SER 148

Extraction apparatus for

the quantitative separation of
a substance from a mixture by
the use of an organic solvent

Operation manual
IMPORTANT:
READ THE INFORMATION CONTAINED IN THE PRESENT MANUAL BEFORE
USING THE UNIT.
THE MANUFACTURER DOES NOT ACKNOWLEDGE ANY RESPONSABILITY
FOR AN IMPROPER USE OF THE EQUIPMENT, NOT RESPONDING TO
DIRECTIONS FOR USE.

The labels applied to the unit warn the user on the dangers to which he is
exposed during the use or the maintenance. The labels must be left on the unit
and substituted if they are no more readable.

Warn of danger
To prevent accidents which could be caused by the unit

Start the operation

Do not start to use the unit before reading directions for use and
maintenance

SAFETY RULES
1) The heating plate can reach a temperature of 260°C during the heating phase
but it is still hot also during the cooling phase. Hang the heat shield before
starting a cycle, to reduce risk.
2) The containers and the materials used during the work must be compatible
with the temperature set on the unit.
3) Conform to the data of the table located on the console and presented in the
operation manual which prescribes the temperature to be set for the different
solvents and which sealing gasket is to be used.

CLEANING
Always unplug the unit before cleaning. The test tube must be cool. Use a damp
cloth and not flammable, not aggressive detergents.
PERSONAL PROTECTION EQUIPMENT
The equipment used for personal protection must be compatible with the possible
dangers deriving from the working materials and from the glass vessels.

MAINTENANCE
Maintenance by the user must be limited to the aspects described by the manual.
Always unplug the unit before performing maintenance.
According to the law on products guarantee our instrument must be repaired at
our factory, save different agreements with our agents.

GUARANTEE
Starts from the date of delivery note and is referred to the number of register of
the single unit.

ALS. MI 240 rev. 2 1

 Velp Scientifica - SER 148 Manual

Contents

Forewords 4
Instrument description 5
Checking the instrument as received 5
Assembling and installing 6
Safety rules 7
Operation controls 9
Programming and starting 10
Programming the electronic timer 10
Choice of heating temperature 12
Flash point 14
Choice of seals 14
Operation cycle 15
Operations at the end of work 17
Maintenance 18
Oxidizing procedure for cleaning glassware 19
Electric scheme 20
Hydraulic scheme 22
Spare parts and accessories 23

ALS. MI 240 rev. 4 2

 Velp Scientifica - SER 148 Manual

Analytical methods

Determination of crude fat in meat and meat products 26

Determination of total fat in meat, fish, cacao and derived products 27
Determination of fat content in vegetable proteic substances 28
Extraction of oil and grease from wastewater 30
Extraction of oil and grease from sludges 31
Extraction of plasticizer mixtures from vinyl chloride plastics 32
Extraction of rosin from paper and paperboard 33
Extraction of oils from leather 34
Determination of unsaponified and unsaponifiable fat in soaps 35
Extraction of ethanol soluble components from detergent formulations 36

ALS. MI 240 rev. 4 3

 Velp Scientifica - SER 148 Manual

Forewords

Extraction by an organic solvent allows in many cases the quantitative separation

of a substance or a group of substances (e.g. fat) from a mixture of solids or
semisolids. The most widely used technique, all over the world, takes name from
the German chemist Franz Soxhlet (1848-1927) professor of chemistry at the
High School of Agricolture of Munich (Bayern); it draws by distillation a pure
solvent into contact with the mixture to be extracted.
In the case of solid or semisolid mixtures the sample is contained in a porous
cellulose or glass fiber extraction thimble which is covered by the pure solvent
which drops from an overhanging condenser. From time to time the solvent with
the extracted substances is drawn to the distillation vessel by an automatic
siphon. The porous thimble retains the fraction of the mixture not soluble in the
solvent. The dissolved substance must not be volatile at the boiling temperature
of the solvent.
The Soxhlet extraction apparatus has been modified in different aspects, some of
which attempting to increase the temperature of solvent coming into contact with
the mixture to be extracted, in order to reduce the extraction time. The Velp
extraction apparatus uses a two stage procedure, in the first one the thimble to
be extracted is directly immersed in the boiling solvent contained in the distillation
vessel; after a defined time the thimble is removed from the solvent and hangs
below the cooling condenser from which the condensed solvent drops and
washes the sample.
This procedure reduces the time required for a complete extraction to 1/4 ÷ 1/6 of
that required by a classical Soxhlet analysis. The fields of application include the
analysis of food and feed, detergents, rubber and plastic formulations,
pharmaceutical products, soil and so on, for soluble compounds as e.g. fats,
tensides, plasticizers, pesticides.
The Velp apparatus allows the use of many different solvents, a large fraction of
which is recovered at the end of the extraction cycle, reducing to a minimum the
related air pollution. The Velp apparatus is manufactured in accordance with the
safety standard IP55 that allows a risk free operation.

ALS. MI 240 rev. 2 4

 Velp Scientifica - SER 148 Manual

Instrument description

The unit is designed for the extraction of solid or semisolid materials using
solvents according to Randall technique. The technique reduces sharply the
extraction times using two phases with a final recovery of the used solvent.
In the first phase the sample containing thimble is immersed in boiling solvent,
while in the second phase it is removed from solvent and reflux washed.
Finally a large fraction of the solvent is distilled and recovered.
The unit with 3 or 6 places is optimized for manuality and safety. The chosen
materials and technical innovation allow the respect of CEI EN 60529 norms with
an IP55 degree of protection allowing a use safe of explosions with a limited
space requirement.

Checking the instrument as received

The SER 148 extractor is delivered in a cardboard box, enveloped in antishock

polyethylene sheet and polystyrene foam. When received, after packing removal
check the integrity of unit.
The instrument is delivered with 3/6 extraction cups, 3/6 Butyl seals, 3/6
extraction thimbles Ø 33 x 80 mm, 3/6 thimble adapters Ø 33 mm, tubes for
hydraulic for water connection, a heat shield and an operation manual.

ALS. MI 240 rev. 2 5

 Velp Scientifica - SER 148 Manual

Assembling and installing

Connect the instrument to a water tap for cooling water and to a water sink
according to the autoadhesive labels on the rear of the instrument (C1):
INLET The plastic tube is made fix by a band fastener. The open water
tap must give a flow at the outlet of about 5 l per minute.
DISCHARGE Cooling water to sink.

C1

Connect the instrument to electric main 230 V (or 115 V) with connections for
power and ground, protected by an earth leakage circuit breaker, tared for 0,5 A
(C2).
The connection to socket by cable and plug must be at least 1.5 m far from the
instrument. Use the plug just installed at factory. The power capacity should be
10 A.
The equipment is to be placed in a forced ventilation environment (e.g. a fume
hood). Do not use it without an adeguate air circulation to vent or in presence of
points of danger like flames or electric sparks.

C2

ALS. MI 240 rev. 2 6

 Velp Scientifica - SER 148 Manual

Safety rules
Safety devices

All electric components are protected according to IP 55 standards, assuring a

complete separation from solvents.
The equipment is protected against overheating by an electronic circuit with an
indipendent safety temperature probe. Moreover there is a continuous cross
control of the values given by the two temperature probes: when the two
temperatures differ more than 20°C thermostatting is interrupted.
The three following cases can occur:
1. Interrupted temperature probe circuit. In this condition the temperature of the
main probe raises to more than 350°C. If the interrupted circuit belongs to the
safety probe, the display shows alternately AL2 and 350.
2. Short circuited probe. The apparent temperature of the probe is less than
2°C. For the main probe the display shows alternately AL1 and 000. If the
safety probe is in short circuit the display shows alternately AL2 and 000.
3. Overheating. In this case the temperature of probes corresponds to the value
of set point +20°C. For the main probe the display shows alternately AL1 and
the temperature of heating plate. If the overheating is noticed by safety probe
the display shows alternately AL2 and the temperature of the heating plate.
In any case of not correct operation the equipment must be repaired by Velp
authorized personnel.

Recommendations for handling organic solvents

As in general when handling chemicals, the following precautions should always

be observed:
a) Control the risk phrases ( R ) as hazard warning on the containers
(e.g. R10 = flammable or R19 = may form explosive peroxides).
b) Always protect eyes and possibly also skin, wearing glasses and gloves.
c) Carry out work as much as possible in a well ventilated environment
(e.g. fume hood).
d) Wash immediately with plenty of water any splash on skin or eyes. If
necessary seek medical attention.

ALS. MI 240 rev. 2 7

 Velp Scientifica - SER 148 Manual

The supply of organic solvents in the laboratory should be reduced to an

indispensable minimum. The solvents not in use should be preserved in metallic
cabinets with holes, outside the laboratory. As far as possible it is recommended
to avoid the use of most volatile solvents if they can be replaced by a less volatile
homologous (e.g. toluene instead of benzene, heptane instead of pentane).
Remember that ethers, unsaturated hydrocarbons, aldehydes and ketones are
peroxide forming solvents and explosions can occur handling them not only by
distillation but also by friction or violent agitation. The control of the presence of
peroxides can be performed by easy to use test kits (e.g. Macherey-Nagel
Quantofix peroxide test strips, Code 913 12 and 913 19).
The destruction of peroxides is not always easy because two phases are formed
with water solutions in most of cases which do not allow polymeric peroxides to
come into contact with the used reducing agents.

WARNING !
The heating plate and the extract collecting vessels can reach a temperature as
high as 260°C (set point). Use tongs to remove the extract collecting vessels at
the end of extraction and solvent recovery phases.

ALS. MI 240 rev. 2 8

 Velp Scientifica - SER 148 Manual

Operation controls

The following controls are operated by push-buttons:

1) SET when pressed allows the display

to show the set point of
temperature for the heating plate.
Also fixes, together with the two
following push-buttons, the
temperature of heating plate from
100 to 260°C.
2) when pressed together with the
SET push-button raises the
temperature set point
3) when pressed together with the
SET push-button lowers the
temperature set point
4) START/STOP when pressed once starts the
heating of the heating plate up to
the temperature set point: when
pressed again the heating is
stopped. When the heating is on
the corresponding LED lights.
5) AIR turns on the pump blowing air
into the water cooled
condensers. The operation is
warned by the corresponding
LED.

ALS. MI 240 rev. 2 9

 Velp Scientifica - SER 148 Manual

Programming and starting

Turn on the unit by the lighted main switch located on the front panel. The display
shows the temperature of heating plate. Set the operation temperature related to
the used solvent pressing the push-buttons SET  or .
Heating starts or stops pressing START/STOP. When a solvent with heavy
vapours is used it is suggested to turn on the air pump during the phase of
solvent recovery, pressing AIR.
If a power black-out happens the unit goes in stand by condition and for starting
again heating the START/STOP push-button must be pressed.

Programming the electronic timer

The unit is equipped by an electronic timer by which it is possible to choose 10

different work programs each composed by 3 time intervals from 0 to 999
minutes corresponding to “Extraction”, “Washing” and “Recovery”.
The time periods t1, t2 and t3, related to each program follow one another. The
time count is warned by a blinking point in correspondence of the first, or second,
or third number shown by the display. The end of each period is warned by a
beeper, intermittent for the first and the second period, continuous for the third.
The sound is stopped pressing the SET key which starts also the count of the
following period and moves the position of the blinking point. At the end of the
last period the beeper is turned off after 10 seconds, the heating is stopped and
the display shows “END”.
To enter the programs the key SET is pressed for about 6 seconds, the display
shows P1. The keys UP  and DOWN  allow to move in succession the
different programs P2, P3, P4 ... P10. To set the time periods of each program,
press the key SET when the display shows the number of the chosen program.
The display shows t1 and then 000. Set the wanted time period in minutes by the
keys UP  and DOWN . Confirm the value pressing START/STOP.
Then the display will show t2 and 000. Set the wanted time and confirm by
pressing START/STOP.
The same procedure is used to set t3. After conferming the last time t3 the
display will show the temperature of the heating block and then it is possible to
start the work cycle.
If the time control is not wanted, set all the times to 000. If the general switch is
turned off, all the programs are retained in the memory and the unit is ready to
perform the last utilized program.

ALS. MI 240 rev. 2 10

 Velp Scientifica - SER 148 Manual

f the key SET is pressed the last used program is shown by the display.
It is possible to choose the wanted program by the keys UP  and DOWN 
confirming by the key START/STOP without returning to time setting.

The time count starts when the chosen heating temperature is reached. Pressing
the key START/STOP during a work cycle the temperature control of the heating
block is stopped and the time count starts from zero.
During a work cycle it is not possible to modify the time settings because the unit
does not allow to enter the program menu. If an electricity black out occurs
during operation, the heating returns pressing START/STOP and time count
starts from zero.

ALS. MI 240 rev. 2 11

 Velp Scientifica - SER 148 Manual

Choice of heating temperature

The temperature of heating plate is to be chosen in relation to the used solvent

(see Table).
The difference between the temperature of heating plate and the boiling point of
solvent varies from 100°C for those boiling at lower temperatures (e.g. diethyl
ether, B.P. 34.6°C) to 260°C for those boiling at higher temperatures.
This is due to the resistance to heat transmission presented by the glass of
extraction vessels. It is to be noted that anyway the temperature of heating plate
does never reach the ignition temperature of the used solvent.
To avoid to overcome the boiling point, which gives rise to a tumultuous start of
boiling, it is indispensable to add to extraction vessels some boiling chips before
weighing for tare determination.
Boiling chips (or bumping stones) = small spheres or pieces of glass, small
pieces of pumice or silicon carbide.

Temperature of heating plate to be set and type of seal required for some of
the mostly used solvents:

Glass cup Recommended Boiling Ignition

Solvent Recommended Seal Set Point °C Point °C
Temperature °C
Acetone 180 Butyl 56.5 538
Carbon tetrachloride 210 Viton 76.7 Not combustible
Chloroform 195 Viton 61-62 Not combustible
Diethyl ether 130 Viton 34.6 180-190
Ethanol 260 Viton/Butyl 78.5 423
Ethyl acetate 235 Butyl 77 427
Methanol 260 Viton/Butyl 64.7 470
n-Hexane 180 Viton 69 234
Petroleum ether 130 Viton 30-80 Not specified
Toluene 260 Viton 110.6 536
Trichloroethylene 250 Viton 86.7 Not combustible

ALS. MI 240 rev. 3 12

 Velp Scientifica - SER 148 Manual

Commonly used solvents listed by increasing boiling point.

TEMPERATURE °C

SPECIFIC HEAT
AUTO IGNITION

EVAPORATION
mm Hg a 25°C

HEAT gcal/g
g/ml at 20°C

PRESSURE
SOLVENT

SPECIFIC
POINT °C

POINT °C

DENSITY

VAPOUR

gcal/g/°C
BOILING

FLASH
Petroleum ether 30 ÷ 80 -40 >200°C 0.662 >400 ~0.500 ~80
Diethyl ether 34.6 -45 180 ÷ 190 0.713 442 0.473 84
Pentane 36.1 -40 (C) 309 0.626 420 0.507 85
Methylene chloride 39.7 - (605) (*) 1.325 340 0.288 60
Acetone 56.5 -18 538 0.788 180 0.538 123
Chloroform 61 ÷ 62 - (*) 1.484 100 0.234 59
Methyl alcohol 64.7 +12 470 0.791 95 0.601 263
Tetrahydrofuran 66 -17 321 0.889 200 0.486 105
n-Exane 69 -40 234 0.660 155 0.600 80
Carbon tetrachloride 76.7 - (*) 1.594 90 0.200 46
Ethyl acetate 77 +7 427 0.902 71 0.476 102
Ethyl alcohol 78.8 +13 423 0.789 42 0.680 204
Methyl ethyl ketone 79.6 -6 516 0.805 71 0.549 106
Benzene (1) 80.1 -11 562 0.879 76 0.419 104
Cyclo hexane 80.7 -18 259 0.778 81 0.520 94
Acetonitrile 81.6 +13 523 0.787 83 0.541 174
Isopropyl alcohol 82.5 +12 456 0.785 32 0.640 159
Trichloroethylene 86.7 - (*) 1.465 60 0.223 57
n-Heptane 98.4 -1 215 0.684 38 0.518 76
Isoctane 99.3 -12 250 0.692 11 0.563 71
Dioxane 101.1 +5 379 1.034 4 0.420 99
Isobutyl alcohol 108 +28 343 0.802 3 0.716 138
Toluene 110.6 +4 536 0.867 24 0.440 87
n-Butyl alcohol 117 +37 343 0.810 13 0.687 141
Acetic acid 118 +39 427 1.050 11 0.522 97
Octane 125.6 +22 210 0.703 12 0.578 73
n-amyl alcohol 138.1 +38 (C) 300 0.817 2 0.711 120
Xilene 138.5 +29 464 0.86 5 0.400 83

(*) not flammable

(C) closed cup
(1) definitely known to be cancinogenic

ALS. MI 240 rev. 2 13

 Velp Scientifica - SER 148 Manual

Flash point

The temperature at which a liquid gives off a quantity of vapour sufficient to form
an ignitable mixture with the air near the surface of the liquid or within the test
vessel.
There are two groups of experimental devices for flash point determination.
For the purposes of the official shipping regulations (ICC, OG, IATA) the flash
point is determined by the Tagliabue open cup method (ASTM D-1310/63)
usually abbreviated TOC. IATA also admits the Abel or the Abel-Pensky closed
cup tester.
Other methods used, generally for the higher flash points, are the Tag closed cup
and Cleveland open cup. The open cup methods approximate better actual
conditions.

Choice of seals

The building materials of retaining seals located between the extraction vessels
and the upper parts of the equipment (water cooled condensers) ask for a
particular attention due to their chemical resistance to different solvents.
Two materials were chosen: Viton (fluorinated polymer) and butyl rubber
(synthetic rubber).
In general terms Viton is easily dissolved by chetones and esthers, while butyl
rubber is attacked by hydrocarbons and halogenated solvents.
Before starting an extraciton it is to be controlled the compatibility of seals for the
used solvent (see Table page 12).

Butyl seal Viton seal

ALS. MI 240 rev. 2 14

 Velp Scientifica - SER 148 Manual

Operation cycle

1. Turn on the unit by the general switch located C3

on control panel.
2. Open cooling water tap.
3. Set the temperature of heating plate in relation
to the used solvent (see Table page 12). Verify the
compatibility of retaining seals.
As delivered the unit is equipped by Viton seals.
If the used solvent requires Butyl seals proceed to
change as described in “Maintenance”.
4. Introduce samples into thimbles. Connect
thimbles to thimble adapters (C3) wearing gloves
to avoid contamination (fat).
Introduce thimbles with thimble adapters into the
unit (C4) with front knobs in “Immersion” position. C4
Then place the knobs in “Washing” position (C5).
5. Place extraction vessels, preweighted with
boiling chips and filled with solvent, into position
on the heating plate (C6).
6. Pull down the lever retaining firmly the
extraction vessels.
7. Place the glass tap located on condensers in
vertical position (open) (C7).
8. Place front knobs in “Immersion” position.
9. Start heating.
10. At the end of extraction in boiling solvent,
move the knobs in “Washing” position.
C5
11. At the end of reflux washing, place the glass
tap located on condensers in horizontal position
(closed) and wait for solvent recovery into lower
part of glass condenser.
12. Disconnect the retaining lever and remove
glass extraction vessels. Remove extraction
thimbles and thimble connectors. Recover solvent
placing empty beakers on the heating plate and
opening the stopcocks.

ALS. MI 240 rev. 2 15

 Velp Scientifica - SER 148 Manual

C6 C7

Randall system

Extraction Washing Recovery

ALS. MI 240 rev. 2 16

 Velp Scientifica - SER 148 Manual

Operations at the end of work

At the end of instrument use:

1. Close the cooling water tap (A6).
2 Turn off the unit by the general switch.

Leave always empty glass beackers on the heating plate to avoid dripping.

A6

ALS. MI 240 rev. 2 17

 Velp Scientifica - SER 148 Manual

Maintenance

Cleaning of cooling tap water filter

Unplug the unit from electric supply and remove the connection to tap water.
Remove the cooling water tube from the unit and pull out the filter using small
pincers. Clean the filter under flowing water or by compressed air (A11).

Replacement of seals
Retaining seals between extraction vessels and glass condensers are to be
changed if required by the used solvent. See the table located on the unit or in
the present operation manual. Anyway seals are to be replaced if wearied or no
more elastic. Get hold of a seal by a small pincer or screwdriver and pull
downward (C9).

A11 C9

ALS. MI 240 rev. 2 18

 Velp Scientifica - SER 148 Manual

Oxidizing procedure for cleaning glassware

Glassware can be washed normally using a solution prepared dissolving 30 g

of sodium hydroxide (NaOH, technical grade), 10 g of sodium tripolyphosphate
(Na5P3O10, technical grade) and 2 g of sodium lauryl sulfate (C 12H25NaSO4,
commercial mixture).
Glassware can be cleaned by chromic acid oxidation (dichromate and
concentrated sulfuric acid), sulfonitric acid oxidation (warm concentrated sulfuric
acid with small amounts of potassium or sodium nitrate or nitric acid), or by
nitrous oxide oxidation (sodium nitrite and concentrated sulfuric or hydrochloric
acids). Chromic-sulfuric acid mixture is prepared adding slowly 1 l of
concentrated sulfuric acid (H2SO4, d.1.84, technical grade) to 35 ml of saturated
sodium dichromate acqueous solution (Na2Cr2O7, 73% w/w at 20°C, technical
grade) while stirring.
The cold mixture must be left into contact with glassware to be cleaned at least
15 minutes, better overnight. The freshly prepared solution is orange red and
becomes green after repeated use due to the reduction of Cr VI to Cr III.
Exausted chromic-sulfuric mixture must be disposed of in a proper manner due to
environmental toxicity of chromium.
Nitrous oxide oxidation of organic residues can be obtained washing the
glassware with hot concentrated sulfuric acid (H 2SO4, 70%, d. 1.62 technical
grade) to which a few drops of saturated sodium nitrite acqueous solution
(NaNO2, technical grade, 100 g into 150 ml of tap water) are added before use.
Another method is based on concentrated nitric acid oxidation: the addition of a
few drops of ethanol produces after minutes an exotermic reaction and the
evolution of oxidizing nitrous oxide.
Attention: all these treatments must be performed under a fume hood due to the
extreme toxicity of evolved nitric-nitrous fumes. After oxidizing treatment the
filters must be thoroughly rinsed with tap water and finally with deionized water
and then dried. Using nitric acid protect hands with PVC gloves and not rubber
which is attacked.

Rusting of components

Chlorinated hydrocarbons contain and release on heating traces of very

aggressive hydrochloric acid. As a consequence thimble holders and thimble
adapters can show some rusting. The very special steel used for these
component is slightly attaked by hydrochloric acid.
The rust can be easily removed by immersion of the components in a solution
composed by nitric acid, technical grade, concentrated, 2 volumes; hydrochloric
acid, technical grade, concentrated, 1 volume; tap water, 5 volumes. By heating
the solution to 50-60°C the rust is removed in a few minutes. After rinsing dry the
components.

ALS. MI 240 rev. 3 19

 Velp Scientifica - SER 148 Manual

Electric scheme 230 V/50 Hz

1) General lighted switch

2) Electric board
3) Air pump
4) Heating plate
5) Working Pt100 probe
6) Safety Pt100 probe
7) Female connector
8) Male connector

ALS. MI 240 rev. 4 20

 Velp Scientifica - SER 148 Manual

Electric scheme 115 V/60 Hz

1) General lighted switch

2) Electric board
3) Air pump
4) Heating plate
5) Working Pt100 probe
6) Safety Pt100 probe
7) Female connector
8) Male connector
9) Transformer

ALS. MI 240 rev. 1 21

 Velp Scientifica - SER 148 Manual

Hydraulic scheme

ALS. MI 240 rev. 2 22

 Velp Scientifica - SER 148 Manual

Spare parts

Cod. Art. Cod. Art. Description:

230V/50 Hz 115V/60 Hz

10.0210.52 10.1210.52 Electronic box IP 55

10.0210.11 10.1210.11 Heating plate IP 55
10.0210.14 Air pump IP 55

Accessories

Cod. Art. Description:

230V/50 Hz

11.1141 Glass extraction vessel

11.1142 Thimble connector Ø 33 mm
11.1143 Viton retaining seal
11.1144 Butyl rubber retaining seal
11.1145 Handling device for extraction cups
11.1146 Thimble weighing cup
11.1147 Pincer for weighing cups
11.1148 Pure cellulose extraction thimbles Ø 33x8 mm
11.1149 Thimbles stand (6 places)
VTCO003 Water cooled glass condenser

ALS. MI 240 rev. 4 23

 Velp Scientifica - SER 148 Manual

Flow sheet for a typical extraction of soluble

matter by an organic solvent from solid
samples

A Grind and homogenize the sample

Weigh the sample, from 3 to 10 g in relation to the presumed

B content of extractable matter

When vegetable or animal protein containing products are

examined a preliminary hydrolysis with hydrochloric acid should
C be performed. Boil one hour in a flask and separate the residue
on a paper filter

Transfer quantitatively the sample or the residue from hydrolysis

D with the paper filter into the extraction thimble and cover it with a
defatted cotton plug

E Introduce the thimble into the extraction equipment

Weigh the extract collecting vessel with some boiling stones,

F beforehand dried to constant weight, and introduce it into the
extraction equipment

G Add the solvent (30-100 ml)

Close the extraction unit, start the cooling water flow and the
H heating. When the solvent starts boiling immerse the thimble by
placing the slider into “Immersion” position

After 15-60 minutes extract the thimble from solvent placing the
I slider into “Washing” position

After 30-60 minutes of reflux washing close the stopcock

located under the water cooled condenser and evaporate
L completely the solvent from the extraction vessel.
If the used solvent has heavy vapour turn on the air pump to
improve the displacement to the condenser

ALS. MI 240 rev. 2 24

 Velp Scientifica - SER 148 Manual

Dry the extract collecting vessel with boiling stones in an oven

M during 30 minutes at a temperature high enough to evaporate
completey solvent residues

N Cool the extract collecting vessel in a dessicator and weigh it

Compute the result. The weight of extract is given by the weight of

O point N less the weight of extract collecting vessel with boiling
stones of point F and is to be referred to sample weight as point B

ALS. MI 240 rev. 2 25

 Velp Scientifica - SER 148 Manual

Determination of crude fat in meat and meat

products

The solvent extraction is performed on dried samples.

1) Sample: 3 g of grounded and homogenized material is weighed in an
extraction thimble with 1 mg proximation. Total weight-thimble weight = wet
sample weight.
2) Chemicals: Petroleum ether 40-60°C, with an evaporation residue not higher
than 10 mg/l.
Sand: Sea sand or siliceous sand with an extractable content lower than 4 mg/
5 g.
Cotton: defatted cotton wadding

Boiling stones.
3) Sample drying: Add some sand to the weighed sample in thimble and mix
with a glass rod. Clean the rod with a small flock of defatted cotton and add this
to thimble. Dry the thimble 1 hour in a 125°C oven. Remove the thimble from
oven and let it cool in a desiccator. Loosen the sample and sand mixture with a
glass rod. Clean the rod with a small flock of defatted cotton and place this on top
of thimble.
4) Extraction: Accurately weigh the glass extraction cup with a few boiling
stones (1 mg proximation). Add 40 ml of petroleum ether. Transfer thimble with
sample and sand, assembled with thimble connector to extraction unit.
30 minutes with thimble immersed in boiling solvent
60 minutes of reflux washing
5) Weighing: Dry the extract containing cup with boiling stones in an oven at
125°C during 30 minutes. Let it cool in a desiccator. Weigh to 1 mg proximation.
6) Expression of results: Refer to wet sample as weighed and give the results
as crude fat/100 g with two decimal figures.
7) Reference: AOAC. Official Methods of Analysis (1990) 15th Ed. 3rd
Supplement 1992. Arlington, Virginia, USA.

ALS. MI 240 rev. 3 26

 Velp Scientifica - SER 148 Manual

Determination of total fat in meat, fish, cacao

and derived products

A preliminary acid hydrolysis is aimed to free occluded or bound lipids

1) Sample: 2 to 10 g of grinded and homogenized matter, weighed in a 250 ml
Erlenmeyer flask (with reflux condenser) with 1 mg proximation
2) Chemicals: Hydrochloric acid (HCl) about 4 N, 100 ml of concentrated
hydrochloric acid, reagent grade, d. 1.19, mixed with 200 ml of water.
Exane or petroleum ether 40-60°C with an evaporation residue not higher than
10 mg/l.

Boiling stones.
3) Sample hydrolysis: Add 100 ml of 4 N hydrochloric acid to Erlenmeyer flask
with sample. Mix well and after assembling the reflux condenser boil 1 hour, add
100 ml of hot water and filter on a double folded paper filter. Wash three times the
Erlenmeyer flask with 60-70 ml of hot water and add the washings to the double
filter with hydrolyzed sample. Wash with water until no more acid reaction is shown
by filtrate.
4) Sample drying: Place the double filter paper in a 300 ml beaker and place
1-2 hours in an oven at 103°C, together with the Erlenmeyer flask used for the
hydrolysis. The dry filters are rolled and introduced in an extraction thimble.
The internal of Erlenmeyer flask and beaker are cleaned with a cotton plug soaked
in solvent, using forceps. The plug is added to thimble
5) Extraction:
30-60 ml of the chosen solvent
Boiling chips
30 minutes with thimble immersed in boiling solvent
60 minutes of reflux washing
6) Weighing: The extract containing vessel with boiling stones is weighed to 1 mg
proximation
7) Expression of result: Refer to wet sample as weighed and give the result as
fat g/100 g with one decimal figure
8) Reference: Amtliche Sammlung von Untersuchungsverfahren (1982). (Official
Collection of German Analytical Procedures). L 06.00-6.
Bestimmung des Gesamtfettgehaltes (Determination of total fat content).

ALS. MI 240 rev. 2 27

 Velp Scientifica - SER 148 Manual

Determination of fat content in vegetable

proteic substances

Fat content is defined as the fraction extractable with a solvent in the conditions
suggested for the utilized method. Occluded or bound lipids are freed before
extraction by acid hydrolysis.
1) Sample: 5 to 10 g of product grinded to less than 1 mm size, weighed in a 250
ml Erlenmeyer flask (with reflux condenser).
2) Chemicals: Hydrochloric acid (HCl), about 4 N, 100 ml of concentrated
hydrochloric acid, d. 1.19, mixed with 200 ml of water.
Diatomaceous flour as filter aid. Exane or petroleum ether 40-60°C with an
evaporation residue not higher than 10 mg/l.
Boiling stones.
3) Sample hydrolysis: From 50 (for a 5 g sample) to 100 ml (for a 10 g sample)
of 4 N hydrochloric acid. After assembling the water cooled reflux condenser the
Erlenmeyer flask is heated to boiling for one hour. At the enf of hydrolysis 150 ml
of water and about 1 g of filter aid are added to the sample still hot.
4) Sample filtration: A double paper filter soaked with water is used to filter the
hydrolyzed sample. The Erlenmeyer flask used for the hydrolysis is washed with
hot water and the washings are poured on the filter. Wash with water until the
washings do not show acid reaction to turmeric indicator paper.
5) Sample drying: The double paper filter is placed on a watch glass and
essiccated in an oven at 102°C for 2 hours together with the flask used for the
hydrolysis. The dry filters are rolled and introduced in an extraction thimble.
The internal of flask and watch glass are wiped with filter paper soaked in solvent
which is also introduced in thimble. Use forceps.
6) Extraction: 30-60 ml of the chosen solvent (according to sample weight)
Boiling stones
15 minutes with thimble immersed in boiling solvent
30 minutes of reflux washing.
7) Extract drying: Extraction vessel with boiling stones in an oven at 100°C for
30 minutes. Cool in a desiccator.
8) Weighing: The extract containing vessel with boiling stones is weighed to 1
mg proximation.
9) Control of extraction: Repeating the extraction on the extracted sample in
the same conditions should give an extract of less than 2 mg; if not, use longer
extraction time.

ALS. MI 240 rev. 2 28

 Velp Scientifica - SER 148 Manual

10) Expression of result: As % of fat, with 2 decimal figures, stating which

solvent was used.

g of extract
Fat % x 100
g of sample

11) Reference: Experimental Station for oil and fat industries (1988).
Determination of fat. Method N°4. Milano, Italy.

ALS. MI 240 rev. 2 29

 Velp Scientifica - SER 148 Manual

Extraction of oil and grease from wastewater

The method determines groups of substances with similar physical

characteristics on the basis of their common solubility in carbon tetrachloride.
Time ago the used solvent was a freon (trichlorotrifluoroethane) dismissed for
ecological reasons. Compounds volatilized at or below 103°C will be lost when
the filter is dried. The method is empirical andd reproducible results are obtained
only following exactly all details.
1) Sample: 1 l of wastewater collected in a wide mouth glass bottle and
immediately acidified to pH 2 or lower with hydrochloric acid (HCl) to hydrolyze
soluble soaps.
2) Chemicals: Carbon tetrachloride (CCl4), reagent grade, with an evaporation
residual not higher than 5 mg/l (0.005‰).
Distilled water
Diatomaceous flour filter aid suspension 10 g/l in distilled water
Boiling stones
3) Sample filtration: Prepare a 12 cm Buchner funnel with a disk of muslin cloth
overlaid with a filter paper disk. Filter under vacuum 100 ml of filter aid
suspension on the Buchner and wash with 1 l of distilled water. Filter acidified
sample until no more water passes the filter. Transfer paper filter to a watch
glass using forceps. Wipe walls and bottom of collecting bottle and Buchner
funnel with pieces of filter paper soaked in solvent and add them to paper filter on
watch glass. Roll the paper filter and introduce it into extraction thimble.
Dry thimble in an oven at 103°C for half an hour.
4) Extraction: 30-60 ml of solvent
Boiling stones
15 minutes with the sample immersed in solvent
30 minutes of reflux
5) Extract drying: Extraction vessel in an oven at 80°C for 15 minutes. Cool in a
desiccator.
6) Weighing: The extract containing vessel with boiling stones is weighed to
0.1 mg proximation.
7) Expression of result: As mg/l with one decimal figure
8) Reference: APHA-AWWA-WPCF (1985) Standard Methods for the
Examination of Water and Wastewater. 16th Ed. Washington D.C., USA.

ALS. MI 240 rev. 2 30

 Velp Scientifica - SER 148 Manual

Extraction of oil and grease from sludges

Acidified sludge essiccated by heating gives low results. Drying is performed

mixing the sludge with magnesium sulfate monohydrate which combines with
water for a 75% of its weight giving the eptahydrate.
1) Sample: About 200 g of wet sludge collected in a wide mouth glass bottle.
Weigh 20 ± 0.5 g of sample in a 150 ml beaker. Acidify to pH 2.0 with
hydrochloric acid (HCl) to hydrolyze soluble soaps.
2) Chemicals: Carbon tetrachloride (CCl4) reagent grade with an evaporation
residue not higher than 5 mg/l (0.005 ‰)
Magnesium sulfate monohydrate. Dry overnight a thin layer of heptahydrate
(MgSO4 . 7 H2O) in an oven at 150°C.
3) Sample dehydration: Add 25 g of magnesium sulfate monohydrate to the
acidified sample in the beaker. Stir to a smooth paste and spread on the sides of
beaker. In 15-30 minutes the paste solidifies. Remove the solids and grind in a
mortar. Add the powder to an extraction thimble. Wipe mortar and beaker with
filter paper wetted with solvent and add to thimble.
4) Extraction: 30-60 ml of solvent
Boiling stones
15 minutes with the sample immersed in solvent
30 minutes of reflux washing
5) Extract drying: Extraction vessel in an oven at 80°C for 15 minutes. Cool in a
desiccator.
6) Weighing: The extract containing vessel with boiling stones is weighed to
0.1 mg proximation.
7) Expression of results: Refer to wet solids, or dry solids if known, as mg/kg.
8) Hydrocarbons: Fatty acids as polar materials can be selectively removed by
silica gel (Si02). Dissolve the residue from oil and grease extraction in 10 ml of
carbon tetrachloride and pass through a small glass column (Ø 1 cm 20 cm high)
filled with 30-60 mesh silica gel, only mineral oils are not retained.
9) Reference: APHA-AWWA-WPCF (1985). Standard Methods for the
Examination of Water and Wastewater. 16th Ed. Washington D.C., USA.

ALS. MI 240 rev. 2 31

 Velp Scientifica - SER 148 Manual

Extraction of plasticizer mixtures from vinyl

chloride plastics

The method is devised for the extraction of monomeric and polymeric plasticizers
containing simple phtalate esters from polyvinyl chloride compositions for
following gas chromatographic analysis.
1) Sample: 1 to 5 g of polymer grinded to 0.02 mm size (20 US mesh) or 5 by
5 mm squares cut from 0.6 to 2 mm thick sheets.
2) Chemicals: 2+1 mixture by volume of carbon tetrachloride (CCl4) and
methanol (MetOH) both reagent grade.
Boiling chips
3) Extraction: The sample weighed to 0.1 mg proximation is placed in an
extraction thimble and extracted 30 minutes immersed in boiling solvent
30 minutes under reflux washing
4) Extract drying: Extraction vessel with boiling stones in an oven at 80°C for
30 minutes
5) Weighing: The extract containing vessel with dried extract and boiling stones
is weighed to 0.1 mg proximation
6) Gas chromatography: For following gas chromatographic analysis the extract
is dissolved with chloroform
7) Reference: The American Society for Testing Materials (1975). Extraction and
Analysis of Plasticizer Mixtures from Vinyl Chloride Plastics. ASTM Standard
Practices. Philadelphia, USA.

ALS. MI 240 rev. 2 32

 Velp Scientifica - SER 148 Manual

Extraction of rosin from paper and

paperboard

The extracted matter is really colophony but in paper manufacture the term
“rosin” is commonly used.
1) Sample: 5-7 g weighted to 0.01 g proximation, cut into 6x40 mm strips. Heavy
paperboard can be cut into thinner layers to improve extraction
2) Sample pretreatment:: If the paper contains carbonates the sample must be
immersed 5 minutes into 1 N hydrochloric acid (HCl), washed with water and
dried at room temperature.
3) Chemicals: Acidified ethanol, 4 ml of concentrated hydrochloric acid (HCl)
into 1 l of 95% ethanol (EtOH)
Boiling stones
4) Extraction: 30-60 ml of solvent
Boiling stones
15 minutes with the sample immersed in solvent
30 minutes of reflux washing.
5) Extract drying: Extraction vessel in an oven at 105°C for 15-20 minutes
6) Weighing: The extraction vessel with the dry extract and boiling stones is
weighed to 0.01 g proximation
7) Calculation:

extract and vessel weight - vessel weight

% rosin  x 100
sampleweight

8) Reference: TAPPI Standard T 408 05-61. Rosin in Paper and Peperboard

TAPPI Standards and Suggested Methods. Technical Association of the Pulp
and Paper Industry. Atlanta, Georgia, USA.

ALS. MI 240 rev. 2 33

 Velp Scientifica - SER 148 Manual

Extraction of oils from leather

During tanning of hides to make leather, vegetable, animal and mineral oils are
used to give water proofness and flexibility
1) Sample: 20 g of leather grinded to less than 1 mm size, weighed to 1 mg
proximation
2) Chemicals: Petroleum ether 40-60°C or dichloromethane (CH2Cl2), reagent
grade
Boiling chips
3) Sample drying: Transfer quantitatively the sample to an extraction thimble,
cover with a defatted cotton plug and keep in an oven at 103°C for 2 hours
4) Extraction: 30-60 ml of solvent
Boiling stones
30 minutes immersed in boiling solvent
60 minutes of reflux washing
5) Extract drying: Extract containing vessel with boiling chips in an oven at
103°C for 30 minutes. Let cool in a desiccator for 30 minutes.
6) Weighing: The extract containing vessel with boiling chips is weighed to 1 mg
proximation.
7) Expression of results: As g/100 g with 2 decimal figures

extract wight (g)

% extractables  x 100
sampleweight (g)

8) Reference: Experimental Station for hides and tanning materials industries

(1992). Leather: determination of substances extractable by dichloromethane.
Neaples, Italy (in Italian). J.Soc. Leather Tech. Chem. (1965). Determination of
substances soluble in dichloromethane. I.U.C./4 49,10.

ALS. MI 240 rev. 3 34

 Velp Scientifica - SER 148 Manual

Determination of unsaponified and

unsaponifiable fat in soaps

Neutral fat and unsaponifiable fat (mostly hydrocarbons) can be determined

together or separated. The method considers the unsaponifiable fat not volatile at
105°C.
1) Sample: 10 to 20 g of soap weighed to 1 mg proximation is kept overnight in
an oven at 105°C.
2) Chemicals: Petroleum ether 40-60°C, reagent grade. Potassium hydroxide
(KOH), ethanol 1 N solution, 56 g in 1 l of 95% ethanol.
Boiling stones.
3) Sample extraction: The dry soap sample is triturated in a mortar with some
washed dry silica sand and transferred quantitatively to an extraction thimble.
15 minutes immersed in boiling solvent
30 minutes of reflux washing
4) Extract drying: Extract containing vessel with boiling stones in an oven at
105°C for 30 minutes. Let cool in a desiccator for 30 minutes.
5) Weighing: The extract containing vessel with boiling stones is weighed to 1
mg proximation. The weight is due to neutral fat and unsaponifiable fat and is to
be referred to the weight of sample before drying.
6) Determination of unsaponifiable fat: The extract of preceding determination
is saponified directly in the extract vessel by boiling one hour with 20 ml of
ethanol KOH 1 N solution. At the end the residue is dissolved with three 20 ml
volumes of petroleum ether which removes the unsaponifiable fat and whose
weight can be determined by evaporation in a weighed vessel.

ALS. MI 240 rev. 2 35

 Velp Scientifica - SER 148 Manual

Extraction of ethanol soluble components

from detergent formulations

Soluble components are represented by organic tensides and by sodium

chloride. Also traces of other salts and raw materials, not transformed, present in
synthetic tensides.
95% (V/V) ethanol does not dissolve inorganic components as sulfates,
carbonates, borates, silicates, phosphates and zeolites.
1) Sample: About 5 g of formulate introduced into an extraction thimble which
was previously tared in a weighing bottle together with a cotton plug.
Weigh with 0.1 mg proximation.
Dry in an oven at 105 ± 2°C.
Close the thimble with the cotton plug and introduce it into the extraction unit.
2) Chemicals: Ethanol 95% (V/V) neutralized with phenolphtalein indicator.
Boiling chips
3) Extraction: 100 ml of solvent
Boiling chips
30 minutes with the sample immersed in solvent
60 minutes of reflux washing
4) Extraction drying: Extraction vessel in an oven at 105 ± 2°C for one hour or
to constant weight
5) Drying of insoluble residue: Introduce the thimble with insolubilized residue
in the same weighing bottle used before. Dry in an oven at 105 ± 2°C during 3
hours.
Let cool in a dessiccator and weigh with 0.1 mg proximation
6) Expression of results:
Percent of soluble components:
g of solublesubstances
x 100
g of sample

Percent of insoluble components:

g of insolublesubstances
x 100
g of sample

7) Reference: Italian norm UNI 24017 (1993). Detergents and tensides.

Determination of ethanol soluble and insoluble substances (in Italian).

ALS. MI 240 rev. 2 36

 Velp Scientifica - SER 148 Manual

Declaration of compliance

We
manufacturer VELP SCIENTIFICA s.r.l.
address Via Stazione, 16
20040 USMATE (MI)
Italy

Under our responsability declare that the product conforms to inforce

prescriptions on safetry and hygiene of machines directive 89/392/CEE, low
voltage directive 73/23/CEE, electromagnetic compatibility directive 89/336/CEE
plus modifications and that the documents listed in the annex V are available at
Velp’s offices as per the machine directive.

ALS. MI 240 rev. 2 37

 Velp Scientifica - SER 148 Manual

Thanks for choosing a VELP’s product !

From more than 15 years Velp offers to the professional in the field a range of
sophisticated and reliable equipment which make available high levels of
know-how and operative capacity at competitive price.
Velp works according to ISO 9001 Certification of Quality.
Instruments are built according to the International norms IEC 1010-1 and to the
rules of CE mark.

We present you our products’ Lines :

Environment
C.O.D.
B.O.D. – B.O.D. Sensor
REFRIGERATED THERMOSTATS
JAR TEST
METALS IN SLUDGES AND SEDIMENTS
NEPHELOMETRIC TURBIDITY
NITROGEN IN DIFFERENT FORMS
PERFORMANCE OF ANAEROBIC DIGESTERS
PHENOLS
LEACHABILITY TEST

Stirring
MAGNETIC STIRRERS
HEATING MAGNETIC STIRRERS
MULTIPLACE MAGNETIC STIRRERS
LIGHTED MAGNETIC STIRRERS
HEATING PLATES
VORTEX
STIRRERS

Food and feed

DIGESTERS
STEAM DISTILLING UNITS
SCRUBBER
CATALYST TABLETS
NITROGEN-FREE WEIGHING BOATS
EXTRACTORS FOR RAW FIBER
EXTRACTORS FOR DIETARY FIBER
EXTRACTORS FOR SELECTIVE EXTRACTION OF SOLUBLE PRODUCTS BY
SOLVENT (FOR FAT)

Pumps
PERISTALTIC PUMPS