dry Sieving (Granule Sizing After Drying) Purpose:

• • To achieve uniform particle size after drying.
  • To eliminate fine dust or oversized granules.
  • To improve flow properties, preventing tablet weight variation.

Equipment Used:

1. 1. 20-Mesh Stainless Steel Sieve (For dry granule sizing)
   2. Vibratory Sifter or Manual Sieving Setup
   3. Stainless Steel Collection Trays
   4. Spatula / Brush for Cleaning Sieve

Procedure:

1. Prepare Sieving Setup:

• • • Place a 20-mesh sieve over a collection tray.
    • Ensure sieve is completely dry before starting the process.

2. Load Dried Granules:

2. • • Transfer fully dried granules onto the sieve.
     • Spread evenly to avoid clogging.
3. Sieving Process:
   • • Gently shake or tap the sieve to allow granules to pass through.
     •  If using a vibratory sifter, set the vibration speed to low-medium to prevent breaking granules.
     • If manually sieving, rotate the material gently using a spatula.
4. Collection of Final Granules:

o Collect uniform-sized granules in the stainless steel tray.

 o If fine powder is excessive (>10% of batch weight), consider regranulation to minimize losses.

5. Handling Oversized Particles:
   • • Any granules that do not pass through the 20-mesh sieve should be reprocessed (reduced in size using a mortar & pestle or granulator) and re-sieved.
     • If too many fines are present, adjust granulation parameters in future batches records of sieve efficiency and batch uniformity.   

Step 8: Lubrication and Final Blending

Equipment Used:

• Blender / Mixer

Procedure:

1. Add Glidant (Colloidal Silicon Dioxide) and Lubricant (Magnesium Stearate) to granules.
2. Mix for 5 minutes to ensure even distribution.

Step 9: Tablet Punching (Compression) Process

Equipment Used:

1. Rotary Tablet Press Machine / Single-Punch Tablet Press
2. Tablet Compression Tooling (Punches & Dies – 10 mm standard round shape)
3. Hopper for granule feeding
4. Compression Force Adjustment Knob
5. Dust Extraction System
6. Tablet De-duster (Optional, for removing excess powder from tablets

Compression Process

• Set the tablet weight and thickness using the machine control panel.
• Adjust the compression force (typically 4–6 kg/cm² for herbal tablets).
• Start the tablet press at a slow speed for trial punches.
• Observe the tablet ejection, ensuring no sticking or capping occurs.

Instruction: Compression force should be optimized to prevent tablet friability (breaking) or lamination (layer separation).

Formulation Images

RESULT

Organoleptic Evaluation of Formulated Tablets

2. Physical Evaluation of Tablets

3. Phytochemical Screening (All Batches)

4. UV-Visible Spectroscopic Analysis