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IS 15570 (2005): Textiles - Method of test - Detection of

banned azo colourants in coloured textiles [TXD 7: Textile
Sizing and Finishing Materials]

“!ान $ एक न' भारत का +नम-ण”

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“Knowledge is such a treasure which cannot be stolen”
 IS 15570:2005

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Indian Standard
TEXTILES — METHOD OF TEST —
DETECTION OF BANNED AZO COLORANTS IN
COLOURED TEXTILES

ICS 59.040;71 .040.50

(3 BIS 2005

BUREAU OF INDIAN STANDARDS

MANAK BHAVAN, 9 BAHADUR SHAH ZAFAR MARG
NEW DELHI 110002 :

April 2005 Price Group 3

.. .. ..
Textile Sizing, Finishing Materials and Dyestuffs Sectional Committee, TX 07

FOREWORD
This Indian Standard was adopted by the Bureau of Indian Standards, after the draft finalized by the Textile
Sizing, Finishing Materials and Dyestuffs Sectional Committee had been approved by the Textile Division
Council.
In India, the Environment (Protection) Rules, 1996 prohibit the manufacture and handling of certain azo colorants
d~rived from harmful amines. In view of the possible occupational hazards in the use of these colorants on
textile commodities, formulation of a standard method for detection of such banned azo colorants in coloured
textiles needs no emphasis.
The composition of the Committee responsible for formulation of this standard is given in Annex B.
For the purpose of deciding whether a particular requirement of this standard is complied with, the final value,
observed or calculated, expressing the result of a test or analysis, shall be rounded off in accordance with
1S 2: 1960 ‘Rules for rounding off numerical values (revised’. The number of significant places retained in
the rounded off value should be the same as that of the specified value in this standard.
 IS 15570:2005

Indian Standard
TEXTILES — METHOD OF TEST —
DETECTION OF BANNED AZO COLORANTS IN
COLOURED TEXTILES
1 SCOPE 3 PRINCIPLE

This standard prescribes a method for detection of
certain azo colorants in coloured textiles.
2 TERMINOLOGY
2.1 Banned Azo Colourants— Banned azo colorants
are such colorants as may be obtained by cleavage
of their azo group to form the following amines:
3.1 A sample of textile under test is treated with
sodium dithionite in a citrate buffered aqueous solution
(pH = 6) at 700Cin a closed vessel. The amines released
in the process of reductive cleavage are transferred to
diethyl ether phase by means of liquid-liquid extraction
using separating funnel. The diethyl ether is then
concentrated under mild conditions in a centrifugal/

S1 No. Name of Banned Amine CAS No.

(1) (2) (3)

i) 4-aminobiphenyl 92-67-1
ii) Benzidine 92-87-5
iii) 4-chloro-o-toluidine 95-69-2
iv) 2-naphthylamine 91-59-8
v) 4-chloro-aniline 106-47-8
vi) 2,4-diaminoanisole 615-05-4
vii) 3,3’-diaminodiphenylmethane 101-77-9
viii) 3,3’-dichlorobenzidine 91-94-1
ix) 3,3’-dimethoxybenzidine 119-90-4
x) 3,3’-dimethylbenzidine 119-93-7
xi) 3,3-dimethyl,4,4’- diaminodi phenylmethane 838-88-O
xii) p-cresidine 120-71-8
xiii) 4,4’ methylene-bis (2-chloroaniline) 101-14-4
xiv) 4,4’-oxydiani1ine 101-80-4
xv) 4,4’-thiodianiline 139-65-1
xvi) o-toluidine 95-53-4
xvii) 2,4-toluenediamine 95-80-7
xviii) 2,4,5-trimethylaniline 137-17-7
xix) o-aminoazotoluene 97-56-3
xx) %nnino-4-nitrotoluene 95-55-8
xxi) p-aminoazobenzene 60-09-03
xxii) o-anisidine 90-04-0

NOTE — According to this method, banned aryl amines o-aminoazotoluene and 2-amino-4-nitrotoluene is detected via
o-toluidine and/or 2,4- diaminotoluene mines.

1
IS 15570:2005

turbo evaporator and the residue is dissolved in from different places covering equally all the colours
methanol. and/or fibres, cut into small pieces (about 5 mm x
5 mm) and mix thoroughly. Condition the sample to
3.2 Detection of the amines is done by means of
moisture equilibrium from the dry side in the standard
high pressure liquid chromatography using a diode
atmosphere of 27 * 2°C temperature and 65 + 2 percent
array detector’ (HPLC/DAD), high performance
relative humidity. From this weigh 1.00 + 0.02 g of
thin layer chromatography (HPTLC) or capillary
the sample accurately (in case of dye, take only 0.01
gas chromatography with mass spectrometer and
to 0.05 gas sample).
quantification is done using HPLC with DAD.
6.2 Separation of Amines
4 REAGENTS
Take the weighed out sample in a 30 ml reaction vessel.
4.1 Methanol (HPLC Grade)
Add 17 ml of citrate buffer solution (preheated to 70”C)
4.2 Diethylether (HPLC Grade) to the reaction vessel and shake well. Keep in a water
bath with continuous shaking for 30 min at 70°C. Then
4.3 Citrate-sodium Hydroxide Buffer Solution, add 3 ml of freshly prepared sodium dithionite solution
pH = 6 Preheated to 70°C (see Annex A). to the reaction vessel and shake well. Keep the reaction
vessel in the water bath shaker maintained at 70”C for
4.4 Aqueous Sodium Dithionite Solution, 200 mg/1,
freshly prepared. 30 min. Take out the reaction vessel and cool to room
temperature in about 2 min by keeping in ice-cold
4,5 Standard Amines of Highest Available Purity water. Carry out the liquid-liquid extraction of
the amines by using diethyl ether as given in 6.2.1
4,6 Standard Amine Solutions to 6.2.6.
4,7 Calibration Solutions, 15 ppm. 6.2.1 Transfer the extract from the reaction vessel to
4.8 Sodium Dithionite (Purified/LR) the separating funnel through a suitable glass funnel.
Squeeze the textile material in the reaction vessel using
4,9 Sodium Sulphate (LR) a glass rod and transfer the solution to the separating
funnel.
5 APPARATUS
6.2.2 Add about 10 ml of diethyl ether to the reaction
5,1 High Performance Thin Layer Chromatograph vessel and stir well. Transfer the extract to the
(HPTLC) separating funnel. Repeat this process once more.
5.2 High Performance Liquid Chromatograph Close the funnel.
(HPLC) with Diode Array Detector (DAD) 6.2.3 Shake the separating fimnel for a few seconds,
5.3 Gas Chromatograph-Mass Spectrometer release the pressure by opening the stopcock. Repeat
(GC-MS) this process 2 to 3 times and keep it on the stand.

5.4 30 ml Reaction Vessel 6.2.4 After the formation of two separate layers,
collect the lower layer (aqueous layer) in a breaker.
5.5 Water Bath Shaker Collect the top layer (organic layer) in a test tube.
5.6 Centrifugal Evaporator (Turbo Evaporator) 6.2.5. Transfer the aqueous layer to the separating
funnel. Add about 10 ml Qf diethyl ether and repeat .
5.710,5,2 and 1 ml pipettes (Class A) .
the procedure given at 6.2.3 and 6.2.4.
5.8 60 ml Separating Funnel
6.2.6 Repeat the procedure given at 6.2.5 once more.
5.9100 ml Beaker
6.3 Concentration of Amines
5.10 25 ml, 50 ml Burette (Class A)
6.3.1 Pass the organic layer collected in the test tube
5.11 Amber Coloured Bottles for Reference through sodium sulphate bed to remove water
Standard Solution molecules. Pass a few ml of diethyl ether through the
sodium sulphate bed. Evaporate the extract to dryness
5.12 Test Tubes at not more than 50°C using centrifugal or turbo
evaporator.
6 PROCEDURE

6.1 Sampling
6.3.2 Dissolve the residue in 1 ml methanol and then
analyse by means of HPLC-DAD/HPTLC/GC-MS
Take 1 to 2 g of sample of textile material under test under conditions described in 7.1 to 7.3.

,.
 IS 15570:2005

7 CHROMATOGRAPHIC ANALYSIS Column temperature : 40°C

7.1 High Performance Thin Layer Chromatography Injection volume : 15-20 pl

(HPT~C)
Detection : Diode array detector
Stationery phase : Silica gel 60 with spectrograph at 240 nm,
fluorescence indicator F254 280 nm and 305 mn.
(Size 20 cm X 10 cm, 10 cm NOTES
X 10 cm)
1 For guidance, details of chromatographic data such as
Mobile Solvent 1 : Di-isoprophyl etherl retention time (R) and k- of banned amines in HPLC is given
in Annex A.
n-pentane/Diethyl amine =
9:6:0.5 2 Method of analysis does not cover the various problems
encountered due to formation/presence of isomers.
Mobile Solvent 2 : Chloroform/ethyl acetate =
9.5:0.5 7.3 Gas Chromatography-Mass Spectrometry
(GC-MS)
Mobile Solvent 3 : Toluene/ethyl acetatel
diethyl amine= 7:2.5:0.5 Capillary column : DB 35 Ms, Length — 30 m,
inside diameter — 0.25 mm,
Development : Saturated chamber film thickness — 0.25pm
Chamber saturation : 10min Injector system : Splitless/split
Development distance : 80 mm Injector temperature : 260”C

7.1.1 Post-chromatographic Derivatization

Carrier gas Helium
Temperature : 100”C (2 rein), 100”C to
Derivative the chromatographic plate by diazoisation programme 310”C (15°C/ rein), 310”C
as given in 7.1.1.1. (2 rein)
7.1.1.1 Spread 1 g of sodium nitrite in 20 x 10 twin Detector MS
trough chamber and pour 1 ml hydrochloric acid over Injection volume : 1pl, splitless 1, fUin
it. Cover the trough immediately with a lid. Wait till
the chamber is filled with nitrous limes. Hold the 8 CALCULATION
chromatographic plate over the ties for a minute.
8.1 Calculate the rrmines level from the peak areas #
Fix the plate on the immersion unit. Take sufficient
of the individual amine components as mass portion,
solution of N-naphthy-lethy lenediamine (GR/AR)
Win mgkg, of the sample according to the following
dissolved in methanol in the ratio of 1 g per 200 ml,
in the dip tank. Dip the chromatographic plate in equation:
the solution for 2 s. Dry the plate. Compare the w=(A,xc, xvy(/4,
xA4)
colour of unknown bands with those of the standard where
bands.
,4, = peak area of the amine in the sample in areas
7.2 High Performance Liquid Chromatography units;
(HPLC) C,= concentration of the amine in the calibration
solution, in pg/ml;
Eluent 1 Methanol
V= volume of the extract, in ml;
Eluent 2 0.575 g Ammonium
A.= peak area of the amine in the calibration
dihydrogen phosphate +
solution, in area units; and
0.7 g
M= mass of the specimen, in g.
Disodium hydrogen
phosphate in 1000 ml water, 9 ANALYSIS OF TEST RESULTS
pH-6.9
9.1 Positive
Stationary phase Li chrosorb 60 RP-Select B
(5pm), 250 mm x 4.6 mm or If quantity of any one or more of the amines detected
any equivalent column is greater than 30 mgilcg, it is concluded that azo dyes
had been used for the production or modification of
Flow rate 0.7 to 1.0 ml/min the textile material tested.
Gradient Start 15 percent eluent 1, 9.2 Negative
increase to 80 percent eluent
1 linearly within 45 min If quantity of banned amines is less than or equal to

3
IS 15570:2005

30 mgfkg, it is concluded that azo dyes which are 12.1 Recovery Percentage
prohibited could not be detected in the textile material
under test. Name of the Banned Percent Recove~
Amine
NOTE — o-toluidine and 2,4-diasninotoluene can also be
formed from o-~inoazotoluene and 2-amino 4-nitrotoluene (1) (2)
respectively under test conditions.
4-aminobiphenyl 79.6
10 REPORT
4,4’-diaminodiphenyImethane 67.8
The report shall include the following information: 3,3’-dimethoxybenzidine 72.7

a) Name of the textile material tested; 3,3’-dimethoxybenzidine 66.2

3,3’-dirnethylbenzidine 74.7
b) Test method used;
3,3’-dimethyl-4,4’-diaminodiphenyl 68.0
c) Quantity of individual amines released, if any, methane
in mg/kg; and
4,4’-methylene-bis-(2-chloroaniline) 74.8
d) Identification of the source of manufacture. 4,4’-oxidianiline 83.5
11 CHECK OF THE ANALYTICAL SYSTEM 12.2 Detection Limit
(RECOVERY PERCENTAGE)
Detection limit was checked at 0.2 ppm limit for the
To check the procedure of analysis, determine the following banned amines and were detected
recovery percentage of amines by adding 1.0 ml of
15 ppm amine standard solution to a reaction vessel a) 4-aminobiphenyl benzidine,
containing 16 ml of the preheated citrate-sodium b) 4-chloro-o-tuluidine,
hydroxide buffer solution. Follow the procedure c) 4,4’-diaminodiphenylmethane,
adopted for extraction of the sample (see 6.2). Analyze
the extract and calculate the recovery percentage. d) 3,3’-dichlorobenzidine,
Amine recovery rates shall comply with the following e) 3,3’-dimethoxybenzidine,
minimum requirements:
f) 3,3’-dirnethylbenzidine,
Amine Nos. 1 to 5,7 to 15 and 18 : 70 percent g) 3,3’-dirnethyl-4,4’ - diaminodiphenylmethane,
Amine No. 6 : 20 percent h) 4,4’-methylene-bis (2 chloroaniline), and
Amine Nos. 16 and 17 : 50 percent j) 4,4’-oxidianiline.

12 METHOD VALIDATION (USING HPLC) 12.3 Repeatability

Method validation is carried out using standard Retention time and peak area of the following standard
solutions of some of the banned amines the details of arnines were determined 5 times for each amine and
which are as under. their CV percent calculated and is given below:

Name of Banned CV Percent of CV Percent of Area

A mines Retention Time(RJ
(1) (2) (3)
4-aminobiphenyl 0.10 1.42
Benzidine 2.1 1.45
4-chloro-o-tuluidine 0.26 0.80
4,4’-diaminodipheny lmethane 0.84 1.11
3,3’-dichlorobenzidine 0.15 2.45
3,3 ‘-dimethoxybenzidine 0.65 2.31
3,3‘-dimethylbenzidine 1.37 0.93
3,3’-dimethyl-4,4’-diaminodiphenyl-methane 0.12 0.45
4,4’-methylene-bis (2 chloroaniline) 0.11 0.44
4,4’-oxidianiline 1.08 1,46

,.. ., ,
 IS 15570:2005

12.4 Reproducibility tory adopting the above test methods and there was
More than 3000 samples have been tested in labora- good agreement in the results obtained.

ANNEX A
(Clause 4.3)
PREPARATION OF CITRATE — SODIUM BYDROXIDE
BUFFER SOLUTION pH =6, PREHEATED to 70”C

A-1 Weigh exactly 12.526 g of citric acid (AR) and
6.32 g of sodium hydroxide (AR) and transfer them
into beaker. Add 50 ml of distilled water and stir well
till a clear solution is obtained. Make up the volume
to 1000 ml using distilled water. Cool the solution to
20° C and check pH of the solution. If pH is more
than 6, then adjust the same to 6, by adding a few
drops of tleshly prepared 0.1 N citric acid solution. If
pH is less than 6, then adjust it to 6 by adding few
drops of freshly prepared 0.1 N sodium hydroxide
solution. Keep the solution in a closed contained in a
cold dry place. Check the pH prior to use.

----
*— .“ ‘-.-+
IS 15570:2005

ANNEX B
(Foreword)
COMMITTEE COMPOSITION
Textile Sizing, Finishing Materials and Dyestttffs Sectional Committee, TX 07
Organization Representative(s)

Ahmedabad Textile Industry’s Research Association, Abmedabad DR M. M.GHARIA (Chrhuur)

Ahmedabad Textile Industry’s Research Association, Ahmedabad DRS. RAHMAN
Smu D. L. PATEL (Alternate)
Anil Products Limited, Ahmedabad Smu K. B. CHOX.WI ,.
Smu AAYIL+crnn(Alternate)
Century Textiles and Industires Ltd, Mumbai SsmrMAHESH Srr,mm
Central Institute for Research on Cotton Technology, Mumbai DRP. V. VARADARAIAN
SrmrA. J. Smmus(Alternate)
Colour-Chem Limited, Mumbai DRG. V. G. Ibo
Di-Ichi Karkaria Pvt Ltd, Mumbai Smu S. VISHWANAmAN
DRBr-rAaAmDs.s.u(Alremate)
Indian Dyestuffhrdustries Limited, Mumbai .%mJ. K. JOSm
Smo V. R. ILuuwww(Alterrrate)
Indian Jute Industries’ Research Association, Kolkata DRT, K. G~ ROY
Jaysanth Dye-Chem, Mumbai SHSUs. c. SHARA4A
Smu P.M.I@r-r.wwm(Alternate)
Man-Made Textiles Research Association, Surat DRS. R.N.mc
SmuM.G. PA= (Alternate)
Maize Products, Ahmedabad SmuN. C. Smt-r
Smu A. H. DHAMEIJA(Afternate)
National Peroxide Limited, Mumbai DRS. R. TENDULXAR
SrmrA. K. PATT-MK
(Alternate)
Office of the Textile Commissioner, Mumbai SmuR.A. Lu
The South India Textiles Research Association, Coimbatore SmoK.P.JANAKIRAMAN
The Bombay Dyeing and Manufacturing Co Ltd, Mumbai km A. S.Gosus
Sm P.R.Katsmio (Alternate)
The Bombay Millowners’ Association, Mumbai SW P. M. BHATT
Smu A. C.RANGANS (Alternate)
The Arvind Mills Ltd, Ahmedabad DRJ. J. SHROFF
Smu A. R.PAm (Alternate)
The Ahd Products Limited, Balsad Smu N. S. K. SWNWASAN
DRSUNILB. Jowrr(Alternate)
The Bombay Textile Research Association, Mumbai DRG. N. Srrsm
The Silk and Art Silk Mills Research Association, Mumbai DR S. A. SHAH
SW K. S. TARAPOREWALA (Alternate)
Textiles Committee, Mumbai DRG. S. NADIGER
Smu S. SUaWMMN (Alternate)
Universal Starch Chem Allied Limited, Dhule Smu D. A. PATIL
Wool Research Association, Thane Smrwm G. P. R.ms
Smr V. C. PWSE(Alternate)
In personal Capacity (Technology Advisor, Textiles and Srau M.D. DIxrr
Engineering, A4umbai)
BIS Directorate General SmuM.S. VsmiA,Director & Head(TXD)
[Representing Director General (E#-oficio)]

Member Secretaty
SsrsuB. L. Bwm
Joint Dkector (TXD), BIS

,., -...’
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This Indian Standard has been developed from DOC: No. TX 07 (0460).

Amendments Issued Since Publication

Amend No. Date of Issue Text Affected

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