Experiment Name: Analysis (acidity, reducing and non reducing sugar content) of

jam/jelly/marmalade.

 Analysis of Acidity

Analysis of Total Sugar

Analysis of Moisture content

Analysis of TSS
Analysis Acidity of jam/jelly/marmalade:
 
Principle: The acidity of food sample can be expressed either as % acetic acid citric acid or lactic acid depending on the food
sample. This is estimated by titrating with (N/10) NaOH using Phenolphthalein indicator.
 

1ml (N/10) NaOH =7mg of Citric acid

 
 
Materials: (i) (N/10) NaOH
(ii) Phenolphthalein indicator (1mg + 50ml ethanol + 50ml DW)
 
 
Procedure: Take 25 ml sample solution and titrate against 0.1(N) NaOH using Phenolphthalein indicator. Determine the
exact strength of NaOH solution by titrate against Standard 0.1(N) Oxalic Acid using Phenolphthalein indicator.
 
 
No. of observation Vol. of aliquot taken Burette Reading Average
Result: (ml) Weight of citric acid= N*V* equivalent weight of citric
     3.5  
acid
 10  
 
     3.6
N= Normality of NaOH
 10 V= Volume of NaOH Consumed (in lit)
     3.6
 10
 Food sample
No. of observation Vol. of aliquot taken Burette Reading Average
(ml)

 1    4.9  
 10  
 
 2    4.6
 10

 3   4.8
 10

Oxalic acid
No. of observation Vol. of aliquot taken Burette Reading Average
(ml)

 1    8.9  
 10  
 
 2    8.9
 10

 3   8.8
 10
Analysis of Reducing sugar: Principle Lane and Eynon method is based on the principle of reduction of Fehling’s solution
by reducing sugars. Fehling’s solution is a mixture of copper sulphate and alkaline Rochelle salt (sodium potassium
tartarate). Rochelle salt complexes with the cupric hydroxide formed in alkaline solution and prevent it from
precipitation. Reducing sugars reduces the complexed cupric hydroxide to red, insoluble cuprous oxide under the
experimental conditions. An oxidation-reduction indicator, usually methylene blue, detects the end point of the
reaction.The first step in the estimation of reducing sugars by Lane and Eynon method is the determination of Factor for
Fehling’s solution. Fehling factor is the quantity of invert sugar in grams required to completely reduce the
Fehling’ssolution (usually 5 ml each of Fehling’s A and B solutions). Total sugars include reducing sugars and non-
reducing di - and oligo-saccharides like sucrose, which on mild acid hydrolysis are converted into reducing sugars.
Starch is hydrolysed by strong acids into glucose.
Requirements Equipment and Apparatus Chemical balance, 1mg sensitivity Hot plate Burette (50 ml cap.) with an off-
set tip Volumetric flask, 250 ml Pipette, 5 ml and 25 ml Conical flask, 250 ml Weighing bottle Funnel (small) Whatman
No. 1 filter circles
Reagents:

Fehling’s solution A: Dissolve 69.28 g copper sulphate (CuSO4.5H2O) in distilled water and dilute to 1000 ml. Filter and
store in amber colour bottle.
Fehling’s solution B: Dissolve 346 g Rochelle salt (Potassium sodium tartrate: KNa C 4H4O6. 4H2 O) and 100 g NaOH in
distilled water. Dilute to 1000 ml. Filter and store in amber colour bottle..
Methylene blue indicator: Prepare 1% methylene blue solution in distilled water.
Preliminary titration: Pipette out 5 ml each of Fehling A and B solutions into 250 ml conical flask. Mix and add about 10 ml
water and a few pumice stone or glass beads. Dispense the sugar solution from the burette. Heat the solution to boiling. Add 3
drops of methylene blue indicator. Continue the addition of the sugar solution drop wise until the blue colour disappears to a
brick-red end point. (The concentration of the sample solution should be such that the titre value is between 15 to 50 ml).
Maintain a total boiling peiod of 3 min. Note down the titre value.
Final titration: Pipette out 5 ml each of Fehling A and B solutions into a 250 ml conical flask. Add sample solution about
0.05 to 1.0 ml less than titre value of the preliminary titration. Heat the flask to boiling. Add3 drops of methylene blue
indicator. Complete the titration within 1 min by adding 2 to 3 drops of sugar solution at a time, until the indicator is
decolourized. At the end point, the boiling liquid assumes the brick redcolour. Note down the titre value. Perform the titration
in duplicate and take the average.
Analysis of Non reducing sugar:
Principle Total reducing sugars represent reducing sugars and non-reducing di and oligo saccharides, which can be
hydrolysed into reducing sugars with dilute acids. 4.3.2 Requirements Same as for experiment 4a. 4.3.3 Procedure
Pipette an aliquot of 50 ml of the clarified, de-leaded filtrate to a 100 ml volumetric flask. Add 5 ml of conc. HCl
and allow to stand at room temperature for 24 hours. Neutralise with conc. NaOH solution followed by 0.1N
NaOH using phenolphthalein as end point indicator. Make up to volume and transfer to 50 ml burette having an
off-set tip. Perform the titration against Fehling's solution similar to the procedure described for reducing sugars,
and determine the total sugars as invert sugars.
Analysis of Total Soluble Solid:
Principle:
Refiactometer measures total soluble solids (TSS) concentration based on the principle
of refraction of light. When a ray of light travels obliquely from one medium to another,
it is bent or refracted. The refraction occurs because light travels at slightly different
velocities in different media, the extent being proportional to the density of the solution or
the soluble solids concentration. The refractive index of a medium is defined as the ratio
of,the sine of the angle of incidence to the sine of the angle of refraction when a ray of
monochromatic light is refracted from a vacuum (or, to a very close approximation, from
air) into the medium. In a Brix refiactometer, the refractive index is calibrated into OBrix
readings. As refractive index is dependent on the density of the solution, the measurements
have to be made at a specific temperature (20 "C) or suitable corrections have to be
applied.
Equipments:
Hand Refractometer or Abbe's Refractometer Thermostatically controlled water bath.
 
Procedure:
Place few drops of the sample in between the prisms of hand refractometer and note the
reading at the demarcation line. Apply temperature correction for readings taken at
temperatures other than 20 OC using the following table.
In the case of Abbe's refractometer, circulate 20 C water in the chamber enclosing the
prism from a thermostatically controlled water bath. Place a few drops of the sample in
between the prisms and allow the temperature to equilibrate and note the Brix reading,
which gives per cent of sucrose sugar or TSS.
If sample is thick, squeeze it through cotton and place the drop in between the prisms.
Result:
The readings are expressed as total soluble solids (TSS) = %.
Temperature corrections for readings of per cent sucrose in sugar solution by either Abbe
or Immersion refractometer at temperatures other than 20 %.
i. Analysis of Moisture Content:
The volatile material lost by a substance on heating the boiling temperature is called the moisture.
Actually it is the free water present in the material.
Method: About 5 gm of sample is taken in a Petridis. Then the sample is kept in the hot air oven at 104 0C
temperature. After a certain period the sample is taken out from the oven and the weight of the sample is
taken. Again it is kept in oven and after 15 mint is weighed. If two weights are the same drying
is complete (M2). Otherwise the drying is to be allowed, till two consecutive readings are same.
 
Calculation:
Moisture (percentage by mass) = 100*(M1-M2)/(M1-M)
Here,
M = mass of the empty dish (in gms)
M1 = Mass of the dish with the material before drying (in gms)
M2 = Mass of the dish with the material after drying (in gms)