Pebypassphiamn da ta ably of ¢ ovbellnce Rew” FATS AND OTHER LIPID! Phiysloat Properties. The physical:properties.of the natural fats and oils fife often used to identify them. Usually’ more, than’ one’ property’ is {Neiiured so that the identification can be made with some assurance sinice ral fats and oils vary somewhat in their properties. Th Osition aha constant’ but,varies slightly 1 ate, so a Ma e jo olls, audi tion season, and’breed foraiimal oils) “Melting Point. Fats do not melt sharply but soften over'a range‘ of tem-. fieratures, and it is therefore impossible to apply the melting point, tech- ilque, used in the identification of pure organic compounds, to them. Ifo fat, a fatty acid, or some esters of fatty acids are, heated. very {MOWIY, they will melt, exist as_a liquid as the temperature rises, and then wolldify again. A second melting will then‘occur at a higher temperature. If {he material is then chilled rapidly; ie\ ill melt at a lower temperature when it is warmed again. This behavjor as been known for. many. years, but it is only in recent’ times the use of modern techniques that th oxplanation has beén possible 1igm, the occurrence of more than. f nc ’ \e crystal for ee hismis found in many long-chain carbon compounds. The: - lymorp! number of crystalline forms possible for each fatty acid or each. ester is Atill a matter of debate’ since rani of the forms show melting. points so close together that it is di f n seve i b are de Wnetante a, and gamma; the melting points of trig S are given in Table 2.6. In compounds with several crystalline forms, one form. is the most stable and tétids to be established. Polymorphism is important to the understanding of the’ melting-point behavior of fats, fatty acids, and their esters. Furthermore, poly- morphism plays‘a’ significant role in any operation where fats are solidi- fied. In this latter instarice environmental conditions must. be controlled so that the: product will be uniform—for example, when chocolates are dipped, the formation of the high-melting forms is produced by control- ling the températuire’of solidification. : When melting’point is used to identify ‘a fat, all aspects of the pro- cedure must be carefully controlled. A number of procedures have been de- veloped and’ few will be described briefly. Solid fats aré-plastic over'a fairly wide Tange of temperatures, By plastic we mean that they are soft and can be deformed, but do not have the ability to flow. The spreading quality of butter is the’ result of plastic nature, ha Not enmeshed but are able to slide’ by one another a aL ne d' micrOscoE ically, We See" that hey arescom- tiny Se inva matrix of liquid fat. The crystals are sequently givegeules:does tiot Possess 1 FOOD CHEMISTRY ROHTS Trilinolein. *Frdm Bailey, A. E, York NiXv 1945. the'mixed fat its plastic’ nature. Alfat fat diminishes and the’ amount of liquid i the number of crystals exceeds a critical brittle aiid will lose plasticity. On’ the ot exceeds a critical level, the fat will flow. eS Natural fats are complex mixtures of glycerides each with. its own char-. © acteristic melting point. As the 1 erature of the fat is faised; the melting. = point of first one and then another of these,glycerides exceeded. Eventu- ally a temperature ig reached at. which all of the glycerides. haye. melted and the fat is liquid. The temperature. at which this occurs. is not sharply. defined. The'problem is further ¢ licated by the fact ithat glycerides are bl type of molecule may dissolve in, the soluble’in one another. A particul rature considerably below. its melting icreases so that, the, fat softens. If amount, the.fat will be, hatd and t hand, if the amount of liquid liquid portion of the fat at a ‘tem poitit. Pe i ret ri Glyceridés alo'tiave a tendency tdstiperedol, eto remain.as liquids at ature below their melting point 'as they are coo] wafromza ghér temperature The colitisioiey ofa fat at say Se eee if it'is heatéd “slowly from room temperature than jf it been quickly... cooled from a higher temperature. t equilibrium arid the effects of superc The Softening’ Point of a fat identification, but’ it cannot be appl filled with oil"and packed in ice o er and come to equilibrium. The capi 4FATS AND OTHER LIPIDS a bedker of Wat which’ thee The’ method’ gives repros ‘others, while on lar compounds, for. ter and submerged in’ and the température h ig called the softening ‘point : xome fats, rather poor’ results ‘with example, it cannot be used. he : ; ; ‘The Slipping Point is another énipitical method used to identify some natural fats and fat compounds: Small brass éylinders, filled with the solid he thermometer, As the bath is fat, are suspended’ in’ a baih ‘close ‘to tl : e h he temperature is slowly raised. The point at which the fat_rises Sig recorded’as the slip point., The slip point is Felated to the water beatén into the fat during its manufacture as well as the composition ‘of the fat. The slip point cannot, therefore, be re- cated on a particularsample-with reprodiicible results. The Shot Melting Point.is the témperature‘at which a ‘mail lead shot will fall through'a sample: This method ‘has'soriie‘usefulness. ~ Why natural fats and oils differ in their ; melting behavior has been the subject of a great deal of study. How variations in the composition of a- fat and oil influence the melting has been studied in many free: fatty acids and pure glycerides. Likewise, analysis of the fatty acids present in fats and a comparison with the melting point gives some information. In gen- oral fats which contain relatively large aiiounts of unsaturated fatty acids have relatively low’meltinig points and are’ usually oils at room tempera- lure, while those’ with relatively large ‘amounts of saturated fatty acids have higher melting points. Whén the melting points of pure, simple tri- glycerides are determined, it is found’that"lengthening the carbon chain of the fatty acids iricreases the melting point. For example, the melting point of trimyristin {alpha form) is 46.5°C, tripalmitin, 56.0°C, and tri- fleurin, 65.0°C. Thus fats which are mixtures of glycerides of long chain, fnturated fatty acids have higher melting points than those which have olther numerous ‘unsaturated fatty acids or short chain ones, or both (see Table 2.6). . are arte * Specific Gravity.‘The specific gravity of oils and fats is determined by the usual methods. The teinperature is‘ carefully controlled since significant eee Leatrtiarid occur in short ranges of temperature, The spe- oe gravity sa is usially nica af a C, but Ss y_ TO USE even igh-melting fe eee the:spetifie gravity from one oil’ or fat to es i ceiileteik atike uy aaaeeehbe miele chains or increase in Refractive Index. The index ‘of is do increase the Spesinc AraY avy. beam of light that déd f refraction is the degree of deflection ofa | iat Occurs when it'passes from one transparent medium to” ji oSFOOD CHEMISTRY another. The refractive'indices of-fats and oils-are often measured, both: be- cause they can be rapidiy and accurately determined and, because they are useful in identification of these substances and the. testing of their. purity. An Abbé Refractometer with temperature control is used and the measure- ment is usually at 25°C. With high melting fats 40°C or even.60°C can be used, but temperature must, be controlled and, noted. The: index of refrac- tion decreases as the temperature rises; however, it increases with increase in the length of the carbon chains and also with the number, of double bonds present. , : “Smoke, Flash,,and Fire Points, The smoke;point is the temperature at which a fat or oil gives off a, thin. bluish.smoke. I measured: by a standard method in an.open dish specified by the American. Society for Testing Materials so that the evolution of smioke.can be readily seen and reproduced. The flash point is the temperature at which-the mixtures of vapor with air will ignite; the fire point is the temperature at which the substance will sustain continued combustion. For a. given sample of oil. or fat, the. temperature is. progressively higher for the smoke point, flash point, and: fire point. Table 2.7 gives data for a few oils with flash points measured in an open cup.. The tem- peratures vary with the amount of free fatty acids present in an oil or fat, decreasing with increased frée fatty acids. Since, the amount. of free fatty acids changes with variations in refining, the history of the oil or fat is important. The smoke point of.a fat used for deep fat frying. de- creases with use of the fat. Fats:and-oils with low molecular weight fatty acids have low smoke, flash, and fire points. The number. of double bonds present has little effect on the temperature required. Smoke, flash, and fire points are’ particularly useful in connection with fats used for any kind of frying. Turbidity Point. The turbidity point.of an oil is determined by cooling a mixture of it -and:a solvent in whjch it has.a.limited solubility. The mix- ture is warmed until complete solution occurs and then slowly cooled until the oil begins to separate and turbidity occurs. The temperature at- which turbidity first is detectable is known as the turbidity point. The first sol- vent employed was glacial acetic acid in the Valenta test;.but since it. is difficult to keep the acid pure and since.moisture has a marked effect on the test, other solvents have been substituted. An the Crismer.test the solvent eis methyl alcohol while in the Fryer and: Weston modification of the Cris- mer test it is an equal mixture of 92 per cent ethyl alcohol and_amyl_al- cohol. Data for some oils are given in Table 2.8. : 7 TheXGrbidity point determined for any one oil does show a range of values. It is particularly sensit the presence of free fatty acids and a 6FATS AND OTHER LIPIDS SMOKE, FLASH, AND FIRE POINTS OF ONS? ‘Smoke Poinis |" Flash Points Fire Points : - (Open Cup)! ‘ Oil RN UR bites 76 SFA, cd °F “c Castor; refined re 392.200, 568 298 635 335 Castor, dehydrated 348 176 570 299-638 337 Corn, crude yo 352, 178 562 294 655 346 Corn, refined 4 227+ + 618 326 678 359 Linseed, raw 325 163 540 287 667 353 Linseed, refined 320 160 588 309 680360 Olive, virgin i 391 199 610 321 682 361, Soybean, expeller, crud 337 181 564 296 664 351 Soybean, extracted, crude 410.210 603 317 670 354 Soybean, refined 2492-256 618 326 673 356, Perilla, raw 321 wl 575 302 628 359 Perilla, refined 3520178 608 320 685 363 Perilla; refined 408209 615 324 685 363 *From Detwiler, S. Band Markley, K. S., “Smoke, Flash, and Fire Points of Soybean and Other Vegetablé Oils,” Oil and Soap, 17, 39 (1940). : correction factor:must be introduced for these acids, Nevertheless, different oils show a wide enough range of values so that the test has value in the differentiation of some oils and in the detection of adulteration. ‘Chemical Properties. A number of chemical tests have been evolved dur- ing the years of study of oils and fats which are based on the partial de- termination of the chemical composition of the oil or fat. These tests serve. both to identify the fat_and to detect the prese! ion, All oils ‘and fats show some range of values; therefore sometimes more-than one. “slis necessary, A few of the most commonly used tests are given below. ‘The Reichert Meiss| Number is a measure of the amount of water-soluble. volatile fatty acids; the Polenske Number measures the amount of volatile insoluble fatty. acids; the Saponification Number, the amount of potassium hydroxide required to saponify.the fat; and the Iodine Number, the amount of unsaturation present. These chemical tests, then, differentiate fats and oils on the basis of the chemical composition of the various triglycerides present in the mixture. The Reichert Meissl: Number is defined as the numberof milliliters of 0.1 alkali (such as potassium hydroxide) required to neutralize the yola-_ tile water-soluble:fatty acids in a 5 g sample of fat. The volatile acids will be those in the range of molecular‘ weights from butyric (C,) t6, myristic (C,,)'acid. The Reichert Meissl test determines the amoypt of bufftic and y 7 caproic acids which are readily soluble in'water and the caprylic and capric Yo acids which are‘slightly soluble. The Polenske Number is the number of 27FOOD CHEMisTRY Acidity cee 2°" (As Oleic) °Crismer Fator Oil - Per Cent _Nalue Perilla PS 55 49.0 4 Linseed z ‘ 20 58.3 Tong 5 09 740 Soybean 12 65.0 Niger 0 22 515 ‘Sunflower 7 f 22 59.5 Com” 5 : 28 62S Cottonseed OL 965.0 Sesame ¥ 40 + 60.5 Rape - . 06 (2/823 Almond : 09 (68.2 Peanut. ; LL 720 Olive 1 07 678 Olive “4 1 655° Olive 3.6 615 Cacao 29 1.0 1.72 Chinese Vegetable Tallow _ 69 40 Pali # oO 72 Lard 09 act Tallow : on 13 : Butter fat 1 LS4 » 46.0 Coconut 00 34.0 2.01 340 Coconut 16 30, 201 332 Palm Kernel 00 400 201 40.0 “Vegetable Fats afd Oils,” 2nd ed., 36, Reinhold Publishing Corp. New ‘*Fram Jamieson, G. York, N. ¥., 1943. milliliters of 0.1N.alkali necessary to-neutralize the volatile, water-insoluble pe fatty acids which are present in a'5 g sample. These two determinations are teadily run on the same sample of fat and the:Kirschner Value, described below, also canbe carried out on the same sample? ap ‘elycerol ‘ (soluble—C, and C, Fat Mt, 'Na,SO, pie sodium soaps #525) 4. -.. -» -volatil free fatty: 5. .- acids ’ acids, C,toC,, Jnsoluble—C, toC,, troduced into a,flask and treated | either with al- coholic sodium h ide or sodium hydroxide in gl cerol,; If.alcohol is used, it must be removed by evaporation before the fatty acids are’ neutral- ized. Saponification occurs rapidly under the conditions of the experiment; 28FATS AND OTHER LIPIDS the soaps are’neutralized' with ‘salfu ig'aeid, ‘aiid condensation Of thedistillat ‘ ingollible’ f a “distillate is filtered, dn “aliquot ‘of*the filtrate’ is_ titrated. with standard otassi i droxide and the Reichert Meiss! Number calcu- lated, The condenser; receiver, aiid filter paper are washed with water, and the insoluble‘fatty ‘acids are di solved in ‘succeeding portions of ethyl al- cohol. This solution is titrated with standard alkali and the Polenske Num- ber calculated, oe eee ee? aa ations are:particularly valuable in differentiating butter from coco nd in detecting adulteration of butter or the sub- atitution of fat mixturés with'the physical constants of butter for it. Only a small number of. other oils;contain appreciable amounts of-volatile fatty acids. The Reichert Meiss! Number is particularly valuable in_detecting. adulteration In butter, Although the Reichert Meissl Number varies’ for bulter with season, nutrition, and tirie'ih’the lactational cycle of the cow, It is usually between'24 and 34, higher than other edible oils. The distilla- tlon procedure as sét'up ‘in the Reichert Meiss! and Polenske methods does not remove all of the volatile acids from. the saponification mixture; but if the procedure is followed accurately ‘and the size of the sample restricted to 5 g, reproducible results and valuable information can be obtained. The chief volatile, water-soluble fatty acid in butter is butyric acid. This In determined by the Kirsefiner Value which measures the potential amount of soluble silver salts in the Reichert Meissl distillate. Silver butyrate is soluble in water while’ the silver salts of the other volatile, water-soluble lity acids, are_reatvely insoluble The ‘neutralized distillate from the ‘cichert Meissl determination is treated with silver sulfate and filtered. The filtrate is acidified with sulfuric acid and distilled. The distillate is care- fully collected and titrated with 0.14 alkali, either sodium, potassium, or ae hydroxide. Kirshner Value is calculated from the following equa- jon: . Wie A x, 121(100 +.B) Kirschner Value = , 10,000 where: A = corrected Kirschner titration, and . B = milliliters of alkali to neutralize the 100 ml distillate from Reichert Meiss. Tee ton Tesults from the fact that the quantity of distillate collected rare gsample.in the Reichert Meissl (first) distillation is 110 ml while ml is titrated, and likewise in the Kirschner (second) distillation. But- 296s x er cae wy a ee ee gids FOOD CHEMISTRY ter gives a Kirschner Value.of 19 10,26, coconut oil a imately 1.9, paler kernel arou; ther fats and oils between 0.1 to 0.2, Butter is there- fore remarkable for the fai ge. t of bur is init, and i can be identified on thi 5 The Saponification Number (the Koettstorfer Number) is defined as the number oF milligrams of potassium hydroxide required to saponify 1 g of fat or oil. When potassium hydroxide reacts with a triglyceride, three moles of potassium hydroxide react with one mole of fat. R,COOCH, R,COOK noe gone 'R,COOCH 4 3KOH — R,COOK + HOcH aS R,COOCH, R COOK HOCH, If the triglyceride contains low molecular weight fatty acids, the number of molecules present in a I_g sample of the fat will be greater than if the fatty acids have Tong carbon chains and high molecular weights. The fat with the. low molecular weight fatty acids will consequently have a high Saponifica— tion Number. We find that butter wile ts unvsuslly hieh pereemiage ot butyric-acid has the highest Saponification Num! 2210-250 The method depends on the saponification of a weighed =i pampl of fat of about 5 g with an excess of standard alcoholic potassium hydroxide. The potassium hydroxide is carefully measured into the flask with a buret or pipet. The mixture is boiled under reflux ‘condensation until-saponification is complete, and the remaining potassium hydroxide is determined by back titration with 0.5N hydrochloric acid. : _- Part used in saponification Total KOH (meq) (Total meq-remaining meq) Remaining meq by titration Thé Saponification Number can then be determined by subtracting the number of milliequivalents of hydrochloric acid- used for back titration from the number of milliequivalents of alcoholic potassium hydroxide in- troduced, multiplying by 56.1-mg (milliequivalent weight of KQH) and di- viding by the weight of the sample in grams. 1,24 iw Semhi Saponifcation Number = Mex! x 4 KOH) - (ml x NHCH] x 56.1 ner it “ae ep etgfek ei g sample ; The Hehner Value measures the amount of fatty acids which are insoluble in water. Thus fatsand oils which have high Reichert Mciss! Numbers willFATS AND OTHER LIPIDS have low Hehner. Values. Since most fatty acids present‘in natural fats are not soluble in water, most fats have relatively high Hehiier Values, The value can be determined either.by weighing a sample of fat‘and saponifying— alo fe or by using the solution from the it with alcoholic potassi droxide or byt 50 titration. The alcohol is evaporated, the soaps dis- aponification Numbe! 6 > qolved in hor water, aI with concentrated hydrochloric acid. Free | Folved in ror water, and treated: fatty acids are f .d: and when:the mixture is cooled, the insoluble fatty icids form a cake on the top of the solution. This is filtered on a weighed filter paper, dried, and weighed. : ; The Jodine Number is the nthe of te onsale pound absorbed by 100 g of fat. The double bonds present in the unsatu- ated fatty acids Teact readily with iodine or certain iodine compounds. -“{o,form an addition compound even while.the fatty acid is combined with, erefore a measure of the “Siycerol_in the fat, The Todine Number is thi of th ‘oxtent of un ion of the fatty acids present in a fat. While oleic acid contains one double bond in its TS carbon chain, linolenic acf8Contains ‘ds in its 18 carbon chain. Thus a ‘nolecule of fat contain- three double bon: Thg one oleic acid can absorb_or react with.only one third as much ine. ning one linolenic acid residue. “Tor TBr or ICI) as a molecule of fat contai The fatty acid ‘assortment present i natural fats is fairly characteristic: of the fat. While there Will be variation in each vegetable oil with climate, soil, and variety and in each animal fat. with nutrition and breed, the variations are small compared to the variations between fats. The Iodine _Number is therefore of great value in identifying fats and.oils. ; The lodine Number is determined by dissolving a weighed sample of fat Hub (0.1 g 10.0.5 g) in chloroform or carbon tetrachloride and adding an ex- cess of halogen. After standing in the dark for a controlled period of time, W, the excess, unreacted iodine is measured by thiosulfate titration. In the kaj Hanus method the standard iodine solution is made up in glacial acetic acid Swsuupt tnd contains not only iodine but iodine bromide which accelerates the Te- oni dle tiction. The Wijs method uses an iodine solution made up‘in glacial acetic tomy ncid but ‘containing iodine chloride as accelerator. The.excess iodine .re- F _ ets ith spgium thiosulfate according to the following equation: "JY & brine ta i 2.NajS,0, + 1, =; 2'Nal + Na,S,O, ip “he end point is determined by the disappearance of the blue starch-iodine color 1 a yprbecteydiaeyh PE The Acety! Value is @ measure of the amount of hydroxy fatt eae Satin a fat Castor Oil .is rich in ricinoleic acid, C,,H,,(OH)COOH, and the determination is of value for this and a few other oils. Most edible oils a Says contain only very small-amounts or no hydro: ic . Y xy fatty acids. H aM ef Ky prinecped Chou HENS Can be Let er urnoal 31Accd Vabua (MW) is a yen : of KOH tm op reaclaof ao tke bt : 6 Hh maaben of TS SEAN tas eotirg wm 1°09 cell Volue Non aoly labed Gf pdebanane SY ae penple ahah > : BL CH COOH Ave pv of acl hot fit Et Dalia i Py a