[QUANTITATIVE TEST OF LIPIDA]

I.
II.
III.
IV.

EXPERIMENT TITLE
EXPERIMENT DATE
END OF EXPERIMENT
EXPERIMENT PURPOSE
and Free Fat Acid

V.

BASIC THEORY

:
:
:
:

Quantitative Test of Lipid

October 20th, 2015
October 20th, 2015
To determining number of peroxide

Lipids are another type of organic molecule. Remember that

organic means they contain carbon (C) atoms. It's not like organic
farming at all. When you think of fats, you should know that they are
lipids. Lipids are also used to make steroids and waxes. So, if you
pick out some earwax and smell it, that's a lipid, too!.
The lipids are a large and diverse group of naturally occurring
organic compounds that are related by their solubility in nonpolar
organic solvents (e.g. ether, chloroform, acetone & benzene) and
general insolubility in water. There is great structural variety among
the lipids, as will be demonstrated in the following sections. You may
click on a topic listed below, or proceed page by page.
Lipids are a group of naturally occurring molecules that include
fats, waxes, sterols, fat-soluble vitamins (such as vitamins A, D, E,
and K), monoglycerides, diglycerides, triglycerides, phospholipids,
and others. The main biological functions of lipids include storing
energy, signaling, and acting as structural components of cell
membranes. Lipids have applications in the cosmetic and food
industries as well as in nanotechnology.
Lipids may be broadly defined as hydrophobic or amphiphilic
small molecules; the amphiphilic nature of some lipids allows them
to form structures such as vesicles, multilamellar/unilamellar
liposomes, or membranes in an aqueous environment. Biological
lipids originate entirely or in part from two distinct types of
biochemical subunits or "building-blocks": ketoacyl and isoprene
groups. Using this approach, lipids may be divided into eight
categories:
fatty
acids,
glycerolipids,
glycerophospholipids,
sphingolipids, saccharolipids, and polyketides (derived from
condensation of ketoacyl subunits); and sterol lipids and prenol
lipids (derived from condensation of isoprene subunits)
Although the term lipid is sometimes used as a synonym for fats,
fats are a subgroup of lipids called triglycerides. Lipids also
encompass molecules such as fatty acids and their derivatives
(including tri-, di-, monoglycerides, and phospholipids), as well as
other sterol-containing metabolites such as cholesterol. Although
humans and other mammals use various biosynthetic pathways to
Bio Chemistry |Quantitative Test of Lipid

[QUANTITATIVE TEST OF LIPIDA]

both break down and synthesize lipids, some essential lipids cannot
be made this way and must be obtained from the diet.
A. Fatty acids
Fatty acids or fatty acid residues when they form part of a lipid,
are a diverse group of molecules synthesized by chain-elongation of
an acetyl-CoA primer with malonyl-CoA or methylmalonyl-CoA
groups in a process called fatty acid synthesis. They are made of a
hydrocarbon chain that terminates with a carboxylic acid group; this
arrangement confers the molecule with a polar, hydrophilic end, and
a nonpolar, hydrophobic end that is insoluble in water. The fatty acid
structure is one of the most fundamental categories of biological
lipids, and is commonly used as a building-block of more structurally
complex lipids. The carbon chain, typically between four and 24
carbons long, may be saturated or unsaturated, and may be
attached to functional groups containing oxygen, halogens,
nitrogen, and sulfur. If a fatty acid contains a double bond, there is
the possibility of either a cis or trans geometric isomerism, which
significantly affects the molecule's configuration. Cis-double bonds
cause the fatty acid chain to bend, an effect that is compounded
with more double bonds in the chain. Three double bonds in 18carbon linolenic acid, the most abundant fatty-acyl chains of plant
thylakoid membranes, render these membranes highly fluid despite
environmental low-temperatures, and also make linolenic acid give
dominating sharp peaks in high resolution 13-C NMR spectra of
chloroplasts. This in turn plays an important role in the structure and
function of cell membranes. Most naturally occurring fatty acids are
of the cis configuration, although the trans form does exist in some
natural and partially hydrogenated fats and oils
B. Type of Fatty Acid
Fatty acids that have carboncarbon double bonds are known as
unsaturated. Fatty acids without double bonds are known as
saturated. They differ in length as well.
Fatty acid chains differ by length, often categorized as short to very
long.
Short-chain fatty acids (SCFA) are fatty acids with aliphatic tails
of fewer than six carbons (e.g. butyric acid).[4]
Medium-chain fatty acids (MCFA) are fatty acids with aliphatic
tails of 612[5] carbons, which can form medium-chain
triglycerides.
Long-chain fatty acids (LCFA) are fatty acids with aliphatic tails
13 to 21 carbons.[6]
Very long chain fatty acids (VLCFA) are fatty acids with aliphatic
tails longer than 22 carbons.
Bio Chemistry |Quantitative Test of Lipid

[QUANTITATIVE TEST OF LIPIDA]

C. Free Fat Acid (FFA)

The biosynthesis of fatty acids involves the condensation of
acetyl-CoA. Since this coenzyme carries a two-carbon-atom group,
almost all natural fatty acids have even numbers of carbon atoms.
The "uncombined fatty acids" or "free fatty acids" found in
organisms come from the breakdown of a triglyceride. Because they
are insoluble in water, these fatty acids are transported (solubilized,
circulated) while bound to plasma protein albumin. The levels of
"free fatty acid" in the blood are limited by the availability of
albumin binding sites.
D. Peroxide Value
Detection of peroxide gives the initial evidence of rancidity in
unsaturated fats and oils. Other methods are available, but peroxide
value is the most widely used. It gives a measure of the extent to
which an oil sample has undergone primary oxidation, extent of
secondary oxidation may be determined from p-anisidine test. The
double bonds found in fats and oils play a role in autoxidation. Oils
with a high degree of unsaturation are most susceptible to
autoxidation. The best test for autoxidation (oxidative rancidity) is
determination of the peroxide value. Peroxides are intermediates in
the autoxidation reaction. Autoxidation is a free radical reaction
involving oxygen that leads to deterioration of fats and oils which
form off-flavours and off-odours. Peroxide value, concentration of
peroxide in an oil or fat, is useful for assessing the extent to which
spoilage has advanced.
The peroxide value is defined as the amount of peroxide oxygen
per 1 kilogram of fat or oil. Traditionally this was expressed in units
of milliequivalents, although if we are using SI units then the
appropriate option would be in millimoles per kilogram (N.B. 1
milliequivalents = 0.5 millimole; because 1 mEq of O2 =1
mmol/2=0.5 mmol of O2, where 2 is valence). Note also that the
unit of milliequivalent has been commonly abbreviated as mequiv or
even as meq.

Bio Chemistry |Quantitative Test of Lipid

[QUANTITATIVE TEST OF LIPIDA]

VI.

TOOLS AND METRIALS

A. Tools
Beaker Glass
Measuring pipette
Buret
Erlenmeyer
B. MATERIALS
Oil/fat
Actetate Acid-chloroform
solution (3:2)

KI solution
Na2S2O3 0.1 N
Starch solution 1%
NaOH solution 0.1 N
Oxalic standard solution
0.1 N
PP indicator 1%
Ethanol 96%

VII.

FLOW CHART

1. Determining Peroxide value

(FFA)
Blank solution
VIII.

2. Determining Free Fat Acid

6 gram of sample

Put in Erlenmeyer flask

Added 10 mL alcohol 96% solution
Added 30 mL X.
chloroform- acetate acid Added 5-8 drops PP indicator
Shaked it well
Titrated with NaOH 0.05 N solution until changes to pink solution
Added 0.5 mLXI.
KI solution
Repeat 2 times
Keep until 20 minutes
XII.
Added 30mL distilled
water
Titrated with Na2S2O3
solution until the yellow color dissapear
XIII.
Added 0.5 mL starch solution 1%
Titrated with Na2S2O3
solution again
XIV.
5 gram
IX. of distilled water

XV.

Result of solution

XVI.
XVII.
XVIII.
XIX.
Result of solution
XX.
XXI.
XXII.
XXIII.
Bio Chemistry |Quantitative Test of Lipid

[QUANTITATIVE TEST OF LIPIDA]

XXIV.

Sample solution
XXV.

5 gram of sample
Added 30 mL chloroform- acetate acid
Shaked it well
Added 0.5 mL KI solution
Keep until 20 minutes
Added 30mL distilled water
Titrated with Na2S2O3 solution until the yellow color dissapear
Added 0.5 mL starch solution 1%
Titrated with Na2S2O3 solution again

Result of solution

Bio Chemistry |Quantitative Test of Lipid

XXVI. RESULT OF EXPERIMENT

XXVII.
XXVIII.
N

XXX. Result
XXXI. Hypothesis/React
XXXII. Conclusi
XXXV. Befor
XXXVI.
Afte
XXIX. Treatment
ion
on
e
r
XXXIX.
LXIII. Determining
number
of- Aqueous =
- Palmitat oil +
CXLVIII. The
CXXXVII.
1 peroxide
colorless
acetate acid
2 KI +CH 3 ( CH 2 )14 COOH + H 2 O CH 3oil
( CH 2 )14 COH +2 KO
- Palmitat oil =
LXIV.
chloroform =
sample
yellow ++
XL. LXV.
light yellow
(palmita
5 gram of palmitat oil
Acetate
acid

+
KI
solution
=
CXXXVIII.
XLI. LXVI.
t oil) is
chloroform
=
yellow
+++
XLII. LXVII.
not
Add 30mL chloroform acetate acid

colorless
+
aqueous
=

XLIII. LXVIII.
suitable
Shake it well
2+2 I
- KI solution =
become
2
layer

XLIV. LXIX.
with
Add 0,5 mL KI solution
2+ I 2 S 4 O 6
colorless
CXIII.Up =

XLV. LXX.Keep until 20 minute

Indonesi
S 2 O3
- Na2S2O3 =
orange
XLVI. LXXI.
a
Add 30 mL aqueous
colorless
CXIV.Down =
XLVII.LXXII.
standar
Titrate with Na2S2O3 solution - Starch solution
peach
CXXXIX.
Peroxide
XLVIII. LXXIII.
d
Add 0,5 mL starch solution 1%
= colorless
- Blanko V = 1.1
number
standard
XLIX.
LXXIV.
number
Titrate again
CII.
mL
of
fat/oil
is
max
1
L.
LXXV.
of
- Sample I V = 6.5
CIII.
meq
LI.
LXXVI.
peroxid
mL
CIV.
LII.
LXXVII.
e. The
- Sample II V = 6.5
CV.
- Number of peroxide 1 =
LIII.
LXXVIII.
oil
mL
CVI.
1.8
LIV.
LXXIX.
sample
CXV.
Result solution
CVII.
- Number of peroxide 2 =
LV.
LXXX.
is 1.8
CXVI.
CVIII.
1.8
LVI.
LXXXI.
CXLIX.Because
CXVII.
- Average = 1.8
CIX.
LVII.
LXXXII.
, the oil
CXVIII.
CX.
CXL.
LVIII.
LXXXIII.
has hit
CXIX.
CXI.
LIX. LXXXIV.
with
CXX.
- Palmitat oil =
CXLI.
LX. LXXXV.
free
CXXI.
yellow ++
LXI. LXXXVI. Determining Free Fat
radical.
- Palmitat oil +

LXII.
Acid (FFA)
2
LXXXVII.
LXXXVIII.
LXXXIX.
XC.
XCI.
XCII.
XCIII.
XCIV.
XCV.
XCVI.
XCVII.
XCVIII.
XCIX.
C.
CI.

- Alcohol 96% =
alcohol = loght
CXLII.
CL.
colorless
yellow
CLI.
CXLIII.
- PP indicator = - + PP indicator =
CLII.
6 gramCLIII.
palmitat oil
colorless
light yellow
CXLIV.
- NaoH =
- Sample 1
CLIV.
Put in
Erlenmeyer
colorless
become 2 layer
CLV.
CXLV.
CXXII.
Up
Add CLVI.
10 mL of alcohol 96%
CXII.
= peach
5-8 drops PP indicator
CXLVI. CH3(CH2)14COOH Add CLVII.
CXXIII.
Dow
Titrate
with 0.05 N NaOH
+ NaOH
CLVIII.
n = light yellow
CH3(CH2)14COON
CLIX.
- V 1 = 0.6 mL
a + H2O
CLX.
- V 2 = 0.5 mL
CLXI.
CXXIV.
CXLVII. Standard of
CLXII.
CXXV.
FFA number of
CLXIII.
CXXVI.
fat/oil is max
CXXVII.
ResultCLXIV.
solution
0.3%
CLXV. The oil
CXXVIII.
sample
- % FFA 1 = 0.128%
CXXIX.
%FFA
2
=
0.107%
is not
CXXX.
- Average = 0.117%
suitable
CXXXI.
with
CXXXII.
Indonesi
CXXXIII.
a
CXXXIV.
standar
CXXXV.
d
CXXXVI.
number
of FFA.
The
number
of FFA
from oil

sample
is
0.117%
CLXVI.
CLXVII.
CLXVIII.
CLXIX.
CLXX.
CLXXI.
CLXXII.
CLXXIII.

CLXXIV.

CLXXV.
EXPLANATION
CLXXVI.
CLXXVII.
The first experiment, to knowing peroxide
value of sample oil, this section we weight 5 gram of palmitat oil
yellow++ solution as the sample. Then, we add chloroformacetate acid colorless solution 30mL and shake it until the both of
solution mixed and the solution become light yellow. The function
of adding chloroform-acetate acid is because the characteristic of
sample oil has a polar group and non-polar group. Where polar
group will be bonded with acetate-acid and non-polar group will
be bonded with chloroform. Then add with 0.5 mL KI solution
colorless solution and the solution become yellow+++, the
function of KI solution is to indicate theres peroxide which form in
oil. If the oil has oxidized and forming peroxide, so peroxide will be
oxidizing I- become I2.
CLXXVIII.
Then keep it until 20 minute, in order to the oil
not happened rancidity and for the both of solution mix well. Then
add 30 mL aqueous colorless solution and the solution become 2
layer where up layer is orange solution and down layer is peach
solution, after that titrate with Na2S2O3 colorless solution the
solution have 2 layer. Then add 0.5 mL starch solution 1%
colorless solution, where the function of starch solution is as an
indicator changed the color and theres I2 formed. Then titrate
again with Na2S2O3 colorless solution id for the compound of
Na2S2O3 can reduce I2 which formed become I-, so the solution
become colorless again. Colorless solution is identification that all
of iod has loosed by potassium iodide. The volume that we get is:
CLXXIX.
V1 = 6.5 mL
CLXXX.
V2 = 6.5 mL
CLXXXI.
With reaction is
CLXXXII.
CLXXXIII.
2 KI +CH 3 ( CH 2 )14 COOH + H 2 O CH 3 ( CH 2 )14 COH +2 KOH + I 2

CLXXXIV.

2+2 I
2+ I 2 S 4 O6
S 2 O 3

CLXXXV.
CLXXXVI.
For the standard solution, we used blank
solution with used aqueous with same procedure with used
sample oil. And we get volume is:
CLXXXVII. V blank = 1.1 mL
CLXXXVIII.
We get for peroxide value of fat and oil for
Indonesia standard is max 1 meq. But we get average of sample

oil is 1.8 meq, so our oil sample is not suitable with standard of
peroxide value. Because the oil sample has hit with free radical
and the oil sample has destroyed because happened oxidized and
hit by sunshine, where can be accelerator to appear rancidity.
CLXXXIX.
CXC.
The second experiment, to knowing the value of
Free Fat Acid (FFA) of sample oil, this section we weight 6 gram
palmitat oil as sample oil yellow++ we put in Erlenmeyer. Then
we add with 10 mL of alcohol 96% colorless solution, the function
for alcohol as a solute. The solution becomes light yellow. Then
add with 5-8 drops PP indicator colorless solution and the solution
becomes light yellow, where the function of PP as an indicator to
knowing color changed when the solution titrate with NaOH so we
can know the final equivalent of titration. After titration the
solution become 2 layer, where up layers is peach and down layer
is light yellow.
CXCI. And the reaction is:
CXCII.
CH 3 (CH 2 )14 COOH + NaOH CH 3 (CH 2 )14 COONa + H 2 O
CXCIII.
CXCIV.
CXCV. After titrate we get for volume from sample oil is:
CXCVI.
V1 = 0.6 mL
CXCVII.
V2 = 0.5 mL
CXCVIII.
For the value of standard Free Fat Acid Indonesia
standard we get max 0.3%. but for FFA sample oil we get:
CXCIX.
FFA 1 = 0.128%
CC.FFA 2 = 0.107%
CCI. Average = 0.117%
CCII.
The oil sample is not suitable with Indonesia Standard
number of FFA, its because too bigger value of acid, so the
degree of free fat acid in sample is higher too and it showed that
the quality of the oil is not good. Free fat acid identified total of
free fat acid in oil is broken, because incident of oxidized and
hydrolysis.
CCIII.
CCIV. CONCLUTION
CCV.
From our experiment we can conclude value of
peroxide and free fat acid, that:
1.For value of peroxide we get 1.8 meq is not suitable with the
standard value of peroxide is max 1 meq
2.For the Free Fat Acid we get 0.117% is not suitable with the
standard value Free Fat Acid is max 0.3%
CCVI.
It showed that the quality of oil is not good for the
healthy and cant use again.
CCVII.

CCVIII.QUESTION AND ANSWER

1. Write all reaction which participates in this experiment
of fatty acid test!
For value of peroxide
CCIX.
2 KI +CH 3 ( CH 2 )14 COOH + H 2 O CH 3 ( CH 2 )14 COH +2 KOH + I 2

CCX.

2+2 I

2+ I 2 S 4 O 6

S 2 O3

For Free Fat Acid

CCXI. CH 3 (CH 2 )14 COOH + NaOH CH 3 (CH 2 )14 COONa + H 2 O
CCXII.
2. Mention which include in Fatty Acid essential in body.
Why arachidonic is not fatty acid essential?
Essential
fatty
acids
that
humans
need:
PUFA
(polyunsaturated fatty acids) cis type, namely: Omega-3
fatty acids, such as ALA, EPA and DHA omega-6 fatty acids,
such as linoleic acid Omega 3 EFAs are "good fats" that are
useful to keep the brain and heart health. Arachidonic not
an essential fatty acid because it can be produced in the
human body comes from the elongation of linoleic acid into
the body
CCXIII.
3. What the differences of Fat Acid Saturated and
unsaturated in oxidation process?
Saturated fatty acids have only single bonds, while
unsaturated fatty acids have a double bond. Saturated
fatty acids are more stable (inert) than unsaturated fatty
acids. Double bonds in unsaturated fatty acids easily react
with oxygen (easily oxidized). Double bonds in the
unsaturated fatty acids into two forms namely cis and
trans.
CCXIV.
4. What the differences between oil and Fat from the
structure of molecule?
The molecular structure of the oil have R groups that
unsaturated bonds (no double bonds 2)
However, on the structure of fatty molecules that have a
saturated bond R groups (do not have 2 double bonds)

CCXV.

CCXVI.
CCXVII.

REFERENCESS

CCXVIII.
Anonym.
2015.
Fatty
Acid.
(online).
(https://en.wikipedia.org/wiki/Fatty_acid accesses at October 25th,
2015)
CCXIX. Anonym. 2015. Lipid. (online). (https://en.wikipedia.org/wiki/Lipid
accesses at October 25th, 2015)
CCXX. Anonym.
2012.
Peroxide
Value.
(online).
(https://en.wikipedia.org/wiki/Peroxide_value accesses at October
25th, 2015)
CCXXI. Anonym. 2013. Lipid. (online).
(https://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/lipids.
htm accesses at October 25th, 2015)
CCXXII.

CCXXIII.

CALCULATION

1. Determining Peroxide Value (PV)

CCXXIV. Vblank = 1.1 mL
CCXXV. V1 = 6.5 mL
CCXXVI. V2 = 6.5 mL
mL Na2 S 2 O3 x N Na 2 S2 O3 x 1000
CCXXVII. PV1 =
gram sample
CCXXVIII.

5.4 x 0.01 x 1000

5 gram

CCXXIX.

54
5

CCXXX. PV2

mL Na2 S 2 O3 x N Na 2 S2 O3 x 1000
gram sample

CCXXXI.

5.4 x 0.01 x 1000

5 gram

CCXXXII.

54
5

CCXXXIII.

Average of PV =

= 1.8 meq

= 1.8 meq
1.8+ 1.8
2

= 1.8 meq

CCXXXIV.
2. Determining Free Fat Acid (FFA)
CCXXXV. V1 = 0.6 mL
CCXXXVI.
V2 = 0.5 mL
mL NaOH x N NaOH x Mr
CCXXXVII.
%FFA1
=
gram sample x 1000
CCXXXVIII.

CCXXXIX.

0.6 x 0.05 x 256.43

6 x 1000

CCXL.%FFA2

= 0.128%
mL NaOH x N NaOH x Mr
gram sample x 1000

CCXLI.

0.5 x 0.05 x 256.43

6 x 1000

CCXLII.

= 0.107%

CCXLIII. Average of %FFA =

CCXLIV.

x 100%

x 100%

x 100%

0.128 +0.107
2

= 0.117%

x 100%

CCXLV. ATTACHMENT
CCXLVI.
CCXLVII.

CCXLVIII.
Acetate
Kloroform solution
CCLIII.

CCLIV. Starch 1% solution

CCXLIX.

CCL.

CCLI.

Concentrated KI
solution
CCLV.

CCLVII.

CCLVI. Alcohol 20%

solution

CCLIX.

CCLII. Na2S2O3
solution

CCLVIII.
PP
indicator

CCLXI.

CCLX. NaOH solution

CCLXII.

Ohauss balance

CCLXIII.

CCLXIV.

Aquadest 5g

CCLXX.

CCLXXI.
Aquadest 5g
+ Acetate-kloroform 30
mL + KI 0.5 mL after 20
minutes

CCLXV.

CCLXVI.
Aquadest
5g + Acetate-kloroform
30 mL
CCLXXII.

CCLXXIII.
Aquadest
5g + Acetate-kloroform
30 mL + KI 0.5 mL after
20 minutes

CCLXVII.

CCLXVIII.
CCLXIX.
Aquad
est 5g + Acetatekloroform 30 mL +
KI 0.5 mL

CCLXXIV.

CCLXXV.
Aquad
est 5g + Acetatekloroform 30 mL +
KI 0.5 mL after 20
minutes set as
blanko

CCLXXVI.

CCLXXVII.
Blanko + 30
mL aquadest
CCLXXXII.

CCLXXXIII. Blanko + 30
mL aquadest after
titrated Na2S2O3

CCLXXVIII.

CCLXXIX.
Blanko +
30 mL aquadest
CCLXXXIV.

Blanko + 30 mL
aquadest after titrated
Na2S2O3
CCLXXXVI. + starch
1% 0.5 mL
CCLXXXV.

CCLXXX.

CCLXXXI.
Blank
o + 30 mL
aquadest
CCLXXXVII.

Blank
o + 30 mL
aquadest after
titrated Na2S2O3
CCLXXXIX. +
starch 1% 0.5 mL

CCLXXXVIII.

CCXCII.
CCXCIII.

CCXC.

CCXCIV.
CCXCV.

CCXCI.
Blanko after
second titration

CCXCVI.
CCXCVII.

CCXCVIII.
5
grams sample 1

CCXCIX.

CCC.

5 grams sample
1

CCCI.

CCCII. 5 grams sample 1

CCCIII.

CCCIV.
5
grams sample 1 +
Acetate-kloroform
30 mL
CCCIX.

CCCX. 5 grams
sample 1 + Acetatekloroform 30 mL +
KI 0.5 mL

CCCV.

CCCVI.
5 grams
sample 1 + Acetatekloroform 30 mL

CCCXI.

CCCXII.
5 grams
sample 1 + Acetatekloroform 30 mL + KI
0.5 mL after 20
minutes

CCCVII.

CCCVIII.
5 grams
sample 1 + Acetatekloroform 30 mL + KI 0.5
mL
CCCXIII.

CCCXIV.
Sample 1 +
30 mL aquadest

CCCXV.

CCCXVI.
Sample
1 + 30 mL aquadest

CCCXXI.

CCCXXII.
Sample
1 + 30 mL aquadest
after titrated
Na2S2O3 + starch 1%

CCCXVII.

Sample 1 + 30
mL aquadest titrated
Na2S2O3

CCCXVIII.

CCCXXIII.

CCCXXIV.
Sample 1
+ 30 mL aquadest
after titrated Na2S2O3
+ starch 1%

CCCXIX.

CCCXX.
Sample 1 +
30 mL aquadest after
titrated Na2S2O3
CCCXXV.

CCCXXVI.
Sample 1 +
30 mL aquadest after
titrated Na2S2O3 + starch
1%

CCCXXVII.

CCCXXVIII. Sample
1 after second
titration

CCCXXIX.

CCCXXXI.
CCCXXXII.

CCCXXX.
Sample 1
after second titration

CCCXXXIII.
CCCXXXIV.
CCCXXXV.

CCCXXXVI. 5
grams sample 2

CCCXXXVII.

CCCXXXVIII. 5 grams
sample 2 + Acetatekloroform 30 mL

CCCXXXIX.

CCCXL.
5 grams
sample 2 + Acetatekloroform 30 mL

CCCXLI.

CCCXLII.
5
grams sample 2 +
Acetate-kloroform
30 mL + KI 0.5 mL

CCCXLVII.

CCCXLVIII. Sample
2 after 20 minutes +
aquadest 30 mL

CCCXLIII.

CCCXLIV.
5 grams
sample 2 + Acetatekloroform 30 mL + KI
0.5 mL

CCCXLIX.

CCCXLV.

CCCXLVI.
Sample 2
after 20 minutes +
aquadest 30 mL
CCCLI.

CCCLII.
Sample 2 +
30 mL aquadest after
titrated Na2S2O3
CCCL. Sample 2 + 30
mL aquadest after
titrated Na2S2O3

CCCLIII.

CCCLIV.
Sample
2 + 30 mL aquadest
after titrated
Na2S2O3 + starch 1%
5 mL

CCCLV.

CCCLVII.

CCCLVI.
Sample
2 at second titration

CCCLVIII.
Second
titration process on
sample 2

CCCLXI.

CCCLIX.

CCCLXII.
CCCLXIV.

CCCLX.
Sample
2 after second
titration

CCCLXIII.
Sample 1 and sample 2 compared after second
titration

CCCLXV.

CCCLXVI.

CCCLXVII.

Sample 1 after second

titration

CCCLXVIII.

Sample 2 after second

titration

CCCLXIX.
CCCLXX.

CCCLXXI.
6 gram
of sample (palmitat
oil) in erlenmeyer

CCCLXXII.

CCCLXXIII. 10 mL of
alcohol 96%

CCCLXXIV.

CCCLXXV.
Sample 1 +
10 mL of alcohol 96%

CCCLXXVI.

CCCLXXVII. Sample
1 + 10 mL of alcohol
96% + 7 drops of pp
indicator
CCCLXXXII.

CCCLXXXIII. Sample
2 + alcohol 96% 10
mL + pp indicator
and after titrated

CCCLXXVIII.

CCCLXXIX. Sample
1 after titrated by
NaOH 0.05 N
CCCLXXXIV.

CCCLXXX.

CCCLXXXI. Sample 2 +
alcohol 96% 10 mL

CCCLXXXV.

Sample 1 and 2 compared after

titrated
CCCLXXXVI.

CCCLXXXVII.
CCCLXXXVIII.