JIMMAA UNIVERSITY

COLLEGE OF NATURAL SCIENCES

DEPARTMENT OF CHEMISTRY

Title:- Soxhlet Extraction

Prepared by: W/senbet Bogale

Submitted to: Abera Gure (PHD)

Apr,27/2021
Jimma ,Ethiopi

Table of contents

0
 Contents page

1. Back ground and History………………………………………………………………1

2. Principle (Mechanism) of Soxhlet Extraction…………………………………………3
3. Parameter Affecting their extraction afficieny…………………………………………4
4. Conventional Soxhlet Extraction………………………………………………..……..4
5. A Soxhlet Extraction Procedure………………………………………………….…….6
6. Sample pretreatment before Soxhlet extraction………………………………………...6
7. Reagent and Extraction Solvents………………………………………………….…… 7
8. Improvements in Soxhlet Extraction……………………………………………...…….7
8.1. Automated Soxhlet Extract’n (Soxtec)……………………………………………….…8
8.2. Focused Microwave-Assisted Soxhlet Extraction (FMASE)……………………...……9
8.3. High-pressure Soxhlet extraction…………………………………………………...….10
8.4. Ultrasound-Assisted Soxhlet Extraction…………………………………………...…...11
9. Application of soxhlet extraction……………………………………………………… 12
10. Concolution…………………………………………………………………….………12
11. Reference……………………………………………………………………………….…..13

LIST OF FIGURE

1
 Fig 1 Schematic diagram of a Soxhlet extractor.
Figure 2. Conventional Soxhlet extractor
Figure 3. Diagram of the Randall extractor (left) compared with the
conventional Soxhlet extractor (right)
Figure 4. Diagram of the first FMASE prototype
Figure 6. Sectional view of the high-pressure Soxhlet extractor
Figure 7. Diagram of an ultrasound assisted Soxhlet extractor

1. Back ground and History

A solvent extraction is any technique that is used to separate or isolate a species from a mixture
of compounds or impurities which relies on the solubility characteristics of the particular species
involved.Is the most commonly used method for solid sample preparationIs solid-liquid
extraction which is also known as leaching or lixiviation in physicochemical terminology.It
enables analytes to be removed and separated from solid matrices and also from other
compounds that could interfere in the analytical process. One form of solid-liquid extraction is
Soxhlet extraction. It was invented by German chemist, Franz Von Soxhlet in 1879

Then in 1970s, Edward Randall modified to cut the extraction time to as little as 30’.In all
solvent extractions an attempt is made to take advantage of differences in solubility between the
species being separated.Von Soxhlet devised a liquid/solid extraction apparatus in which a
sample is placed in a cellulose thimble and stationed over boiling solvent.Condensed solvent
would then drip into the sample, solubilizing extractable material and then siphon back into the
boiling solvent, where this cycle would then repeat.After several cycles over many hours, the

2
apparatus is disassembled and the solvent, now containing extract (fat), is evaporated off, leaving
the residue for further analysis.

2. Principle (Mechanism) of Soxhlet Extraction

The extraction of organic compounds, including pesticides, polycyclic aromatic hydrocarbons
and phenols from matrices (soils, sewage sludges, vegetables, plants), has historically been
carried out by using Soxhlet extraction.An alternate methods include the use of:mechanical
shaking, often referred to as shake-flask extraction, orultrasound, in the form of a sonic bath or
probe (sonication)are undoubtedly faster than Soxhlet extraction but it is the former which is
regarded as the benchmark against which all other approaches are often compared.

The mode (principle) of operation of all extraction systems is that organic solvent under the
influence of heat (and pressure) will desorb, solvate and diffuse the organic compounds from the
sample matrix allowing them to transfer into the bulk (organic) solvent.

1: Stirrer bar/anti-bumping granules,2: Still pot (extraction pot) - still pot should not be overfilled
and the volume of solvent in the still pot should be 3 to 4 times the volume of the soxhlet
chamber.,3: Distillation path,4: Soxhlet Thimble,5: Extraction solid (residue solid),6: Syphon

3
arm inlet,7: Syphon arm outlet,8: Expansion adapter,9: Condenser,10: Cooling water in,11:

Cooling water out

Fig 1 Schematic diagram of a Soxhlet extractor.

3. Parameter Affecting their extraction afficieny

Extraction efficiency is influenced by three interrelated factors:- solubility, mass transfer, and
matrix effects

Solubility of an analyte:

Depends largely on the type of the solvent, and for a selected solvent, its solubility is affected by
temperature and pressure

Mass transfer:

Refers to analyte transport from the interior of the matrix to the solvent.It involves solvent
penetration into the matrix and removal of solutes from the adsorbed sites.It depends on the
diffusion coefficient as well as on the particle size and structure of the matrix. High temperature
and pressure, low solvent viscosity, small particle size, and agitation facilitate mass transfer.

Matrix effect:

The solvent–analyte interactions should be accounted forIt influences solvent selection which in
turn depends largely on the nature of the analytes and the matrix.Polar for polar analytes (e. g.
methylene chloride)Nonpolar for nonpolar analytes (e. g. hexane).The water-miscible solvents
can penetrate the layer of moisture on the surface of the solid particles, facilitating the extraction
of hydrophilic organics.The hydrophobic solvents then extract organic compounds of like
polarity.

Temperature and pressure are also considered.Classical methods including Soxhlet extraction,
automated Soxhlet extraction, and ultrasonic extraction are operated under atmospheric pressure,
with heating or ultrasonic irradiation.

4. Conventional Soxhlet Extraction

4
Classic Tool for the Extraction of Solid Samples.The analysis of solid samples is difficult and challenging
owing to the complexity of sample matrices.The need for sample pretreatment, homogenization, and
preparation. Soxhlet extraction has long been used for the extraction and recovery of valuable analytes
from different solid matrices.The apparatus for Soxhlet extraction consists of:solvent reservoir –
Distillation flask,extractor body, Extraction thimble,an electric heat source (e.g. an isomantle) &a water-
cooled reflux condenser.

Two variations of the apparatus are possible: (A) Solvent vapour passes external to the sample containing
thimble, which results in cooled organic solvent passing through the sample; this extraction process is
relatively slow. (B) Solvent vapour surrounds the sample containing thimble; the hot organic solvent
allows more rapid extraction.but the mode of operation of both is the same

A B

1. Condenser:
Its function is to cool the solvent vapour and cause it to condense(turn back into a liquid)

2. Extraction thimble:
Used to hold the solid sample and allow for the condensed solvent to saturate and pass through thereby
extracting active material.This can be likened to a filter in that it retains the insoluble components and
allows whatever is dissolved to pass.

3. Distillation flask:
Function is to hold the solvent pool and serve as a reservoir for the concentrated material.

5
 Figure 2. Conventional Soxhlet extractor

5. A Soxhlet Extraction Procedure

A Soxhlet extraction procedure usually involves the following steps.

First, the sample material is packed in filter paper and placed in the thimble.Next, vapors of fresh
solvent, produced in a distillation flask, pass through the thimble containing the material to be
extracted and are liquefied in the condenser.When the liquid reaches the overflow level in the
thimble, a siphon aspirates the solution, and the liquid falls back into the distillation flask,
carrying the extracted solutes into the bulk liquid.The separation of solute from solvent takes
place in the distillation flask.Then solute is left in the flask and fresh solvent vapors pass back
into the solid bed of sample material.The operation is repeated until complete extraction is
achieved.

6. Sample pretreatment before Soxhlet extraction

The type and nature of the sample determine the number of different steps that have to be
incorporated during sample preparation procedure.For non-polar, non(semi)volatile organic
compounds:water-containing solid matrices such as soils, plant tissues, foodstuffs, sediments,
sewage sludge etc. should be dried with a desiccant (e.g., sodium sulfate).The presence of water
in the sample:can decrease solvent extraction efficiency, leading to low recoveries of non-polar
compounds.May cause water-soluble compounds be co-extracted.For fat or other non-polar

6
analytes bound to proteins, carbohydrates and/or minerals:hydrolysis has to be carried out in
order to break bonds (solubilize).When samples include a lot of water-soluble compounds, pre-
extraction with water followed by desiccation is essential

7. Reagent and extraction Solvents

Should be based on its ability to dissolve target analytes while not affecting the sample matrix.
the type of sample and the strength of analyte–matrix interactions should also be
considered.Polarity of the extraction solvent should be similar to that of the target analytes.For
polar analytes, ethanol or methanol are mostly used.For lipophilic components, two solvents of
different polarity, i.e., n-hexane and methanol, which are the solvents most commonly used to
extract the lipophilic and total (lipophilic and hydrophilic) fractions respectively.

REAGENTS AND STANDARDS

 Reagent grade methylene chloride.

 Organic-free reagent water.
 Sodium sulfate (granular, anhydrous), Na2SO4. Purify by baking at 400ºC for 4
hours, cooling and rinsing the sodium sulfate with methylene chloride, . A method
blank must be analyzed, demonstrating that there is no interference from the
sodium sulfate.
 Extraction solvents - All solvents must be pesticide quality or equivalent.
 Soil/sediment and aqueous sludge samples shall be extracted using either of the
following solvent systems:

 Methylene chloride, CH2Cl2.

 Exchange solvents - All solvents must be pesticide quality or equivalent.
 Hexane, C6H14.
 Acetonitrile, CH3CN.
8. Improvements in Soxhlet Extraction
The most significant drawbacks of Soxhlet extraction are the long time required for the
extraction and the large amount of organic solvent wasted, which is not only expensive to
dispose of but which can cause environmental pollution itself.Moreover, the conventional device
is not easily automated.

7
There are two different ways in which to circumvent the drawbacks of conventional Soxhlet
extraction, namely: the use of one of the new alternatives (such as Supercritical Fluid Extraction
(SFE), Microwave Assisted Extraction (MAE), Accelerated solvent extraction (ASE), etc.); and
the improvement of conventional Soxhlet.

8.1. Automated Soxhlet Extract’n (Soxtec)

Soxtec, which applies Randall method, was improved by EPA in 1994Leaching time is reduced,
and is fully compatible with classical Soxhlet extraction, and precision is much better in
comparison with the traditional approach.The simple principle is that the material to be
extracted, in this case, fats and waxes, is more soluble in hot solvent than in cold or room
temperature solvent.The extraction is carried out in three stages: boiling, rinsing, and solvent
recovery

Boiling is a thimble containing the sample is immersed in the boiling solvent for about 60
minutes.The solvent vapor refluxes in the condenser and the condensed solvent returns through
the sample to the boiling solvent.Extraction here is faster than Soxhlet, because the contact
between the solvent and the sample is more vigorous, and the mass transfer in a high-temperature
boiling solvent is more rapid.

Rinsing is use to remove residual extract from the sample.the thimble is lifted above the boiling
solvent and suspended for a period of time until residual traces of the extractable material are
flushed out of the sample and fall back into the solvent reservoir.the condensed solvent drips into
the sample, extracts the organics, and falls back into the solvent reservoir.this rinse–extract
process is similar to Soxhlet and is usually set for 60 minutes.If the dissolved extract is to be
subjected to further processing, the extraction is now complete. Otherwise, the solvent is
removed in the third step. Recovery(Evaporation)Is a concentration step for 10 to 20 minutes.The
solvent is evaporated to 1 to 2 mLSince the concentration step is integrated in Soxtec, the extract
is ready for cleanup and analysis.The solvent is evaporated and when the internal valve is closed,
the condensed solvent is redirected out and collected in reservoir for possible reuse or
disposal.The condensate collects in the lower part of the condenser; it can be reused for the next
extraction.Thanks to the short path to the apparatus, the sample can be evaporated until it is
almost dry.

8
Comparison of the Soxtec extraction and Soxhlet extraction include:Soxhlet is benchmark and
nearly universal.Low solvent requirement,Soxtec reduces the extraction time to 2 to 3 hours as
compared to 6 to 48 hours in Soxhlet.It also decreases solvent use from 250 to 500 mL per
extraction to 40 to 50 mL per extraction.Higher yield and recovery were reported.But higher cost
of the system

Figure 3. Diagram of the Randall extractor (left) compared with the conventional Soxhlet
extractor (right)

8.2. Focused Microwave-Assisted Soxhlet Extraction (FMASE)

The system performs the same three steps but with two significant differences:A different
heating source (microwave instead of electricity) is used; and The sample is also irradiated with
microwave energy, making it easier to rupture the analyte-matrix bonds

9
Two aspects are considered here:

Dielectric constant of the solvent – Solvents with high dielectric constant such as water,
methanol or acetonitrile have an ability to absorb microwave energy due to reorientation of
dipoles by the electric field.

Dielectric loss coefficient – is ability of the solvent to convert microwave energy into heat and
describes the efficiency of microwave energy conversion into thermal energy.Polar solvents can
be heated rapidly.

Nonpolar solvents cannot be heated due to a lack of dipoles and they are transparent for
microwave energy.In order to perform the extraction process assisted by microwaves three types
of solvents can be used:Solvents of high dielectric loss coefficient,A mixture of solvents of high
and low dielectric loss coefficient, andMicrowave transparent solvents used with a sample of
high dielectric loss coefficient.

In 1998, the first prototype focused micro-wave assisted extractor was designed by Luque de
Castro et al. and constructed by Prolabo (Paris, France).This prototype was based on standard
Soxhlet apparatus combined with a microwave device.

Figure 4. Diagram of the first FMASE prototype

8.3. High-pressure Soxhlet extraction

High pressure in Soxhlet devices is achieved by placing the extractor in a cylindrical heavy
Pyrex glass or stainless steel.It ensures high extraction efficiencies, unattended operation, a low
extraction cycle temperature (avoiding the loss of low boiling compounds), and no solvent

10
disposal.It enables a solvent-free extractable matter/residue to be obtained that can be easily
dissolved using a suitable solvent for gas chromatography studies.The main drawback of this
alternative is the change from supercritical to liquid state of the extractant, which affects Soxhlet
performance.It uses low boiling solvents or gases that can be liquefied at a temperature between
0 0C and 20 0C and a pressure up to 1500 psi (e.g., CO2, N2O, SF6, pentane)

Figure 6. Sectional view of the high-pressure Soxhlet extractor.

8.4. Ultrasound-Assisted Soxhlet Extraction

In this method the Soxhlet chamber is placed in a thermostatted bath to which ultrasound is
applied by means of an ultrasonic probe;Efficiency and precision similar to those of conventional
Soxhlet extraction and FMASE;In addition, it provides decomposition effect – sample grinding
between Soxhlet cycles is eliminated;However, its application is limited application due to
possible sample decomposition by ultrasound

11
 Figure 7. Diagram of an ultrasound assisted Soxhlet extractor

In this method the number of cycles and the ultrasonic radiation amplitude are the main variables
affecting the extraction process

9. Application of soxhlet extraction

It is the most use ful application for solid-liquid extraction in various fields such as:-

 pharmaceutic
 Envaromental
 Food stuffs
Now days soxhlet apparatus is still common and widely used as a references and standard method in
many laboratory for the extraction of oil from various material.for example Microwave-Assisted
Soxhlet Extraction fat extraction from:-prefried and fried meat,fish,bakerproduct.MIS extrsction oil
from:-olive seed,olenginous seed,meat,and bakery product.

10. Concolution
For a number of decades, conventional Soxhlet extraction has been the most widely used
extraction technique for solid samples.

There are many official methods based on this technique that have become standards against
which performance of new leaching methods based on modern extraction techniques is compared
and established.

The advantages and drawbacks of traditional Soxhlet extraction have opened up ways of
developing and introducing modifications that provide excellent extraction efficiencies of a wide
range of analytes.

12
 11. Reference

1) Pawliszyn, J. In Comprehensive Sampling And Sample Preparation: Soxhlet Extraction

and New Developments Such as Soxtec, 2012, 2, 65-82
2) J. D. Wineford, Sample Preparation Techniques in Analytical Chemistry: chemical
Analysis A Series Of Monographs On Analytical Chemistry And Its Applications, John
11.Wiley & Sons, Inc., Hoboken,NewJersey,2003,162,142-145
3) Naudé, Y., van Aardt, M., and Rohwer,E.R. (2009) Multi-channel open tubular traps for
headspace sampling, gas chromatographic fraction collection and olfactory assessment of
milk volatiles.J.Chromatogr.A,2009,1216,,2798–2804
4) F. Soxhlet, “Die gewichtsanalytische Bestimmung des Milchfettes,” Dingler’s
Polytechnisches Journal, 1879, 232, 461-465.

13