Determination of Free Fatty Acids in Ghee



Ghee, a popular and flavorful type of clarified butter, is an essential ingredient in many cuisines worldwide. It is rich in fat, mainly composed of triglycerides, and contains free fatty acids (FFAs) in trace amounts. The determination of free fatty acids in ghee is crucial as it affects its quality, taste, and shelf life. In this article, we present a comprehensive laboratory procedure to quantify FFAs in ghee accurately.


  1. Ethanol (95%)
  2. Phenolphthalein indicator solution
  3. Sodium hydroxide (NaOH) solution (0.1 M)
  4. 1% phenolphthalein in ethanol solution
  5. Ghee samples


Step 1: Preparation of Ethanol Solution

Prepare a 95% ethanol solution by mixing 95 mL of ethanol with 5 mL of distilled water. Use a clean, dry glass beaker for this purpose.

Step 2: Sample Preparation

Weigh accurately about 10 grams of ghee into a clean, dry conical flask using an analytical balance. Record the weight of the sample.

Step 3: Saponification and Titration of Ghee

Add 50 ml of ethanol solution to the Boling point and while still above 70°C neutralize it to phenolphthalein  with 0.1N NaOH Add the neutralized alcohol to flask containing ghee sample and mix the contents of the flask Bring the mixture to boil and while it is  still hot Titrate with 0.1N NaOH. Saking vigorously during the Titration. The end point of the Titration is reached when the addition of single drop produces a slight but a definite colour change persistenting for at least 15 seconds

Step 4: Calculation

Calculate the amount of free fatty acids (expressed as oleic acid, C18:1) in the ghee using the following formula:

% FFAs = [V * N * 282.5 * 100] / W


V1 = Volume of NaOH used in the titration of the ghee sample (mL)

N = Normality of the NaOH solution

282.5 = Equivalent weight of oleic acid

W = Weight of the ghee sample taken (grams)


The determination of free fatty acids in ghee is a critical aspect of assessing its quality and shelf life. By following the comprehensive laboratory procedure outlined above, one can accurately quantify FFAs in ghee samples. Maintaining quality standards and ensuring low FFA levels in ghee will contribute to its desirability and enhance its culinary applications in various cuisines.