Bruce R. D’Arcy Geoff Hawes Ian Bentley

A University of Queensland Publication

Copyright © 2006 by The University of Queensland All rights reserved

The University of Queensland


Food Chemistry – A Practical Manual


Food Chemistry

Course References

Page No.

1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11.

pH and Titratable Acidity of Orange Juices Salt Determination Carbohydrate Analysis Moisture Determination Protein Determination Properties of Enzymes Properties of Lipids Nonenzymic Browning – Maillard Reaction Ascorbic Acid Determination Ash Determination Enzymic Browning: Activity of Fruit Homogenates

7 11 15 19 23 29 33 37 41 45 49

Appendix 1 Tables for Converting Specific Gravity to % TSS (°Brix) Appendix 2 Lane Eynon Titration Tables

53 55

All practical exercises take one week except No. 4. A number of practicals will be done concurrently, ie. two 1 week practical exercises are done concurrently over 2 weeks. The practicals exercises will not be done in the order above but as assigned each year by the lecturer in charge.

Food Chemistry – A Practical Manual

The University of Queensland


Course References
The following references are a list of chosen references available in UQ libraries that may be useful for preparing for practicals and for practical report preparation. Some of the references listed below are specifically highlighted as part of particular practical exercises. AOAC International. (2001). ‘Official Methods of Analysis’. 17th Ed. (AOAC International: Gaithersburg, MD) Aurand, L. W., Woods, A. E. and Well, M. R. (1987). ‘Food Composition and Analysis’. (AVI: New York). Australian and New Zealand Food Standards Code (available in UQ Biological Sciences Library). Australian Food Composition Tables (available in UQ Biological Sciences Library). Belitz, H. D. and Grosch, W. (1999) ‘Food Chemistry’ 2nd Ed. (Springer-Verlag : New York). Charalambous, G. (1990) ‘Flavors and Off-Flavours’. (Elsevier: New York). Christian, G.D. (1994) ‘Analytical Chemistry’ 5th Edition (John Wiley and Sons Inc. : New York, USA) Christen, G.L. and Smith, J.S. (Ed.) (2000) “Food Chemistry: Principles and Applications” (Science Technology Systems, West Sacramento, CA, USA) Clark, N. (1992). ‘Food Chemistry’. (Food Trade press : Westerham, England) Coultate, T. P. (2002). ‘Food - The Chemistry of its Components’ 4th Ed. (Royal Society of Chemistry: London). de Mann, J. M. (1999) ‘Principles of Food Chemistry’ 3rd Ed. (Aspen Publishers, Inc.: Gaithersburg, Maryland, USA). Dewdney, P. A., Burke, C. S. and Crawford, C. (1975). ‘Commercial Use of Enzymes in the Food Industry’. (BFMIRA : Leatherhead, Surrey) Dickinson, E. and Stainsby, G. (1982). ‘Colloids in Foods’. (Applied Science : London). Fennema, O. (1996). ‘Food Chemistry’ 3rd ed. (Marcel Dekker: N.Y.)

Food Chemistry – A Practical Manual


Food Chemistry

Gruenwedel, D. W. and Whitaker, J. R. (1984) ‘Food Analysis: Principles and Techniques, Vol. 1 and 2’. (Marcel Dekker: New York). Heimann, W. (1980). ‘Fundamentals of Food Chemistry’. (Ellis Horwood: Chichester, England). Holland, B., Welch, A. A., Unwin, I. D., Buss. D. H., Paul, A. A. and Southgate, D. A. T. (2002) ‘McCance and Widdowson’s The Composition of Foods’. (Royal Society of Chemistry: Cambridge). James, C.S. (1999) ‘Analytical Chemistry of Foods’ (Blackie Academic and Professional: London) Kirk, R. S and Sawyer, R. (1991) ‘Pearson's Chemical Analysis of Foods’ 9th Ed. (Longman Scientific and Technical; Harlow, Essex, UK). Lawrence, J. R. (1984). ‘Food Constituents and Food Residues: Their Chromatographic Determination’. (Marcel Dekker: New York). Lee, F. A. (1983). ‘Basic Food Chemistry’ 2nd Ed. (AVI: Westport, Conn.). Maarse, H. and Van Der Heij, D. G. (1994) ‘Trends in Flavour Research’. (Elsevier: New York). Maynard, (1970). ‘Methods in Food Analysis’, 2nd Ed. (Academic Press, London). Miller, D. D. (1998) ‘Food Chemistry: A laboratory manual’. (John Wiley & Sons. Inc. : New York, USA). Morton, I. D. and McLeod, A. J. (1982). ‘Food Flavours - Part A – Introduction’. (Elsevier: Amsterdam). Nielsen, S. S. (ed.) (1998) ‘Food Analysis’ 2nd Ed. (Aspen Publishers, Inc.: Gaithersburg, Maryland, USA). Nollet, L. M. (ed.) (1992) Food Analysis by HPLC (Food Science and Technology Vol 52) Paul, A. A. and Southgate, D. A. T. (1988). ‘The Composition of Foods’ 4th Rev. Ed. (Elsevier: Amsterdam). Pomeranz, Y. and Meloan, C. E. (1994) ‘Food Analysis: Theory and Practice’ 3rd Ed. (Chapman and Hall: New York, USA). Robinson, D. S. (1987). ‘Food-biochemistry and nutritional value’. (Longman: London).

Food Chemistry – A Practical Manual

M. ‘Mechanism and Theory in Food Chemistry’. (Holt. (online).The University of Queensland 6 Schwimmer. (Medpharm Scientific Publishers: Stuttgart). Conn. (A. S. I. D. United States Department of Agriculture. ‘Source Book of Food Enzymology’. (2002) ‘Nutrient Data Laboratory’. S. and Leary.: New York). (1998).gov. S. D. Z. J. (1981).A. Sikorski. and West. Fauchman. D. (1992). 6th Edition. Inc. ‘Principles of Instrumental Analysis’. (www. (Van Nostrand Reinhold : New York).). A. (Saunders College Publishing: New York) Skoog. 4th Edition. E.usda. Food Chemistry – A Practical Manual . (1989).). ‘Principles of Instrumental Analysis’.J. H. and Kraut. V. : Westport. D. W. Souci. (2000) ‘Food Composition and Nutrition Tables’. Rinehart and Winston. W. W. Wong. (1996) ‘Chemical and Functional Properties of Food’ (Technomic Publishing Co. (Tech)) Skoog.

PART A PREPARATION AND STANDARDIZATION OF 0. Add + HCl (Hydrochloric Acid) NaCl (Salt) + H2O (Water) Food Chemistry – A Practical Manual . is made visible by means of an indicator. the sodium hydroxide used to titrate the natural acidity must be standardized. Preparation of Approx. analyses used to give an indication of the quality of the fruit juice during processing. the indicator is phenolphthalein. This knowledge can then be used to prepare solutions of standard concentration which are used then in other volumetric analyses. The purpose of a titration is to measure the volume of one solution required to react completely with a known volume of some other solution. Titration is an important volumetric technique. If the colour is bright pink or pink-purple. the solution will remain colourless.1M NaOH Weigh approximately 1 g of the sodium hydroxide pellets in a 100 mL beaker. The major acid in orange juice is critic acid and this can be measured titrimetrically. Because of the natural buffering capacity of many food materials. before titratable acidity of a fruit juice can be determined. approximately 50 mL of distilled water and dissolve. Two drops of this indicator are added to the hydrochloric acid solution before titration. In this experiment. the titration has gone too far and it will be necessary to repeat the titration.7 Food Chemistry PRACTICAL EXERCISE NO 1 pH AND TITRATABLE ACIDITY OF ORANGE JUICES INTRODUCTION The determination of pH and titratable acidity are routine Q. In this experiment. NaOH (Sodium Hydroxide) METHOD 1. However.C. 0. pH is only a very mediocre measure of changes in the acidity of a substance. At the end-point the solution will be a pale pink colour. The end-point of the titration ie. A more accurate measure is a quantity called the titratable acidity. An indicator is a substance which changes colour at the end-point of a titration. the point at which exactly enough acid is added to neutralise the sodium hydroxide.1 M SODIUM HYDROXIDE The techniques used in quantitative chemistry to measure out volumes of liquids are called volumetric techniques. the volume of sodium hydroxide (NaOH) required to react with 20 mL of hydrochloric acid (HCl) will be measured.

1M using the formula: MAVA a where MB VA VB a b MA = = = = = MBVB b = Molarity Acid Molarity Base Volume Acid Volume Base Balancing factor Acid Balancing factor Base Food Chemistry – A Practical Manual . Rapidly add about 18 mL of sodium hydroxide (about 2 mL less than the amount needed) to the conical flask. Calculation: Calculate the molarity of the sodium hydroxide solution given that the molarity of the hydrochloric acid is 0.(8) at least two more times or until 2 consecutive values are within 0. Clamp the burette in a vertical position and fill it with the sodium hydroxide solution using a small funnel. 8. when the pink colour just appears and remains for 15-20 seconds). 5.1 mL. Rinse the beaker several times with distilled water and transfer the washings to the volumetric flask.1 M sodium hydroxide solution. Check that the burette is clean and the tip clear. Rinse out a 20 mL pipette with the 0. Adjust the burette tap to deliver the acid dropwise. take the final burette reading. Take the initial burette reading (it does not have to be zero). Rinse it out with the approximately 0. Make up to the mark with distilled water and mix thoroughly. When the end point is reached (ie. Standardization of the 0. Repeat steps (4) . 7. Remove the funnel when the burette is full but do not let the burette overflow. Pipette 20 mL of 0.The University of Queensland 8 Quantitatively transfer the solution to a 250 mL volumetric flask via a funnel. Add 2 drops of phenolphthalein indicator to the flask. 2. 9.1 M hydrochloric acid solution and a 250 mL conical flask with distilled water from the wash bottle.1 M hydrochloric acid solution into the conical flask. 3.1 M NaOH 1. Note: This is only true if M1V1 _____ 1 = M2V2 _____ 1 2. 4. 6.

COONa CH2 . Be sure to calibrate the pH meter before use using the supplied buffers.COONa (Sodium Citrate) + 3NaOH + 3H2O (Water) (Sodium Hydroxide) Ensure you do not forget about the dilution factor. prepare freshly squeezed orange juice. (iii) Calculate the per cent citric acid in the various juice samples using your NaOH molarity determined in Part A and given that the balanced equation of the reaction between citric acid and sodium hydroxide is: CH2 .1 M NaOH until a slight pink end-point is reached which persists for about 15-20 seconds. Repeat until titration volumes are within 0. In addition.COONa OH -C . (a) pH Determination Determine the pH (hydrogen ion concentration) of the various orange juice samples using the pH meter.COOH (Citric Acid) CH2 .C . (ii) Titrate with the standardized solution of 0. and then pipette 25 mL of the diluted juice into a 250 mL conical flask and add a few drops of phenolphthalein indicator.COOH HO .1 mL.9 Food Chemistry QUESTIONS 1.COOH CH2 . 2. (b) Titratable Acidity Determination Determine the titratable acidity of these juices using your standardized 0. Food Chemistry – A Practical Manual . Why are the burette and pipette rinsed out with the solution to be used in them? Why does it not matter if the conical flask is wet with water? PART B METHOD ANALYSIS OF ORANGE JUICES A number of commercial orange juices are supplied.1 M NaOH from Part A and the procedure below: (i) Dilute the juice sample(s) (1 : 4) using a 100 mL volumetric flask.

& Bauman. Bauman. Define titratable acidity.. G.S. R. 4. 2. and Sawyer. UK) Food Chemistry – A Practical Manual . R. Outline the procedure for correct maintenance of pH electrodes.J.The University of Queensland 10 QUESTIONS 1. What is an indicator? Why is it important to select the correct indicator for an acid-base titration? REFERENCES Shugar. Shugar. 3. (1991) ‘Pearson's Chemical Analysis of Foods’ 9th Edition. R. R.A.. L. (1981) ‘Chemical Technicians Ready Reference Handbook’ 2nd ed. (McGraw Hill: New York) Kirk. (Longman Scientific and Technical: Harlow. Essex.

cheese. However. Silver nitrate is the most common reagent used in precipitimetry. the usual practice is to prepare approximate solutions from commercial reagent and then standardize using sodium or potassium chloride.R. because of the extreme cost of A. The method varies slightly for the analysis of different products.1 M AgNO3 solutions. Avoid contact with silver nitrate as it produces brown stains on skin.11 Food Chemistry PRACTICAL EXERCISE NO. ClBrISCN+ + + + Ag+ Ag+ Ag+ Ag+ AgC1 AgBr AgI AgSCN As well as direct titrimetric methods. margarine and bottle spring water. Samples that can be analysed include butter. 2 SALT DETERMINATION INTRODUCTION Salt is added to foods to enhance the flavour. reagent. Food Chemistry – A Practical Manual . To overcome the former problem when using sodium chloride for the standardization of 0. bromide (Br-).B. act as a preservative or in some cases both. (thus leading to possible weighing errors) and a slightly hygroscopic nature. halides are precipitated by the addition of excess silver nitrate solution and the excess Ag+ back titrated with standard thiocyanate. a standard NaCl solution is used instead of the direct titration technique. The latter may be overcome by drying for 1-2 hours at 250-350°C before using. End point detection is achieved using either potassium chromate (Mohr's method) or a fluorescein-type indicator (Fajan's Method).46) is a good primary standard but has the disadvantage of possessing a low molecular weight. In the latter. iodide (I-) and thiocyanate (SCN-). Silver nitrate may be used as a primary standard provided it is freshly recrystallised and has not been allowed to photo-decompose. being suitable for the estimation of chloride (Cl-). N. PART A STANDARDIZATION OF AgNO3 WITH SODIUM CHLORIDE Sodium chloride (NaCl = 58. silver nitrate solutions may be used in conjunction with thiocyanate solutions using a back titration technique eg. the Volhard's Method.

The obvious sign of this effect is a blue-black appearance in the precipitate. Avoid areas of intense natural light for completion of this exercise since photodecomposition in the AgCl will results and cause problems.The University of Queensland 12 METHOD Obtain the solution of approx. 2. then thoroughly washing with deionised water. Calculate the molarity of this solution. 3. Determine the molarity of the silver nitrate solution. Silver chloride deposits on the sides of working flasks may be removed by rinsing with a few mL of bench ammonium hydroxide. Notes: (i) Constant agitation is necessary throughout the titration to promote coagulation of the colloidal AgCl. 6. 1. Transfer 20 mL aliquots using a pipette to 250 mL conical flasks and add 3 drops of fluorescein indicator. The reason for this is that the organic fluorescein molecule is absorbed preferentially on to the AgCl particle at the end point where it produces a colour change which is best observed in the coagulated precipitate. Dissolve and quantitatively transfer the solution to a 250 mL volumetric flask then make up to the mark with distilled water. Prepare the standard NaCl solution by accurately weighing about 0. 0. Repeat the titration to 0. 4. Titrate with 0.40 g of AR NaCl into a clean dry beaker.1 mL agreement. All silver residues from titrations must be returned to the Silver Residues Bottles located in the fume cupboard. (ii) (iii) (iv) Food Chemistry – A Practical Manual . The end point lies within 1 mL of this point and is detected by the first observable pink colouration in the precipitate (after settling). 5.1 M AgNO3 in a clean conical flask that has been washed chloride-free with distilled water.1 M AgNO3 solution. until AgCl formed coagulates and settles to the bottom of the flask. with constant agitation.

05 M AgNO3 until a faint permanent red-brown colour is produced. Food Chemistry – A Practical Manual . normal margarine. METHOD Accurately weigh approximately 2 g of butter or margarine or low salt butter or margarine into a 250 mL conical flask and add 100 mL boiling distilled water. Determine the % salt content for butter. AgNO3 + NaCl → AgCl + NaNO3 (white) 2Ag+ + CrO4 –2 → Ag2CrO4 (red brown) Titre: 1 mL 0. 2.55 ºC and add 2 mL K2CrO4 indicator. and a low salt margarine or butter. Shake to melt sample. What is a standard solution? What are the major characteristics of a primary standard and what is the importance of using a primary standard? What is a meniscus and what is the correct procedure for reading a meniscus? What does it mean to accurately weigh approximately 4 g? 3.1 M AgNO3 = 0.13 Food Chemistry PART B SALT CONTENT OF BUTTER AND MARGARINE This determination will use potassium chromate as indicator.005846 g NaCl. Titrate with the standardized 0. cool to 50 . 4. QUESTIONS 1.

. L. (1991) ‘Pearson's Chemical Analysis of Foods’ 9th Edition. Bauman. and Sawyer.The University of Queensland 14 5.J. R. 6. Compare the chloride level (mg/mL) in bottled spring water found in this exercise with that on the package label. G. REFERENCES Shugar.. (1981) ‘Chemical Technicians Ready Reference Handbook’ 2nd ed. What are the limits for salt stated by the ANZFA Food Standards Code for butter and margarine? Do the samples comply with these limits? Compare the calculated salt content of butter and margarine to the value in the Australian Food Composition Tables. & Bauman. R. (Longman Scientific and Technical: Harlow.A.. UK) Food Chemistry – A Practical Manual .S. R. Essex. Shugar. R. (McGraw Hill: New York) Kirk. 7.

TSS (%) can be read off directly. The flask is then made up to the mark with distilled water. Similarly prepare the 10% and 20% w/v and w/w solutions. Food Chemistry – A Practical Manual . the specific gravity can be converted to °Brix. However. Once the 20% w/w solution has been prepared. for the 5% w/w sucrose solution. Additionally. This refers to the amount of solute (eg. sucrose (5 g) in a 100 mL beaker is dissolved in distilled water (30-40 mL) and then transferred quantitatively to a 100 mL volumetric flask using 10 mL washings of distilled water. followed by distilled water to make up to 100 g total mass. 10 °Brix = 10% total soluble solids. For w/w examples. sugar. °Brix corresponds directly to the total soluble solids expressed as a per cent. 10% and 20% w/w and w/v sucrose solutions using the following method. Refractometer. pour it into a 100 mL volumetric flask and note your observation of its volume relative to the final volume of the 20% w/v solution. and two of the more important ones include: (i) Hydrometer. this is true only for pure sugar solutions. Do not use your finger as a stopper. for the 5% w/v sucrose solution. In some refractometers. There are a number of ways of measuring the total soluble solids. Generally. Mix the sucrose solution well by swirling before analysis. eg. (ii) METHOD (i) Prepare 5%.15 Food Chemistry PRACTICAL EXERCISE NO 3 CARBOHYDRATE ANALYSIS PART A. the TSS (%) can be obtained. The specific gravity may be measured using a specific gravity hydrometer. and the flask inverted several times to mix. organic acids) dissolved in a solvent (most often water). DETERMINATION OF TOTAL SOLUBLE SOLIDS (TSS) INTRODUCTION A term that will often be encountered in a QC laboratory is total soluble solids or per cent soluble solids. a plastic stopper added. and then by using conversion tables. In others. tare a 250 mL conical flask and weigh in sucrose (5 g). the refractive index is measured and by reference to conversion tables. soluble solids refer to sugar(s) dissolved in water. and label each of these carefully: For w/v examples. Brix hydrometers can be used to measure the °Brix directly.

L. Bauman. Record the °Brix. Shugar. Inc. The hydrometer must not touch the side of the cylinder. New York).. REFERENCES AOAC International. ‘Official Methods of Analysis’. Food Chemistry – A Practical Manual .) (1998) ‘Food Analysis’ 2nd Ed. 16th Ed.. MD) Kirk. and Brix hydrometers. 3.G.A. Depress slightly and then note the reading at the top of meniscus. (Aspen Publishers. The Brix hydrometers give the per cent soluble solids directly. (iii) Using the refractometer. Essex. Lower the hydrometer into the sample contained in a sufficiently large measuring cylinder. clean the prism with tissue wool soaked in distilled water.T. measure the soluble solids contents of the three (3) unknown solutions provided. Shugar. (Longman Scientific and Technical: Harlow.S. S. measure the TSS (%) of each solution using the following procedure: Place a few drops of the liquid on the main prism surface. and Sawyer. G. (1981) ‘Chemical Technicians Ready Reference Handbook’ 2nd Ed. S. R. use the table provided in the Appendix 1. UK) Nielsen. measure the TSS (%) of the three samples of fruit juice provided. (iv) Measure the temperature of the solutions. (AOAC International: Gaithersburg. & Bauman. QUESTIONS 1. R. (1991) ‘Pearson's Chemical Analysis of Foods’ 9th Ed. (1995).The University of Queensland 16 Using the electronic refractometer only. To convert the specific gravity readings provided by the SG hydrometer to per cent soluble solids. Why are the soluble solids contents of the w/w and w/v solutions different? Why is temperature important when measuring the °Brix of a solution using a refractometer? Comment on the advantages and disadvantages of both instruments in measuring TSS. Maryland.: Gaithersburg. USA). R. DO NOT TOUCH PRISM WITH SHARP OBJECTS. R. 2. (McGraw Hill. After determination. (ed. (ii) Using BOTH the electronic refractometer and the S.

and used in a retitration. If the colour is discharged before the addition of 15 mL of dilute jam solution. and calculate the concentration of invert sugar from tables (see Appendix 2). and then make the solution up to the mark with distilled water. this time adding before heating almost all of the diluted jam solution required to effect near-complete reaction (the preliminary titration minus 3 mL). Add a small volume (30 mL) of distilled water to dissolve the jam. At the end point. Preliminary Titration Pipette 5 mL each of Fehling’s A and B solution into a clean 250 mL conical flask.17 Food Chemistry PART B METHOD DETERMINATION OF % INVERT SUGAR Weigh accurately (to 2 decimal places) about 0. It is important to keep the solution boiling during the titration. since it can revert to the blue colour on cooling.8 – 1 g (any weight in this range is suitable) of jam into a clean 100 mL beaker. Use this diluted jam solution for the following titrations (Lane and Eynon method). Record the volume of diluted jam solution required to titrate. Filter the solution before placing it in the burette.5 drops of the methylene blue indicator and complete the titration within a total boiling time of 3 min. If the colour is not completely discharged after the addition of 50 mL of dilute jam solution.5 drops of 1% aqueous methylene blue solution and continue titrating as above until the blue colour is completely discharged giving a brick-red colour. then a new jam solution diluted by a factor of 2 must be prepared. This will give an approximate measure of the titration value. Boil gently for 2 min. N. which is then used in a retitration. This is done to ensure the titration is completed quickly. If this concentration = y mg/100 mL and the original mass of jam taken was x g. and add from a burette the diluted jam solution (15 mL). then quantitatively transfer (using several 20 mL washings with distilled water after initial transfer of the 30 mL jam solution from the beaker) the solution to a 250 mL volumetric flask. then a new jam solution must be prepared with its strength increased by a factor of 2. Accurate Final Titration Repeat the titration. Boil the liquid using a hot plate and add further quantities of the diluted jam solution (1 mL at a time) to the boiling liquid until the blue colour is nearly discharged. The titration must be completed quickly (maximum time is 3 min) while keeping the liquid boiling.B. Food Chemistry – A Practical Manual . the blue colour should be completely discharged and the liquid should be brick-red. then % invert sugar in jam = y / 4x %. add 3 . Then add 3 .

(ed.The University of Queensland 18 REFERENCES Kirk. (1991) ‘Pearson's Chemical Analysis of Foods’ 9th Ed. Food Chemistry – A Practical Manual . D. (1965). (Leonard Hill: London). and Sawyer.. R. (1998) ‘Food Chemistry: A laboratory manual’. Maryland.: Gaithersburg. (John Wiley & Sons.H. R. : New York. Nielsen. D.) (1998) ‘Food Analysis’ 2nd Ed. Essex. UK). (Aspen Publishers. Rauch. G. S. S. ‘Jam Manufacture’. (Longman Scientific and Technical: Harlow. Inc. Inc. Miller. USA). USA).

Food Chemistry – A Practical Manual . The moisture content is frequently an index of stability and quality. It is closely concerned with the economics and legal aspects of food processing.1% per 1 h drying period.12% for dried vegetables to 27 -35% for jams and jellies. DO NOT COMPLETELY SEAL THE DESICCATOR. 1.75%. a rapid but not very accurate determination is usually preferable. METHOD All methods below are to be done in duplicate. for fresh vegetables. from 50 . since during the removal of all water present.19 Food Chemistry PRACTICAL EXERCISE NO 4 MOISTURE DETERMINATION INTRODUCTION The determination of moisture is one of the most important and widely used analytical measurements in the processing and testing of food products. The particular methods used for determining moisture content will depend on the nature of the food product and on whether speed of execution or accuracy of result is desired. The accurate determination of moisture in foods is extremely difficult. Thus. During processing. such as dehydration or concentration.102°. while the marketing of perishable products requires more accurate moisture determination to meet legal and storage requirements. Repeat until the loss in mass does not exceed 0.3 g (to 2 decimal places) of grated carrot (in duplicate) and skim milk powder (in duplicate) into numbered weighed moisture dishes (do not using taring). Direct Drying in an Oven Weigh accurately 2 . from 65% in ripe avocados to 95% in rhubarb. for most foods there is a standard method from which other methods are calibrated. NOTE: Only use tongs when handling dishes during moisture determination. from 66% in green lima beans to 96% in cucumbers. Remove and cool to room temperature in a desiccator and reweigh. Samples should be finely grated (where possible) and kept in a covered container before use. which have been previously dried in a hot air oven and cooled in a desiccator. The moisture content of processed foods varies even more from about 7 . for fresh meat and fish. Place in a hot air oven and dry over 1 h intervals at a temperature 100° . many other volatile substances may also removed. The moisture content of foods varies considerably: for fresh fruits. and is also a measure of yield and quantity of food solids.

eg. the method is time consuming and should only be used for accurate determination. Two foods are to be analysed in duplicate by each student. Transfer dish to the oven for a further 1 h. METHOD Place about 20 g of purified sand and a short glass stirring rod in a numbered dish made of nickel or aluminium. This is to ensure that a large amount of water is not drawn into the vacuum pump. and reweigh. into the clear space. cool and reweigh. stirring at intervals in order to obtain a well ventilated mass. condensed milk. cool in a desiccator to room temperature. Transfer dish and rod to the oven and dry at 100º for 1 h. Tilt the sand to one side of the dish and accurately weigh. cheese. If the food sample has a high moisture content. Dry to constant mass. sausages. METHOD a. between 1. 30 min in an oven at 100 . and is often used as the official method. Cool in a desiccator (do not seal completely) and reweigh. Repeat until loss in mass does not exceed 0. particularly if the pump does not have a trap between it and the vacuum oven. Dry in a vacuum oven at approximately 75 ºC. Weigh accurately about 5 g of well mixed dry skim milk powder (in duplicate) and 5 g of grated carrot in duplicate) into separate weighed numbered moisture dishes (do not use taring) which has been previously dried in an oven and cooled in a desiccator.5 . b. Record the total weight. Stir. Food Chemistry – A Practical Manual .102°C. Place on a warm hotplate for 20 min.3g (to 2 decimal places) of butter. However.The University of Queensland 20 Calculation: Moisture and volatile matter % w/w = Loss in mass x 100 Mass of sample 2. etc. remove. the samples should be first dried in a hot air oven at approximately 60-70 °C for a short time to remove the majority of the moisture. Direct Drying in an Oven with Sand This method is suitable for butter. margarine. Moisture and volatile matter % w/w = 3. Loss in mass x 100 Mass of sample Determination of Moisture by the Vacuum Oven Method This is an accurate method suitable for nearly all foods. Remove from the oven.1%. Cool in a desiccator to room temperature and weigh. c. Calculation: .

5 h drying periods. Another good method of judging the end-point is to place a clean dry watchglass on top of the beaker. Connect the flask to the appropriate Dean-Stark receiver and connect to the condenser. Gently heat the sample in the beaker on a hot-plate. The evolution of steam is indicated by condensation on the watch glass. The approach of the end-point may be judged by the cessation of rising bubbles of steam. Calculation Moisture and volatile matter % w/w = Loss in mass x 100 Mass of sample 6. one of which is skim milk. Determination of Moisture by the Hot-Plate Method The two foods.1% in successive 1. Constant mass is attained when the loss is not more than 0. margarine or similar fat in duplicate. Determination of Moisture by the Toluene Distillation Method Weigh into a 250 mL round-bottom flask 30 g of dry skim milk powder. Add sufficient toluene to cover the sample completely (about half full) by pouring toluene through the condenser. as well as by the advance of foam. Insert a loose cotton plug in the top of the condenser to prevent condensation of atmospheric moisture within the tube. Determination of Moisture by the Infrared Balance Method This is a rapid method suitable for drying powders. The beaker and thermometer must be weighed together before adding the food sample. Weigh accurately 5-10 g (to 2 decimal places) of butter. These calibrated instruments give the % moisture directly with no calculations required.21 Food Chemistry d. which may result from too rapid ebullition of moisture. Stir the contents gently with the thermometer to avoid spluttering. into a dry 100 mL beaker containing a small thermometer which reads to 240 °C. The temperature of the sample should at no time be permitted to exceed 130°C. butter and margarine must be analysed in duplicate. except at the end of the test. 5. Each student will determine the moisture content of 5 foods of choice in duplicate. Calculation: Moisture and volatile matter % w/w = Loss in mass x 100 Mass of sample 4. Instruction is the use of the two automated infrared balances will be given by the tutors. Food Chemistry – A Practical Manual . a. such as skim milk powder. which has been thoroughly mixed.

Essex. increase the distillation rate to 4 drops per s until no more water comes over. and allow the collecting tube to come to room temperature. QUESTIONS 1. S. REFERENCES AOAC International. Next. 2. (Aspen Publishers. heat the flask to boiling in a heating mantle.The University of Queensland 22 b. (ed. Remove from heat. Ask for assistance at this point of the experiment. Rinse thoroughly and dry completely before using. R. then repeat the washing. Inc. Continue the distillation for a short time to determine if any more water comes over. (AOAC International: Gaithersburg. Wash down the condenser (brush down with a burette brush saturated with toluene while at the same time pouring toluene through the top of the condenser). MD) Kirk. S. (1991) ‘Pearson's Chemical Analysis of Foods’ 9th Ed. 16th Ed. ‘Official Methods of Analysis’.: Gaithersburg. (Longman Scientific and Technical: Harlow. USA). and distil slowly (about 2 drops per s) until most of the water has come over. Calculation Moisture % w/w = Volume of water x 99. Maryland. Check your suggestions against the normal official methods used. Food Chemistry – A Practical Manual . Why keep samples for moisture determination in a sealed container before use? Include a diagram of the Dean-Stark apparatus used in this experiment.7 Mass of sample Note For the Whole Practical Exercise: The entire apparatus to be used should be cleaned with an acid cleaning solution to minimize the adherence of water droplets to the sides of the condenser and receiver. R. If drops of water adhere to the sides of the collecting tube. and Sawyer. UK) Nielsen. All this has been done prior to the practical class.) (1998) ‘Food Analysis’ 2nd Ed. In a fume cupboard. 3. force them down by means of a rubber tubing slipped over the end of a glass rod. (1995). Correlate your results for each food and suggest the most suitable method in each case.

Set the thermostat to 400 ºC and turn on for 45 min. which is the most universally used and probably the most accurate. After the stated time. fish and cheese about 25%. Digestion Perform duplicate analyses. or colorimetrically with ninhydrin reagent. Secondly. Leave the water pump and manifold connected. The Kjeldahl method is carried out in two distinct phases. 5 PROTEIN DETERMINATION INTRODUCTION Nitrogen is the most distinguishing chemical element present in proteins. METHOD A. Firstly. PART A SEMI-MICRO KJELDAHL DETERMINATION OF TOTAL ORGANIC NITROGEN The Kjeldahl (1883) and Dumas (1831) methods determine the total organic nitrogen. H2SO4 to produce (NH4)2SO4 with the aid of catalysts such as mercury or selenium and K2SO4 to raise the boiling point. There have been many modifications to the Kjeldahl method. Transfer to a numbered digestion tube and add approx. which is used to calculate the protein present through the use of previously determined factors. The protein content of foods varies widely: most fresh fruits and vegetables have less than 6%. ammonia is formed after neutralization of (NH4)2SO4 with NaOH. and most meats. with the assistance of the tutor and while wearing the safety shield provided.1 g) that contains approximately 10 mg of protein. Food Chemistry – A Practical Manual . which is then analysed by steam distilling it into a weak acid (boric acid) and titrating against standard HCl. Weigh out accurately (to 2 decimal places) on a low ash paper (cigarette paper). The determination of nitrogen is thus often used to give protein level. white flour and fresh eggs about 12%.23 Food Chemistry PRACTICAL EXERCISE NO. and a blank (reagents only but no flour). Connect the exhaust manifold onto the tubes and turn on the water vacuum pump. as many other nonprotein nitrogenous substances are also present in foods. but this is only a crude estimate. the nitrogen compounds are digested with conc. Add 3 mL of concentrated H2SO4 and place the rack and tubes in the digestion apparatus. 2 g K2SO4 and a catalyst tablet. lift the rack out of the digestion block and place on the stand to cool. an amount of each flour sample (0. which are the most common organic nitrogen compounds among foodstuffs.

Place digestion tube in the distillation apparatus. Add boric acid (10 mL) plus indicator to a 250 mL conical flask and place it in the right hand side of the apparatus. OPERATING INSTRUCTIONS FOR GERHARDT DISTILLATION UNIT 1. Depress start button. Distillation There are many distillation assembles suitable for recovering ammonia from the digest.The University of Queensland 24 If the samples still contain black particles. remove the exhaust manifold. and finally place a mark on the digestion tube about 40 mm above surface of the liquid. In essence. 9. 2. 10. The apparatus cannot be used until the green start light is on. Close the safety screen. Depress the NaOH control in small amounts to add NaOH to the digest up to the required mark. 7. Turn on cooling water. Ensure that the appropriate tubes are in the distilled water and sodium hydroxide containers.0. most of these employ steam distillation. 11. depress the stop button. 3. Steam setting switch set to low. 8. ensuring the exit tubing is below the level of the liquid in this collecting flask. 5. Turn on white power switch. B. wash down the sides of the tubes with distilled water and digest for a further 15 min. When the tubes are cool. a buzzer indicates that distillation has been completed. add approximately 5 mL distilled water carefully to each tube. 12. Food Chemistry – A Practical Manual . Check there is a mark on the digestion tubes approximately 40 mm above the surface of the liquid. To cancel the buzzer. 6. Timer set to 2. 4. 13. After 3 min distillation. This automatically resets the timer.

Add about 10 mL of NaOH by pressing the NaOH control. Add in approximately an equal amount of 0.25 Food Chemistry 14. This is quantitative since the colour depends on the number of peptide bonds.38 PART B BIURET DETERMINATION OF SOLUBLE PROTEIN Compounds containing the peptide bond (-CO-NH-) give a violet colour with NaOH and dilute copper sulphate (Biuret reagent). Liquid egg white contains 10-11 % protein. Press the start button to clean out the system. PREPARATION OF BULK EGG WHITE PROTEIN SOLUTION Separate the liquid white from the yolk of an egg into a pre-weighed beaker.1 M NaOH and dissolve the egg white using a magnetic stirrer.02 M HCl from a semi-micro burette (10 mL). Calculation % (w/w) nitrogen = (mL std acid for sample . In this experiment. ie. add distilled water (20 mL) to a clean digestion tube and place in the apparatus. Shake the solution to mix. Using distilled water. corn Wheat flour Milk and products 6. rinse the beaker several times. Use an analytical balance.70 6. and dilute to the volume with distilled water. The green-yellow distillate is then titrated to a pale pink/red colour with 0. beans. Calculate the approximate concentration of this protein solution in mg protein / mL for use below. 100-110 mg protein / g egg white. meat. peas. and determine the mass of liquid egg white separated. PREPARATION OF PROTEIN STANDARDS Standard Gelatin Protein Solution: 10 mg protein/mL distilled water. Note. gelatin is 88% pure so this has to taken into account when calculating the amount to prepare 100 mL of the protein solution.25 5. Food Chemistry – A Practical Manual . Liquid egg white contains 10-11 % protein of which albumins (ovalbumin and conalbumin) constitute 67%. 15. Weigh the beaker and egg white. When all distillations have been completed. wash the dissolved egg white into a 100 mL volumetric flask (filter if cloudy).mL of std acid for blank) x molarity x 14 x 100 mg of sample Multiply by the appropriate factor to obtain % protein Eggs. the amount of albumin in liquid egg white will be determined.

Gentle heat will be required to dissolve the gelatin. When dissolved.4 mL 1.8 0.8 mL 1. between the minimum and maximum gelatine protein concentrations). The biuret reagent MUST NOT be added until all of the solutions are in the twelve test tubes. Add this amount and the appropriate volume of distilled water (to bring to a total of 2 mL) for Solution 6 in the right hand column of the table above.2 mL 8 mL 1.8 mL 8 mL 0. Then. calculate the volume (mL) of this bulk egg white solution required to fit on the calibration curve (ie. make up to the 100 mL mark with distilled water.2 0. Food Chemistry – A Practical Manual . cool the solution. Please check this calculation with the tutor before proceeding.4 mL 8 mL 0. Set up twelve test tubes in six pairs.6 All the test tubes must have a total of 10 mL so the maximum amount of the bulk egg white solution is 2 mL. This can be achieved by rinsing the beaker several times with small amounts of distilled water (10 mL) and adding the washings to the volumetric flask. and transfer quantitatively to a 100 mL volumetric flask.The University of Queensland 26 Weigh the calculated amount into a beaker and dissolve in distilled water (60 mL). PREPARATION OF FINAL EGG WHITE PROTEIN SAMPLE (SOLUTION 6) Using the concentration of the bulk egg white protein solution calculated above.4 1.6 mL 0. Then add this amount to the final pair of test tubes (Solution 6).6 mL 8 mL 1.2 mL 0. 6) ? mL ? mL 8 mL ?? H2O Protein Solution (Gelatin) Biuret Reagent Conc. Diluted Gelatin Protein Solution (mg/mL) 2 mL 0 mL 8 mL 0 1. Do not over heat. BLANK (Std 1) Std 2 Std 3 Std 4 Std 5 Final Egg White Sample (Soln.

S.27 Food Chemistry PROTEIN DETERMINATION Now add biuret reagent (8 mL) to all of the tubes and mix each well on a vortex mixer. From this concentration. (Aspen Publishers.: Gaithersburg.) (1998) ‘Food Analysis’ 2nd Ed. USA). Food Chemistry – A Practical Manual . calculate the albumin concentration (% albumin) of the liquid egg white used in this practical exercise. and compare to that in the literature. R. taking account of the dilution factor for this experimental step.: Gaithersburg. Nielsen. determine the protein concentration (% protein) of the original liquid egg white used. M. Finally. Using this graph. 16th Ed. UK) 9th Edition. Inc. J. Kirk. and Sawyer. Plot a best fit straight line (going through zero-zero) of protein concentration (x axis) versus absorbance (y axis) at 550 nm for the gelatin protein standards. S. (ed. Leave the tubes to stand for 30 min and read absorbance on a spectrophotometer at 550 nm. Maryland. Inc. Essex. USA). determine the protein concentration (mg/mL) of the diluted egg white solution (Solution 6). (1999) ‘Principles of Food Chemistry’ 3rd Ed. ‘Official Methods of Analysis’. QUESTION Why is it necessary to cool the standard protein solution before making it up to volume in a volumetric flask? REFERENCES AOAC International. R. (1991) ‘Pearson's Chemical Analysis of Foods’ (Longman Scientific and Technical: Harlow. MD) de Mann. followed by the concentration of the bulk egg white solution (100 mL) taking account of the dilution factor. Maryland. (Aspen Publishers. (1995). (AOAC International: Gaithersburg.

The University of Queensland 28 Food Chemistry – A Practical Manual .

(use same size tubes). pectinases. Set up the following test tubes . Food Chemistry – A Practical Manual . If the calcium ions are removed from the milk. Tube A with 10 mL of milk + 1 mL of rennet solution. if allowed to act upon milk. A solution of rennet is available. Shake the tubes and place in a water bath at 37 ºC (place each set of tubes in the water bath at the same time). eg. and sugar splitting enzymes. Observe changes in both tubes with gentle shaking every min. 1. hydrolysis. Casein is a phosphoprotein which is soluble in the presence of calcium ions. Explain the results. These are by far the most important group of enzymes. but it now remains in solution. and include prototypic enzymes or proteases. the casein is still converted into para-casein. as far as foods and food processing are concerned. also present in milk. lipases. RENNIN The stomach of a calf and other young mammals contains a proteolytic enzyme (hydrolysed proteins) called rennin. will cause it to clot (in the presence of calcium ions). The chief proteins in milk are casein and lactalbumin. and the oxalated milk is acted on by rennin.29 Food Chemistry PRACTICAL EXERCISE NO 6 PROPERTIES OF ENZYMES Part A HYDROLYTIC ENZYMES INTRODUCTION Hydrolytic enzymes are those which break down large polymer molecules into smaller molecules by insertion of water. by precipitation with oxalic acid as calcium oxalate. The enzyme rennin is present in commercial rennet for making cheese. Calcium ions are required for clotting. carbohydrases. Tube B with 10 mL of milk + 1 mL of boiled rennet solution. in the absence of the calcium ions and no clotting occurs. which. 1. ie. METHOD Commercial rennet containing the enzyme rennin is required.

rennin on milk showing chemical structures where appropriate. Cool and add 15 drops of 1% aqueous phenol red indicator (changes colour in the presence of acid). in pancreatic juice and milk) to glycerol and fatty acids. Add just enough 2% Na2CO3 to give a definite red colour eg. boil solutions B and C then cool and add 1 mL of 1% CaC12 solution to both. mix on a vortex mixer. Lipase catalyses fat hydrolysis (lipolysis) in milk leading to the development of off odours. METHOD Take about 20 mL of milk and boil in order to destroy bacteria that might produce lactic acid from lactose present. LIPASE Neutral fats are esters of glycerol and are hydrolysed by lipases (eg. Next set up three tubes as below. and incubate at 37 ºC. Make up the following mixtures. Observe changes in each tube every min for 5 min. QUESTION Give a general overview of the mechanism of action of the enzyme. colour of strawberry milk. Oxalated Milk (mL) 2. Add 1% potassium oxalate (6 mL) to milk (20 mL) before preparing these three solutions.The University of Queensland 30 2. 2. This reaction is reversible and is helped by the presence of bile salts. (mL) 1 1 1 boiled 1% CaC12 (mL) 1 Nil Nil A B C 4 4 4 After 5 min. The lipase should be added last. Food Chemistry – A Practical Manual . this process is called hydrolytic rancidity. Observe and explain.5% Rennet Soln. and so be more accessible to the enzyme. which enable the fat to form smaller droplets.

Peroxidase is one of the most heat resistant plant tissue enzymes and its extent of thermal inactivation has been shown to parallel that of enzymes responsible for off-flavour formation and undesirable browning that occur in unblanched or under blanched.2% Lipase extract (mL) 2 2 boiled 2 Water (mL) 1 1 Nil 2% Bile Salts (mL) Nil Nil 1 A B C 5 5 5 Shake the tubes and place in a water bath at 37 ºC and examine the colour changes at 30 sec intervals. Outline the mechanism of lipase activity using chemical structures where appropriate. The likelihood that enzymic browning will occur in these products can be determined using the peroxidase test. The method is based on the oxidation of guaiacol by peroxidase to give a reddish-brown pigment. The peroxidase test is used to determine the adequacy of blanching of frozen vegetables by determining the activity of the enzyme peroxidase. Food Chemistry – A Practical Manual .31 Food Chemistry Phenol red/milk mixture (mL) 0. 2. QUESTIONS 1. frozen or dehydrated vegetables and fruits. What would happen if too much 2% Na2CO3 is added during the lipase test? PART B PEROXIDASE ACTIVITY DETERMINATION INTRODUCTION Enzyme activity is widely used as a test of adequate heat treatment (blanching) during processing.

B. then blanching is adequate. Disregard colour developing after 3. J. D. (Marcel Dekker: N.5 min. To ensure adequate sampling. Blend with 150 mL distilled water for 1 min at moderate speed. Nielsen. : New York. S.: Gaithersburg. Mix well and watch for any colour difference from blank.05% guaiacol solution (in 50% alcohol) and 1 mL of 0. Maryland. QUESTIONS 1. REFERENCES D’Arcy. Maryland. USA).) (1998) ‘Food Analysis’ 2nd Ed. (1998) ‘Food Chemistry: A laboratory manual’.R. If no difference after 3. (ed. fresh) and blanched vegetables (ie frozen). D. (1996). (John Wiley & Sons. To the sample add 1 mL of 0. Sample: Add 2 mL of filtrate to 20 mL distilled water for sample. (Aspen Publishers. Inc. Fennema. Why are some enzymes more stable to thermal treatment than others? Name three enzymes that cause problems in frozen vegetables if they are not inactivated. Inc. ‘Web/CD Notes for Food Chemistry’. (1999) ‘Principles of Food Chemistry’ 3rd Ed. peroxidase with the substrate. Blank: Add 2 mL of filtrate to 22 mL distilled water for blank. M. Outline the mechanism of the reaction of the enzyme. Food Chemistry – A Practical Manual . de Mann. take 50 g of the largest pieces or parts of the vegetables since these are the most likely to be under blanched. O. 3.5 min. 2.08% hydrogen peroxide solution. ‘Food Chemistry’ 3rd Ed. guaiacol that leads to the formation of the red pigment. Describe the reactions involved.) Miller. (Aspen Publishers. and then filter through cotton wool.The University of Queensland 32 METHOD Perform test on unblanched (ie.: Gaithersburg. Inc. S. USA).Y. USA).

6% fat while some dairy products. together with fats and oils. taste and storage stability of the food. METHOD A.B. Food Chemistry – A Practical Manual .33 Food Chemistry PRACTICAL EXERCISE NO. spreads. Nuts are traditionally high in fat (35 . while steroids. grinding and thorough mixing gives a better extraction and a more representative sample. Fats. steroids. such as cheese and butter. The fat content of fresh food varies greatly. From a food science point of view. contain up to 82% fat. but a greasy diethyl ether extract residue and the absorption of the red dye Sudan III by an diethyl ether extract/water emulsion indicates their presence. They consist of fats. Most other compounds in foods are insoluble in these solvents and thus the term "crude fat" or "ether extract" is employed to cover all the compounds that are solvent extracted. resins. 7 PROPERTIES OF LIPIDS INTRODUCTION The lipids are a naturally occurring group of substances insoluble in water. etc. phospholipids and carotene have individual identification tests. but soluble in organic solvents. phospholipids. thus. These include small amounts of waxes. a hot plate is sufficient.70%). There is no general chemical test for the presence of lipids. Most fruits and vegetables. Take great care when using solvents especially diethyl ether and petroleum spirit as a flame is not necessary for their ignition. steroids etc. Milk contains about 3. oils. the most important group is that of the fats and oils (differing only in their melting point) which not only occur widely in food but are also extracted and incorporated in the production of other foods such as baked goods. SOLVENT EXTRACTION METHOD All methods use extraction with organic solvents. waxes. Fats and oils and higher lipids are characterized by their extreme insolubility in water and their solubility in organic solvents such as diethyl ether or petroleum spirit (alkanes). Lipids of all types are widely distributed both in plant and animal material. plant pigments. etc. except avocados (26%) and olives (17%). The physical and chemical properties of these lipids differ widely and are of great importance in food processing as they influence the texture.. fatty acids. fatty acids and lecithin give a translucent stain when smeared on paper (Grease Spot Test). have less than 1% crude fat while fish and meat vary from less than 1% in cod and haddock to over 20% in some fish and most meat cuts. N.

Extract the thimble in the soxhlet apparatus for 2 h with petroleum spirit (50º . and then add 10 mL of chloroform to dissolve the oil. IODINE VALUE (IV) The iodine value of an oil or fat is defined as the mass of iodine absorbed by 100 g of the sample. Animal fats (butter.110 80 .The University of Queensland 34 Soxhlet Extraction Weigh out approx. add 10 mL of Hanus iodine solution to both flasks.70º) contained in a round-bottom flask.2 g fat or 0. and thus the iodine value indicates the degree of unsaturation of the fatty acid residues of the glycerides.200 Iodine Value Iodine Value Iodine Value Iodine Value The iodine value is often most useful in identifying the source of an oil.1 M Na2S2O3 = 0. cool to room temperature. and weigh the flask and the extracted oil. Iodine values are normally determined using Wigs or Hanus methods. the higher iodine values indicate oils and the lower values fats. but depends on the method used.70 80 . almond) Semi-drying oils (cottonseed. distil off the solvent from the RB flask using the rotary evaporator. Close the thimble top with cotton wool. Then add 15 mL of 15% potassium iodide solution and 40 mL of distilled water. It is constant for a particular oil or fat. sunflower) 30 . and allow to stand for exactly 30 min with occasional shaking. 10 g of the biscuit sample accurately (to 2 decimal places) into a soxhlet thimble. Weigh again. adding it gradually with constant shaking until the yellow colour has nearly disappeared. add a few drops of starch indicator (Vitex) and continue titration until the blue colour has disappeared. then dry the oil in the flask in an oven at 70 C for a maximum of 10 min.1 g oil) into a 250 mL conical flask. In a second 250 mL conical flask prepare a blank of 10 mL of chloroform (no oil). Then. CHEMICAL QUALITY TESTS FOR FATS AND OILS 1. HANUS METHOD Weigh out accurately a suitable quantity of oil or fat (0. remove from the oven.1 M Na2S2O3.B. soya) Drying oils (linseed. dripping. The unsaturated fatty acid residues of the glycerides react with iodine.0127 g iodine) Food Chemistry – A Practical Manual . Generally. Calculate Hanus Iodine Value as g iodine taken up per 100 g oil. N. shaking well all the time to take up residual iodine.140 120 . The difference between the blank (largest titration volume) and the test sample gives the amount of iodine absorbed by the oil. sesame. Repeat with blank (no oil or fat present). Titrate the unreacted iodine with 0. Weigh the round-bottom (RB) flask containing a small amount of anti-bumping granules (2-3 boiling chips) before the extraction. lard) Non-drying oils (olive. (Titre: 1 mL 0. After extraction. Using a safety pipette. B.

using 0. Weigh out 6 g oil or 20 g of fat into a 250 mL conical flask. palm-kernel oil (SV 247) and butter fat (SV 225) which contain a high proportion of the lower molecular weight fatty acids may be detected.5 M alcoholic KOH with no fat or oil present). 3. Most oils have similar values (eg.1 M KOH (from a 10 mL burette) using vigorous stirring until a definite pink colour persists for 15 s.5 M HCl = 28. boil and titrate hot with 0.196) but the presence of coconut oil (SV 255). the FFA should be very low but oils having a FFA (as oleic acid) of about 0. olive oil series 188 . heat using a hot plate (refluxing as for the oil sample is not required).5 M HCl. Express result as: Acid Value: mg KOH required per g of oil. by first determining the difference between the blank (largest titration volume) and the test sample.5 M alcoholic potassium hydroxide solution and 2-3 boiling chips.5 mL of 1% phenolphthalein indicator.6 mg KOH) Food Chemistry – A Practical Manual . if not. With freshly refined oils and fats. Add 25 mL (using a volumetric pipette) of 0. while the iodine value/number gives an indication of the degree of unsaturation (the number of double bonds) of the fatty acid residues of the glycerides in the oil.05 mg KOH The saponification value/number gives an indication of the average molecular mass of the glycerides.3 . Add the hot neutralized alcohol (50 mL) to the fat.35 Food Chemistry 2. In a second 250 mL conical flask heat and neutralize 100 mL of 95% ethanol with 0. FREE FATTY ACIDS (FFA) OR ACID VALUE The acid value of an oil and fat is defined as the number of mg of KOH required to neutralize the free fatty acid in 1 g sample. Next. Add 1 mL of phenolphthalein (1%) solution and titrate while hot (in the round-bottom flask) the excess alkali with 0.1 M KOH = 5. continue refluxing. Weigh 2 g of oil into a 250 mL quick fit round-bottom flask.1 M KOH (1-2 drops of KOH should be enough). Calculate the saponification value expressed as mg KOH required per g of oil. Its value is inversely proportional to the mean molecular mass of the fatty acids present. Boil gently under reflux for 40 min. Titre: 1 mL of 0.1. or the % by mass (g/100 g) of free fatty acid present. Make sure there is one phase in the contents. SAPONIFICATION VALUE (SV) The saponification value of an oil or fat is defined as the number of mg of potassium hydroxide (KOH) required to neutralize the fatty acids resulting from the complete hydrolysis of 1 g of the sample.5% are noticeably rancid to the palate. carry out a blank determination (use only 25 mL 0. (Titre: 1 mL 0.

16th Ed. QUESTION Comment on the quality of the oil tested by referring to the literature for the values for foods studied. (Aspen Publishers. Maryland. % FFA = Acid Value / 2 (for oleic acid) Determine if the oil or fat is rancid enough to be discarded by using the level of % FFA (as oleic acid). (1995). MD) Fennema. ‘Official Methods of Analysis’.The University of Queensland 36 or as % Free Fatty Acids calculated as oleic acid (g oleic acid per 100 g of oil) (Titre: 1 mL 0.Y. Food Chemistry – A Practical Manual .) Nielsen.1 M KOH = 0. Inc. S. REFERENCES AOAC International. S. USA).028 g oleic acid) ie.: Gaithersburg. (Marcel Dekker: N. O. (ed.) (1998) ‘Food Analysis’ 2nd Ed. ‘Food Chemistry’ 3rd ed. (1996). (AOAC International: Gaithersburg.

amino acids. If this is necessary. a food). Mix well. METHOD 1. The colour produced may be read at 420 nm on the spectrophotometer. vegetables. complex "melanoidin" browning polymers are formed. about 10 mL distilled water is usually needed. flavour and nutritive value of the processed food product. concentration of reactants. Final Experiments Using the Model System Using the above model system. In both cases. pH. glucose. crust on baked goods. The tubes may have to be diluted so that a reading can be obtained on the spectrophotometer. Food Chemistry – A Practical Manual . colour of maple syrup) or detrimental (darkening of dehydrated fruits. These may have a significant effect upon the colour. flavour. lactose. add 4 mL 50 °Brix invert sugar + 1 mL 10% glycine + 1 mL pH 8 buffer. etc. cover the tops of the tubes with foil and incubate in a water bath at 100 °C for 30 min. The extent of this browning will increase with concurrent increase in process pH and/or temperature. Trial of the Model System A model browning system is as follows: To a test tube. canned or dried milk) to the food. Browning reactions may be desirable (eg. set up the following 10 tubes (all of the same size) and record your results in a table. colour of caramel. The following experiments attempt to gauge the effect of several variables and the rate of browning in a model sugar-amine browning system. lysine). maltose) and amino compounds (eg. amines. add the same amount of dilutent to all tubes. browning reactions may occur. Nonenzymic browning is found in food processing operations involving dehydration/concentration (powdered / condensed milk). inhibitors or preservatives. fructose. When reducing sugars (eg. proteins) are present together in a system (eg.37 Food Chemistry PRACTICAL EXERCISE NO 8 NONENZYMIC BROWNING – MAILLARD REACTION INTRODUCTION Nonenzymic browning is purely chemical and occurs by several mechanisms involving reactive sugars and amino compounds (eg. 2. eggs. These reactions are not enzyme-catalysed but are affected by temperature. The above is only a model for the 10 mini experiments to be performed below.

The blank is used to calibrate the absorbance to zero on the UV/Vis spectrophotometer before each measurement. All the buffers used were prepared using phosphate solutions. 3. at 100 °C for 30 min.1 g sodium metabisulphite. and explain the effect of pH. 4. Tubes 1-16. discuss the effect of the variables tested on nonenzymic browning. quickly place in a beaker of cold water for 5 mins than in an ice bath for 15 mins to bring the temperature to room temperature as quickly as possible. 18 and 19 should go in the one large 2 L beaker of boiling water on a hotplate.The University of Queensland 38 Tubes 1-16: Effect of pH: 3.5 at 100 °C for 30 min in duplicate. QUESTIONS 1. and the final absorption measured in the same randomized manner using the spectrophotometer at 420 nm. the tubes should be removed from the hot water. 2. After heating for 30 min. Tube 20 (blank): 4 mL 50 °Brix invert sugar + 1 mL of distilled water (instead of glycine) + 1 mL pH 8 buffer.5. Following the 30 min heating at 100 °C. 4. Determine at what pH the Maillard reaction rate reaches a constant level. 8.) Plot absorbance versus pH. The preparation of the reagents for Tubes 1-16 should be done in a randomized manner. 6. Tubes 17: Effect of temperature: 70°C °C at pH 8 for 30 min. placed in tap water in a large beaker for 5 mins and then in an ice bath for 15 min to cool quickly to room temperature. Use chemical structures to outline the major mechanisms of non-enzymic browning. 5. What are the possible nutritional implications of non-enzymic browning? (Use chemical structures. Tube 19: 4 mL 50 °Brix invert sugar + 1 mL of distilled water (instead of glycine) + 1 mL pH 8 buffer. 8. 7. at 25 °C for 30 min. at 100 °C for 30 min. Food Chemistry – A Practical Manual . Place tube 17 in a constant temperature water bath set at 70 °C. Tube 18: 4 mL 50 °Brix invert sugar + 1 mL of 10 % glycine + 1 mL pH 8 buffer + 0. Do not cool to below room temperature as this will lead to problems with the spectrophotometric measurements (fogging of the cuvette). RESULTS From your results and from the literature.

Inc. M. de Mann. ‘Food Chemistry’ 3rd ed. USA). Maryland. (1999) ‘Principles of Food Chemistry’ 3rd Ed. (Marcel Dekker: N. Inc. ‘Web/CD Notes for Food Chemistry’. P. USA). (Aspen Publishers. D. (1995). : New York. O. D. Fennema. (John Wiley & Sons. ‘Food: The Chemistry of its Components’. (1998) ‘Food Chemistry: A laboratory manual’. Give three examples each of desirable and undesirable nonenzymic browning reactions in food systems. D’Arcy. 3rd ed. (1996).39 Food Chemistry 4.) Miller. REFERENCES Coultate.Y. (Royal Society of Chemistry: London).: Gaithersburg.R. B. T. J. Food Chemistry – A Practical Manual .

The University of Queensland 40 Food Chemistry – A Practical Manual .

41 Food Chemistry PRACTICAL EXERCISE NO. 9 ASCORBIC ACID DETERMINATION DETERMINATION OF ASCORBIC ACID BY REDOX TITRATION Besides being an important vitamin.50 mg in 200 mL distilled water (this solution is prepared). subtract a blank titration (no food sample).1 mg / 1 mL freshly made up. 1.10 days. Titrate in triplicate 2 mL aliquots of the fruit juices (which should contain between 10 100 mg ascorbic acid / 100 mL) plus 5 mL extracting solution using the standardized indophenol solution. and calculate the ascorbic acid as mg / 100 mL by the following method: (mean sample titration volume – blank titration volume) x (standardized mg AA/mL dye) x 100 2 2. Take the average of the three titrations. b.10% acetic acid. Express standardized indophenol solution as mg ascorbic acid equivalent per 1 mL of dye. a. Each student prepares their own 100 mL of standard. ascorbic acid is a sugar acid and a powerful reducing agent. This will keep in the refrigerator for 7 .6-dichlorophenol (indophenol) which is blue in neutral solution and pink in acid. The most common assay of ascorbic acid in foods is based on its oxidation to dehydroascorbic acid by the redox dye 2. The titration is carried out in acid conditions and at the end point the dye appears rose pink. Ascorbic Acid Standard . Food Chemistry – A Practical Manual . Standardize in triplicate the indophenol solution by rapidly titrating against 2 mL aliquots of standard ascorbic acid solution plus 5 mL of extracting solution until a distinct rose-pink persists for at least 5 s. VITAMIN C DETERMINATION OF FRUIT JUICES Fruit juice samples (2 mL) are to be analysed. but a modified method will be used for determining the Vitamin C content in a potato. METHOD Extracting Solution . Indophenol Solution . VITAMIN C DETERMINATION OF POTATOES The above method for Vitamin C determination is very useful for fruit juices etc.

Weigh and cook one piece in distilled water until soft (15 . CALCULATION OF ASCORBIC ACID (AA) IN POTATOES Volume of dye = 5 mL (from titration) ∴ mass of ascorbic acid in final sample solution (20 mL) = 5 x 0. Note. Filter using a buchner funnel and cotton wool. * If the volume of indophenol used = 17 mL then 1 mL of indophenol reacts with 2 mg ascorbic acid 17 = 0. ∴ in 2 mL there are 2 mg of ascorbic acid (the exact amount will depend on how much you actually weighed out). if the potato weighed 32. It is important to know the exact amount of extracting solution used.The University of Queensland 42 METHOD Standardize the indophenol dye as per above method (no need to repeat if already done in (1) above at the same practical session).1176 = 0.65 g.1176 mg ascorbic acid ie. Continue as above for the raw potato. Blend the cooked potato in extracting solution equal to 4 times its mass. and titrate a 20 mL aliquot as above for the raw potato. CALCULATIONS FOR ASCORBIC ACID DETERMINATION 1. Titrate 20 mL aliquots of the extract using the standardized indophenol solution.1176 mg ascorbic acid 2. eg.588 mg If 20 mL sample taken from a solution of 30 g potatoes + 60 mL acetic acid extracting solution (total 90 mL): Food Chemistry – A Practical Manual . * STANDARDIZATION OF DYE (TITRE DETERMINATION) STD ascorbic acid solution is prepared = 1 mg / mL. Titre: 1 mL indophenol = 0.20 min). * 2 mL aliquot of STD ascorbic acid solution is titrated with indophenol. perform a blank. Peel a potato and cut into two pieces. Weigh the other piece and blend for 1 min with approximately twice its mass of extracting solution. blend with 65 mL or 70 mL of extracting solution.

(1999) ‘Principles of Food Chemistry’ 3rd Ed. USA). (1998) ‘Food Chemistry: A laboratory manual’. D. (1988). Welch. A. Buss. USA). % Vitamin C Loss = mg/100 g RAW . Maryland. Food Chemistry – A Practical Manual .R. J.588 x 90 mg 20 = 2. B. and Southgate. Ed. A. A.83 mg/100g Calculate the ascorbic acid content of raw and cooked potatoes in units of mg / 100 g 3. T. Holland. and Southgate.. D’Arcy.A. REFERENCES Australian Food Composition Tables. ‘The Composition of Foods’ 4th Rev. How does your values for ascorbic acid content of raw and cooked potatoes compare to food composition tables? Comment on the effect of boiling potatoes in water on the Vitamin C content.65 x 100 mg in 100 g potato 30 = 8. g COOKED x 100 mg/100 g RAW Calculate the % loss in vitamin C on cooking potatoes. (Aspen Publishers. (1991) ‘McCance and Widdowson’s The Composition of Foods’. I. B. D. A.65 mg But this 2. D. D. Miller. A. Inc. 2. QUESTIONS 1. : New York. A. A. M. and its possible nutritional importance..65 mg is actually in 30 g of potato: ∴ 2. de Mann.. D. H. Paul. D. T. (Royal Society of Chemistry: Cambridge).43 Food Chemistry Then mass of AA in original solution = 0. (Elsevier: Amsterdam). ‘Web/CD Notes for Food Chemistry’. Unwin.: Gaithersburg.. Inc. (John Wiley & Sons.

The University of Queensland 44 Food Chemistry – A Practical Manual .

600°C) is used to avoid loss of volatile chlorides. Use two flour samples in duplicate (four samples) 1. syrups. DETERMINATION OF TOTAL ASH CONTENT AOAC recommends using a quantity of material representing at least: 2 g of dry matter for fish products. Wet ashing is used when determining mineral constituents since less loss of volatiles occurs. to as high as 6. NOTE: Use the four decimal place analytical balances throughout this practical exercise. 3 – 5 g of cereal food. Ash content can be an index of the quality of a product such as in flour. sulphates. METHOD You are provided with samples of two different types of common flour. For these purposes. The average ash contents of fresh food ranges from 0. incineration at fairly low temperatures (550°C . Food Chemistry – A Practical Manual .0% for eggs. grain and stock feed. 10 ASH DETERMINATION INTRODUCTION Ash may be regarded as the inorganic residue remaining after oxidation of the organic matter in a sample. preserves. in a muffle furnace or by wet combustion with mixtures of sulphuric.45 Food Chemistry PRACTICAL EXERCISE NO. phosphates. Oxidation may be conducted by incineration over an open flame. meat or vegetable products. Use the methods below to find the adulterated samples and the type of adulteration. 10 g for jellies. milk or cheese. The composition of ash will vary according to the nature of the food material but usually consists of the more common metals ions in the form of their oxides. sugar products. 25 g of fruit juice. 5 – 10 g of sugar. fresh or canned fruit. or indicate adulteration of the sample. nitric and perchloric acids. The official method states that ashing is required on meat products before a salt analysis can be performed. silicates and chlorides.0% for green olives and bacon. The weight differences are too small and will not be measurable if a two decimal place balance is used. white sugar and oils. as in spent tea.

particularly in the analysis of fruit and sugar products. Place the dishes in the muffle furnace in a known order in case the numbers are erased. Once charred. Finally. 4. Food Chemistry – A Practical Manual . 3. Place the filter paper in the silica crucible and ignite in the fume cupboard. number four silica crucibles. Add 25 mL of 10% HCl (not 0. Weigh into the numbered crucibles the flour sample (3 g) in duplicate. WATER SOLUBLE ASH CONTENT This is of value in determining added mineral matter. DO NOT place the hot crucibles on the painted surface of the fume cupboard. From the mass. Determine the percentage of ash in the sample.1 M HCl) to the water insoluble ash. and wash with an equal volume of hot distilled water. Next determine the % water soluble ash in the flour sample(s) used. Once the papers have ashed. cool the dish and moisten the ash with distilled water prior to reheating. cover the crucible with a watch glass (or other suitable container). Filter through ashless filter paper and wash with hot distilled water. Filter through an ashless filter paper into a numbered conical flask. Cover with a watch glass and boil gently over low heat for 5 min. place the crucibles on a heat resistant mat and transfer to a muffle furnace heated to 550 °C. Char the sample (until no visible smoke is present) on a hot plate in the fume cupboard. Remove the watch glasses before placing the crucibles in the furnace. Note. After incineration.The University of Queensland 46 Using a marker pen. Cool and reweigh. calculate the % acid insoluble ash in the flour(s). 2.1 M HCl. ALKALINITY OF ASH This is of value in detecting adulteration of foods with minerals and in determining the acidbase balance of the food. Follow the procedure for water soluble ash for igniting the paper and determine the amount of acid insoluble ash. Keep the filtrate for determining alkalinity of ash. Care must be taken to ensure that the ash does not float out of the crucible. calculate the amount of water insoluble ash. Leave in the muffle furnace for 4 h until the ash is white or greyish-white. 10% HCl is not the same as 0. and by difference. If the paper catches fire. Add 20 mL of distilled water to the flour ash and heat until nearly boiling. cool in a desiccator and reweigh (use tongs). ACID INSOLUBLE ASH CONTENT This is of value in detecting adulteration of spices and confectionery with dirt and talc etc. If the material fails to ash properly. the amount of water soluble ash. cover the crucible with a watch glass and transfer them to the muffle furnace for 10 min heating at 550 °C.

(AOAC International: Gaithersburg. 2nd Ed. (1970). QUESTIONS 1. USA). Add a few drops of methyl orange and back titrate the excess acid with standard 0. Why use standard solutions and volumetric glassware for alkalinity of ash? Determine the AOAC Method Number for the ash determination of flour using the most up-to-date AOAC reference in the UQ libraries. 2.1 M HCl (usually 25 mL and use a volumetric pipette) to the filtrate from the water soluble ash.47 Food Chemistry Add a measured excess of standard 0. (Aspen Publishers. By difference.1 M acid required to neutralize the ash from 100 g of sample. (1995).1 M HCl). Inc.1 M HCl required to neutralize the alkali in the flour ash (1 mL of NaOH neutralizes 1 mL of 0. determine the volume (ml) of 0. MD) Maynard.: Gaithersburg. Express the alkalinity as the number of mL of 0. (Academic Press.1 M NaOH. Food Chemistry – A Practical Manual . ‘Methods in Food Analysis’. London). ‘Official Methods of Analysis’.) (1998) ‘Food Analysis’ 2nd Ed. REFERENCES AOAC International. Nielsen. S. (ed. Maryland. S. 16th Ed.

The University of Queensland 48 Food Chemistry – A Practical Manual .

and the use of chemical inhibitors. pear. substrate and oxygen.. by blending 10 g of fruit with 100 mL of distilled water. 2. control of pH. namely: enzyme.01 M citrate buffer pH 6. For oxidative browning to occur. These include destruction of the enzyme by heat. Extracts should be clear and MUST be stored in an ice bath for a short time (same practical session only). Switch on a spectrophotometer and allow to warm up for 10 minutes. prepare the sample test tubes near the spectrophotometer so that there is no delay in placing the sample in the instrument. Tissue Extract Preparation Prepare tissue extracts of fresh fruits.5 M catechol. It is not necessary to filter the whole extract since only a few mL are required. It is important to add the fruit extract last. Place in a clean test tube. then filtering using a buchner funnel. three components must be present. Once the instrument is zeroed. Measurement Oxidation Rate The method is based on the oxidation of catechol at pH 6 and 25°C. and zero the instrument using a blank consisting of 10 mL 0. catechol as the substrate. Set the wavelength to 420 nm. These enzymes catalyse the oxidation of phenolic substrates to quinones which subsequently polymerize to give brown-black pigments. the reaction occurs at room temperature.01 M citrate buffer.5 mL of the enzyme solution of clear tissue extract.49 Food Chemistry PRACTICAL EXERCISE NO 11 ENZYMIC BROWNING: ACTIVITY OF FRUIT HOMOGENATES INTRODUCTION Enzymatic browning of cut or damaged fruits and vegetables is due to the action of polyphenol oxidases (collectively called phenolase) on phenolic substrates. elimination of oxygen.g.5 mL distilled water. eg. 1 mL 0. METHOD Each student is to do tests on one fruit extract at three different pH’s. with the colour development being assessed by absorption at 420 nm in a spectrophotometer. and 0. 1 mL catechol and 0. A number of methods are used both commercially and domestically to inhibit this reaction. 1. Food Chemistry – A Practical Manual . or banana etc. All buffer solutions need to be removed from the refrigerator at least 3 h prior to use. The standard method for examining enzymic browning in-vitro is to use a model system consisting of fruit homogenate as the enzyme preparation and the phenolic compound. a ‘pink lady’). Apple (e. 10 mL 0. and then allowed to warm to room temperature.

New separate blanks (with each pH) are required. with one graph (ie. once with added sodium bisulphite (50 ppm) and once with sodium chloride (1000 ppm). All solutions must be freshly prepared.The University of Queensland 50 Mix well.5 min initial readings.2 mL of the concentrated sodium bisulphite solution (5750 ppm) as part of the preparation of the pH 6 blank detailed above. Perform this exercise for one fruit.000 ppm) to the model system sample to achieve 2000 ppm Cl ions.5 min for the first 3 min. add 0. add 0.000 ppm) as part of the preparation of the pH 6 blank detailed earlier. 3. apple. 5 and 7. pH’s 3-7) for each fruit extract.2 mL of the concentrated NaCl solution (115. Effect of Added Sodium Bisulphite and Sodium Chloride on Enzyme Activity Next. A2 min – A1 min) from the graphs (results for each fruit homogenate are to be shared between group members). 4. i. Start timing from when the enzyme solution is first added to the test tube (time zero). 4. place the cuvette in the spectrophotometer. Then record every minute until 10 min when activity should have ceased.e. RESULTS 1. Perform these parts of the practical exercise for one fruits only.e. determine and tabulate the polyphenol oxidase activity (where the slope is greatest) of the three fruit extracts at pH’s 3-7 (express as increase in absorption / minute for the first 2 minute period. Plot absorbance (y axis) versus time (x axis) for each pH and each fruit homogenate (each fruit on a different graph). including at 0 min. You must be quick with the transfer to the cuvette to record the 0 min and 0. pear and banana). Effect of pH Change on Enzyme Activity Repeat the exercise using pH’s 3. Perform this exercise for one fruit. Also include the plots for the NaCl and bisulphite trials with the plots of the 5 different pH’s.2 mL of the concentrated sodium bisulphite solution (5750 ppm) to the model system sample to achieve 100 ppm sulphite. repeat the exercise twice at pH 6. 8 buffer solutions instead of the pH 6 one. a new blank (100 ppm sulphite) needs to be prepared by adding 0. (b) Effect of added sodium bisulphite on enzyme activity: In another tube. a new blank (1000 ppm chloride) needs to be prepared by adding 0. Additionally. Food Chemistry – A Practical Manual . Next. (a) Effect of added sodium chloride on enzyme activity: In one tube. and read the absorption every 0.2 mL of the concentrated NaCl solution (115. quickly transfer the solution to a cuvette. Next share the results with other members of your practical group so you have all the data for the three different fruit homogenates (i. Plot these results on the same graphs as those for the pH 6 part. Additionally.

‘Food Chemistry’ 3rd ed. 'Principles of Food Chemistry'. USA). Is bisulphite an effective inhibitor of enzymic browning. ‘Web/CD Notes for Food Chemistry’. (Marcel Dekker: N. (1996). D’Arcy. 3. eg.51 Food Chemistry 2. D. 'Handbook of Food Additives'. Show the mechanism of action of polyphenol oxidase in the formation of enzymic browning pigments. Quote the Australian and New Zealand Food Standards Code for the permitted levels of sodium bisulphite in various food stuffs. : New York. ‘Food: The Chemistry of its Components’. effect of pH by plotting activity (A2 min – A1 min) versus pH (x axis) on another graph for each fruit. B. D. de Man. (1995). 3rd ed.R. Show the chemical structure of sodium bisulphite and the mechanism of action as an anti-browning agent. Determine the effect of increased acidity on polyphenol oxidase activity. Miller. Fennema. (1972).Y.) Furia. 4. REFERENCES Coultate. QUESTIONS 1. P. Comment on your results.(Royal Society of Chemistry: London). (1990). E. 2nd ed.C. and why? 2. Why is there a difference in the inhibition of sulphite ions relative to chloride ions? Discuss why the graph obtained in (1) above flattens off with time. 5. T. J. T. Inc. What is the optimum pH for each fruit homogenate? Determine the effect of added sulphite and chloride ions on polyphenol oxidase activity. (C. (Van Nostrand Reinhold: New York). (John Wiley & Sons. Press: Ohio). O. Food Chemistry – A Practical Manual .R. 3. 2nd ed. (1998) ‘Food Chemistry: A laboratory manual’.

The University of Queensland 52 Food Chemistry – A Practical Manual .


The University of Queensland 54 Food Chemistry – A Practical Manual .


The University of Queensland 56 Food Chemistry – A Practical Manual .

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