FOR COMMENTS ONLY FINAL  DRAFT UGANDA 
STANDARD  
  FDUS EAS 766-1 
Second  Edition  
2013-mm-dd 
 
 Reference number  
FDUS EAS 766-1: 2013 
© UNBS 201 3 
  
Antibacterial toilet soap — Specification — Part 1: Solid  
 FOR COMMENTS ONLYFDUS EAS 766-1: 2013  
ii                © UNBS 2013 - All rights reserved 
 Compliance with this standard does not, of itself confer immunity from legal obligations  
A Uganda Standard does not purport to include all necessary provisions of a contract. Users are 
responsible for its correct application  
© UNBS 2013 
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilised in any 
form or by any means, electronic or mechanical, including photocopying and microfilm, without prior written 
permission from UNBS. 
Requests for permission to reproduce this document should be addressed to 
The Executive Director 
Uganda National Bureau of Standards 
P.O. Box 6329 
Kampala 
Uganda 
Tel: 256 41 505 995 
Fax: 256 41 286 123 
E-mail: unbs@infocom.co.ug  
Web: www.unbs.go.ug  
 FOR COMMENTS ONLYFDUS EAS 766-1: 2013  
© UNBS  2013 - All rights reserved                iii National foreword 
Uganda National Bureau of Standards (UNBS) is a parastatal under the Ministry of Tourism, Trade and 
Industry established under Cap 327 , of the Laws of Uganda. UNBS is mandated to co-ordinate the elaboration 
of standards and is  
(a) a member of International Organisation for Standardisation (ISO) and  
(b) a contact point for the WHO/FAO Codex Alimentarius Commission on Food Standards, and  
(c) the National Enquiry Point on TBT/SPS Agreements of the World Trade Organisation (WTO). 
The work of preparing Uganda Standards is carried out through Technical Committees. A Technical 
Committee is established to deliberate on standards in a given field or area and consists of representatives of 
consumers, traders, academicians, manufacturers, government and other stakeholders. 
Draft Uganda Standards adopted by the Technical Committee are widely circulated to stakeholders and the 
general public for comments. The committee reviews the comments before recommending the draft standards 
for approval and declaration as Uganda Standards by the National Standards Council. 
This Final Draft Uganda Standard, FDUS EAS 766-1 : 2013 , Antibacterial toilet soap — Specification — Part 1: 
Solid , is identical with  and has been reproduced from an East African  Standard , EAS 766-1: 2013,  
Antibacterial toilet soap — Specification — Part 1: Solid , and is being proposed for adoption as a Uganda 
Standard.
 
 
  This standard cancels and replaces US EAS 766:2011 which has been technically revised. 
 
Wherever the words, “ East African Standard" appear, they should be replaced by "Uganda Standard."  
  FOR COMMENTS ONLY EAS 766-1 : 2013 
ICS 77.100. 40 
© EAC 2013                                                                                                                         First Edition 2013  
 
EAST AFRICAN STANDARD 
Antibacterial toilet soap — Specification — Part 1: Solid 
EAST AFRICAN COMMUNITY FOR COMMENTS ONLYFOR COMMENTS ONLYEAS 766-1 : 2013  
ii © EAC 2013  – All rights reserved  
 Copyright notice 
This EAC document is copyright-protected by EAC. While the reproduction of this document by 
participants in the EAC standards development process is permitted without prior permission from EAC, 
neither this document nor any extract from it may be reproduced, stored or transmitted in any form for 
any other purpose without prior written permission from EAC. 
Requests for permission to reproduce this document for the purpose of selling it should be addressed 
as shown below or to EAC’s member body in the country of the requester:  
© East African Community 2013  — All rights reserved 
East African Community  
P.O.Box 1096 
Arusha 
Tanzania 
Tel: 255 27 2504253/8 
Fax: 255 27 2504481/2504255 
E-mail: eac@eachq.org 
       Web: www.eac-quality.net 
Reproduction for sales purposes may be subject to royalty payments or a licensing agreement. 
Violators may be persecuted FOR COMMENTS ONLYEAS 766-1 : 2013  
© EAC 2013  – All rights reserved  iii 
 Contents Page 
Foreword ............................................................................................................................................................  iv 
1 Scope  ................................ ................................ ................................ ................................ ...................... 1 
2 Normative ref erences  ................................ ................................ ............................................................  1 
3 Terms and definitions  ................................ ................................ ...........................................................  1 
4 Requirements  ................................................................ ................................ ................................ ......... 2 
4.1 General  requirements  ................................ ................................ ...........................................................  2 
4.2 Specific quality requirements  ................................ ................................ ................................ .............. 2 
5 Packaging and marking  ................................ ................................ ................................ ........................ 3 
5.1 Packaging  ................................ ................................ ................................ ................................ ............... 3 
5.2 Marking  ................................ ................................ ................................ ................................ ................... 3 
6 Sampling  ................................ ................................ ................................ ................................ ................ 3 
7 Crite ria for conformity ................................ ................................ ...........................................................  3 
Annex A (normative)  Permitted antibacterial agents ..................................................................................... 4  
Annex B (normative)  Determination of Trichlorocarbanilide (TCC)and TricIosan (TCN) in soaps 
by HPLC .................................................................................................................................................. 5  
B.1 Principle  ................................ ................................ ................................ ................................ ................. 5 
B.2 Normal phase HPLC  ................................ ................................ ..............................................................  5 
B.2.1  Reagents  ................................ ................................ ................................ ................................ ................ 5 
B.2.2  Apparatus  ................................ ................................ ................................ ................................ ............... 5 
B.2.3  Procedure  ................................ ................................ ................................ ................................ ............... 6 
B.2.4  Calculation  ................................................................ ................................ ................................ ............. 6 
B.3 Reverse phase  ................................................................ ................................ ................................ ....... 7 
B.3.1  Reagents  ................................ ................................ ................................ ................................ ................ 7 
B.3.2  Apparatus  ................................ ................................ ................................ ................................ ............... 7 
B.3.3  Procedure  ................................ ................................ ................................ ................................ ............... 7 
B.3.4  Calculations  ................................................................ ................................ ................................ ........... 8 
Annex C (normative)  Determination of chloroaniline .................................................................................... 9  
C.1 Principle  ................................ ................................ ................................ ................................ ................. 9 
C.2 Safety precautions  ................................ ................................ ................................................................  9 
C.3 Reagents  ................................ ................................ ................................ ................................ ................ 9 
C.4 Apparatus  ................................ ................................ ................................ ................................ ............. 10 
C.5 Procedure  ................................ ................................ ................................ ................................ ............. 10 
C.6 Determination of chloroanilines  ................................ ................................ ................................ ........ 11 
C.7 Calculations  ................................................................ ................................ ................................ ......... 11 
Annex D (normative)  Sampling ...................................................................................................................... 13 
D.1 Procedure  ................................ ................................ ................................ ................................ ............. 13 
D.2 Preparation of test samples  ................................ ................................ ................................ ............... 13 
D.2.1  Composite sample  ................................ ................................................................ ...............................  13 
D.2.2  Samples for testing  ................................ ................................ .............................................................  13 
 FOR COMMENTS ONLYEAS 766-1 : 2013  
iv © EAC 2013  – All rights reserved  
 Foreword 
Development of the East African Standards has been necessitated by the need for harmonizing requirements 
governing quality of products and services in the East African Community. It is envisaged that through 
harmonized standardization, trade barriers that are encountered when goods and services are exchanged 
within the Community will be removed.  
In order to achieve this objective, the Community established an East African Standards Committee mandated 
to develop and issue East African Standards.   
The Committee is composed of representatives of the National Standards Bodies in Partner States, together 
with the representatives from the private sectors and consumer organizations. Draft East African Standards 
are circulated to stakeholders through the National Standards Bodies in the Partner States. The comments 
received are discussed and incorporated before finalization of standards, in accordance with the procedures 
of the Community. 
East African Standards are subject to review, to keep pace with technological advances. Users of the East 
African Standards are therefore expected to ensure that they always have the latest versions of the standards 
they are implementing .  
EAS 766-1 was prepared by Technical Committee EAS/TC 074, Surface active agents . 
EAS 766 consists of the following parts, under the general title Antibacterial toilet soap — Specification —: 
Part 1: Solid.  
Part 2: Liquid.  
 FOR COMMENTS ONLYEAS 766-1 : 2013  
© EAC 2013  – All rights reserved  v 
 Introduction 
Human skin provides a favourable environment for the existence and multiplication of a variety of microbes. 
The conventional toilet soap washes away the germs but does not kill them. The function of an antibacterial or 
antiseptic toilet soap is not only to clean the skin, but also to reduce drastically the bacterial count on the skin. 
This prevents skin infections and perspiration odour caused by the decomposition of perspiration by bacteria. 
The antibacterial toilet soap is specially effective against staphylococcus  and similar bacteria which have the 
habit of residing in the under layers of skin. The antibacterials are substantive to the skin and this tackles the 
microbes between two washes. Antibacterial toilet soaps shall be used regularly to be effective. 
In the present edition hexachloroprene has not been permitted to use as antibacterial agent. 
Trichlorocarcanilide (TCC) on heating decomposes to chloroanilines which can be harmful to skin and hence 
the limit and method for determination of chloroaniline is incorporated.  
 
 
 FOR COMMENTS ONLYFOR COMMENTS ONLYEAST AFRICAN STANDARD  EAS 766-1: 2013  
 
© EAC 2013  – All rights reserved  1 
 Antibacterial toilet soap — Specification — Part 1: Solid 
1 Scope 
This East African Standard specifies the requirements and methods of sampling and test for solid antibacterial 
toilet soap. 
2 Normative references 
The following referenced documents are indispensable for the application of this document. For dated 
references, only the edition cited applies. For undated references, the latest edition of the referenced 
document (including any amendments) applies. 
EAS 127, Synthetic laundry detergents for household use — Specification  
EAS 377-1 , Cosmetics and cosmetics products — Part 1: List of substances prohibited in cosmetic products 
EAS 377-2, Cosmetics and cosmetics products — Part 2: List of substances which cosmetic products must 
not contain except subject to restrictions laid down  
EAS 377-3 , Cosmetics and cosmetics products — Part 3: List of colourants allowed in cosmetic products 
EAS 377-4 , Cosmetics and cosmetics products — Part 4: List of preservatives allowed in cosmetic products 
EAS 377-5 , Cosmetics and cosmetics products — Part 5: Use of UV filters in cosmetic products  
EAS 794, Determination of the microbial inhibition of cosmetic soap bars and liquid hand and body washes — 
Test method  
ISO 456, Surface active agents — Analysis of soaps — Determination of free caustic alkali  
ISO 685, Analysis of soap — Determination of alkali content and total fatty matter content  
ISO 673, Analysis of soap — Determination of matter insoluble in ethanol  
ISO 862, Surface active agents — Vocabulary  
3 Terms and definitions 
For the purposes of this standard, the terms and definitions given in ISO 862. FOR COMMENTS ONLYEAS 766-1: 2013  
2 © EAC 2013  – All rights reserved  
 4 Requirements 
 4.1 General requirements 
4.1.1  Solid antibacterial toilet soap shall be, thoroughly saponified, milled soap or homogenized soap or 
both, white or coloured, perfumed, and compressed in the form of firm and smooth cakes, tablets or bars and 
shall possess good cleaning, lathering and antibacterial properties. 
4.1.2 Perfume, moisture, normal colouring matters and preservatives acceptable in toilet soaps may be 
added. 
4.1.3 The solid antibacterial soap shall contain permitted antibacterial agent in accordance with Annex A.   
4.1.4 The solid antibacterial soap shall pass the antibacterial activity test when determined by the method 
given in EAS 794. 
4.1.5 All the substances used in the solid antibacterial soap shall comply with the requirements of all parts 
of EAS 377. 
4.1.6 Solid antibacterial toilet soap shall pass the test for dermatological safety.   
4.2 Specific quality requirements 
Solid antibacterial toilet soap shall comply with the specific quality requirements specified in Table 1. 
Table 1 — Specific quality requirement for solid antibacterial toilet soap 
SI No.  Characteristic  Requ irement  Test method  
i.  Total fatty matter, % by mass,  min 76.0 ISO 685  
ii.  Free caustic alkali, as sodium hydroxide 
(NaOH), % by mass,  max 0.1 ISO 456  
iii.  Free carbonated alkali, as sodium carbonate 
(Na 2CO 3), % by mass,  max 1.0 ISO 685  
iv.  Matter insoluble in al cohol, % by mass,  max 2.5 ISO 673  
v.  Antibacterial agent TricIosan (TCN) and 
Trichlorocarbanilide (TCC) , % by mass , max.  1 
either singly or in combination  Annex B  
vi.  Chloroaniline content , ppm, max  10 Annex C  
vii.  Synthetic detergent  Shall be absent  EAS 127  
viii.  Phosphate  Shall be absent  EAS 127  
NOTE  1 Solid antibacterial Toilet soap is liable to lose moisture on storage. The results of analysis in 
respect to free caustic alkali, free carbonated alkali and matter insoluble in alcohol should be recalculated in 
relation to the minimum specified total fatty matter by means of the following equation:   
matterfatty  total actualmatterfatty  total specified minimum  result  actualresult Corrected
 
The corrected results should  be used to determine whether the product is up to standard.  
NOTE 2  Trichlorocarbanilide (TCC) is not heat stable and decomposes into chloro anilines on prolonged 
heating above 60 °C. If TCC is used in soap, the manufacturer should take care that such soap is not 
subjected to temperature above 60°C during the entire manufacturing process or during storage.  
 FOR COMMENTS ONLYEAS 766-1: 2013  
© EAC 2013  – All rights reserved  3 
 5 Pack aging and marking 
5.1 Pack aging 
The product shall be packed in a suitable, well-closed container, to protect the integrity of the product during 
transportation and sale. 
5.2 Marking 
The packages shall be securely closed and marked with the following particulars: 
a) name of the product as “antibacterial toilet soap” ;  
b) manufacturer ’s name and physical address;  
NOTE The name, physical address of the distributor/supplier and trade mark may be added as required. 
c) net content; 
d) batch number or lot number ;  
e) date of manufacture and best before date ;  
f) Total Fatty Matter content (TFM); 
g) country of origin; 
h) the antibacterial agent used and its level; and 
i) list of ingredients in descending order of quantity 
6 Sampling 
Sampling shall be done in accordance with Annex D. 
7 Criteria for conformity 
The lot shall be deemed to comply with the requirements of this standard if, after inspection and testing, the 
requirements of Clause 4 and 5 are satisfied.  FOR COMMENTS ONLYEAS 766-1: 2013  
4 © EAC 2013  – All rights reserved  
 Annex A 
(normative) 
 
Permitted antibacterial agents 
The following is the list of antibacterial agents used generally in antibacterial soap:  
a) Triclosan (TCN); 
b) Trichlorocarbanilide (TCC); 
c) Zinc oxide;  
d) Chloro xylenols; 
e) Plant extracts; 
f) Any other internationally accepted antibacterial agent. FOR COMMENTS ONLYEAS 766-1: 2013  
© EAC 2013  – All rights reserved  5 
 Annex B 
(normative) 
 
Determination of Trichlorocarbanilide (TCC)and TricIosan (TCN ) in soaps 
by HPLC 
B.1 Principle 
TCC and TCN are antibacterial agents, which are separated from other components in soap by normal phase 
or reverse phase liquid chromatography, detected spectrophotometrically and quantified by comparison with 
standard TCC and TCN. The method can estimate as low as 1 ppm of the above compounds: 
Procedures for both normal and reverse HPLC has been described and provide the option to use either 
method whichever is available to the users. Both methods are comparable. 
B.2 Normal phase HPLC 
B.2.1 Reagents 
B.2.1.1 Iso-octane , HPLC grade 
B.2.1.2 Iso-propanol (2-propanol) , HPLC grade 
B.2.1.3 Hexane , HPLC grade 
B.2.1.4 Standard TCC , 99 % pure 
B.2.1.5 Standard TCN , 99 % pure 
B.2.2 Apparatus 
B.2.2.1 High Performance Liquid Chromatograph consisting of a pump, a sample injector of fixed 
volume with UV detector having variable wavelengths and a recorder 
B.2.2.2 Standard volumetric flasks  
B.2.2.3 Pipettes 
B.2.2.4 Magnetic stirrer 
B.2.2.5 Millipore filter appara tus with 0.5 micron filter  
B.2.2.6 Column , comprising:  
a) Silica column , stainless steel 25 cm x 0.46 cm packed with Normal phase-silica 5 micron 
(Lichrosorb Si -60) ; or 
b) Cyano column , stainless steel 25 cm x 0.40 cm packed with (Lichrospher 100) cyano 5 
micron. 
NOTE Either, of the above columns can be used depending on the availability. FOR COMMENTS ONLYEAS 766-1: 2013  
6 © EAC 2013  – All rights reserved  
 B.2.2.7 Mobile phase: 
a) for silica column, transfer 20 mL of iso-propanol into a 500-mL volumetric flask and make up to mark 
with iso-octane and mix well. Assemble millipore filter apparatus and filter the solvent system prior to 
use; and 
b) for cyano column, transfer 50 mL of HPLC grade iso-propanol (2-propanol) into a 500-mL volumetric 
flask, fill up to the mark with hexane and mix well. Assemble millipore filter apparatus and filter the 
solvent system prior to use. 
B.2.2.8  HPLC conditions  which include the following: 
a) Detector wavelength flow rate: 280 nm; 
b) Flow rate:      0.5 m L/min; 
c) Injection volume:    20 µL; 
d) Retention time; 
e) Silica column: 
 TCN - 7.5 min ; and   
 TCC - 19.2 min; 
f) Cyano column: 
 TCN - 4.0 min; and  
 TCC - 7.5 min. 
B.2.3 Procedure 
B.2.3.1 Standard preparation (see note under B.3.4) 
Weigh accurately 25 mg of Triclosan (TCN) and 25 mg of TCC into a 100-mL volumetric flask and make up to 
volume with the mobile phase and mix well. Pipette 1.0 mL of this solution in a 50 mL volumetric flask and 
dilute with mobile phase. Final concentration of TCC and TCN is 250 µg/50 mL (5.0 ppm). 
B.2.3.2 Sample preparation 
Weigh accurately 1 g of homogenized sample into a 100-mL standard flask, and dilute to the mark with mobile 
phase. Pipette 10 mL of the supernatant liquid to a 50-mL volumetric flask, dilute with mobile phase, to the 
mark, and filter through 0.45 m filter. 
B.2.3.3 Chromatography 
Equilibrate the column, maintained at a temperature of 30 °C, with the mobile phase with a flow rate of 0.5 mL 
/min for iso-octane - iso-propanol mobile phase and 1.0 mL/min for Hexane - iso-propanol mobile phase for 30 
min. Set the wavelength at 280 nm. Inject 20 µL of standard solution and then sample solutions. 
Measure area of the peaks of respective retention time for standard and sample. 
B.2.4 Calculation 
TCN shall be expressed as follows: FOR COMMENTS ONLYEAS 766-1: 2013  
© EAC 2013  – All rights reserved  7 
 
100x
sample of  ion Concentratx TCN standard of AreaTCN standard of ion Concentratx TCN for sample of Areamassby  percent TCN,  
100x
sample of  ion Concentratx TCC standard of AreaTCC standard of ion Concentratx TCC for sample of Areamassby  percent TCC, 
 
B.3 Reverse phase 
B.3.1 Reagents 
B.3.1.1 Methanol, HPLC grade 
B.3.1.2 Sodium Dihydrogen Phosphate Monohydrate , chemical grade  
B.3.1.3 Standard TCC 
B.3.1.4 Standard TCN (TCS) 
B.3.2 Apparatus 
B.3.2.1 Column 
B.3.2.1.1 Octyldimethylsilyl (C- DB) 
B.3.2.1.2 Supercosil LC-8- DB, 15 cm x 4.6 mm . 5 micron 
B.3.2.2 Mobile phase 
MeOH/0.01 M Phosphate buffer 62:38 v/v 
0.01 M Phosphate buffer: Dissolve 1.38 g sodium dihydrogen phosphate monohydrate in 1 000 mL of distilled 
water. Prepare to pH 3.0 by 10 % phosphate solutions. 
B.3.3 Procedure 
B.3.3.1 Standard preparation (see Note under B.3.4) 
B.3.3.1.1 Weigh accurately about 90 mg of TCN. Dissolve in methanol and make up to 1 000 mL volumetric 
flask with methanol. 
B.3.3.1.2 Weigh about 110 mg of TCC, dissolve well with methanol, and make up the volume to 1 000 mL. 
B.3.3.1.3 Accurately pipette 10 mL of the solution prepared in B.3.3.1.1 into the volumetric flask containing 
TCC (B.3.3.1.2). And make up to the volume with methanol. Then accurately pipette 5 ml of the solution into a 
50-mL volumetric flask. Make up to the volume with methanol. Filter this standard solution through 0.45 µm 
filter. 
B.3.3.2 Sample preparation 
Weigh accurately about 1.0 g of product, dissolve in methanol and make up to 100 mL in a volumetric flask 
with methanol. Filter this sample solution through 0.45 m filter. FOR COMMENTS ONLYEAS 766-1: 2013  
8 © EAC 2013  – All rights reserved  
 B.3.3.3 HPLC conditions 
The HPLC conditions include the following: 
a) Detector wavelength: 280 nm; 
b) Column temperature: 35 ºC; 
c) Flow rate:    1.0 m L/min; and 
d) Injection volume:  10 µL. 
Prepare the standard solution and the sample solution at the same time. Inject the standard solution three 
times and calculate the average of each ingredients peak count. Inject 10 µg the sample solution and 
determine each ingredients percentage by the calculation shown. 
B.3.4 Calculations 
The TCN and TCC shall be expressed as follows: 
TCN, percent by mass = 
 
 100 t sr s

MAFA M  
TCC, percent by mass =  
 
 100 t sr s

MAFA M  
where 
Ar is the peak area of the test sample; 
As is the averaged peak area of the standard;  
F is the purity, expressed as percent, of the standard; 
Ms is the mass, in grams, of the standard; and 
Mt is the mass, in grams, of the test sample. 
NOTE Both TCC and TCN are photosensitive, hence standards should be freshly prepared. FOR COMMENTS ONLYEAS 766-1: 2013  
© EAC 2013  – All rights reserved  9 
 Annex C 
(normative) 
 
Determination of chloroaniline 
C.1 Principle 
The chloroanilines are extracted from soap with dimethyl sulfoxide and diazotized with nitrous acid. The 
reaction products are then coupled with N-1-(naphthyl) ethylenediamine hydrochloride to produce coloured 
compounds which are estimated spectrophotometrically. 
C.2 Safety precautions 
Dimethyl sulfoxide (DMSO) is readily absorbed into the skin. Inhalation or skin penetration must be avoided. 
DMSO should never be pi petted using mouth. Always use pipette bulb. The standard chloroanilines and N-1-
(naphthyl)-ethylenediamine hydrochloride shall not be allowed to come into contact with the skin. If they 
should, then wash the contaminated parts thoroughly with soap and water. 
A supply of diluted sodium hypochlorite should be at hand at all times to deal with accidental spillages of 
chloroaniline solution. Spillage on laboratory surface should be treated immediately with the sodium 
hypochlorite solution, followed by water. 
C.3 Reagents 
C.3.1 Dimethyl Sulphoxide (DMSO) , AR grade 
C.3.2 Hydrochloric a cid, concentrated (specific gravity, 1.18) 
C.3.3 Sodium nitrite , 0.4 % w/v  analytical grade, freshly prepared (aqueous) 
C.3.4 Ammonium sulphamate , 2 % w/v  solution freshly prepared (aqueous) 
C.3.5 N-1-(naphthyl) ethylene , 0.1 % w/v  solution diamine hydrochloride freshly prepared (aqueous) 
C.3.6 n-Butanol , AR grade 
C.3.7 Sand , acid purified 40 - 100 micron mesh 
C.3.8 Solvent mixture comprising:  
 DMSO   5 volumes 
 n-Butanol   2 volumes 
 Distilled water  2 volumes 
 Hydrochloric acid 1 volume 
Mix n -butanol, water and HCI. Cool the mixture and add DMSO. 
C.3.9 4-Chloroaniline and 3, 4-Dichloroaniline, AR grade FOR COMMENTS ONLYEAS 766-1: 2013  
10 © EAC 2013  – All rights reserved  
 C.4 Apparatus 
C.4.1 Spectrophotometer,  suitable for use at 554 nm 
C.4.2 Cuvettes, glass (matched pair) 10 mm 
C.4.3 Water bath,  thermostatically controlled at 25 °C 
C.4.4 S topwatch 
C.4.5 Standard laboratory glassware  
C.4.6 Filter Paper,  Whatman No. 541 
C.5 Procedure 
C.5.1  Dissolve 0.349 8 g of 3,4-dichloroaniline and 0.2753 g of 4-chloroaniline in solvent mixture (see  C.3.8) 
in a 250 mL amber volumetric flask. 
Dilute to mark with solvent mixture [1 mL is 2.5 mg mixed chloroanilines (stock solution)]. 
C.5.2 Dilute this stock solution with solvent mixture as given below: 
a) take 5 mL of stock solution and dilute it to 250 mL with solvent mixture (1 mL = 50 µg mixed 
chloroanilines); and 
b) take 5 mL of the above solution [see  (a)] and further dilute to 250 mL with solvent mixture. [1 m L = 1 
µg mixed chloroanilines]. 
Use this solution for preparation of calibration curve. 
Transfer using a burette 0, 1 mL , 2 m L, 5 mL , 10 m L, 20 m L, 40 m L into 50 mL amber volumetric flasks. 
C.5.3 From a burette, add sufficient solvent mixture to make total volume to 40-mL in each flask. The flasks 
are incubated in a water bath at 25 °C for 20 min: After exactly 20 min, add 2-mL of reagent (see  C.3.3) into 
each flask and return them to the water bath for exactly 10 min (measure with a stop watch). 
Then add 2 mL of reagent (see  C.3.4) into each flask and return them to the water bath for exactly 10 min. 
Swirl the flask occasionally. 
Then add 2 mL of reagent (see  C.3.5) into each flask and remove them from the water bath. Dilute to volume 
with distilled water, mix and allow to stand for 30 min. Measure absorbance at 554 nm against the blank 
solution as prepared in C.5.4. 
C.5.4 In preparing the blank solution, take 40 mL of solvent mixture in a 50 mL amber volumetric flask. 
Incubate the flask in a water bath at 25 °C for 20 min. After exactly 20 min, add 2 mL of reagent (see C.3.3 ) 
into the flask and return it to the water bath for exactly 10 min. Then add 2 mL of reagent (see C.3.4) into the 
flask and return it to the water bath for exactly 10 min (swirl the flask occasionally). Then add 2 m L of reagent 
(see C.3.5 ) into the flask and remove it from the water bath. Dilute to volume with distilled water, mix and 
allow to stand for 30 min. Use this blank solution for preparation of calibration curve only. 
C.5.5 Prepare a graph by plotting weight ( g) of chloroanilines contained in each 50 mL-flask against 
absorbance. The linear calibration will pass through the origin/or determine the average absorbance ( AA) of 1 
g of mixed chloroanilines by dividing sum of absorbances of all different aliquots of the standard by sum of 
g of chloroanilines in all different aliquots of standard. FOR COMMENTS ONLYEAS 766-1: 2013  
© EAC 2013  – All rights reserved  11 
 C.6 Determination of chloroanilines 
C.6.1 Weigh to the nearest mg 3.0 g - 15 g of finely grated soap and add 10.0 g - 15.0 g of acid purified 
sand. Transfer quantitatively the sample and the sand into a mortar and grind the mixture thoroughly with a 
pestle to give a homogenous mass. Transfer the mass to a previously weighed 250-m L flat bottom flask 
quantitatively and reweigh. Add DMSO (100 mL), stopper firmly and attach the flask to an automatic shaker. 
Shake for 1 h. Filter the DMSO extract through Whatman No. 541 into a 250 m L amber volumetric flask. Wash 
the flask and filter paper with small aliquots of DMSO. Allow the filtrate to drain completely, dilute to volume 
with DMSO and mix. Transfer 20 mL DMSO extract into a 50-mL amber volumetric flask. Add 20 m L of 
solvent mixture. The flask is incubated in a water bath at 25 °C for 20 min. After exactly 20 min, add 2 mL of 
reagent (see  C.3.3) into the flask and return it to the water bath for exactly 10 min (measure with a stop 
watch). Then add 2 mL of reagent (see  C.3.4) into the flask and return it to the water bath for exactly 10 min 
(swirl the flask occasionally). Then add 2 mL of reagent (see  C.3.5) into the flask and remove it from the water 
bath. Dilute to volume with distilled water, mix and allow to stand for 30 min. Read the absorbance at 554 nm 
against blank (prepared as below). 
C.6.2 Prepare the blank solution by mixing 20 mL of DMSO extract of sample and 20 mL of solvent mixture 
in a 50 mL amber volumetric flask. Incubate the flask in a water bath at 25 °C for 20 min.  
After exactly 20 min, add 2 mL of distilled water into the flask and return it to the water bath for exactly 10 min. 
Then add 2 mL of reagent (see  C.3.4) into the flask and return it to the water bath for exactly 10 min (swirl the 
flask occasionally). Then add 2 mL of reagent (see C.3.5) into the flask and remove it from the water bath. 
Dilute to volume with distilled water, mix and allow to stand for 30 min. Use this solution as a blank for reading 
sample only. 
C.6.3 Deduce the amount of chloroanilines (µg) from the calibration graph curve. 
NOTE The determination should be completed in one day. 
C.7 Calculations 
Determine the amount of mixed chloroanilines in the aliquot of test solution from the calibration graph. 
Chloroaniline content (in ppm) = 

MMMMM
23 1
20250  
where 
M is the mass, in grams, of soap; 
M1 is the mass, in grams, of sand; 
M2 is the mass, in grams, of soap and sand transferred to the flask ; and  
M3 is the mass, in micrograms, of mixed chloroanilines found from calibration graph/or it can be 
calculated as given below: 
M3 = 
(AA) ines chloroanil mixed g  1 of absorbance Averagesample the of Mass
μ  
where 
AA = 
g in ines chloroanil standard of ion concentrat of Sumstandards the of OD the of Sum
μ  
 
 FOR COMMENTS ONLYEAS 766-1: 2013  
12 © EAC 2013  – All rights reserved  
 Weight of soap actually used, in g  = 
12
MMMM
  
 FOR COMMENTS ONLYEAS 766-1: 2013  
© EAC 2013  – All rights reserved  13 
 Annex D 
(normative) 
 
Sampling  
D.1 Procedure  
D.1.1 In a single consignment, all packages (cartons) containing toilet soap cakes drawn from the same 
batch of production shall constitute a lot. For ascertaining the conformity of the lot to the requirements of this 
standard, tests shall be carried out on each lot separately. The number of packages to be selected for drawing 
the sample shall be in accordance with Table D .1.  
Table D .1 — Scale of sampling 
Number of packages (cartons) in 
the lot  
N Number of packages (cartons) to be 
selected  
n Number  of samples  
4 to 15  3 3 
16 to 40  4 4 
41 to 65  5 2 
66 to 110  7 2 
111 an d above  10 1 
 
D.1.2 The packages shall be selected at random, using tables of random numbers. If these are not 
available, the following procedure shall be applied:  
Starting from any package, count all the packages in one order as 1, 2, 3.... N, selecting every k th package, 
where k is the integral part of N  n.  
D.1.3 From each package thus selected, draw at random an equal number of cakes so as to obtain a total 
mass of at least 2 kg.  
D.2 Preparation of test samples 
D.2.1 Composite sample  
Weigh each cake separately (including any material that may have adhered to the wrapper), and calculate the 
average mass. Cut each of the remaining cakes into eight parts by means of three cuts at right angles to each 
other through the middle. Grate finely the whole of two diagonally opposite eighths of each specimen. Mix the 
gratings and place in a clean, dry, airtight glass container.  
D.2.2 Samples for testing 
Immediately after preparation of composite sample (D .2.1), take at one time all test samples required for the 
tests in 4.2. Weigh out the test sample required for determination of free alkali or acid content, and use it 
immediately.  
 FOR COMMENTS ONLYFOR COMMENTS ONLYEAS 766-1: 2013    
 
© EAC 2013  – All rights reserved