PUBLIC REVIEW DRAFT DRAFT UGANDA STANDARD  
  DUS DEAS 96-1 
First  Edition  
2016-mm-dd 
 
 Reference number  
DUS DEAS 96-1: 2016 
© UNBS 201 6 
  
Sanitary towels —Specification — Part 1: Disposable  
 PUBLIC REVIEW DRAFTDUS DEAS 96-1: 2016  
ii                © UNBS  2016  - 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 2016 
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: 256 417 333 2 50/1/2/3 
Fax: 256 414 286 123 
E-mail: info@unbs.go.ug  
Web: www.unbs.go.ug  
 PUBLIC REVIEW DRAFTDUS DEAS 96-1:2016 
© UNBS  2016 - All rights reserved                iii National foreword 
Uganda National Bureau of Standards (UNBS) is a parastatal under the Ministry of Trade, Industry and 
Cooperatives established under Cap 327, of the Laws of Uganda, as amended.  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 Agreement 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 Draft Uganda Standard, DUS DEAS 96-1: 2016, Sanitary towels —Specification — Part 1: Disposable,  is 
identical with  and has been reproduced from a Draft East African Standard , DEAS 96-1 : 2016, Sanitary towels 
—Specification— Part 1: Disposable,  and is being proposed for adoption as a Uganda Standard. 
This standard cancel and replaces US EAS 96:2008 , Sanitary towels - Specification , which has been 
technically revised. 
This standard was developed by the Subcommittee on Textiles and textile products (SC 1) under the Textiles, 
leather, paper and related products’ Standards Technical Committee (UNBS/TC 7).  
Wherever the words, “ East Afric an Standard" appear, they should be replaced by "Uganda Standard." 
 
   DEAS 96 -1: 2016  
ICS 59.080.30  
© EA C 2016 ……………………………………………………………………………………… Second Edition 2016   
 
DRAFT EAST AFRICAN STANDARD  
Sanitary towels —Specification — Part 1: Disposable  
EAST AFRICAN COMMUNITY  DRAFT  EAST AFRICAN STANDARD  DEAS 96:2016  
 
© EA C 2016 ……………………………………………………………………………………… Second Edition 2016   
 Copyright notice  
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participants in the EAC standards developmen t process is permitted without prior permission from EAC, 
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© East African Community 2016  — All rights reserved  
East African Community   
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Reproduction for sales purposes may be subject to royalty payments or a licensing agreement. 
Violators may be persecuted   
© EAC 2016  – All rights reserved  Contents  Page 
Foreword  ................................ ................................ ................................ ................................ .............................. 3 
1 Scope  ................................ ................................ ................................ ................................ ...................... 4 
2 Normative references  ................................ ................................ ................................ ............................ 4 
3 Definitions  ................................ ................................ ................................ ................................ .............. 4 
4 Requirements  ................................ ................................ ................................ ................................ ......... 5 
4.1 Materials  ................................ ................................ ................................ ................................ ................. 5 
4.1.1  Absorbent filler  ................................ ................................ ................................ ................................ ......5 
4.1.2  Covering  ................................ ................................ ................................ ................................ ................. 5 
4.1.3  Protective barrier  ................................ ................................ ................................ ................................ ...5 
4.2 Workmanship and finish  ................................ ................................ ................................ ....................... 5 
4.2.1  Absorbent filler  ................................ ................................ ................................ ................................ ......5 
4.2.2  Securing mechanism  ................................ ................................ ................................ ............................. 5 
4.4 Microbiological requirements  ................................ ................................ ................................ ............... 6 
Annex  A (normative)  Determination of water soluble colouring matter  ................................ ...................... 9 
Annex  B (normative)  Determination of water -resistance of protective barrier (cone test method)  ....... 10 
Annex  C (normative)  Method for determination of absorbe ncy capacity)  ................................ ................  12 
Annex  D (normative)  Method for determination of absorbency rate  ................................ .........................  13 
Annex  E (normative)  Determination of moist ure content  ................................ ................................ ...........  14 
Annex  F (normative)  Determination of water soluble extract  ................................ ................................ .... 16 
Annex  G (normative)  Determination of fluorescence  in sanitary towels  ................................ ..................  18 
Annex  H (normative)  Determination of flushability  ................................ ................................ .....................  19 
Annex  J (normative)  Microbiological examination  ................................ ................................ ......................  20 
Bibliography  ................................ ................................ ................................ ................................ .....................  24 
 
  
 Foreword  
Development of the East African Standards has been necessitated by the need for harmonizing requirements 
governing quality of produc ts 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 pri vate 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 acco rdance 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 stand ards 
they are implementing . 
 
EAS 96 was prepared by Technical Committee EAS/TC 061, Textiles and Textile Products . 
 
This second edition cancels and replaces the first edition (EAS  96:2008), which has been technically revised  
  
© EAC 2016  – All rights reserved  Sanitary towels —Specificatio n — Part 1: Disposable  
1 Scope  
This Draft East African Standard specifies the requirements for disposable  sanitary towels  also called sanitary 
pads/s anitary napkins . This standard does not apply to reusable sanitary towels .  
2 Normative references  
The foll owing referenced documents are indispensable for the application of this standard. For dated 
references, only the edition cited applies. For undated references, the latest edition of the referenced 
document (including any amendments) applies  
ISO 139, Texti les — Standard atmospheres for conditioning and testing  
ISO 3071 , Textiles -- Determination of pH of aqueous extract  
ISO 4833 Microbiology of the food chain -- Horizontal method for the enumerati on of microorganisms c olony 
count at 30 degrees C  
ISO 7218, M icrobiology of food and animal feeding stuffs -- General requirements and guidance for 
microbiological examinations  
ISO 6888 -2:1999 Microbiology of food and animal feeding stuffs -- Horizontal method for the enumeration of 
coagulase -positive staphylococci (Staphylococcus aureus and other species) -- Part 2: Technique using rabbit 
plasma fibrinogen agar medium  
3 Definitions  
For the purposes of this East African Standard, the following definitions shall apply:  
3.1  
sanitary towel  
a hygienic composite produc t with a porous outer covering and a highly absorbent filler  
3.2  
package  
the smallest unit of sanitary towels as declared by a manufacturer that can be purchased by a consumer   
 4 Requirements  
4.1 General  
All materials, dyes and chemicals used in the manu facture of sanitary towels  shall not cause any 
undesirable effects  to the skin.  
4.2 Materials  
4.1.1  Absorbent filler  
When visually examined, the absorbent filler shall be white or light in colour and shall be free from any water 
soluble colouring matter wh en tested in accordance with Annex A. It shall not contain extraneous materials, 
which are not designed to enhance performance.  
4.1.2  Covering  
The absorbent filler covering shall be made of good quality fabric with sufficient porosity to permit the 
assemb led towel to meet absorbency requirement.  
4.1.3  Protective barrier  
The protective barrier shall be water resistant (no wetting of outer surface and no water penetration) when 
tested in accordance with Annex B.  
4.2 Workmanship and finish  
4.2.1  Absorbent f iller 
The absorbent filler shall be continuous and neatly cut to the required size. It shall be free from hard lumps. It 
shall be completely covered and free from wrinkles that are not a design feature.  
4.2.2  Securing mechanism  
Any o f the following may be  used:  
a) Loops or tabs which shall extend beyond the le ngth of the filler material;  
b) Adhesive strip or patch;  
c) Wings  with adhesive which shall be of sufficient length in such a manner as to form folds around the 
panty/brief for securing the sanitary towel when  in use.  
4.2.3 Protective barrier  
The sanitary towel shall have a protective barrier on one side, which shall be clearly identifiable with a mark, 
colour or some other means.  
4.2.4 Freedom from defects  
The sanitary towel when visually examined shall be free from defects, which affect the appearance and utility 
such as oil stains, dirt, soil particles and hard lumps.  
4.2.5 Odour  
The sanitary towel shall have no unpleasant odour either in dry state immediately after sampling from the 
packages or after we tting the sample with distilled water.   
© EAC 2016  – All rights reserved  4.2.6 Texture  
The sanitary towel shall be smooth and soft when felt by hand.  
Note  If harsh absorbent fillers or cover fabrics are used in the manufacture of sanitary towels, these may cause 
discomfort and body rash es on the delicate skin due to undesired friction.  
4.3 Performance requirements  
The sanitary towel shall comply with the performance requirements given in Table 1.  
Table 1 — Performance requirements for sanitary towels  
Characteristics  Requirement  Test method  
Absorbency capacity  No leakage  Annex C  
Absorbency rate ,s, max.  10 Annex D  
pH of aqueous extract”  5.5 – 8.5 ISO 307 1, Method B1 
Moisture content of filler material , % 
m/m, max.  8 Annex E  
Water sol uble extract of filler material, 
(%), m/m,  max.  1.0 Annex F  
Fluorescence of filler material  None  Annex G  
Size (mm) min.  Width 60  
Length 180  Measure by a ruler  
1 Incase a jelly forms, dilute with more distilled water before determining the pH. 
 
4.5 Microbiological requirements  
4.5.1 When  packed in st erile conditions as declared by the manufacturer (see 6.1 c)) sanitary towels shall 
pass tests for sterility when tested in accordance with ISO 6887 -1. 
4.5.2  When packed in non -sterile condition:  
a) The total viable bacterial count, when determined in accorda nce with J.4 a) shall not exceed 1000 per 
gram of  sanitary towel;  
b) When tested in accordance with J.4 b), c) and d), sanitary towels shall be free from 
Enterobacteriaceae, Staphylococcus aureus, and Pseudomonas aeruginosa respectively.  
4.5 Flushability  
When declared to be flushable in water closets, sanitary towels shall be manufactured from dispersible  
material, which shall pass the test described in Annex H.  
5  Packaging  
5.1  Package  
Sanitary towels shall be supplied in packages made of suitable mater ials, which are sealed so as to protect 
them from moisture, soiling and contamination during storage and transportat ion.  
 5.2  Bulk packaging  
When supplied in bulk, the bulk package shall be made of materials, which are strong enough to hold the 
number of d eclared packages and protect the quality of the product during handling, transportation and 
storage. It shall be properly sealed to prevent the packages from spilling. Only packages with the same  
batch number and containing the same size shall be packed to gether in a bulk package.  
6  Marking  
6.1  Packages  
The following information shall appear legibly and indelibly on the outside of each package:  
a) the manufacturer’s nam e and/or registered trade mark;  
b) the words “Sanitary towels/ sanitary napkins/ sanitary pads”;  
c) words ‘Sterile’ if applicable  (4.4.1);  
d) words or symbol indicating whether flushable ( if so declared/or claimed 4.5);  
e) secu ring mechanism (as per 4.2.2.);  
f) number o f sanitary towels in a package;  
g) batch identification number;  
h) country or region of manufa cture;  
i) disposal instructions;  
j) storage instructions  
k) date of manufacture.  
l) expiry date  
m) size of the sanitary towel  
6.2 B ulk packages  
The following information shall appear legibly and indelibly on the outside of each bulk package : 
a) the manufacturer’s name and/ or registered trade mark;  
b) the words “ Sanitary towels/sanitary napkins/sanitary pads ”. 
c) Number of packages in a bulk package   
d) expiry date  
7 Sampling  
7.1 Lot 
In any consignment all packages of the sanitary towels of the same size and type belonging to one ba tch of 
manufacture or supply shall constitute a lot.   
© EAC 2016  – All rights reserved  7.2 Scale of sampling  
7.2.1  Samples shall be tested from each lot ascertaining its conformity to the requirements of this 
specification.  
7.2.2  The number of packages to be selected from a lot shall be in  accordance with Table 3. 
Table 3 –scale of sampling  
Number of packages in a lot  Number of packages to be selected  
Up to 250  6 
251-500 8 
501-1000  11 
1001 -2500  15 
2501 -5000  20 
5001 and above  30 
 
7.2.3 The bulk packages and packages shall be selected at random.  
7.3 Number of tests  
7.3.1  Each package selected as  per table 3  shall be inspected for packaging and marking requirements  
7.3.3  Sanitary  towel s selected as  per table 3  shall be examined for requirements  stipulated in clause 4  
7  
 Annex  A 
(normativ e) 
 
Determination of water soluble colouring matter  
A.1 Principle  
Absorbent filler material is extracted in ethanol and then viewed for any colouring matter.  
A.2 Apparatus  
A.2.1  Weighing balance  
A.2.2  Narrow percolator  
A.2.3  Cylindrical glass tube  
A.3 Procedure  
Extract 10 g of absorbent filler material in 100 mL ethanol in a narrow percolator until 50 mL of the extract are 
obtained. Pour the liquid into a clean cylindrical glass tube at least 20 cm wide and view the layer on a white 
background.  
A.4 Test report  
Bluish or greenish shade indicates the presence of colouring substance  
 
 
 
  
© EAC 2016  – All rights reserved  Annex  B 
(normative)  
 
Determination of water -resistance of protective barrier (cone test 
method)  
B.1 Apparatus  
B.1.1  Funnel, metallic, glass or plastic of sufficient si ze for holding the test piece with water.  
B.1.2  Glass, container for collecting water under the glass funnel.  
B.1.3  Burette, for introducing water into the test piece  
B.2  Test piece preparation  
Cut a square test piece of approximately 6.5 mm in length  from the protective barrier and fold into a cone 
without creas ing the folds (see Figure B.1).  
 
Figure B.1 — Folding of specimens   
 
 
Figure B.2 — Test apparatus  
B.3 Procedure  
Assemble the apparatus as shown in Figure B.2. Pour slowly approximately 5 mL of distilled water into the 
cone assembly. Let it stand for 24 h  
A.4 Test report  
Observe for water in the glass container and wetness of the outer surface of the cone  
 
 
 
 
 
 
 
  
© EAC 2016  – All rights reserved  Annex  C 
(normative)  
 
Method for determination of absorbency capacity)  
C.1 Apparatus  
C.1.1 Flat level surface  
C.1.2 Burette  
C.1.3 Metallic block , of mass 1 kg and dimensions 150 mm x 50 mm x 15 mm  
C.2 Reagents  
1 % solution of potassium dichromate made by dissolving 1 g K2Cr2O7 in 100 mL distilled water.  
C.3 Procedure  
C.3.1 Lay the s anitary towels on a flat level surface.  
C.3.2 Drip at the rate of 15 mL per minute, 30 mL of the fluid (see C.2) on to the centre of sanitary towel from a 
height of approximately 2 mm.  
C.3.3 After the towel has absorbed the full amount of fluid, place a metallic block of mass 1 kg (C.1.3) for one 
minute on the portion where the fluid was absorbed.  
C.4 Test report  
Observe the back and sides of the sanitary towel for any leakage.  
 
 
 
 
  
 Annex  D 
(normative)  
 
Method for determination of absorbency rate  
D.1 Apparatus  
D.1.1  Water tub , tub of depth at least 100 mm and maintained at room temperature.  
D.1.2  Stop watch , with an accuracy of 0.2 s.  
D.1.3  Cylindrical basket , weighing 2.7 ± 0.3 g, of height 80 mm, diameter 50 mm with square opening of  
15 mm to 20 mm,  made of copper wire of 0.4 mm diameter.  
D.1.4  Weighing machine  
D.2 Preparation of test specimens  
Carefully isolate the absorbent filler material and weigh 5 g; insert into the basket.  
D.3 Procedure  
Drop the test specimen in a horizontal position into the water tub Using the stopwatch, measure the time it 
takes the basket and its contents to sink below the surface of water in seconds. Record the absorption period 
to the nearest 0.1 s. Repeat the test for at least two test specimens.  
D.4 Calculation  
Calculate the arithmetic mean of the absorbency rate of the absorbent filler material tested.  
 
 
 
  
© EAC 2016  – All rights reserved  Annex  E 
(normative)  
 
Determination of moisture content  
1 Principle  
A specimen of specified mass of filler material of Sanitary towel is dried in an oven at sp ecified temperature 
and the moisture content is determined.  
E.2 Apparatus  
E.2.1  Balance , with an accuracy of 0.05 % of the weighed mass  
E.2.2  Sample container  
Waterproof when sealed, will be used for transfer of analyzed material and during weighing.  
E.2.3  Oven , well ventilated with a temperature of 102 °C to 105 °C  
E.3 Sample preparation  
E.3.1  Take a sufficient number of dry sample containers, number them and take their masses after they 
are held open for a short period of time so that they will hav e the same air pressure as the surrounding 
atmosphere. Then leave them open until you take the test piece.  
E.3.2  Take 5 random pieces from the absorbent filler material of sanitary towel.  T he test pieces shall 
weigh 5 g.  
E.3.3  If the surrounding atmosphe re is hot and humid, prevent water condensation on the internal and 
external surfaces of the container.  
E.3.4  Handle the test pieces gently to prevent dirt or changes in water content.  Don’t touch the test pieces 
with your bare hands. Put the test pieces  in a container just after taking them and close the container 
immediately.  
E.4 Procedure  
E.4.1  Dry the test pieces in an oven with a temperature of 102 ºC to 105 °C. Open the containers lid and 
dry the specimen inside the container. Open the container f or a moment, to balance the air pressure inside the 
container with the surrounding pressure, weigh the container that holds the specimen again and calculate the 
weight of the specimen.  
E.4.2  First cycle of drying will last at least 30 minutes.  Return the  container with the test pieces to the 
oven, for at least half the first cycles drying time. Take the container out and take the mass with the test 
pieces inside.  Repeat the drying and weighing cycles.  When the drying time on every cycle is at least half  of 
the total previous drying cycle times. Continue the process until the difference between two consecutive 
masses does not exceed 0.1 % of the original mass of the specimen.  
E.5 Calculations  
Calculate the moisture content using the following formula an d round the results up to the nearest 0.1 %.   
 
 
Where,   
a is weight of the container with the specimen before drying (in grams);  
b is weight of the container with the specimen after drying (in grams);  
c is weight of the container (in grams); and  
V  is water content (in weight %).   
© EAC 2016  – All rights reserved  Annex  F 
(normative)  
 
Determination of water soluble extract  
F.1 Apparatus  
F.1.1  Weighing machine, sensitive to 1 mg  
F.1.2  Conditioning chamber  
F.1.3  Beaker, of more than 200 mL capacity  
F.1.4  Measuring flask  
F.1.5  Steam ba th  
F.1.6 Oven  
F.2 Procedure  
F.2.1  Weigh, approximately 12 g from the sample and expose to the standard atmosphere for testing 
textile (EAS 240).  
F.2.2  Weigh, to the nearest milligram, the conditioned test specimen.    
F.2.3  Cut the test specimen into s mall pieces and boil the pieces in 200 mL of distilled water in a beaker 
for half an hour.   
F.2.4  Filter into a 500 mL measuring flask. Extract the test specimen twice again for 15 minutes and filter 
the aqueous extract into the same flask. Pour the solut ion into a beaker and concentrate it to a small volume.  
Then transfer it to a dish of known mass, washing the beaker with a little distilled water.  
F.2.5  Evaporate the contents of the dish on a steam bath and dry in an air oven at 105 0C to 110 0C. Cool 
the dish in a desiccator and weigh. Heat again at 105 0C to 110 0C in the dry oven for 30 minutes. Cool the 
dish in the desiccator and weigh.  
F.2.6  Repeat this process of heating, cooling and weighing until the difference in mass between two 
successive we ighings is less than one milligram.  
F.3. Calculation  
Water so luble extract, % by mass = 
  
where,  
m0 is the mass, in g, of the empty dish;  
m1 is the mass, in g, of the dish with the residue ; and  
m2 is the mass, in g, of the dish with the m aterial taken for the test.  
  
   
© EAC 2016  – All rights reserved  Annex  G 
(normative)  
 
Determination of fluorescence in sanitary towels  
G.1 Principle  
A layer of absorbent filler material is examined under ultra violet radiation for the presence of fluorescent brightening 
agents  
G.2 Apparat us 
G.2.1.  Ultra -violet source  
G.2.2  Graduated S cale, in mm  
G.3 Procedure  
Examine a layer of absorbent filler material of approximately 5 mm thick under ultra violet radiation of wave 
length 365 mm.  
G.4 Test report  
Bright fluorescence indicates the pres ence of fluorescent brightening agents.  
 
 
  
 Annex  H 
(normative)  
 
Determination of flushability  
H.1 Apparatus  
Use the flushing toilet equipment as described in Figure H.1. The pipes to the septic tank should be 
transparent acryl tubes to allow the sanitary  end of the tube to an open tank so as to collect the products.  
H.2 Test method  
H.2.1 Throw three sanitary towels of test samples into the chamber pot, and flush water (8 to 12) litres per 
time from the flush tank.  
Dimensions in millimetres  
 
H.3 Test r eport  
Report whether the sanitary towels are completely flushed down or not.  
  
© EAC 2016  – All rights reserved  Annex  J 
(normative)  
 
Microbiological examination  
J.1 Apparatus and equipment  
Use apparatus and equipment complying with the relevant requirements of ISO 7218:2007  
J.2 Media an d reagents  
J.2.1  General  
Ensure compliance with the general requirements for the ingredients and for the preparation of media and 
reagents given in ISO 7218   
J.2.2  Bacteriological peptone  
Peptone               10 g  
Disodium phosphate dodecahydrate         1 g  
Sodium chloride             5 g  
Mono -potassium phosphate          1.5 g  
Dissolve the ingredients in distilled water and make up to 1 L. Adjust the pH value to be 7.0 ± 0.1 after 
sterilization. Dispense 300 mL volumes into flasks of capacity 500 mL a nd sterilize by autoclaving at 121 °C ± 
2 °C for 20 min.  
J.2.3  Plate count agar  
Agar               15 g  
Glucose               1 g  
Tryptone               5 g  
Yeast extract              2.5 g  
Dissolve the ingredients in distilled water, made up to 1litre, a nd adjust the pH value to 7.2 ± 0.2. Dispense 15 
mL volumes into bottles and sterilize by autoclaving at 121 °C ± 2 °C for 20 min.  
J.2.4  Neutral red -bile salt peptone glucose medium  
Peptone               20 g  
Glucose               10 g  
Bile salts No. 3             1.5 g  
Sodium chloride           5 g  
Neutral red            0.03 g   
 Crystal violet            0.002 g  
Dissolve the ingredients in 400 mL of distilled water and make up to 500 mL boiling to aid solution. Adjust the 
pH value to 7.4 and filter to a cl ear solution. Dispense 10 mL volumes into bottles each containing a Durham 
tube and sterilize by autoclaving at 121 °C ± 2 °C for 20 min  
J.2.5  Fluid soybean -casein digest medium  
Pancreatic digest of casein         17 g 
Papaic digest of soybean meal        3 g  
Sodium chloride           5 g  
Dibasic potassium phosphate        2.5 g  
Dextrose             2.5 g  
Dissolve the ingredients in distilled water and make up to 1 litre, warming slightly to aid solution. Cool the 
solution to room temperature and adjust the p H value to be 7.3 ± 0.2 after sterilization. Filter to clarify (if 
necessary), dispense into suitable containers, and sterilize by autoclaving at 121 ± 2 °C for 20 min.  
J.2.6  Centrimide agar medium  
Pancreatic digest of gelatine        20 g  
Magnesium chlo ride         1.4 g  
Potassium sulphate          10 g  
Agar             13.6 g  
Cetyltrimethylammonium bromide (Cetrimide)    0.3 g  
Glycerine            10 mL  
Dissolve all the solid ingredients in distilled water, make up to 1 L, and then add the glycerine. Heat, agitating 
frequently, and boil for 1 min. Adjust the pH value to be 7.2 ± 0.2 after sterilization. Dispense into suitable 
containers and sterilize by autoclaving at 121 °C ± 2 °C for 20 min.  
J.2.7  Pseudomonas agar  medium for detection of fluorescein  
Pancreatic digest of casein         10 g  
Peptic digest of animal tissue        10 g  
Anhydrous dibasic potassium phosphate      1.5 g  
Magnesium sulphate (MgSO4.7H2O)       1.5 g  
Glycerine            10 mL  
Agar             15 g  
Dissolve all the solid ingred ients in distilled water, make up to 1 L, and then add the glycerine. Heat, agitating 
frequently, and boil for 1 min. Adjust the pH value to be 7.2 ± 0.2 after sterilization. Dispense into suitable 
containers and sterilize by autoclaving at 121 °C ± 2 °C f or 20 min.  
J.2.8  Pseudomonas agar  medium for detection of pyocyanin  
Pancreatic digest of casein           20 g   
© EAC 2016  – All rights reserved  Anhydrous magnesium chloride          1.4 g  
Anhydrous potassium sulphate          10 g  
Agar               15 g  
Glycerine              10 Ml  
3  Preparation of Test Suspension  
Transfer 300 ml of the sterile solution of bacteriological peptone (J.2.2) to a sterile wide -mouthed jar of 
capacity not less than 1 litre and not more than 2litre. The jar shall have a mouth of diameter not less than 150 
mm an d not more than 250 mm, and is fitted with a hermetically closing glass or metal -and-glass lid.  
Aseptically place the towel under test in the solution in the jar, fit the lid, agitate the contents of the jar for 2 
min and then allow the jar to stand for 1 0 min.  Repeat this agitating and standing procedure twice more.  
Aseptically remove about 100 ml of the test suspension for testing as described in J.4 below.  
J.4  Procedure  
J.4.1  Total viable bacterial count  
Into each of three sterile petri dishes as eptically pipette a 1 mL portion of the test suspension. To each dish 
add 15 mL of freshly melted plate count agar (J.2.3) that has been cooled to 45 °C, and mix well. Incubate, 
count and calculate the total count as described in ISO 4833  Part 2. From the total viable bacterial count and 
the mass of the sanitary towel, calculate the total viable bacterial count per gram of sanitary towel.  
J.4.2  Examination for the presence of Enterobacteriaceae.  
Aseptically add 10 mL of the test suspension to a bottle tha t contains neutral red -bile salt peptone glucose 
medium (J.2.4). Incubate the bottle for 24 h to 36 h at 37 ± 0.5°C and examine for the presence of 
Enterobacteriaceae as evidenced by the formation of acid and gas.  
J.4.3  Examination for the presence of Staphylococcus aureus .  
Use the media, reagents and procedure described in ISO 6888 -2 to examine the test suspension (see J.3).  
As a control, pipette 0.1 mL of a 1:1000 dilution of an 18 h to 24 h culture of Staphylococcus aureus  SATCC 
Sta 10 into Staphyloco ccus medium and proceed as with the test suspension.  
J.4.4  Examination for the presence of Pseudomonas aeruginosa  
a) Aseptically pipette 10 mL of the test suspension into 90 mL of fluid soybean -casein digest medium 
(J.2.5) and mix well.  Incubate for 24 h at 30 °C to 35 °C. By means of an inoculating loop transfer a 
portion from the 24 h incubated sample tube of fluid soybean -casein digest medium to the dry surface 
of petri dishes each containing approximately 20 mL of Cetrimide agar medium (J.2.6). Incubat e at 30 
°C to 35 °C and examine after 24h, and again after 48 h incubation, for suspect colonies, bearing in 
mind that in general greenish fluorescent colonies are typical of Pseudomonas aureginosa and that in 
its presence a gram stain examined microscopica lly will reveal gram -negative slender rod -shaped 
cells.  
b) As a control, add 0.1 ml of a 1:1 000 dilution of an 18 h to 24 h culture of Pseudomonas aeruginosa  
SATCC Pse 11 mL to 100 mL of fluid soybean -casein digest medium (J.2.5), and proceed as with the 
test suspension.  
c) If none of the colonies obtained from the test suspension conforms to the description given in i) above 
and the control culture has been satisfactorily recovered, deem the test sample to be free from 
Pseudomonas aeruginosa.   
 d) If colonies con forming to the description given in i) above are found, streak representative suspect 
colonies from the Cetrimide agar onto the surfaces of Pseudomonas agar  medium for the detection of 
flourescein (J.2.7) and Pseudomonas agar medium for the detection of py ocyanin (J.2.8) to obtain 
isolated colonies.  Cover and invert the petri dishes and incubate at 30 – 35 °C for at least 3 days.  
Examine the streaked surfaces under ultraviolet light for suspect colonies, as described in Table J.1.  
 
 
Table J.1 — Descripti on of colonies  
Medium  Description of colonies  
Pseudomonas agar for the detection of fluorescein  Generally colourless to yellowish  
Yellowish fluorescence in ultra violet light  
Pseudomonas agar for the detection of pyocyanin  Generally greenish. Blue fluor escence in ultraviolet light  
 
If any further doubt exists as to the identity of the colonies, obtain final confirmation by inoculating the suspect 
colonies to the wells on commercially available diagnostic kits in accordance with the manufacturer’s 
instru ctions.   
© EAC 2016  – All rights reserved  Bibliography  
[1] SLS 111:1989, Specification for sanitary towels -  
 EAS 96: 2016   
 
© DEAC 2016 – All rights reserved