NON THERMAL EXTRACTION OF BIOACTIVE COMPOUNDS Bioactive Compounds Extraction of Bioactive Compounds Microwave Assisted Extraction Supercritical Fluid Extraction  Ultrasound Assisted Extraction Enzyme-assisted extraction Pressurized liquid extraction Flow Diagram Equipment Mechanism of extraction Conventional techniques non-Conventional techniques Advantages Disadvantages

1. Topic:- NON THERMAL EXTRACTION OF
BIOACTIVE COMPOUNDS
Made by:- MANAV PREET
Follow on:-
https://www.facebook.com/profile.php?id=100006887986785
https://www.linkedin.com/in/manav-preet-b25a2b11a/
https://www.instagram.com/manav_bansal98
https://bit.ly/dealarea

2. Bioactive Compounds
 Bioactive compounds are defined as components of food that
influence physiological or cellular activities in the humans that
consume them.
 Bioactive compounds are phytochemicals, that are capable of
modulating metabolic processes and resulting in the promotion
of better health.
 hey exhibit beneficial effects such as antioxidant activity,
inhibition or induction of enzymes, inhibition of receptor
activities, and induction and inhibition of gene expression.
 They are being used in the prevention of cancer, heart disease,
and other diseases.
 Eg.- lycopene, resveratrol, lignan, tannins, and indoles.

3. Extraction of Bioactive Compounds
 The extraction of bioactive compounds depends on several
factors, such as the extraction technique, raw materials,
and the extraction solvent that are used.
 Two types of techniques are used for the extraction of
bioactive compounds:-
1. Conventional techniques require the use of organic
solvents, temperature, and agitation. Eg.- Soxhlet,
maceration, and hydrodistillation.
2. Non-conventional/Modern techniques are green
techniques due to reduced use of energy and the
implementation of organic solvent, which are beneficial
in relation to the environment.

5. Microwave Assisted Extraction
 Microwaves are electromagnetic fields in the frequency range from 300
MHz to 300 GHz. They are made up of two oscillating fields that are
perpendicular such as electric field and magnetic field. The principle of
heating using microwave is based on its direct impacts on polar
materials.
 The advantage of microwave heating is the disruption of weak hydrogen
bounds promoted by the dipole rotation of the molecules. A higher
viscosity of the medium lowers this mechanism by affecting molecular
rotation. The migration of dissolved ions increases solvent penetration
into the matrix and thus facilitates the solvation of the analyte.
Mechanism of extraction by MAE:-
 Separation of solutes from active sites of sample matrix under increased
temperature and pressure
 Diffusion of solvent across sample matrix
 Release of solutes from sample matrix to solvent.

6. Equipment:-
 The MAE systems are classified into multi-mode system and mono-mode
system.
 Multi-mode system allows random dispersion of microwave radiation in
cavity by a mode stirrer, while focused system (mono-mode) allows focused
microwave radiation on a restricted zone in cavity.
 Usually, the multi-mode system is associated with high pressure (HP), while
the mono-mode system is employed under atmospheric operating pressure.
However, mono-mode system can also run at HP.
Multi-mode System Mono-mode System

7. Supercritical Fluid Extraction
 Supercritical extraction is characterized by changes in temperature and
pressure, which transforms the gas in the supercritical fluid, where the gas
and liquid phases are indistinguishable. It is a mass transfer operation, in
which the convection in the supercritical solvent phase is generally the
main transport mechanism. The extraction is rapid, selective, and does not
require further cleaning; furthermore, it can be performed on small
amounts of sample.
 This technique can be summarized in two steps: the solubilization of the
chemical compounds present in the solid matrix, followed by their
separation in the supercritical solvent. The solvent flows through the
packed bed and solubilizes compounds that are present in the matrix. The
solvent subsequently leaves the extractor and by a reduction in pressure
and an increase in temperature it becomes a solvent-free extract.
 The solubility of the extracts depends on the density of the solvent. Thus,
the following different supercritical fluids have been described in studies:
CO2, propane, cooking gas, ethane, ethene, methanol, nitrous oxide, n-
butene, n-pentene, sulfur hexafluoride, and water.

8. The SFE process is governed by four key steps: extraction, expansion, separation,
and solvent conditioning. The steps are accompanied by four generic primary
components: extractor column (high-pressure vessel), pressure control valves,
separator column, and pressure intensifier (pump) for the recyclable solvent. The
system has other built-in accessories, such as heat exchangers for providing a
source of heating; condensers for condensing supercritical fluids to liquid; storage
vessels; and a supercritical fluid source. Raw materials are usually ground and
charged into a temperature-controlled extractor column forming a fixed bed,
which is usually the case for batch and single-stage mode.
Single-Stage Extraction Process

9. Ultrasound Assisted Extraction
 Ultrasound is a special type of sound wave, and its frequency ranges from
20 kHz to 100 MHz. Like other waves, it can pass through a medium by
creating compression and expansion. Therefore, this process produces a
phenomenon known as cavitation, which further leads in production,
growth, and collapse of bubbles. A large amount of energy can be produced
during the conversion of kinetic energy of motion, and thereby, it helps in
heating the contents of the bubble.
 Bubbles have temperature about 5000K, pressure of 1000 atmosphere,
heating, and cooling rate above 1010K/s. Based on this principle, UAE has
been developed.
 The effects caused by the ultrasonic waves are compression and expansion
cycles during the passage through the fluid. The expansion can create
bubbles or cavities in a liquid. The combination of these factors (pressure,
heat, and turbulence) is used to accelerate mass transfer in the extraction
process.

10. Equipment:-
 UAE experiments can be performed with a sonotrode and a glass reaction tank. The
double-layered mantle of the reactor, which allows the control of extraction
temperature with a cooling system by means of water circulation. The transducer is
connected to the horn with a ‘booster’ installed in amplification mode And finally
the sonotrode, which needs to be immersed into the middle of the liquid and
samples have to be filled in the tank. Continuous UAE is carried out with an
apparatus, which is made up of a circulatory pump and the inlet is placed in a large
beaker, which contains water and samples.
Batch ultrasonic-assisted extraction

11. Enzyme-assisted extraction
 The addition of specific enzymes such as cellulase, α-amylase,
and pectinase during extraction enhances recovery by breaking
the cell wall and hydrolyzing the structural polysaccharides
and lipid bodies.
 There are two approaches for EAE process:- 1. Enzyme-assisted
aqueous extraction (EAAE), 2. Enzyme-assisted cold pressing
(EACP).
 EAAE methods have been developed mainly for the extraction
of oils from various seeds, whereas, EACP technique is used to
hydrolyze the seed cell wall because in this system
polysaccharide-protein colloid is not available.
 Various factors including enzyme composition and
concentration, particle size of plant materials, solid to water
ratio, and hydrolysis time are recognized as key factors for
extraction.

12. Flow Diagram for the Enzyme Assisted Extraction

13. Pressurized liquid extraction
 The concept of PLE is the application of HP to remain solvent liquid
beyond their normal boiling point. HP facilitates the extraction process.
Automation techniques are the main reason for the greater development
of PLE-based techniques along with the decreased extraction time and
solvents requirement.
 PLE technique requires small amounts of solvents because of the
combination of HP and temperatures, which provides faster extraction.
This technology operates under pressures generally ranging from 100 to
1,000 Mpa.
 High pressures generate the deprotonation of the charged groups and
disrupt hydrophobic bonds and salt bridges, resulting in changes in
form and the denaturation of proteins. Additional solvent can enter the
cells and further compounds can permeate the cell membrane,
increasing the yield.

14.  The solvent was pumped into the extraction cell, which was placed in an electrical
heating jacket at a desired temperature, until the required pressure was obtained.
Extraction samples were placed in a 6.57cm3 extraction cell containing a sintered
metal filter at the bottom and upper parts. The cell containing the sample was
heated, filled with extraction solvent, and then pressurized. The sample was placed
in the heating system for 5min to ensure that the extraction cell would be at the
desired temperature (313–393 K) during the filling and pressurization procedure.
After pressurization, the sample with pressurized solvent was kept statically at the
desired pressure (5–10MPa) for the desired time (3–15min). After PLE, the extracts
were rapidly cooled to 5°C in ice water using amber flasks to prevent anthocyanin
degradation.
Pressurized liquid extraction set-up