Determination of saponification value of oils and fats

I. Introduction
Saponification value (SV) is defined as the number of milligrams of KOH required to
completely saponify one gram of the oil/fat. The procedure involves the use of excess
alcoholic KOH for a complete saponification of the sample in which both free and
esterified fatty acids are converted to salts (soap), followed by back titration of the excess
base with standard HCl solution in the presence of phenolphthalein as an indicator (Figure
1). SV is an index of mean molecular weight of the fatty acids of the constituting
triacylglycerols. Lower the saponification value, larger the molecular weight of fatty acids
in the glycerides and vice-versa.
RCOOH + KOH → RCOOK + H2O

KOH + HCl → KCl + H2O

Figure 1. Principle of method of determination of saponification value.
It would be important to note that most of the common oils/fats of vegetable or animal
origin (sunflower, soybean, rapeseed, pork lard, beef tallow, chicken fat, etc.) contain
almost only long chain fatty acids (C18 and C16), resulting in similar SV ranging from
168–196. Milk fat differs substantially from other fats and oils in terms of the fatty acid
profile, including relevant amounts of short chain (C4–C6) and medium chain (C8–C12)
fatty acids, which is reflected in its high SV (213–227). Consequently, SV may be helpful
in the detection of the adulteration of dairy products with cheaper fats and oils, because the
addition of an oil/fat rich in C18 to a dairy product will result in a decrease in the SV
(Ivanova et al., 2022).
Ivanova et al., 2022. Saponification Value of Fats and Oils as Determined from 1H-NMR Data: The Case of Dairy
Fats. Foods. 11(10):1466. doi: 10.3390/foods11101466.

II. Materials
- 250 mL Florence flask.
 - Technical balance, microburette.
- 0.5 N KOH in ethyl alcohol.
- 1% phenolphthalein in ethyl alcohol.
- 0.5 N HCl.
- Water bath

III. Procedure
1. Test sample
- Weight 2-3 grams of sample into a 250 mL Florence flask.
- Add exactly 20 mL of 0.5 N KOH solution using a microburette.
- Connect the flask with an air condenser.
- Incubate the flask in a boiling water bath for an hour (complete saponification is
indicated by an absence of any oily matter and the appearance of a clear solution).
- Cool with tap water.
- Add 5 drops of phenolphthalein.
- Titrate with 0.5 N HCl until the solution turns colorless.

2. Blank sample
- Add 2-3 ml of distilled water into 250 ml Florence flask.
- Add exactly 20 mL of 0.5 N KOH solution using a microburette.
- Connect the flask with an air condenser.
- Incubate the flask in a boiling water bath for an hour.
- Cool with tap water.
- Add 5 drops of phenolphthalein.
- Titrate with 0.5 N HCl until the solution turns colorless.

IV. Calculation
SV is calculated by the following formula:
(a−b) x 28.05
SV = m
a: Volume in mL of 0.5 N HCl used for blank sample.
b: Volume in mL of 0.5 N HCl used for test sample.
28.05: Weight in mg of KOH present in one ml of 0.5 N KOH solution.
m: Weight in g of the sample.