SAMPLING AND ANALYSIS OF COMMERCIAL FATS AND O1LS AOCS Official Method Cd 3-25 Reapproved 1997 + Revied 2001 + Revised 2002 Saponification Value DEFINITION 1 gram of the sample. SCOPE Applicable to all nosmal fats and oils ‘The saponification value is the amount of alkali necessary to saponify a definite quantity of the sam- ple. It is expressed as the number of milligrams of potassium hydroxide (KOH) required to saponify APPARATUS 1. Erlenmeyer flasks—alkali resistent, 250 or 300 mL, ‘with T 24/40 ground-glass joint. 2. Condensers—with T 24/40 ground-glass joint to fit Erlenmeyer flasks. 3. Water bath, or a hot plate with variable heat control 4. Distillation flask—2 L, with 724/40 ground-glass joint, fitted with water-cooled condenser, for refluxing and distilling 95% ethyl alcohol as noted in Reagents, 2. REAGENTS 1. Hydrochloric acid, 0.5 M—accurately standardized See AOCS Specification H 14-52. 2. Alcoholic potassium hydroxide (KOH)—prepared by placing 5-10 g of reagent-grade KOH (see Notes, Caution) pellets in a 2-L flask, adding 1-155 L of 95% ethyl alcohol (see Notes, Caution) and boiling in a water bath under reflux for 30-60 min. Distill and collect the alcohol in a clean, glass-stoppered receiver. Dissolve 40 g of potassium hydroxide, low in carbon- ate, in | L of the distilled alcohol (see Notes, Caution) with cooling to keep the temperature at or below 15°C. ‘This solution should remain clea. (See Notes, 1). 3, Phenolphthalein indicator solution— 1.0% in 95% ethyl alcool. PROCEDURE 1. Melt the sample (see Notes, 1) if itis not already liquid ‘and filter through dry filter paper to remove any impuri- ties and moisture. The sample must be completely dry. 2. Weigh a test portion of such size thatthe back titration is 45-55% of the blank. This usually requires a test portion of 1.0-2.2 g if 25 mL of KOH are used (Table Dor 4-5 g if 50 mL of KOH are used. Add the alco- holic KOH with a pipet and allow the pipet to drain for a definite period of time (see Notes, 2). 3, Prepare and conduct blank determinations simultane- ‘ously with the sample and similar in all respects, except ‘omitting the fat or ol, Tablet : Weight of test portion. 4, Connect the condenser and boil gently, but steadily, until the sample is completely saponified. This usually requires about 1 hr for normal samples (see Notes, 3). Make certain that the vapor ring in the condenser does not rise to the top of the condenser, or loss may occur. 5. After the flask and condenser have cooled somewhat, but not sufficiently to form a gel, wash the inside of the condenser with a small quantity of distilled water. Disconnect the condenser, add about 1 mL of phenolph- thalein indicator and titrate with 0.5 M HCI until the pink color just disappears. Record the volume of 0.5 M HC required for the titration CALCULATIONS 1. Saponification value: Where— . volume of 0.5 N HC! required to titrate blank, mL. volume of 0.5 N HC! required to titrate sample, mL M = molarity of HCI solution W= mass of sample in grams PRECISION Repeatability and reproducibility values from an interna- tional collaborative study are given in Table 1 NOTES Caution Potassium hydroxide, like all alkalies, can burn skin, eyes and respiratory tract severely. Wear heavy rubber gloves and face shield to protect against concentrated alkali liquids. Use effective fume-removal device or gas mask to protect respi- ratory tract against alkali dusts or vapors. When working with extremely caustic materials such as potassium hydrox- ide, always add pellets to water and not the reverse. Alkalies are extremely exothermic when mixed with water. Take pre- cautions to contain the caustic solution in the event that the mixing container breaks from the extreme heat generated, Ethyl alcohol (ethanol) is flammable. Use a fume hood when heating or evaporating this solvent. Hydrochloric acid is strong acid and will cause severe ‘burns. Protective clothing should be worn when working. with this acid. It is toxic by ingestion and inhalation and a trong irritant to eyes and skin. The use of a properly operat- ing fume hood is recommended. When diluting the acid, always add the acid to the water, never the reverse. Page 1 of 2SRWPLING AND ANALYSIS OF COMMERCIAL FATS AND OWS Ca 3-25 » Saponification Value Table 2 Results of a 2000 collaborative study on the determination of saponification value. - ~ (60% Coconut oil ~ ‘Sample Rapeseed oil Palm oll Coconutoil + 40%MCT oil MCToil ‘Total numberof laboratories accepted 19 7 20 18 16 Mean of the laboratory values (mg/g) 19020 19950 256.80 287.50 334.10 Repeatability ‘Standard deviation, s, on 056 om on 141 Relative standard deviation, RSD,% oa 03 03 02 oa Repeatability value, 28 x 3, 201 136 2.02 199) 3.95 Reproducibility ‘Standard deviation, 5g 17 204 447 236 286 Relative standard deviation, RSD,% 09 10 16 os 09 Reproducibility value, 2.8 5p 491 5m 11.67 661 8.09 NUMBERED NOTES 1. Ethanol may be stabilized by either of the following ‘methods: a) the addition of 5 g aluminum pellets to the reflux flask prior to refluxing and distillation or b) the addition of 4 g aluminum tert-butylate and allowing it to stand for several days. In both cases decant the clear supematant from the precipitated potassium carbonate formed after dissolving the potassium hydroxide. 2, When the saponification value is required on the fatty Page 2 of 2 acids, the preparation and seperation are performed as directed in AOCS Official Method TI 1a-64 For consistent results, a 25- or 50-mL. class A volumetric pipet calibrated “to deliver” should be used. The pipet should be allowed to drain for the time specified Some samples, particularly those difficult to saponify, may require up to 2 hr refluxing. This can only be determined by trial, Clarity and homogeneity of the test solution are partial indicators of complete saponifi- cation, but they are not necessarily absolute criteria os