INTRODUCTION:

Extraction^{1, 2}

In the extraction process, substances are separated by dissolving each component with specific solvents, yielding the Raffinate Phase (rich in the original feed solvent) and the Extract Phase (rich in solute). When component volatility equals 1, traditional distillation is ineffective, making Extraction the preferred, efficient method. Extraction also serves as a cost-effective alternative when distillation becomes prohibitively expensive.

Curcumin^{3, 4,5}

Curcumin, the primary phytochemical in Curcuma longa L., lends its yellow color. India, a major turmeric producer, has a history of using it in food, dyeing, and traditional medicine.

Found in the rhizome, curcumin manages inflammation, offers antibacterial effects, and shows therapeutic potential in diabetes, inflammatory disorders, and cancers. Its content variations stress selecting high-curcumin plants. Extraction involves ethanol and column chromatography, revealing diverse pharmacological activities: antimicrobial, antidiabetic, anti-inflammatory, anticancer, and antioxidant.

Methodology and Techniques:

Choice of Solvent

The choice of solvent for the extraction are Hexane, acetone, Ethanol etc. The acetone and alcohol are good exctracting solvent and the extractive yield can also be high from the extraction. The acetone is Slightly superior to the alcohol. The results shows that the yield from the acetone is higher than the Other solvent.

Extraction of Curcumin

Curcumin can be extracted from the following Methods:

• Conventional Extraction Using Soxhlet^6,7

Solvent extraction, commonly known as "solid-liquid extraction" or "maceration," is widely used. Various solvents, including non-polar organic solvents and combinations of organic solvents and water, are employed for curcumin extraction. Turmeric rhizomes or roots are dried at 105°C for 3 hours, ground to a fine powder, and stored in a refrigerator. The Soxhlet method involves weighing 60 mg of turmeric powder and placing it in a filter paper thimble. Ethanol is chosen as the extracting solvent. Extraction occurs at 60-70°C for 8 hours in the Soxhlet apparatus.

• Extraction Process:- The oleoresin was extracted from the provided sample using hexane, and the resultant mixture was processed in a rotary vacuum evaporator. Approximately 0.5 ml of the oleoresin was added to 10 ml of hexane. Spectrometric readings were then recorded, with hexane serving as the reference blank. The curcumin content was found to be predominantly located in the lower layer of the separating funnel, while the oleoresin was concentrated in the upper layer. The obtained curcumin was subsequently subjected to a drying process and subsequently powdered.

Fig. 1- Extraction process

• Microwave-assisted extraction of curcumin^8,9,11:

Microwave-Assisted Extraction (MAE) is an established and eco-friendly method that uses microwave energy (300 MHz to 300 GHz, commonly 2.45 GHz) to induce heating, expediting mass and heat transfer processes. This efficiency enhances the extraction of bioactive compounds, like curcumin, from plant materials into the solvent medium.

• Ultrasound-assisted extraction of curcumin^8,10,11:

Ultrasound-assisted extraction (UAE) relies on sound waves (20 kHz to 100 MHz) causing microbubble formation and subsequent cavitation. This leads to shear forces, shock waves, and acoustic streaming, converting kinetic energy into heat for physicochemical reactions. UAE enhances mass transfer, diffusion rates, and solvent interaction with plant compounds. Crucial variables like frequency, temperature, duration, solvent, and plant characteristics impact curcumin yield

• Enzyme-assisted extraction of curcumin¹²:

In the Enzyme-Assisted Extraction (EAE) method for curcumin, we use enzymes like α-amylase, glucoamylase, and aminoglycoside. This method is gaining attention for its efficiency and eco-friendliness. We start with 1 g of turmeric powder in a 250ml flask, mix it with 100ml of water, and add 50 ml of pH 5 McIlvaine's buffer. Enzyme concentrations ranging from 1-5% w/w of turmeric are introduced. The flask is sealed and agitated at 130 rpm, incubated at 65°C for 6 hours. Curcumin extraction with 10 ml of acetone takes 1-5 hours, followed by solvent separation and evaporation.

Analysis and characterization Techniques for curcumin:

• Thin layer chromatography¹³

Thin-layer chromatography (TLC) uses a glass plate coated with aluminum oxide or silica gel as the stationary phase. The mobile phase, a solvent, is chosen based on the mixture's components. TLC relies on compound partitioning between the stationary solid phase and the mobile liquid phase. A small quantity of a compound is applied above the base of the TLC plate, which goes into a developing chamber with solvent below the sample. Capillary action drives solvent movement through the plate. Compounds either adhere to the solid phase or dissolve in the solvent, moving up the plate based on their physical properties and adherence to the "Like Dissolves Like" principle. Compounds resembling the mobile phase move farthest, while those less soluble remain near the starting point.

• Preparation of plate¹³

Commercially available TLC plates are typically manufactured with specific particle size ranges to enhance reproducibility. These plates are produced by blending an adsorbent, such as Silica gel, with a minute quantity of an inert binding agent, such as calcium sulfate (gypsum), along with water. This amalgamation is carefully applied as a dense slurry onto an unreactive substrate sheet, typically composed of materials like glass, thick aluminum foil, or plastic. Subsequently, the resulting plate undergoes a drying process and is activated through heating in an oven for a period of thirty minutes at 110°C. The thickness of the adsorbent layer generally ranges from 0.1 to 0.25 mm for analytical applications and extends from 0.5 to 2.0 mm for preparative TLC purposes.

• Thin Layer Chromatography of Curcumin:

Preparation of stationary phase:

1. Take few grams of silica gel G in a beaker then add some amount of distilled water to make slurry. and Stir the solution properly to attain uniform viscosity. Next step is Pour the slurry on a glass slide. Keep the glass slide in hot air oven for 30 minutes at 1050C.then Dried TLC plate is obtained and handled properly.

Preparation of mobile phase -: A mixture of 10 volumes of chloroform, 1.0 volume of methanol, 0.5 volume of glacial acetic acid. Cover the beaker with the lead and keep it for 15 mins for saturation.

Spotting of TLC plate-: Mark the TLC plate 1 cm from the top and 1 cm from bottom. Next step is application of spot of curcumin methanol solution with the help of Capillary tubes which is obsolete and gives circular spotting.

Fig. 2- Curcumin Plate Fig. 3– Mobile Phase

Fig. 4 Fig. 5 Fig. 6

Figure 4, 5, shows that TLC plate under visible light under UV chamber.

Figure 6- Shows TLC plate under the wavelength of 365 nm.

distance traveled by the compound

Rf (retention factor) = ---------------------------------------

distance traveled by the solvent fornt

Calculation of Rf value:

The average Rf value of curcumin is 0.90.

From the experiment, the Rf value was obtained as follows

Distance travelled by the solute: 6.1cm

Distance travelled by the solute:7.5

From the formula it is obtained that

Rf value: 0.81

CONCLUSION:

Curcumin, also known as diferuloylmethane, stands as the predominant phytochemical within Curcuma longa L. from the Zingiberaceae family. This polyphenolic compound is responsible for the herb's distinctive yellow hue. The primary cultivation of turmeric occurs in tropical and subtropical regions, with India being the leading producer. Our investigation focused on the extraction of curcumin from turmeric, employing various advanced methods, with results compared to the conventional Soxhlet method, serving as the reference standard. While modern extraction techniques such as microwave-assisted, ultrasound-assisted, and enzyme-assisted extractions did not yield extraction efficiencies as high as the Soxhlet method, their notable advantages, including lower extraction temperatures, shorter extraction durations, and minimal solvent usage, render them more favorable extraction approaches.

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Received on 28.10.2023 Modified on 21.12.2023

Asian J. Pharm. Tech. 2024; 14(1):13-15.

DOI: 10.52711/2231-5713.2024.00004