Fats and Oils analysis

1. Acid number
Acid number is a number that indicates the amount of free fatty acids
contained in fats / oils that usually associated with hydrolysis process of fat /
oil. Hydrolysis of fat / oil by water with an enzyme catalyst / heat on
triglyceride ester bond will form free fatty acids. The presence of free fatty
acids in fats / oils usually use as indicator of damage fats / oils by hydrolysis
process. Formation of free fatty acids will accelerate oxidative damage to fats /
oils because free fatty acids are easier to oxidize than in the form of esters.
The amount of free fatty acids in sample shown by the acid number and
usually expressed as number of milligrams KOH needed to neutralize free fatty
acids in 1 gram of oil or fat. Acid number is determined by saponification
reaction, by reacting fatty oil with a base such as KOH or NaOH.






2. Saponification Number
The saponification number is the number of milligrams of potassium
hydroxide required to convert one gram of the fat completely into glycerin and
potassium soap. It gives information concerning the character of the fatty acids
of the fat and in particular concerning the solubility of their soaps in water. The
higher the saponification number of a fat free from moisture and
unsaponifiable matter, the more soluble the soap that can be made from it.
The information is of special importance to soap makers. Table 2 gives the
saponification numbers of the commoner commercial fats and oils.
CH
2
O
2
C(CH
2
)16CH
3
CH
2
OH

CHO
2
C(CH
2
)16CH
3
+ 3 NaOH CH
2
OH + 3CH
3
(CH
2
)16CO
2

-
Na
+


CH
2
O
2
C(CH2)16CH
3
CH
2
OH

Triestearin base glycerol sodium
stearate






3. Esterification Number
Esterification number is amount of an ester organic acid. It has relation
with the acidic number and Saponification number. Esterification number can
be calculated with this equation below



4. Unsaponified Number
Unsaponified materials are compounds which are often soluble in oil
and can not be saponified by alkali, for examples: alcohol, sterol, dyes, and
hydrocarbons. We can do this determination for all food oils but marine
animal oils.

R-H + KOH tidak tersabunkan
NaOH(sisa)/KOH + HCl NaCl /KCl + H
2
O

)



5. Determination of Total Fatty Acids
Total Fatty Acids is the total fatty acids in the food oil and can be tessted
by saponification method. After drying, soap from the saponification of oil in
alcohol with 0.5 N NaOH or KOH is dissolved in water.

O

R C Na + HOH R-COOH + NaOH
H
+
R-COOH + C
8
H
14
larutan campuran
C
8
H
14
dipanaskan menjadi tersulingkan






6. Determination of number Hehner.
Hehner number is used to determine amount of Insoluble fatty acid in
water. Fats with a high molecular weight will have a low Hehner number.
Filtrate from saponification test is heated and release the alcohol. Soap is
dissolved in hot water and HCl to form free fatty acids. When the mixture is
immediately cooled, fatty acids will obtain an insoluble layer in water. This
layer was filtered and weighed.
7. Bilangan Reichert-Meissl
Reichert-Meissl number shows the amount of fatty acids that can
dissolve in water and easily evaporates. This test express as the number of ml
of 0.1 N NaOH used to neutralize the fatty acids from the distillation of 5 grams
of fat or oil. Fatty acids with 4-6 carbon atoms will easier to evaporate and
dissolve.
Perhitungan :


A= ml NaOH 0,1N for sample
B= ml NaOH 0,1N for blank

8. Bilangan Polenske
These numbers determine the volatile fatty acid levels, but insoluble in
water (fatty acids C8 to C14). Polenske number is the number of ml of 0.1 N of
base to neutralize C8-C14 fatty acids in 5 grams of sample. Polenske also can
be used to test the counterfeiting butter.

9. New Kirschner Numbers (New Kirschner Value = NKV)

NKV is the number of ml of 0.1 N alkali for every 5 grams of fat or oil to
neutralize the volatile fatty acid silver salts which is soluble in water-ethanol
mixtures. Determination NKV used to different margarine and butter, and
determine counterfeiting. Add ag2so4 to the distillate of NKV determination,
and it will form the silver salt is soluble in water, then do distillation process.


10. Determination of Iodine Number
Determination of iodine showed unsaturated fatty acid in of fats and
oils. Unsaturated fatty acids are able to form saturated compounds. Number of
iodine bound shows that there are many double bonds in the fatty acid. Iodine
number shows amount of iodine (gram) in 100 g fat or oil.

H H
I I
H H
I
2






Table 1. Iodin Numbers of Common
Fats*
Fat or Oil lodin number
Linseed oil 173 - 201
Tung Oil 170.6
Menhaden oil 139 - 173
Whale oil 121 - 146.6
Soy bean oil 137 - 143
Sunflower oil 119 - 135
Corn oil 111 - 130
Cottonseed oil 108 - 110
Sesame oil 103 - 108
Rapeseed oil 94 - 102
Peanut oil (arachis) 83 - 100
Olive oil 79 - 88
Horse oil 71 - 86
Lard 46 - 70
Palm oil 51.5 - 57
Milk fat 26 - 50
Beef tallow 38 - 46
Mutton tallow 35 - 46
Cacao butter 32 - 41
11. Determination of Diene Numbers
Diene number is a measure that is calculated euivalen dienophilik
reagent with the amount of iodine absorbed by 100 grams of oils or fats. This
test is based on the reaction of fatty acid with reagents dienophilik. Synthesis
of diene also underlies this test, ie one double bond fatty acids would bind one
molecule dienophilik. Which is used as the reagent dienophilik namely maleic
anhydride as easily calculated by titration. Effect of temperature affect the
same result. The temperature is too high resulting free hydroxyl group (in
hydroxy fatty acid hydroperoxide and the oxidation or other) reacts with
maleic anhydride so bilanagan diene obtained thus imprecise.

12. Determination of Acetyl and Hydroxyl Numbers
Acetyl and Hydroxy chemical properties of oil or fat to determine the
free hydroxyl group that is often present in oil or fat, either natural or
synthetic (mainly on castor oil, croton oil and monoglycerides). Acetyl numbers
are expressed as milligrams KOH needed to neutralize the acetic acid obtained
from the saponification of 1 gram of fat or oil or wax that already in
acetylation. Numbers hydroxyl itself is the amount of acetic acid which is used
to esterificating 1 gram of fart or oil that equivalents with amount of KOH.

13. Bilangan Peroksida
Bilangan Peroksia merupakan indikator terpenting dalam menentukan
kualitas suatu bahan minyak. Asam lemak tidak jenuh dalam minyak bisa
mengikat oksigen pada ikatan rangkapnya dan membentuk peroksida.
Keberadaan senyawa peeroksida ini akan menurunkan kualitas bahan minyak.
fatty acid
OH
acetic anhydride
fatty acid
O C
O
CH
3
fatty acid
OH
+
COOH H
3
C
titrate with standardized
KOH
acetylated fatty acid


14. Numbers p-anisidin
Decomposition of peroxide produces many compounds, especially aldehyde
group. The number of aldehyde in sample expressed by the value (p-value).
The reaction between an aldehyde compound with a reagent para anisidin in
acetic acid solvent will produce a yellow color that absorbance can be
measured at a wavelength of 350 nm. Number of paraanisidin and peroxide
numbers can be used to determine the total number of oxidation (oksidation
total value) which is equivalent with two times peroxide number plus the
paraanisidin number. Numbers of total oxidation is often used as parameters
of oxidative damage levels of fat / oil
15. Degrees ketengikan
Degrees ketengikan fat / oil shows how much damage the fat / oil has
occurred. Ketengikan test is a test used to measure fat oxidation stability.
Stability of fat oxidation can be measured quickly by using Methrom Rancimat.
Measurement of oil and grease damage done based on the results of oxidation
of volatile compounds that cause the smell of rancid fats like sour dikarboksil.
Rancid-smelling compounds can increase the electrical conductivity so that it
can be integrated in the form of the relationship between the time of
induction to the conductivity.
16. Numbers TBA
Test number Thio Barbituric acid (TBA) is commonly used to measure the level
of ketengikan fat / oil or food products containing fat / oil. In the oxidation of
fat, fat decomposition result component that can form the aldehyde derivative
compounds, namely malonaldehyde. The presence of malonaldehyde in
samples of fat / oil showed that the sample has undergone further oxidation.
Malonaldehyde formed compounds will react with TBA reagent and produce
red pigment. The higher TBA numbers then the level of oxidation of fat / oil is
getting higher.

Chemical Properties of fat and oil

1. Esterifikasi
Esterification process is used to convert free fatty acids from triglycerides,
and form form esters. Esterification reaction can be performed by interifikation
and ester exchange reaction (transesterification)




2. Hydrolysis
In hydrolysis reaction, fats and oils will be converted into free fatty acids
and glycerol. This reaction can affect fats and oils. It is because of the amount
of water in fat and oil.
CH2 O C R1 R1COOH CH2O

CH O C R2 + 3 H2O R1COOH + CH2O

CH O C R3 R1COOH CH2O
Trigliserida asam lemak gliserol

3. Saponification
This reaction is carried out with the addition of base solution to the
triglycerides. When the saponification reaction had been completed, then a
layer of water containing glycerol can be separated by distillation.

CH2O2C(CH2)16CH3 CH2OH

CH2O2C(CH2)16CH3 + 3 NaOH CH2OH + 3CH3(CH2)16CO2
-
Na
+


CH2O2C(CH2)16CH3 CH2OH

4. Hydrogenation
The hydrogenation process used to clarify fatty acids or oils bonds when
the process is completed, the oil is cooled and the catalyst separated by a
filter. The result plastic or hard oil, depends on the degree of saturation.

CH3 - (CH2)7 - CH = CH - (CH2)7 - COOH

CH3 - (CH2)16
COOH

5. Ketone forming
Ketones are produced through the decomposition of esters by hydrolysis.




6. Oxidation
Oxidation can take place if there is contact between the number of oxygen
with the fat or oil. The occurrence of this oxidation reaction will cause rancidity
in fats or oils.

Correlation between the parameters:

1. Saponification Number Decreasing
Saponification of palm oil will lower the acid number and level of the
fatty acid hydrolysis.

2. The relationship between the number Saponification with the number of
C atoms in the fatty acid
In glycerides, fatty acids that have short-chain C it will get a higher BP
than fatty acids with chain length C.

3. The relationship between the number Saponification, acid number with
the number of ester
We can calculate esterification number by formula: saponification
number minus acid number, so it can be concluded that higher fatty
acids have lower number of esters.

4. The relationship between numbers peroxide with iodine number
Based on the definition that the iodine number indicates the number of
double bonds contained in a fat or oil, so higher level unsaturated of fat
/ oil means the iodine number is high.
And peroxides number express amount of free fatty acid in samples
through oxidation reactions. Stability of fatty acids oxidation based on
the amount of double bond..
Then we can conclude higher iodine number have higher peroxide
number too.

5. Correlations between acetyl and hydroxyl number
Acetyl Numbers expressed as the amount of KOH which is needed to
neutralize the acetic acid obtained from the saponification of 1 gram of
oil, fat or wax that has been distilled. And hydroxyl number is the
number of acetic acid used to esterificate 1 gram of fat or oil which is
equivalent to milligrams of KOH.
Both numbers were used to determine the free hydroxyl group that is
often found in natural or synthesis fats and oils, especially in castor oil,
croton oil and monoglycerides.

6. Relations between Numbers Diene with iodine Numbers
Numbers diene is amount of reagent dieophilik equivalent with amount
of iodine absorbed by 100 grams oils or fats. This is based on the
synthesis of the diene, in one double bond of fatty acids will bind one
molecule dienophilik, not only iodine.

7. Relations between Numbers Hehner with Reichert Meissel Numbers
Hehner number is used to measure the amount of fatty acids that are
not soluble in water. And Reichert-Meissl number shows the amount of
fatty acids that can dissolve in water and easily evaporates. So, oils or
fats that have high number Reichert Meissl will have low Hehner
numbers.

8. Relations between Numbers Krischner with Numbers Riechert Meissel
Krischner Numbers are used to determine the amount of volatile fatty
acids that have dissolving silver salt in water and volatile. And Reichert-
Meissl number shows the amount of fatty acids that can dissolve in
water and easily evaporates (volatile). So the measurement of Reichert-
Meissl number is based on measurement of silver salts that dissolve in
the determination of the number Reichert Meissel.

9. Relations between Numbers Thiocynagen with iodine Numbers
Numbers Thiocynogen determined based on selective and partial
addition of the pseudo-halogen-thiocynogen by unsaturated fatty acids.
Because of thiocynogen amount absorbed by unsaturated fatty acids,
such as linoleic and linolenic, it is not same with the amount of absorbed
iodine, then by measuring the amount of iodine absorbed and can be
determined thiocynogen oil or fatty acid composition.

10. Relations between Numbers Hehner with unsaponified material number
Because of Hehner numbers are percentage of total fatty acids which is
not soluble in water and the unsaponified material. That means
unsaponified materials also can be calculated with Hehner number.

11. Relations between Numbers Saponification with unsaponified number.
unsaponified fat is oil that is not saponified because trapped by a
substance and make it difficult to saponified, then add and excess of
alkali measured so Saponification number can be set.

12. Relations between Numbers Saponification with molecular weight
Saponification number is the amount of alkali required to saponified a
number of oil samples, and expressed the number of milligrams of KOH
needed to saponified 1 gram of oil / fat.
Oil with low molecular weight will have a high saponification number,
whereas if high molecular weight means low saponification Numbers.