oe ar HALLSTAR”* aa an efecive date: } ee ee es a) 02/18/2020 Modification : Validation : Fonction, ‘Guality-Control Technician ‘Quality-Control Manager Nom: Claire DIANA ‘Aurélie COUEDON a a a Création le 12/19/2019 Modification le 02/05/2020 Validation le : 02/05/2020 ] THIS DOCUMENT IS THE INTELLECTUAL PROPERTY OF HALLSTAR FRANCE PLEASE DO NOT COPY / DISTRIBUTE IT This document and files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. CO-57-P.C If you have received this document in error, please destroy it. Oleic acidity titration 1131 PURPOSE This analysis allows to measure the content of free fatty acids contained in a sample. The acidity is, expressed in weight percentage. Il, DOMAIN OF APPLICATION This procedure is to follow in these cases: ~ Control of a new produced batch - Control of a new batch of specific solvent, or matrix - Reevaluation of a validated batch (Non-exhaustive listing) Il, RELATED DOCUMENTS/ REFERENCE IV. DEFINITIONS/ ABBREVIATIONS V. CONCERNED PERSONS Quality-Control laboratory staff. VI. DESCRIPTION OF THE PROCESS 1) MATERIAL - Graduated burette Precision balance ~ Heating and stirring plate -Magnetic bar - Plastic pipettes 2) REAGENTS AND SOLVENTS - Absolute ethanol - Potassium hydroxide = Thymolphtalein 3) SOLUTIONS PREPARATION a) Thymolphtalein (20 g/L in an ethanolic solution) Weight precisely 0.20g of Thymolphtalein in 10 mL of absolute ethanol. This solution can be kept during one month in the fridge. Check regularly the aspect and db it again when observing a deposit. b) Potassium hydroxide Put one pellet of potassium hydroxide in 80 mL of water. Note the weight of the pellet. 4) COLORIMETRIC DETERMINATION a) Neutralization In a beaker of 100 mL (n*1), heat until boiling, 25 mL of ethanol and 0.25 mL of the prepared thymol blue ethanolic solution. This solution is heating under constant stirring. CO-57-P.C Oleic acidity titration 213While the temperature of the ethanolic solution is more than 70°C, neutralize it, carefully, with the potassium hydroxide solution, under stirring and heating. Add the potassium hydroxide solution, drop by drop. The neutralization is completed when adding one drop, significantly change the color of the solution in green/blue, for at least 15 seconds. b) Reference Titrate a very stable oil, which oleic acidity value is known. The value will be reference sample. Check that the percentage of the reference is equal to the percentage of oleic acidity obtained during the last determination c) Sample titration In a beaker of 100 mL. (n°2), weight 1.0 g of the sample to analyze. Add a magnetic bar. Add the content of the beaker n°1 (containing the neutralized solution). Stir the solution, under heating, Boil it (around 70°C). After the complete solubilization of the sample, titrate with the potassium hydroxide solution, stirring continuously. The reaction is completed when adding one drop, significantly change the color of the solution in greeniblue, for at least 15 seconds. Perform the both step as many times as the tested samples 5) CALCULATION AND EXPRESSION OF THE RESULTS Oleic avidity = (v*C*282)((10"w) % V:: volume, in mL, of potassium hydroxide solution used for the sample titration CC: concentration, in mol/L, of the potassium hydroxide solution w: weight, in g, of the titrated sample. VI. HISTORICAL Old referencing Modification date New referencing CO-57-P.A (04/16/2019 CO-57-P.B CO-57-P.B (02/05/2020 CQ-57-P.C CQ-57-P.C Oleic acidity titration 3/3.