इंटरनेट मानक Disclosure to Promote the Right To Information Whereas the Parliament of India has set out to provide a practical regime of right to information for citizens to secure access to information under the control of public authorities, in order to promote transparency and accountability in the working of every public authority, and whereas the attached publication of the Bureau of Indian Standards is of particular interest to the public, particularly disadvantaged communities and those engaged in the pursuit of education and knowledge, the attached public safety standard is made available to promote the timely dissemination of this information in an accurate manner to the public. “जान1 का अ+धकार, जी1 का अ+धकार” “प0रा1 को छोड न’ 5 तरफ” Mazdoor Kisan Shakti Sangathan Jawaharlal Nehru “The Right to Information, The Right to Live” “Step Out From the Old to the New” 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” Satyanarayan Gangaram Pitroda ““IInnvveenntt aa NNeeww IInnddiiaa UUssiinngg KKnnoowwlleeddggee”” “!ान एक ऐसा खजाना > जो कभी च0राया नहB जा सकता हहहहै””ै” Bhartṛhari—Nītiśatakam “Knowledge is such a treasure which cannot be stolen” IS 15570:2005 W’wfhm m5-1-R–wMtR-vmdail-h--i-RJh WN7hmmTaml Indian Standard TEXTILES — METHOD OF TEST — DETECTION OF BANNED AZO COLORANTS IN COLOURED TEXTILES ICS 59.040;71.040.50 (3BIS 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. InIndia, theEnvironment (Protection) Rules, 1996prohibit themanufacture andhandling ofcertainazocolorants 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 incoloured textiles needs no emphasis. The composition of the Committee responsible for formulation of this standard isgiven in Annex B. For the purpose of deciding whether aparticular requirement of this standard iscomplied 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 asthat of the specified value inthis 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 3.1 A sample of textile under test is treated with certain azo colorants in coloured textiles. sodiumdithionite inacitratebuffered aqueous solution (pH=6)at700Cinaclosedvessel.Theaminesreleased 2 TERMINOLOGY inthe process ofreductive cleavage are transferred to 2.1Banned Azo Colourants— Bannedazocolorants diethyletherphasebymeansofliquid-liquid extraction are such colorants as may be obtained by cleavage using separating funnel. The diethyl ether is then of their azo group to form the following amines: concentrated under mild conditions in a centrifugal/ S1No. 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, bannedaryl amines o-aminoazotoluene and2-amino-4-nitrotoluene isdetected via o-toluidineand/or2,4-diaminotoluenemines. 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 thedry side inthe standard high pressure liquid chromatography using a diode atmosphere of27*2°Ctemperature and65+2percent 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 Taketheweighedoutsampleina30mlreaction vessel. 4.1 Methanol (HPLC Grade) Add 17mlofcitratebuffer solution(preheated to70”C) 4.2 Diethylether (HPLC Grade) tothereaction vessel andshake well. Keep inawater bathwithcontinuous shaking for30min at70°C.Then 4.3 Citrate-sodium Hydroxide Buffer Solution, add3mloffreshlyprepared sodium dithionite solution pH = 6 Preheated to 70°C (see Annex A). tothereactionvesselandshakewell. Keepthereaction vessel inthewater bath shaker maintained at70”Cfor 4.4 Aqueous Sodium Dithionite Solution, 200 mg/1, 30min. Take outthe reaction vessel and cool to room freshly prepared. 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 the separating funnel through a suitable glass funnel. 4.8 Sodium Dithionite (Purified/LR) Squeezethetextilematerial inthereaction vesselusing 4,9 Sodium Sulphate (LR) aglass rod and transfer the solution to the separating funnel. 5 APPARATUS 6.2.2 Add about 10ml ofdiethyl ether tothe 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. Close the funnel. 5.2 High Performance Liquid Chromatograph (HPLC) with Diode Array Detector (DAD) 6.2.3 Shake the separating fimnel for a few seconds, release the pressure by opening the stopcock. Repeat 5.3 Gas Chromatograph-Mass Spectrometer this process 2to 3times and keep it on the stand. (GC-MS) 6.2.4 After the formation of two separate layers, 5.4 30 ml Reaction Vessel collect the lower layer (aqueous layer) in a breaker. 5.5 Water Bath Shaker Collect the top layer (organic layer) in atest tube. 5.6 Centrifugal Evaporator (Turbo Evaporator) 6.2.5. Transfer the aqueous layer to the separating funnel. Add about 10ml Qfdiethyl 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 at6.2.5 once more. 5.9100 ml Beaker 6.3 Concentration of Amines 5.10 25 ml, 50 ml Burette (Class A) 6.3.1 Passthe organic layer collected inthe test tube through sodium sulphate bed to remove water 5.11 Amber Coloured Bottles for Reference molecules. Pass afew ml of diethyl ether through the Standard Solution sodium sulphate bed. Evaporate theextract todryness 5.12 Test Tubes at not more than 50°C using centrifugal or turbo evaporator. 6 PROCEDURE 6.3.2 Dissolve the residue in 1ml methanol and then 6.1 Sampling analyse by means of HPLC-DAD/HPTLC/GC-MS Take 1to 2 g of sample of textile material under test under conditions described in 7.1 to 7.3. 2 ,. 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 X10cm, 10cm NOTES X10cm) 1 For guidance, details of chromatographic data such as Mobile Solvent 1 : Di-isoprophyl etherl retentiontime(R)andk- ofbannedaminesinHPLCisgiven inAnnexA. n-pentane/Diethyl amine = 9:6:0.5 2 Methodof analysis doesnot coverthe various problems encounteredduetoformation/presenceofisomers. Mobile Solvent 2 : Chloroform/ethyl acetate = 7.3 Gas Chromatography-Mass Spectrometry 9.5:0.5 (GC-MS) Mobile Solvent 3 : Toluene/ethyl acetatel Capillary column : DB 35 Ms, Length — 30 m, diethyl amine= 7:2.5:0.5 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 Carrier gas Helium 7.1.1 Post-chromatographic Derivatization 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 1g of sodium nitrite in 20 x 10twin Detector MS trough chamber andpour 1mlhydrochloric acid over Injection volume : 1pl, splitless 1,fUin it. Cover the trough immediately with alid. 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 rrmineslevel 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, ofthe sample according to the following dissolved in methanol in the ratio of 1g per 200 ml, equation: in the dip tank. Dip the chromatographic plate in the solution for 2 s. Dry th e plate. Compare the w=(A,xc,xvy(/x4A, 4) colour of unknown bands with th ose 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 mlwater, 9 ANALYSIS OF TEST RESULTS pH-6.9 9.1 Positive Stationary phase Li chrosorb 60 RP-Select B If quantity of any one ormore ofthe amines detected (5pm), 250mm x4.6mm or isgreater than 30mgilcg,itisconcluded that azodyes any equivalent column had been used for the production or modification of Flow rate 0.7 to 1.0ml/min the textile material tested. Gradient Start 15 percent eluent 1, 9.2 Negative increase to 80percent eluent 1linearly 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 inthetextile material Name of the Banned Percent Recove~ under test. Amine NOTE — o-toluidine and 2,4-diasninotoluene can also be formedfromo-~inoazotoluene and2-amino4-nitrotoluene (1) (2) respectivelyundertestconditions. 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 a) 4-aminobiphenyl benzidine, 15ppm amine standard solution to a reaction vessel containing 16 ml of the preheated citrate-sodium b) 4-chloro-o-tuluidine, hydroxide buffer solution. Follow the procedure c) 4,4’-diaminodiphenylmethane, adopted forextraction ofthesample(see6.2). Analyze d) 3,3’-dichlorobenzidine, the extract and calculate the recovery percentage. Amine recovery rates shall comply withthe following e) 3,3’-dimethoxybenzidine, minimum requirements: f) 3,3’-dirnethylbenzidine, Amine Nos. 1to 5,7 to 15and 18 : 70 percent g) 3,3’-dirnethyl-4,4’ - diaminodiphenylmethane, Amine No. 6 : 20 percent h) 4,4’-methylene-bis (2 chloroaniline), and j) 4,4’-oxidianiline. Amine Nos. 16and 17 : 50 percent 12 METHOD VALIDATION (USING HPLC) 12.3 Repeatability Method validation is carried out using standard Retentiontimeandpeakareaofthefollowing standard solutions of some of the banned am ines the details of arnines were determined 5 times for each amine and which are as under. their CV percent calculated and isgiven below: Name of Banned CV Percent of CV Percent ofArea Amines 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 4 ,.. .,, IS 15570:2005 12.4 Reproducibility tory adopting the above test methods and there was good agreement inthe results obtained. More than 3000 samples have been tested in labora- 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 than 6, then adjust the same to 6, by adding a few 6.32 g of sodium hydroxide (AR) and transfer them drops oftleshly prepared 0.1N citric acid solution. If intobeaker. Add 50ml ofdistilledwater andstirwell pH is less than 6, then adjust it to 6 by adding few till a clear solution isobtained. Make up the volume drops of freshly prepared 0.1 N sodium hydroxide to 1000 mlusing distilled water. Coolthe solution to solution. Keep the solution in aclosed contained in a 20°Cand checkpH of the solution. IfpH ismore cold dry place. Check thepH prior to use. ---- *— .“ ‘-.-+