FOOD ADDITIVES

FOOD ADDITIVES

USEFUL FOR Mpharm 1st year pharmaceutical analysis

1. FOOD ADDITIVES

2.   INTRODUCTION TO FOOD ANALYSIS Food analysis is a very important branch of analytical chemistry, able to provide information about chemical composition, processing, quality control (QC) and contamination of foodstuffs, ensuring compliance with food and trade laws.

3. INTRODUCTION TO FOOD ADDITIVES Food additives are substances added to food to preserve flavour or enhance its taste, appearance, or other qualities. Food additives are used for the purpose of maintaining or improving the keeping quality, texture, consistency, appearance and other technological requirements.

4. Classification of Food additives COLOURS PRESERVATIVES ACIDITY REGULATORS ANTIOXIDANTS ANTICAKING AGENTS ANTIFOAMING AGENTS ARTIFICIAL SWEETENERS ENZYMES EMULSIFIERS EMULSIFYING AGENTS

5. Classification of Food additives FLAVOURS FLAVOUR ENHANCERS MODIFIED STARCHES STABILIZERS THICKENING AND GELLYING AGENTS. FOAMING AGENTS RAISING AGENTS HUMECTANTS BULKING AGENT COLOUR RETENTION AGENTS FIRMING AGENT ETC.

6. PRESERVATIVES Preservatives are the compounds used to prevent and retard the microbial spoilage of food. “Substance which when added to food is capable of inhibiting, retarding or arresting the process of fermentation, acidification or other decomposition of food”. They are classified into Class I and Class II preservatives

7. Class I preservatives 1. Common salt 2. Sugar 3. Dextrose 4. Glucose 5. Spices 6. Vinegar or acetic acid 7. Honey 8. Edible vegetable oils

8. Class II Preservatives 1. Benzoic acid including salts thereof 2. Sulphurous acid including salts thereof 3. Nitrates or Nitrites and/or Sodium and Potassium in respect of foods like ham, Pickled meat 4. Sorbic acid and its sodium, 5. Potassium and calcium salts 6. Propionates of Calcium or sodium, 7. Sodium, Potassium and Calcium salts of Lactic acid. 8. Methyl or Propyl parahydroxy Benzoates 9. Sodium Diacetate .

9. Benzoic Acid: Qualitative Methods Ferric Chloride Test: Acidify the food product with hydrochloric acid (1+3) and extract with diethyl ether. Evaporate the solvent on a hot water bath removing last traces of solvent under a current of air. Dissolve the residue in few ml of hot water and add few drops of 0.5% ferric chloride solution. Salmon colour precipitate of ferric benzoate indicates the presence of benzoic acid.

10. Quantitative Methods Titrimetric Method: Principle : Benzoic acid is separated from a known quantity of the sample by saturating with sodium chloride and then acidifying with dilute hydrochloric acid and extracting with chloroform. The chloroform layer is made mineral acid free and the solvent is removed by evaporation. The residue is dissolved in neutral alcohol and the amount of benzoic acid is determined by titration against standard alkali .

11. Calculate the benzoic acid contents as follows: Benzoic acid (ppm) = 122× Titre × Dilution× 1000 × ml of 0.05N sodium hydroxide Weight of sample × aliquot taken (100 or 200ml of filtrate)

12. NITRATE AND NITRITE IN FOODS Sodium and potassium salts of nitrate and nitrite are added mainly to preserve meat and meat products such as cured meat and meat pickles. Determination of Nitrate and Nitrite in Foods Principle: The sample is clarified with alumina cream and the amount of nitrate present determined by allowing it to diazotise Arsenilic acid and coupling the diazonium salt with n-1-naphthylethylene diamine. The colour so formed is e-absorbance is measured at 545 nm. An aliquot of the sample is mixed with spongy cadmium in order to reduce any nitrate present and the nitrite so produced is determined in the same way.

13. NITRATE AND NITRITE IN FOODS Principle The amount of nitrate present is then calculated by subtracting the nitrite from the total. Extracted into n-butanol and the nitrite so produced is determined in the same way . The amount of nitrate present is then calculated by subtracting the nitrite from the total. The amount of nitrate present is then calculated by subtracting the nitrite from the total .

14. ANTIOXIDANTS Antioxidants are added to oils and fats to prevent oxidative rancidity. Examples Ethyl gallate Propyl gallate Octyl gallate Dodecyl gallate Butylated hydroxy anisole (BHA) Tocopherol Ascorbic acid

15. Qualitative method Thin Layer Chromatographic Detection of Antioxidants Principle: The sample oil is dissolved in petroleum ether and extracted with acetonitrile. Acetonitrile extract is evaporated in vacuum in a rotary evaporator at a temperature not exceeding 40ºC. The residue is dissolved in alcohol, applied to TLC plates and after development, spots are visualized by spraying with Gibb’s reagent.

16. Quantitative method Antioxidants by High Performance Liquid Chromatography: Applicable to propyl gallate (PG), Trihydroxybutyrophenone ( THBP) Ter - butyl hydroquinone (TBHQ), Nor dihydroguaritic acid (NDGA), butylated hydroxyl anisole (BHA), butylated hydroxyl toluene(BHT) at 20- 200 ug / gm in oils and fats 10-100 ug /gm in butteroil and to octyl and dodecyl gallate at 10 – 100 ug / gm in butter oil) Principle: Antioxidants are extracted into acetonitrile. Extract is concentrated and diluted with 2 propanol. Antioxidants are separated by liquid chromatography and measured by UV detection at 280 nm .

17. ARTIFICIAL SWEETENERS SACCHARIN : Qualitative methods Detection: Phenol sulphuric acid test: To the residue obtained after removing solvent, add 5 ml of phenol sulphuric acid reagent (pure colourless crystals dissolved in equal weight of sulphuric acid) and heat for 2 hrs at 135-140ºC. Dissolve in small amount of hot water and make it alkaline with 10% sodium hydroxide. Magenta or reddish-purple colour develops if saccharin is present. Resorcinol sulphuric acid test: To the residue add 5 drops of resorcinol- sulphuric acid (1:1) and heat on a low flame until the product turns red. Dissolve in 10 ml of water and make it alkaline using 10% sodium hydroxide solution and add few drops of iodine solution. A green fluorescence is developed if saccharin is present.

18. Quantitative Methods General method for non-alcoholic beverages Principle Saccharin is extracted from a known quantity of acidified sample with diethyl ether. The solvent is removed and the residue is digested with hydrochloric acid and made to a known volume. An aliquot is treated with Nessler’s reagent and the absorbance of the coloured product is measured at 425 nm.

19. DULCIN Qualitative methods Deniges-Tourrou Test: Moisten dry residue with nitric acid and add one drop of water. Presence of Dulcin is indicated by orange red coloured precipitate. Quantitative Method: UV-Spectrophotometric Method: Principle: Dulcin is extracted from the prepared sample under alkaline conditions with diethyl ether. The residue after removal of solvent is taken in ethyl acetate. The absorbance of the solution is read at 294 nm.

20. FLAVORS & FLAVOR ENHANCERS Flavors and flavor enhancers form a divergent group of organic compounds both natural and synthetic in nature. They are used in trace amounts to impart a characteristic flavor. Menthol, vanillin and monosodium glutamate are of interest as they are extensively used in various foods. Menthol is used mainly to flavor confectionery and pan masala. Vanillin is extensively used in ice creams and monosodium glutamate to enhance flavor of meat, soups etc., and Gas chromatography is extensively used in determination of various flavoring compounds. EXAMPLES Menthol Vanillin Monosodium glutamate

21. Gas Chromatographic determination of menthol in mentholated sweets and Pan Masala Principle: Menthol along with other flavouring matter is steam distilled into chloroform. The dry chloroform extract is directly subjected to gas chromatography on 10% carboway 20M using FID. The amount of menthol present in the sample is determined using peak height/area of sample and standard.

22. Determination of Monosodium Glutamate in Food Principle: Glutamic acid is extracted from foods using water, separated from other amino acids by using an ion-exchange resin chromatography and titrated potentimetrically using 0.1N sodium hydroxide .

23. STABILIZERS Stabilizer is an additive to food which helps to stabilize food emulsions. EXAMPLES: Gums Alginate Agar Guar gum Starch Pectin Gum arabic

24. THICKENING AGENTS Thickening agent is an substance which can increase the viscosity of a liquid without substantially changing its other properties. EXAMPLES Starch Gum pectin

25. Quantitative method Determination of pectin contents The carbazole test was carried out essentially as described by Filipov & Vlasyeva (1973). The rapid procedure was carried out as follows. Sample sizes were: 0-2-0-5 g dried pectin, 0-5-1 0 g dry rough material or 20-25 g fresh weight of fruits, vegetables, berries or canned fruits. From solutes, neutralized pectic hydrolysates or juices, 20-50 ml was used. Before use, the samples of the dried material were weighed in vials moistened with 1-2 ml ethanol to prevent clotting, and subsequently dissolved or suspended in 10-20 ml distilled water each. Per 10ml of solution or suspension, 1-2 ml of 1 N NaOH was added and thoroughly mixed De-esterification occurred for 20 min at 20-25°C .

26. the suspension was acidified by the addition of 1 N HC1, 1-5 x the volume of 1 N NaOH added before, and mixed thoroughly To precipitate the pectin, 50 ml of 01 N HC1 was added. The suspension was stirred and kept for 5 min at room temperature to equilibrate the concentrations in medium and pectin flakes The final volume or weight of each sample was measured

27. The mixture was then filtered through a wide pore filter (Whatman 1) From the filtrate 10-20ml were pipetted into a 250 ml flask The residue of the filter was pooled with the remaining filtrate. Funnel and vial were washed twice with distilled water and the wash solutions were added to the residue-filtrate mixture and thoroughly stirred. The filtrate in the flask and the mixture were separately titrated with 0-1 N NaOH using Hinton’s indicator. From the result of the titration of the 10-20 ml filtrate, the HC1 content of the original volume was calculated. Together with the results of the titration of the pooled filtrate, residue and washings, the total amount of all pectic acids of the original sample can be estimated.

28. The amount of poly- galacturonic acid (pectin) was calculated as follows: P %=(V2-V1).176.0-1.K/1000 W*1OO where V1-the calculated volume (ml) of 0T N NaOH for the titration of HC1 in the entire reaction mixture; V2-the volume (ml) of 0T N NaOH for the titration of the entire reaction mixture; 176-the gram-equivalent of pectic acid; 0-1 is the concentrationof NaOH , used for titration; K-the coefficient to the NaOH concentration; and W-the weight of the sample (g).

29. JELLING AGENT Jelling agents is any substance added to a food product to provide the texture of a gel. EXAMPLES Gums Acacia Alginic acid Bentonite Gelatin Tragacanth

30. Identification of Gums: Wet 0.25 to 0.50 gm of the material to be identified with 1 to 2 ml of 95% alcohol and add 50 ml distilled water. Suspend the solid material in the water by shaking or stirring and heat when the sample dissolves. Discontinue heating, otherwise hold at 85 to 95ºC for 15 min

31. Confirmatory Tests: Alginates and de- esterfied pectin’s: 0.2 ml of 3N HCL / other mineral acid + 3 to 4 ml of the sample A gelatinous precipitate ( alginates or de-esterified pectin) Irish Moss: Add 2 to 3 drops of 0.5% methylene blue solution in water + 1 ml of the sample solution Precipitation of purple stained fibres confirms Irish Moss. .

32. Methyl cellulose: 5 ml of sample with 25 ml of 95% alcohol + 2 to 3 drops saturated sodium chloride No precipitate confirms methylcellulose Agar: Precipitate gum from 5 ml of sample with alcohol stain with tincture of iodine Blue color is formed ( Starch is also stained blue by the reagents) Starch: Add 1 to 2 drops of iodine solution + 1 ml of sample A blue or purple color confirms starch (Some samples of gum tragacanth may give a faint blue test)