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भारतीय मानक IS 17010 : 2018

Indian Standard

तम्बाकू एवं तम्बाकू उत्पाद —

अपचायक एवं कुल शर्क रा का निर्धारण

Tobacco and Tobacco Products —

Determination of Reducing
and Total Sugars

ICS 65.160

© BIS 2018

भारतीय मानक ब्रयू ो

B U R E A U O F I N D I A N S TA N D A R D S
मानक भवन, 9 बहादरु शाह ज़फर मार्ग, नई िदल्ली – 110002
MANAK BHAVAN, 9 BAHADUR SHAH ZAFAR MARG
NEW DELHI-110002
         www.bis.gov.in  
www.standardsbis.in

July 2018  Price Group 2

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Tobacco and Tobacco Products Sectional Committee, FAD 04

FOREWORD
This Indian Standard was adopted by the Bureau of Indian Standards, after the draft finalized by the Tobacco and
Tobacco Products Sectional Committee had been approved by the Food and Agriculture Divisional Council.
The chemical properties of leaf tobacco are influenced by agricultural practices, soil type and nutrients, weather
conditions, stalk position and curing procedures. A change in any of these factors can markedly alter the chemical
composition of tobacco leaf and thus affect the quality of tobacco products.
The total and reducing sugar contents of tobacco are important indicators of the quality of tobacco raw materials and
finished tobacco products. This standard will help in getting uniform and reproducible results while determining
the total and reducing sugar contents in tobacco and tobacco products and thereby a consistent quality product.
In reporting the result of a test or analysis made in accordance with this standard, if the final value, observed or
calculated, is to be rounded off, it shall be done in accordance with IS 2 : 1960 ‘Rules for rounding off numerical
values ( revised  )’.
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IS 17010 : 2018

Indian Standard
TOBACCO AND TOBACCO PRODUCTS —
DETERMINATION OF REDUCING
AND TOTAL SUGARS

1 SCOPE 4 QUALITY OF REAGENTS

This standard specifies a method for the determination Unless specified otherwise, pure chemicals shall be
of reducing and total sugars in tobacco and tobacco employed in tests and distilled water shall be used
products by iodometric titration method. where the use of water as reagent is intended.
NOTE — ‘Pure chemicals’ shall mean chemicals that do not
2 REFERENCE contain impurities which affect the test results.
The standard given below contains provision which, 5 APPARATUS
through reference in this text, constitute provisions of
this standard. At the time of publication, the editions Usual laboratory apparatus and in particular the
indicated were valid. All standards are subject to following:
revision, and parties to agreements based on this
standard are encouraged to investigate the possibility 5.1 Fast Boiling Water Bath
of applying the most recent editions of the standard 5.2 Hot Plate or Heating Mantle
indicated below.
5.3 Whatman Filter Paper No. 40
IS No. Title
IS 5643 : 1999 Tobacco and tobacco products — 5.4 Laboratory Mill, fitted with 1 mm sieve.
Method of test for tobacco (second
5.5 Timer
revision)
6 REAGENTS
3 TERMINOLOGY
For the purpose of this standard, the following 6.1 Lead Acetate ((CH3COO)2Pb.3H2O)
definitions shall apply.
6.2 Shaffer-Somogyi Carbonate Reagents
3.1 Reducing Sugars a) Sodium carbonate, (Na2CO3) anhydrous
Sugar that, in its native form, can reduce the alkaline b) Potassium sodium tartarate (Rochelle Salt),
copper oxide into cuprous oxide, is classified as (KNaC4H4O6.4H2O)
reducing sugar. The ability of an aqueous extract of c) Copper sulphate (CuSO4.5H2O) 10 percent
tobacco to cause this reduction is indicative of the solution — Dissolve 7.5 g in 75 ml of distilled
presence of reducing sugar and can be quantified by the water.
method prescribed in this standard.
d) Sodium bicarbonate, anhydrous (NaHCO3)
3.2 Non-Reducing Sugars e) Potassium iodate solution — Dissolve 3.567 g of
Water soluble carbohydrates, which do not possess the potassium iodate in 1 litre of distilled water.
reducing property, but acquires this property after acid f) Potassium iodide
hydrolysis. The difference in the sugar contents before
and after hydrolysis is a measure of non-reducing sugar 6.2.1 Preparation of Shaffer-Somogyi Reagent
content in tobacco. Dissolve 25 g of anhydrous sodium carbonate [6.2 (a)]
and 25 g of Rochelle salt [6.2 (b)] in 500 ml of distilled
3.3 Total Sugars water in 1 litre volumetric flask. Add with stirring 75
The total of water-soluble reducing and non-reducing ml of the copper sulphate solution [6.2 (c)] slowly by
sugars is known as total sugars, which are determined a pipette well below the surface of the liquid. Add 20
after acid hydrolysis according to the procedure g sodium bicarbonate [6.2 (d)], dissolve by stirring,
described in this standard. then add 5 g of potassium iodide [6.2 (f)]. When all the
solids are dissolved, add 200 ml of potassium iodate

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IS 17010 : 2018

solution [6.2 (e)]. Make up the volume. Mix thoroughly 8 PROCEDURE

and allow the reagent to stand for two days and filter
before use. The reagent is stable for one year if stored 8.1 Preparation of Sample
in an amber coloured bottle. Prepare the tobacco sample for analysis by grinding
(the sample should totally pass through 1 mm sieve)
6.3 Potassium Iodate Solution, 0.1 N.
and determine the moisture content. Ideally moisture
6.4 Sodium Thiosulphate (Na2S2O3.5H2O) Solution, content shall be less than 15 percent for grinding. If the
0.1 N tobacco is wet for grinding, it may be air dried.
Dissolve 24.82 g of sodium thiosulphate and 0.2 g of 8.2 Test Portion
sodium carbonate in 200 ml of distilled water and dilute
Determine the oven moisture content of the ground
to one litre.
sample by the method as prescribed in IS 5643.
6.4.1 Standardization Transfer about 1 g of ground sample, weighed to the
nearest 0.01 g, to a 250 ml Erlenmeyer flask.
Standardize the sodium thiosulphate solution (see 6.4)
against 0.1 N potassium iodate solution (see 6.3). For 8.3 Extraction
titration, take 25 ml of 0.1 N potassium iodate solution
Add exactly 100 ml of distilled water to the flask
in a 250 ml conical flask, add 3 g of potassium iodide
(see 8.2) and shake for 1 h on a flask shaker and filter
and 10 ml of 1 N sulphuric acid. Titrate against 0.1 N
through Whatman No. 40 filter paper. Add 0.5 g of lead
sodium thiosulphate until pale yellow colour. Add
acetate to the filtrate, shake well and allow to stand for
approximately 65 ml of water and add 2 ml of starch
30 min.
indicator solution (see 6.6) and continue titration till
dark blue colour changes to colourless. 8.4 Removal of Interfering Substances and
6.5 Sodium Thiosulphate Solution, 0.005N Deleading
Prepare 0.005 N solution by diluting 25 ml of 0.1 N Filter the solution through a Whatman no. 40 filter
sodium thiosulphate solution into 500 ml with water. paper into a dry 250 ml conical flask containing 1g
The solution is unstable and hence should be discarded potassium oxalate. Shake well and allow to stand for
after 4 h. 30 min. Filter the solution through Whatman No. 40
filter paper into a dry 100 ml conical flask.
6.6 Starch Indicator Solution
Disperse 1 g soluble starch in 20 ml water and pour 8.5 Preparation for Total Sugars − Hydrolysis of
into about 80 ml boiling water. Boil for 1 min and cool. Total Sugars
Prepare a fresh starch solution daily. Pipette 10 ml of solution from 8.4 into a 50 ml conical
flask, add 5 ml of 0.5 N hydrochloric acid and boil on
6.7 Sodium Hydroxide Solution, 0.5 N.
hot plate/heating mantle for 3-5 min. Cool to room
6.8 Sulphuric Acid Solution, 2 N. temperature and add 5 ml of 0.5 N sodium hydroxide
solution. Transfer quantitatively to 50 ml volumetric
6.9 Hydrochloric Acid, 0.5 N. flask and dilute to volume. Mix thoroughly.
6.10 Potassium Iodide Solution, 2.5 percent 8.6 Preparation for Reducing Sugars
Weigh 2.5 g in 100 ml volumetric flask and dilute to Pipette 10 ml of the filtrate (8.4) into a 50 ml volumetric
volume with distilled water. flask. Make up the volume.
6.11 Potassium Oxalate 9 DETERMINATION OF SUGAR
7 PRINCIPLE 9.1 Pipette 5 ml aliquot from 8.6 (for reducing sugars)
An aqueous extract of the tobacco sample is prepared and 5 ml aliquot from 8.5 (for total sugars) into separate
and the reducing sugars (as glucose) is determined by 100 ml conical flasks, add accurately 5 ml of the Shaffer
Shaffer-Somogyi method. Cuprous oxide produced Somogyi reagent to each flask and mix thoroughly. Add
by the reaction of a reducing sugar with the Somogyi anti-bumping granules and cover flasks with a glass
reagent is reacted with potassium iodate in the bulb or funnel. Heat for exactly 15 min by immersing
presence of sulphuric acid. Excess potassium iodate is in the boiling water bath. Ensure that the water level
determined by adding potassium iodide and titrating in the bath is above the reaction mixture. Remove the
the liberated iodine with sodium thiosulphate solution flasks from the water bath, cool in cold water (20-25°C)
using starch as an indicator. The non-reducing sugars and add 2 ml of 2.5 percent KI solution by running
are hydrolysed with dilute hydrochloric acid and the it from a pipette down the walls of the flask without
total reducing sugars (as glucose) is determined as stirring or agitation, add 3.0 ml of 2 N sulphuric acid
above. and mix thoroughly. Titrate the liberated iodine with

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IS 17010 : 2018

0.005 N thiosulphate solution till the solution becomes V1 = volume of original sample extract (100 ml);
pale yellow. Add 100 µl starch indicator solution and V2 = volume of diluted sample extract (50 ml);
complete titration. The end point is blue to colourless.
V3 = volume of original sample extract used for
9.2 Carry out 3 blank determinations using 5 ml of dilution (10 ml); and
water in place of sample extract 9.1. V4 = volume of sample used for titration (5 ml).
9.3 Subtract titre values of test solution from that of 10.2 In case the titre values exceeds 17.9 ml, the
blank and calculate the amount of glucose corresponding following formula may be used for calculating mg
to the titre difference from the formula. glucose
Glucose, mg = 0.112 × (ml of 0.005 N sodium
10 CALCULATIONS
thiosulphate) + 0.04 ml of 0.005 N Sodium Thiosulphate
10.1 Calculate sugars (as glucose), percent by dry mass = (Blank titre value – Sample titre value)
from the following formula:
11 REPORTING
Sugars (as glucose), percent by
The report will carry the following:
G × V1 × V2 × 100 100
=
dry mass × a) Full identity of the sample;
W × 1000 × V3 × V4 (100 − M )
b) Date of test;
Where, c) Moisture content, percent of sample as received;
G = glucose, in mg (see Table 1); d) Reducing sugar content, percent on dry basis;
W = weight of sample, in g; e) Total sugar content, percent on dry basis; and
M = percent moisture content; f) Non-reducing sugar, percent on dry basis.

Table 1 Milligrams of Glucose Corresponding to the Difference Between

the Titration Values for the Blank and the Unknown
(Clause 10.1)
0.005 N Milligram of Glucose Corresponding to Titration Difference of
Na2S2O3
(ml)
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
(1) (2) (3) (4) (5) (6) (7) (8) (9) (10) (11)
0 0.05 0.06 0.08 0.09 0.10 0.11 0.13 0.14
1 0.15 0.16 0.17 0.19 0.20 0.21 0.22 0.21 0.24 0.35
2 0.27 0.28 0.29 0.30 0.31 0.32 0.33 0.35 0.36 0.37
3 0.38 0.39 0.41 0.41 0.42 0.43 0.45 0.46 0.47 0.48
4 0.49 0.50 0.53 0.54 0.54 0.55 0.56 0.57 0.58 0.59
5 0.60 0.61 0.64 0.65 0.65 0.66 0.67 0.68 0.69 0.70
6 0.72 0.73 0.74 0.75 0.76 0.77 0.78 0.79 0.80 0.82
7 0.83 0.84 0.85 0.86 0.87 0.88 0.89 0.90 0.91 0.83
8 0.94 0.95 0.96 0.97 0.98 0.99 1.00 1.01 1.02 1.04
9 1.05 1.06 1.07 1.08 1.09 1.10 1.11 1.12 1.13 1.15
10 1.16 1.17 1.18 1.19 1.20 1.21 1.22 1.23 1.24 1.26
11 1.27 1.28 1.29 1.30 1.32 1.33 1.34 1.35 1.36 1.37
12 1.38 1.39 1.40 1.42 1.43 1.44 1.45 1.46 1.47 1.48
13 1.49 1.50 1.52 1.53 1.54 1.55 1.56 1.57 1.58 1.59
14 1.60 1.61 1.63 1.64 1.65 1.66 1.67 1.68 1.69 1.70
15 1.71 1.72 1.74 1.75 1.76 1.77 1.78 1.79 1.80 1.81
16 1.82 1.83 1.85 1.86 1.87 1.88 1.89 1.90 1.91 1.92
17 1.93 1.94 1.95 1.96 1.97 1.98 1.99 2.01 2.02 2.03

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This Indian Standard has been developed from Doc No.: FAD 04 (2786).

Amendments Issued Since Publication

Amend No. Date of Issue Text Affected

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