You are on page 1of 24

Analysis of food

Lipids: Solution of unsaponifiable matter in saponifiable matter (glycerides). Classification of lipids: a) Simple b) Compound c) Derived

a) Simple lipids: Esters of fatty acids and glycerol → fats and oils Esters of fatty acids & higher molecular weight monohydric alcohol → wax.
1-

Simple lipids: are composed of: Wax Mixed > One type of fatty acid

Triglycerides Simple One type of fatty acid 2-

Compound lipids or conjugate lipids:

1. Phospholipids Consists of Fatty acids + glycerol + phosphoric acid + nitrogenous base Ex.: Lecithin present in egg yolk

2. Glycolipids Consists of fatty acids + carbohydrates + nitrogenous base as called: Glucolipids cerebrozides

3. Sphingomilines Consist of fatty acids + nitrogenous base + phosphoric acid but no glycerol

3. Derived Lipids: it is the hydrolysis product of the previous 2 types (e.g., fatty acids, glycerol, ……….)

N.B. 2: Phoshpolipids are also called phosphatides or phosphoglycerides.

Q.1.Compare between fats &oils? Answer: Both fats and oils have the same chemical properties (esters of fatty acids and glycerol) but they differ in their physical properties (Fats  Solids or semisolids & Oils  Liquids) Composition of lipids Lipids are solutions, its solvent is the glyceride part (saponifiable part 98%) and its solute is non glyceride (non – saponifiable part 2%). Saponifiable matter (98%) Glyceride part (98%), solvent Liable to alkaline hydrolysis (Saponification): giving glycerol & K salt of fatty acid (Soap)
O C R O O O O C R O C R K salt of fatty acid (soap) OH Glycerol Alkaline Hydrolysis O 3 R C OK + OH OH

Unsaponifiable matter (2%) Non- glyceride part (2%) Does not undergo alkaline hydrolysis because they are composed of: 1. Fat soluble vitamins (A, D, E, K). 2. Chromolipids ex. carotene responsible for the yellow color. 3. Hydrocarbons ex. squalene. 4. Steroids or sterols. Determine whether the oil or fat is genuine or not

+ 3 KOH

The physical and chemical properties of lipids vary with variations of the glyceride part. Types: TG DG MG

Important Notes:
1.

C4  is present only in butter fat.

linolenic.Butter fat contains C4. C8 and C10  are steam volatile but water insoluble and alcohol soluble. Play an important role in vital activities as repairing and reproduction. Hydroxy fatty acids Naturally occurring Omega Synthetic . Saturated fatty acids 2. skin permeability. normal growth. 7. 4. 6. C18  stearic acid  is present in milk and animal fat and it is mostly used as a base in suppositories  but commonly used as its salts because it is very irritant as an acid. C10  are present in butter fat and coconut oil. Unsaturated fatty acids 3. Why Linoleic. C6. C10 which are all short chain fatty acids of which: 2. Arachidonic acids are called essential fatty acids? Answer: 1. C8. so they have to be supplied externally. C6. C12  20  are long chain fatty acids. 2. Cyclic fatty acids 4. 5. Q. Fatty Acids 1. this confirms adulteration. C20  Arachidic acid present in arachids oil which is very close to olive oil and almond oil and therefore it can be used in adulteration of these oils because it is chapter but upon analysis when arachidic acid is found in these oils and normally it' not present except in arachis oil. 3. 8. C8. They have more than are double bond and the body cannot synthesize them. C4 and C6  are water soluble fatty acid and steam volatile.

Unsaturated fatty acids may contain up to 6 double bonds. Abbreviations: Position of double bond . Generic name: 1. In case of SATURATED fatty acid the name has the suffix (noic) and this suffix is added after the number of carbon atoms. 5. they have the suffix (enoic) added after the number of carbon atoms (if there are 2 double bonds (dienoic). if three trienoic and so on.. sometimes the fatty acid contains an additional functional group e. The name of the fatty acid is derived from the number of carbon atoms. They are either saturated or unsaturated.General Characteristics of naturally occurring fatty acids: Carbon atoms in the fatty acids/ or the number of carbon atoms in the fatty acids chain is even no. In case of unsaturated fatty acids. 1. 4. 2. 3.g. Naturally present fatty acids are cis-non conjugated. 2. Hydrocarbon chain is not straight but zigzag. 3. General formula: CnH2n O2 Nomenclature of fatty acids: The name of fatty acid could be: 1. (4-24) starting from the carbon atom of the carboxylic a group. (-OH) as in case of hydroxy acids. 2. The main functional group is (-COOH).

or more) 2Omeg a f. Hydroxy fatty acids: a. Their presence indicates adulteration. Trans 2. 3. Unsaturated Fatty Acids Naturally occurring unsaturated fatty acids Omega fatty acid 1Highly unsaturated (5 d. Synthetic (not naturally occurring) Ex. 4. b. of No.b. Trivial name: check from table of unsaturated fatty acids. . C18 Chaulmoegric acid (chaulmoegra oil).Those are fatty acids having hydroxyl groups and they could be present in saturated or unsaturated fatty acids. of double Carbon atoms bonds 3. Chaulmoegra oil = rapeseed oil. Δ No.C……………. 3Very strong antioxidants.Present in castor oil  has a hydroxyl value of 150. Conjugated 3.a are present only in marines not in plants. oleic. Cyclic fatty acids: C16 Hydrocapric acid (chaulmoegra oil). …………….Formed during hydrogenation of oils.. linoleic. . linolenic a (Iso-acids) Characteristics of iso acids: 1. 4.

Ester linkage 3.Examination of Lipids It involves 2 steps: 1. 1. Reactions with carboxylic acid (-COOH) group: Presence of free carboxylic acids in lipids is due to hydrolysis of lipids in presence of water. Chemical examination II. Acid value: Definition: It is the no.Hydroxyl of the glycerides groups of the hydroxy acids 4. CH2 R2OCO CH CH2 OCOR3 OCOR1 CH2 OH R1 COOH + 3 H2 O HOCH CH2OH + R2 COOH R3 COOH To determine the extent of hydrolysis or rancidity. The hydrolysis is accelerated by acids. we should determine the acid value of oil/fat (lipid). . bases or enzymes. Double bonds of hydrocarbon chain of fatty acids. Physical examination 2. of mg. Chemical Examination Chemical examination of lipids involves reactions with: 1. Free carboxylic acids present in the lipid 2. of KOH required to neutralize the free fatty acids present in 1 g oil or fat (lipid).

B. N.A available for the reaction).: Acid valve can't differentiate between genuine and adulterated oils. Why warming? To facilitate the reaction. . 3. ↑ S. Acid valve gives an indication about the hydrolytic rancidity of oils and edibility. We use (mg KOH) in order to express acidity because oils/ lipids contain more than one fatty acid and so since we don't know the mol. and ↓ surface tension subdivides the oil into fine globules.B. Some remarks: 1. 2.1 N KOH 3 10 drops ph ph 5 ml neutral alcohol 1 g lipid End point = colourless – 1st pink. Why not use an organic solvent? Because the oil will completely be soluble in the organic solvent and the reaction will not take place. Why alcohol? For extraction of free fatty acids from lipid (dissolves the free f. 4.a.Procedure: ≠ 0.: Acid valve shouldn't exceed 5 if > 5 ∴ sample is rejected Significance: 1. 2. Why n-alcohol To remove its acidity (since alcohol is liable to oxidation yielding acid). N. wt.

Principle: Back titration" Known xss of KOH Lipid Residual (Pink) ≠ St. acid (HCl) E.a.P. wt. ple Eq. of oleic acid in calculations. w of sam t.of all fatty acids forming the oil so we depend on the fact that in the majority of oil the main f. wt x N 1000 In this case the mol.) .a. wt used in calculations: in case of palm oil is palmitic acid Coconut oil is : Lauric acid Significance: > 1 % acidity ∴ oil is rancid ∴ rejected 2. Except of roils in which the main f. Reactions of the ester linkage of the glycerides ASaponification value: Definition: It's the number of mg KOH required to neutralize the free fatty acids and saponify the glycerides present in 1 g lipid (it's a measure of both free fatty acids and saponify the glycerides present in 1 g lipid (it's a measure of both free and combined fatty acids). is oleic acid so we use the mol. wt. of this main f. is known in this case we use the mol.P.: Pink – colourless Saponifiable part = Known Xss – residual KOH Known Xss – mls of HCl (E. and this is called: Percent activity: % acidity = m of standard x F x 100 ls.a.

2) (2) Why alcoholic alkali is used? Answer: Because alcohol subdivides the lipid into fine globules.5 N KOH not 0.Procedure: Air condenser 1.ph. Titrate ≠ 0. (pink). 4. 1g lipid 2. of alkali used. 25 ml 0.1 N? Answer: 1) To reduce the volume taken of KOH To accelerate the reactions by providing more drastic conditions by ↑ conc. thus facilitates the interaction between the lipid and alkali during saponification.B for 30 min. Reflux on boiling W. while NaOH produces hard soap (producing clumps). (4) Why should we do blank? Blank must be done as part of KOH may be converted into K2 CO3 by reaction with CO2 in air which reacts with HCl to the .5 N KOH (alcoholic KOH) 3. 5. not NaOH? KOH is more preferred since it produces soft soap.5 N HCL E.P: Pink-colourless Important Questions (1) Why 0. (3) Why KOH. Add 10 drops ph.

ph. changes its colour at pH suitable for determination of residual KOH (upon consumption of residual KOH) at pH range (8-10) while M.a.5 CO32.bicarbonate step (0.5 KOH) in presence of phenol phthalin as indicator.) = total KOH. CO32. Why air condenser? To avoid volatilization of alcohol.   C→l K salt of f.4  and 1st Xss of HCl 6. (soap).ph & not M.a.O? At the end of the Rx.+ H+  HCO3– + H+  H2O + CO2 11 8. Principle: Back titration" Known xss of KOH Lipid Residual (Pink) ≠ St. the flask contains 3 substances: Residual KOH & Glycerol & K salt of f.= 0.O (5) Why ph.O.a. 3.ph M. changes its colour (not suitable)  pH. H H Residual KOH + K salt of f.3 3-8 pink Colourless Colourless Red Red Yellow pH ph.   C→l complete neutralize pH 8-10 (ph. acid (HCl) E.ph.a. changes its color at pH range (3-4) upon complete consumption of residual KOH and K salt of f. changes its color) M.P.: Pink – colourless Saponifiable part = Known XSS – residual KOH .O. The end point should be used to determine only residual KOH and the ph.

B. Ester value = Sap value .a. wt. Value of mineral oil = zero ∴ N.a.Ester valve Definition: No.: Sap. Butter Short chain f. of KOH required to saponify the glyceride present in 1 g lipid.: Sap.P. Reactions related to the hydroxyl (-OH) group: A) Hydroxyl value: Definition: it is the number of mg. KOH required to neutralize the acetic acid capable of combining by acetylation with 1 g lipid.) Significance: Gives an indication about the mean molecular weight and the chain length of f.a.acid value 3. Value of butter = 220-225 Sap. α 1 chain length Oils Long chain f.B. . Value of vegetable oils = 180-195 Sap. B.Known XSS – mls of HCL (E. of mg.Gs: 1 mean mol. N. comprising the lipid Sap value α In: T. value is not significant in detection of adulteration of oil by oil but can detect the adulteration of oil with liquid paraffin.

Reflux for 30 min on boiling WB. 1g lipid 2. why? 1. as indicator. Procedure: Air condenser 1.P. Acetic anhydride in pyridine 3.Principle: Principle: Back titration" Known xss of acetic anhydride Lipid Residual ↓ + H2O Liberation of acetic acid ≠ titration against alcoholic KOH using ph. Why pyridine? Answer: Provide non aqueous conditions → prevent conversion of acetic anhydride into acetic acid from the beginning. 2. Blank (B): is done due to the reaction conditions (heat and time) to find out the actual amt.ph. Blanks are to be done. of acetic anhydride available for the reaction. Add H2O then boil 5.P) can be determined by titration of lipid sample without acetic anhydride against alcoholic KOH. E. 2.: Colorless – 1st pink. . 4. Back titrate the residual acetic acid ≠ KOH (alcoholic KOH) Equations: OH OCOCH3 OCO HO OCO + Acetic anhdride CH3OCO OCO OCO Important Questions: 1. Blank (A) ACID VALUE: Free fatty acids present in the sample react with KOH (leads to false +ve increase in E.

Why H2O is added? 1. value of non acetylated oil (Blank) . Hydroxyl value > 15 except for H. Acetylated oil prepared by reaction of oil sample with acetic anhydride. Stop the reaction. How to differentiate? Answer: Carry out acid value If ↑↑ If normal B) Acetyl value: Def: Number of mg of KOH required to neutralize the acetic acid resulting from 1 g of acetylated oil during soponification. Principle: 1. 2. Acetyl value = Saponification value of acetylated oil – sap. ∴ Rancid ∴ Castor oil.Hydroxyl value = (B + A) – (Sample + A)  B + A – S – A So the error is cancelled. 2. 3. what should we suspect ? Answer: The oil is Castor oil or rancid Q. To decompose the residual acetic anhydride into acetic acid. Saponification value of non acetylated oil is determined. of castor oil = 150 Q: If the hydroxyl value was high. Significance: Used as a measure of rancidity and hydroxyl fatty acids (present in recinoleic acid present in castor oil).V. Saponification value of acetylated oil is determined. 3.

and sterols. + M.G. Significance: Hydroxyl value and acetyl value are the measure of rancidity and hydroxyl acids present in lipids. + hydroxy acids OH OCO HO OCO + KOH T. The halogenating agent: halogens are arranged in the following descending order according to their reactivity (F2 > Cl2 > Br2 > I2). + M. Halogenation is an addition reaction and it depends on: The rate of addition of halogens depends on: 1. of double bonds Distance between double bonds and carboxylic acid group Conjugation (inverse) 3. . 2. D. + Hydroxy acids. A measure of degree of unsaturation of lipids.G.B.G. + D. in a dark place. = Sap value of non acetylated oil By difference = acetyl value = D.G. 4.G. 2.G.G. Reactions related to double bonds: 1. N.G. Type of unsaturation: No.OCOCH3 OCO CH3OCO OCO + KOH T.: The acetyl value of most lipids reaches 20 due to the presence of M. Conditions of halogenation: the reaction must be carried out in presence of an organic solvent.

B.C. Halogenating agent Hanus Composition 0. The following reagents can be used as a halogenating agent during determination of I. 1.+ S4 O62- Procedures: Leave to stand in dark for 1 hr 25 ml Hanus reagent 5 ml CHCl3 0. In bromine – Dioxane reagent: loose coordination of Br2 enhances the addition py.The following chemical constants can be used to differentiate different lipids according to the degree of unsaturation.S. Hanus reagent: consists of I2/ Br2 / glacial acetic acid Principle: Known xss of Hanus reagent Lipid Residual IBr ↓ + KI I2 ≠ Na2S2O3 using starch as indicator I2 + 2 S2 O32.V.2 N IBr in glacial acetic acid Bromine – dioxane reagent 0.2 N Br2 + Dioxane in chloroform Br Br O + O Br Br + Time of addition 60-120 min 1 minute N. Iodine value: • Definition: Number of parts of iodine absorbed by 100 parts of lipid by weight.F.2 g lipid In G. 2 I. • I2 is usually accompanied by a carrier in order to facilitate the quantitative addition. .

V.V.V = 270. 3.V. a large amount of I2 is present. a. I. = zero) so adulteration with liq. stable complex with iodine which cannot be broken by Na2S2O3.V.b.  I.1 N) ↓ (Brown  yellow  straw yellow) ↓ 8 add 1 ml starch O Blue  N a S → colourless 2 4 Important Questions: 1. forming a v. . a. 1.Why starch is added after straw yellow color is obtained? Because from the beginning of the reaction.Add 10 ml KI Dilute with H2O Titrate the liberated I2 (equivalent to residual hanus ≠ St.V. of fat or oil ↑ by increasing the degree of unsaturation of f.V.(0. S2O32. of double bonds. while linoleic acid with 2 d. 2. = 180 and linolenic acid with 3 d. = 90. where F. Significance: I. with one double bond as oleic acid has I. is used for detection of adulteration of fats or oils with liquid paraffin (I. Paraffin ↓ I.  I.V..b. Why blank must be done ? Due to volatility of I2 2. I. α No.

80-100 100-140 140-200 Main F.V. I. a. I.V. arachis Cotton seed.V. Ex.V. I. Principle: As I2 value but the reagent used is thiocyanogen (SCN)2 or (SCN-SCN). .). it will dry due to auto-oxidation of highly unsaturated f. In general: I. α 1/ Melting point of lipid.V. Why drying oils? Drying means when the oil is spread to a thin film and exposed to air. sunflower Since olive and almond oils are both non drying oils I. but if they are from the same group  I. Oleic acid Linoleic acid Linolenic acid Examples Olive. Adulteration of olive oil with sesame oil (↑ I. almond. 6.V.V.4. can't detect adulteration of each one with the other but it can detect adulteration between oils of different groups. and No.a. can differentiate between 2 oils if they are from different groups. with polymerization and formation of an elastic film. 4. Drying I. 5. 3. is useless. Thiocyanogen value: Definition: It is the number of parts of thiocyanogen absorbed by 100 parts of lipid calculated as I2. I. sesame Linseed.V.V α Liability to oxidative rancidity. Semidrying 3. non drying 2. classified oils into 3 groups: Group 1. of double bonds Melting point of lipid Oxidative rancidity (Liability to) 2.

Can detect hydrogenation of oil. of linoleic acid (2 d. and d. so it can react with one d. linoleic linolenic acids can be calculated from I. of oleic acid. Bromine value: Definition: it's the number of parts of Br2 absorbed by 100 parts of lipid. how? Br2 value = Conjugated + Unconjugated.b).B.V. As for linolenic.b.V and (SCN)2 v. 90 180 270 (SCN)2 V 90 90 180 Significance: the proportions of saturated. Is useful for determination of total unsaturation as it measures both conjugated and unconjugated f.b.: We have to take in consideration that (SCN)2 leaves one double bond without addition when there is more than one double bond. of insaturated f.b. Significance: 1.N.b. oleic. of oleic acid.b. Fatty acid Oleic Linoleic Linolenic I. of linoleic acid (3 d.a.a. I2 value = conjugated only . of linoleic. of the two d. 2. in other words. it reacts with 2 double bonds of the three double bonds (1st and 3rd) and this is confirmed through comparison between I.b. 3. (SCN)2: Reacts with only d.V and (SCN)2 V. Reacts with two d.b. Reacts with one d.) But why? Due to the steric hinderance caused by the bulky reagent.

as indicator. Disadvantages: 1. # 0. 4. Dissolve 60 g of maleic anhydride in toluene. Lack of stability of Br2 as a reagent. Maleic anhydride value (Diene value): Definition: It's the number of parts of maleic anhydride absorbed by 100 parts of lipid (calculated as I2). Significance: Used for detection of conjugated double bonds only therefore used as an indication for hydrogenation. warm then cool and dilute to I L (6% maleic anhydride reagent). Can't differentiate between conjugated and unconjugated f. ∴ No hydrogenation If Br2 V. Br2 V. 3 g oil + 25 ml maleic anhydride reagent → Residual maleic anhydride  dilute with H2O and extract with ether  aqeous solu.a. 2. 2. Equations: . > I2 V ∴ hydrogenation. 3. can be calculated (through the increase in weight) gravimetrically. Procedure: 1.ph. Reflux for 3 hrs  E.1 N NaOH using ph.P: colorless – 1st pink.If Br2 V = I2 v.

P is inversely proportional to the no. Hydrogenation reaction is selective? Answer: Because the more unsaturated fatty acid undergoes saturation at first. of double bonds. ↑ catalyst conc. ↓ pressure. . 2.: Raney Ni or Pt can be used as catalyst in hydrogenation process. oils which are liquids are converted to semisolid or solid lipids which melt at mouth temperature. press. This leads to an increase in M. ↑ Pressure ↑ 170oC 4 atm. and it's called Diene – aldol or Dies Alder reaction.B. Important Questions: 1. Temp. To control the selectivity of the reaction: ↑ Temperature.HC HC CH + CH2 HC HC CO O CO HC HC CH CH CH CH CO CO O Conjugated diene (dienophile) Maleic anhydride (diene) Reaction between dienophile and dine occurs only in conjugated systems. of oils as M. What is the role of raney Ni in hydrogenation process ? Answer: Raney Ni can act as a catalyst by attraction of both H2 and double bonds by adsorption.P. Hydrogenation of Oil: Definition: It is the addition of hydrogen to the double bonds in presence of a catalyst. It is prepared by reduction of NiCO3 or Ni formate. Unsaturated lipid + H2 Raney Ni →   o partially hydrogenated lipids + isoacids. N.

4. The degree of hydrogenation is adjusted so that the oil is converted to semisolid product of suitable consistency for human consumption & melts easily at mouth temperature. 3. What are the advantages & disadvantages of hydrogenation? Answer: Advantages Disadvantages .3. Give hard fat (used in waxes. close to that of mouth temperature (prefered). Accompanied by isomerism which could be : Geometrical (Trans acids) (Isoacids) 3. candles. Part of the unsaturated fatty acids undergo hydrogenation 2. Partial hydrogenation 1. soap).P. What is the importance of hydrogenation? Answer: Hydrogenation is an industrial process used for production of edible fats from oils to simulate butter fat. All unsaturated fatty acid are converted into stearic acid. Compare between complete & partial hydrogenation ? Complete hydrogenation 1.Give lipids which has M. Positional Cis but conjugated 4.

=(114-114.Add alcoholic solution of digitonin to separate the unsaponifiable part. cancer.P. 4.More faint in colour.. obesity. diabetes. ↑ risk of coronary artery disease.(which ↑ LDL conc. 2.P. 4.Less liable to rancidity. How to detect the adulteration of butter fat with hydrogenated oil? Answer: 1. =(125-127oC) ∴ Cholesterol acetate ∴ Ergosterol acetate 2. If M. 1. 5-The product of hydrogenation has 2-Lack of the essential fatty acids improved stability & better keeping which cannot be synthesized by the quality. Alzheimer's disease . 3.Rearrangement of the double bond so conversion of the cis configuration into trans. body & must be supplied from diet.1. 3.Sterol acetate test.Low price. 1.Test for Ni: . & ↓ HDL conc. 5. 2.8 oC) If M.Acetylation by acetic anhydride  sterol acetate. liver dysfunction. infertility.Saponification by KOH: yielding a soluble part which is saponifiable part (98%) and insoluble partwhich is unsaponifiable part (2%).Check the melting point of the product.offensive odour of marine oil is removed.

a.a. Saturated Insoluble Iso-acid ∴ Iso-acid Then measure I2 value. is formed. if +ve .a.Test for iso-acids (lead salt test): a) Saponification of the sample to liberate the K salt of f.a. if +ve ∴ red gelatinous ppt.Butter fat or oil + H2SO4/HNO3 (to extract Ni traces) then Ni is oxidized into Ni2+ then the medium is made alkaline by ammonia dimethyl glyoxime. 3. b) Acidification to liberate the free f. Addition of ether. followed by extraction by organic solvent. c) d) Addition of Pb acetate to form the Pb salt of f. Soluble ∴ Natural unsaturated f.