1.6 Determination of soluble solids 1.6.1 Principle: Measurement of the refractive index of the test solution at 20 °C, using a refractometer, and use of tables correlating refractive index with soluble solids content (expressed as Sucrose), or direct reading of the soluble solids content on the refractometer. 1.6.2 Apparatus a) Refractometer - indicating the refractive index by means of a scale graduated in 0.001, in order to allow readings to be estimated to 0.0002. Refractometer - indicates the percentage by mass of Sucrose by means of a scale graduated in 0.5 %, in order to allow readings to be estimated to 0.25 %, This refractometer shall be adjusted so that at 20 °C it registers for distilled water a soluble solid (Sucrose) content of zero. {+} FRUIT AND VEGETABLE PRODUCTS | 2012 b) Water circulating apparatus - to maintain the temperature of the prisms of the refractometer constant to within 05 |C in the neighbourhood of 20 % which is the reference temperature. If the temper ature of circulating water is different from 20 °C use temperature correction as per table on page no. ©) Beaker- capacity 250 ml 1.6.3 Procedure: 1.6.3.1 Preparation of test solution (a) Clear liquid products Thoroughly mix the sample and use it directly for determination. (b) Semi thick products ( purees ete) Thoroughly mix the sample. Press a part of the sample through a gauge /muslin cloth folded in four, rejecting the first drops of the liquid and reserving the remainder of the liquid for the determination (©) Thick products ( jams, Jellies ete) Weigh into the tared beaker to the nearest 0.01 gm, a suitable quantity (upto 40 gm) of the sample and add 100 - 150 ml of distilled water. Heat the contents of the beaker to boiling and allow to boil gently for 2- 3 minutes, stirring with a glass rod. Cool the contents and mix thoroughly. After 20 minutes weigh to the nearest 0.01gm, then filter through a fluted filter paper or a Buchner funnel into a dry vessel. Reserve the filterate for determination 1.6.3.2 Determination Adjust the water circulation in order to operate at the required temperature and allow it to flow to bring the prisms of the refractometer to the same temperature which shall remain constant to within 05°C during the determination.1.6.3.2 Determination Adjust the water circulation in order to operate at the required temperature and allow it to flow to bring the prisms of the refractometer to the same temperature which shall remain constant to within 0.5°C during the determination. Put a small quantity of the test solution (2- 3 drops are sufficient ) on the fixed prism of the refractometer and immediately adjust the movable prism. Suitably illuminate the field of view. Bring the line divi ding the light and dark parts of the surface in the field of view to the crossing of the threads and read the value of refractive index. SF S FRUIT AND VEGETABLE PRODUCTS | 2012 Determine percent sugar from the table If the determination has been carried out at a temperature other than 20°C + 0.59C the following corrections are required (a) For the scale indicating refractive index apply the formula ne = nf + 0,0013(r - 20) where n3® is the refractive index at 20 nb _ is the refractive index at the temperature of measurement: t is the temperature of measurement, in degrees Celsius. (b) For the scale indicating percentage by mass or Sucrose correct the result according tothetable1 1.6.3.3 Calculation (a) Refractometer with refractive index scale Read from table 2 the percentage mass of sucrose corresponding to the value of refractive index corrected for temperature if necessary. In the case of liquid or semi thick products the soluble solid content is equal to the number found. If the determination has been carried out on a diluted sample the soluble solid content is equal to Pxml mo Where, Pis the percentage by mass of soluble solids in the diluted solution m0 is the mass, in gm ofthe sample before dilution m 1 is the mass in gm of the sample after dilution ‘Take the result as the arithmetic mean of two determinations. Express the result to one decimal place1.7 Determination of Sodium Chloride (salt content) in brine: 1.7.1Principle: {+} FRUIT AND VEGETABLE PRODUCTS | 2012 Direct titration of NaCl in brine with standard silver nitrate solution based on the Mohr method is adequate for routine analysis. 2Reagents: i) 5% (w/w) aqueous potassium chromate solution. fi) 0.1N aqueous silver nitrate solution. ili) N Sodium Hydroxide 1.7.3 Procedure: ‘Take 5 to 10 gm of liquid portion from the drained weight determination. If itis acidic, neutralize it with standard Sodium Hydroxide using phenopthlein as indicator. Add 1 ml of 5% aqueous potassium chromate solution and titrate with 0.1N AgNOs solution to produce red-brown end point. Titre value x Normality of AgNO; x 58.4 x 100 NaCl % Weight of the sample x 1000 (Ref: - $1 Handbook of Food Analysis (Part VIII) - 1984 page 5) 1.8Metallic contaminants: Follow procedures given in the Manual of Methods of Analysis - Metals 1.9 Pesticide rresiduc Follow procedures given in the Manual on Pesticide residues. 1.10 Microbiological examination: Follow procedures given in the relevant Manual. {>} FRUIT AND VEGETABLE PRODUCTS | 2012FRUIT AND VEGETABLE PRODUCTS | 2012 2.0 Thermally processed fruit and vegetable juices fruit beverages / fruit drinks and fruit nectars (canned / bottled) / flexibly/ aseptically packaged and non-thermally processed fruit beverages / drinks. 2.1Total Solids (a) Insoluble matter absent (applicable to jellies and syrups also) Digest pure quartz sand that passes No 40 but not No 60 sieve with HCI, wash acid free, dry and ignite. Preserve in stoppered bottle. Place 26- 30 gm sand and a short stirring rod in the dish about 55 mm in diameter and 40 mm deep, fitted with cover. Dry the dish thoroughly cool in a dessicator and weigh immediately. Add enough sample to yield about 1 gm dry matter and mix thoroughly with sand. Heat on steam bath 15 - 20 minutes, stirring at 2 - 3 minutes interval or until mass becomes too stiff to manipulate readily. Dry at less than 70 ° C under pressure of about 50 mm Hg in vaccum oven passing a current of dry air (dried over CaSO, or P20;). Make trial weighings at 2 hours interval after 18 hrs until change in weight is equal to 2 mg. Calculation: % Total Solids = (W3-W1) X100 (w2-w1) (b) Insoluble matter present (applicable to jams, marmalades and preserves also) Accurately weigh into a large flat bottomed dish sufficient sample that will give 2. 3 gm dry matter. If necessary to secure thin layer of material, add few ml water and mix thoroughly. Dry at 70 ° C under pressure less than 100 mm Hg until consecutive weighing’s made at 2 hr intervals vary less than 3mg. (Ref: - A.O.AC 17% edition, 2000, Official method 920.151 Solids (Total) in Fruits and Fruit Products). {»} FRUIT AND VEGETABLE PRODUCTS | 2012 2.2 Total soluble solids: Follow method given in Clause1.6 above 2.3 Determination of pH Value: - PH is the measurement of H+ ion activity; It measures active acidity. pH may be determined by measuring the electrode potential between glass and reference2.3 Determination of pH Value: - pH is the measurement of H+ ion activity; It measures active acidity. pH may be determined by measuring the electrode potential between glass and reference electrodes; pH meter is standardised using standard pH buffers. Use homogenized sample for the determination of pH. Preparation of sample for pH measurement: Liquids Immerse the standardized electrode tip into the solution and stir the sample gently by means of a rod or “flea” to give a constant pli value. Non-homogeneous products [fit is useful to know the pH of different components of the sample or differences between the pH at several points of the test portion, separate these as best as possible, homogenize and read them separately. For a bulk pH measurement, homogenize a representative aliquot to give a moist homogeneous mixture. Treat bulk as for moist homogeneous products (7.1.4). Dry products Have a standard practice for the dilution of dried materials - particularly when comparing the pH of sub-samples of the same product, as pH may change with the extent of dilution. For example homogenize with an equal volume of distilled or deionized water. Immerse the electrode in the sample and mix gently until a constant pH reading is obtained. »} FRUIT AND VEGETABLE PRODUCTS | 2012 Moist homogeneous products Homogenise the sample, immerse or embed the electrode and ensure that there is adequate contact between probe and sample. Read when the meter reading is stable. Do three separate measurements on the test sample - the extreme readings should not differ by more than 0.15 pH units. Take as the result the arithmetic mean of the three readings. 2.4Determination of acidity (Applicable to jams , jellies also) Titrable acidity can be expressed conveniently in gms acid per 100 gm or per 100 ml as appropriate, by using the factor appropriate to the acid as follows: 1 ml of 0.1. N NaOH equals Malic acid - 0.0067 gms Oxalic acid - 0.0045 ome2.4 Determination of acidity (Applicable to jams , jellies also) Titrable acidity can be expressed conveniently in gms acid per 100 gm or per 100 ml as appropriate, by using the factor appropriate to the acid as follows: 1 ml of 0.1 N NaOH equals Malic acid - 0.0067 gms Oxalicacid - 0.0045 gms Citric acid monohydrate - 0.0070 gms Citric acid anhydrous - 0.0064 gms Tartaric acid - 0.0075 gms Lacticacid - 0.0090 gms Aceticacid - 0.0060 gms Oleic acid - 0.00282 gms (@) Colourless or slightly coloured solutions Take 10 gm well mixed juice, dilute to 250 ml with neutralised or recently boiled water. Titrate with 0.1 N NaOH using 0.3 ml phenolpthlein for each 100 ml of the solution to pink end point persisting for 30 seconds. Report acidity as ml 0.1 N NaOH per 100 gm or 100 ml as required. (b) Highly coloured solutions ‘Sample preparation: 1) Dilute known weight of sample with neutralized water and titrate to just before end point with 0.1 N alkali using 0.3 ml phenolphthalein for each { 2 } FRUIT AND VEGETABLE PRODUCTS | 2012 100 mi solution being titrated. Transfer measured volume (2-3 ml) of solution into about 20 ml of neutral water in small beaker. (in this extra dilution fruit juice becomes so pale that phenolphthalein colour is easily seen). If test shows that end point is not reached, pour extra diluted solution back into original solution, add more alkali and continue titration to end point. By comparing dilutions in small beakers differences produced by a few drops of 0.1 N alkali can be easily observed. 2) In case of jams and jellies mix sample thoroughly. Weigh 300 gm mixed sample into a 2 litre flask and dissolve in water heating on steam bath if necessary. Apply as little heat as possible to minimize inversion of sucrose. Cool dilute to volume, mix thoroughly by shaking and use aliquots for various determinations. If insoluble material is present mix thoroughly and filter first. Electromeric method can be used to raise the pH slowly to 8.15 to remove colour interference. (Ref: - A.0.A.C 17% edn, 2000, Official method 942.15 Acidity (Titrable) of fruit produets read with AOAC official method 920. 149 Preparation of test sample).Electromeric method can be used to raise the pH slowly to 8.15 to remove colour interference. (Ref: - A.O.A.C 17% edn, 2000, Official method 942.15 Acidity (Titrable) of fruit products read with A.O.AC official method 920. 149 Preparation of test sample). Potentiometric method can also be used for the same. Slowly add 0.1N alkali till its pH raises to 8.1.Calculate acidity as predominant acid present in the sample by the formula: Acidity= Eq.wt of acid x N NaOH x 100. ‘Sample weight/vol. of alcohol x 1000 2.5 Determination of Volatile acids 2.5.1 Apparatus (2) Steam Distillation Apparatus as shown below { » } FRUIT AND VEGETABLE PRODUCTS | 2012 2.8.2Procedure ‘Add about 600 ml boiled water to outer chamber of still. Dissolve 10 gm test sample in water, dilute to 25 ml, Pour this into inner chamber and stopper. Boil water 3 minutes with side arm opening. Close and distill about 300 ml into Erlenmeyer flask. Add 0.5 ml phenolphthalein to distillate and titrate rapidly with 0.1 N NaOH until pink colour persists 15 seconds Express results as gm acetic acid per 100 ml or gm 1 ml.1 NNaQH = 0.0060 gm acetic acid x 10 (10 gm test sample taken)2.6 Determination of total Sugars: The presence of added sucrose can be detected by determining sugars before and after inversion by copper- reduction methods. {Standardization of Febling’s solution: Prepare standard dextrose solution into a 50ml. burette. Find the titre (volume of dextrose solution required to reduce all the copper in 10 ml. of Fehling solution) corresponding to the standard dextrose solution (Refer table below),Pipette 10 ml of Fehling’s solution into a 300 ml of conical flask and run in from the burette almost the whole of the standard dextrose solution required to effect reduction of all the copper, so that more than one millilitre will be required later to complete the titration. Heat the flask containing mixture over wire gauze. Gently boil the contents of the flask for 2 minutes. At the end of two minutes of boiling add without interrupting boiling, one mL of methylene blue indicator solution. While the contents of the flask begins to boil, begin to add standard dextrose solution (one or two drops at a time) from the burette till blue color of indicator disappears [The titration should be completed within one minute so that the contents of the flask boil together for 3 minutes without interpretation. Note the titre (that is total volume in mi. of std. dextrose solution used for the reduction of all the copper in 10 mil. of Febling’s solution) Multiply the titre (obtd. by direct titration) by the number of milligrams of anhydrous dextrose in one millilitre of standard dextrose solution to obtain the dextrose factor. Compare this factor with the dextrose factor and determine correction. Dextrose factors for 10 ml. of Fehling’s Solution Titre (ml) | Dextrose factor | Dextrose content per 100 ml of solution {mg) 15 49.1 327 16 49.2 307 17 49.3 289 18 49.3 274 19 49.4 260 20 49.5 247.4 {=} FRUIT AND VEGETABLE PRODUCTS | 2012 21 235.8 22. 2255, 23. 216.1 24 207.4 25. 199.3 26 1918 27, 184.9 28 1 29 172.5 30 167.0 31 161.8 32 156.9 33, 152.4 34 148.0 35, 1489 36 140.0 37. 136.4 38, 132.9 39, 129.6 40 126548 51.0 106.2 49, 51.0 104.1 50 SL1 102.2 Miligrams of anhydrous dextrose corresponding to 10 mi of Fehlings solution Reference: Table 2: IS 6287:1985, Methods for sampling and analysis for sugar confectionery, Pg.11 ‘Transfer test sample representing about 2- 2.5 gm sugar to 200 mi volumetric flask, dilute to about 100 ml and add excess of saturated neutral Lead acetate solution {about 2 ml is usually enough). Mix, dilute to volume and filter, discarding the first few ml filterate. Add dry Pot. or Sod. Oxalate to precipitate excess lead used in clarification, mix and filter, discarding the first few ml filterate. {«} FRUIT AND VEGETABLE PRODUCTS | 2012 Note: Use of Potassium Ferrocyanide and Zinc acetate is preferable instead of Lead acetate and Sodium oxalate, due to safety issues. Take 25 mi filterate or aliquot containing (if possible) 50 - 200 mg reducing sugars and titrate with mixed Fehling A and B solution using Lane and Eynon Volumetric method. (1) Fehling A: Dissolve 69.28-g copper sulphate (CuSO,.5H,0) in distilled water. Dilute to 1000 mi. Filter and store in amber coloured bottle. (2) Fehling B: Dissolve 346 g Rochelle salt (potassium sodium tartrate) (K Na C)H1s06 4120) and 100 g NaOH in distilled water. Dilute to 1000 ml. Filter and store inamber coloured bottle. For inversion at room temperature, transfer 50 ml aliquot clarified and deleaded solution to a 100 ml volumetric flask, add 10 ml HCI (1+ 1) and let stand at room temperature for 24 hours. (For inversion, the sample with HCI can be heated at 70° C for 1 br. This saves time and makes the whole process shorter). Neutralise exactly with conc. NaOH solution using phenolphthalein and dilute to 100 mi. Titrate against mixed Fehling Aand B solution (25 ml of Fehlings Solution can be considered for the purpose) and determine total sugar as invert sugar (Calculate added sugar by deducting reducing sugars from total sugars). Reducing and total reducing sugar can be calculated as, Reducing sugar (%) = mg, of invert sugar x vol. made up x 100 TRx Wt of sample x 1000 ‘Total reducing sugar (%) = mg, of invert sugar x final vol. made up x original volume x. 100 ‘TR x We of sample x aliquot taken for inversion x1000 Total sugar (as sucrose) (%) = (Total reducing sugar - Reducing sugar) x 0.95 +2.8 Determination of Vitamin C (Ascorbic Acid): ‘The ascorbic acid content in fruits and vegetables can be estimated by macerating the sample with stabilising agents such as 20 % metaphosphoric acid. 2.8.1 Principle: 2, 6 -dichlorophenol indophenol is reduced to a colourless form by ascorbic acid. ‘The reaction is specific for ascorbic acid at pH 1 to 3.5. The dye is blue in alkaline solution and pink in acid. 2.8.2 Reagents: (1) Standard Indophenol Solution - Dissolve 0.05 gm 2, 6 dichlorophenol indophenol in 50 mi, water, to which 42 mg, sodium carbonate is added, and make upto 200 ml with water and filter. Sodium carbonate is added for stability purpose. The dye solution keeps for a few weeks if stored in refrigerator. Prepare fresh if possible and standardize before use. Blank correction: Dissolve 50 mg 2,6-dichloroindophenol Na salt that has been stored in desiccator over soda lime, in 50 mL H20 to which has been added 42 mg NaHCO3; shake vigorously, and when dye dis solves, dilute to 200 mL with H20. Filter through fluted Paper into amber glass-stoppered bottle. Keep stoppered, out of direct sunlight, and {#} FRUIT AND VEGET: PRODUCTS | 2012 store in refrigerator. (Decomposition prod ucts that make end point in distinct occur in some batches of dry indophenol and also develop with time in stock solution. Add 5.0 mL extracting solution con taining excess ascorbicacid to 15 mL dye re agent. Ifreduced solution is not practically colorless, discard, and prepare new stock solution. If dry dye isat fault, obtain new supply.) ‘Transfer three 2.0 mL. aliquots ascorbic acid standard solution to each of three 50 mL Exfenmeyers containing 5.0 mL HPO3-CH3COOH solution, B(a)(Z). _Titrate rapidly with indophenol solution from 50 mL. burette until light but distinct rose pink persists °5 s. (Each titration should require ca 15 ml. indophenol solution, and titrations should check within 0.1 mL).Similarly titrate 3 blanks com posed of 7.0 mL HPO3- CH3COOH solution, Bfa)(1), plus volume H20 ca equal to volume indophenol solution used in direct titrations. After substracting average blanks (usually ca 0,1 mL) from standardization titrations, calculate and express concentration of indophenol solution as mg ascorbic acid equivalent to 1.0 mL. re agent. Standardize indophenol solution daily with freshly pre pared ascorbic acid standard solution. (2) Standard Ascorbic acid solution - Dissolve 0.05 gm pure ascorbic acid in 60 ml of 20 % metaphosphoric acid (HPO3) and dilute with water to exactly 250 ml in a volumetric flask.(2) Standard Ascorbic acid solution - Dissolve 0.05 gm pure ascorbic acid in 60 ml of 20 % metaphosphoric acid (HPO3) and dilute with water to exactly 250 ml in a volumetric flask. (3) Metaphosphoric acid - 20% (4) Acetone 2.8.3 Standardisation of Dye: Pipette 10 ml of standard Ascorbic acid solution in a small flask and titrate with indophenol solution until a faint pink colour persists for 15 seconds. Express the concentration as mg Ascorbic acid equivalent to 1 ml of dye solution ie 10 ml of (»} FRUIT AND VEGETABLE PRODUCTS | 2012 Ascorbic acid solution = 0.002 gm ascorbic acid If 0,002 gm ascorbic acid requires V ml dye solution to neutralize it then 1 ml dye solution = 0.002 /V gmascorbic acid. 2.8.4Procedure Pipette 50 mi of unconcentrated juice (or the equivalent of concentrated juice) into a 100 ml volumetric flask, add 25 ml of 20 % metaphosphoric acid as stabilizing agent and dilute to volume. Pipette 10 ml in a small flaskand add 2.5 ml acetone. Titrate with indophenol solution until a faint pink colour persists for 15 seconds. 2.8.5 Calculation Vitamin of Vitamin per 100g/ml = Titer value x Dye factor X Vol made up X 100 Where, Aliquot x : is wt. or vol. of sample mg Ascorbic acid /g, tablet, ml, etc. = (X - B) x (F/E) x (V/¥) Where, X= average ml for test solution titration, B = average ml for test blank titration, F = mgascorbic acid equivalent to one mi iodophenol standard solution, E = no. of g, tablets, ml, etc, assayed V= volume initial test solution and ‘Y= volume test solution titrated Note:- Acetone may be omitted if sulphur dioxide is known to be absent. Its function is to formVitamin of Vitamin C per 100g/ml = Titer value x Dye factor X Vol made up X 100° Where, Aliquot x : is wt. or vol. of sample mg Ascorbic acid /g, tablet, ml, ete. = (X -B) x (F/E) x(V/Y) Where, X= average ml for test solution titration, B= average ml for test blank titration, F = mgascorbic acid equivalent toone ml iodophenol standard solution, E = no. of, tablets, ml, ete. assayed \V= volume initial test solution and ‘Y= volume test solution titrated Note:- Acetone may be omitted if sulphur dioxide is known to be absent. Its function is to form the acetone bisulphate complex with sulphur dioxide which otherwise interferes with the titration. Sometime a small proportion of the ascorbic acid in foods becomes reversibly oxidized during aging and forms dehydroascorbic acid. If this is suspected, first estimate the ascorbic acid as above, then through another portion of the solution pass a stream of Hydrogen sulphide for 10 minutes. Stopper the flask and allow it to {»} FRUIT AND VEGETABLE PRODUCTS | 2012 stand overnight in a refrigerator. Then remove hydrogen sulphide by bubbling nitrogen through the mixture and titrated as before. The difference between the two titrations gives a measure of the dehydroascorbic acid. One international unit of vitamin C = 50 ug ascorbic acid. (Ref = F.A.0 Manuals of Food Quality Control 14 / 8, page 194 / Pearson's Composition and Analysis of Foods 9th edn,1991, page 264 and AOAC Official Method 967.21 Ascorbic acid in Vitamin preparation and juices ) 2,9 Determination of Ethanol Content 2.9.1 Principle Note: + This test method covers only the product which does not contain ethanol as an ingredient. + The method is not applicable to products containing more than 5 % (m/m) of ethanol. Separation of ethanol by distillation followed by oxidation by Potassium dichromate in a sulphuric acid medium and determination of excess dichromate by Ferrous ammonium sulphate in the presence of Ferrous 1, 10 phenathroline as indicatorFRUIT AND VEGETABLE PRODUCTS | 2012 16.0 JAM, JELLY AND MARMALADE 16.1 Total Soluble Solids: - Follow method given in clause 1.6 16.2 Acidity: - Follow method given in clause 2.4 Take 10 gms. of sample; mix thoroughly with about 50 mt water and titrate with 0.1N NaOH using phenolphthalein indicator. Report acidity as citric acid and as malic acid if apple predominates. With highly coloured jams such as blackberry and black current, titrate potentiometrically to pH 8.1, 16.3 Fruit Content: Refer Method No.2.11 16.4 Preservatives: Refer to the Manual on Food Additives 16.5 Added Colouring Matter: Refer to the manual on food additives 17.0 DEHYDRATED FRUITS / DEHYDRATED VEGETABLES: 17.1 Preparation of sample: Observe the sample closely for mould, insect, larvae, extraneous matter etc. Take about 25 to 5O gms of sample, grind quickly to pass through a 30 mesh sieve. 17.2 Determination of Moisture: - Follow method given in clause 4.1 17.3 Determination of Total Ash - Follow method given in clause 14.4. Weigh accurately about 5 gm sample for determination of ash. 17.4 Determination of Acid insoluble ash - Follow method given in clause 5.3. Use the ash obtained in 17.3 for determining acid insoluble ash. 17.5 Test for presence of Peroxidase 17.51 Reagents (2) FRUIT AND VEGETABLE PRODUCTS | 2012 (a) Guaiacol solution - 1 % prepared by dissolving 1 gm of 0.9 ml guaiacol in 50 ml ethyl alcohol and adding 50 ml! water