FINAL DRAFT UGANDA 
STANDARD  
  FDUS EAS 786 
First  Edition  
2013-mm-dd 
 
 Reference number  
FDUS EAS 786: 2013 
© UNBS 201 3 
  
Skin care creams, lotions and gels — Specif ication  
 FDUS EAS 786: 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 f or its correct application  
© UNBS 20 13 
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 wri tten 
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  
 FDUS EAS 786: 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 mandate d 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 Agreemen ts 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 
consum ers, 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 rec ommending the draft standards 
for approval and declaration as Uganda Standards by the National Standards Council.  
This Final Draft Uganda Standard, FDUS EAS 786: 2013, Skin care creams, lotions and gels — Specification , 
is identical with  and has been r eproduced from an East African  Standard, EAS 786: 2013, Skin care creams, 
lotions and gels — Specification , and is being proposed for adoption as a Uganda Standard.  
Wherever the words, “ East African  Standard" appear, they should be replaced by "Uganda Standard."   EAS 786 : 2013 
ICS 71.100.70  
© EAS 2013                                                                                                                              First Edition 2013  
 
 
EAST AFRICAN STANDARD  
 
Skin care creams, lotions and gel s — Specification  
EAST AFRICAN COMMUNITY  
 EAS 786 : 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 d evelopment 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 t o 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  EAS 786 : 2013 
© EAC 2013 – All rights reserved  iii 
 Contents  Page 
Foreword  ................................ ................................ ................................ ................................ .............................  v 
1 Scope  ................................ ................................ ................................ ................................ ......................  1 
2 Normat ive references  ................................ ................................ ................................ ............................  1 
3 Terms and definitions  ................................ ................................ ................................ ...........................  1 
4 Requirements  ................................ ................................ ................................ ................................ .........  2 
4.1 General requirements  ................................ ................................ ................................ ...........................  2 
4.2 Specific requirements  ................................ ................................ ................................ ...........................  2 
5 Packaging and labelling ................................ ................................ ................................ ........................  3 
5.1 Packaging  ................................ ................................ ................................ ................................ ...............  3 
5.2 Labelling  ................................ ................................ ................................ ................................ .................  3 
6 Sampling  ................................ ................................ ................................ ................................ ................  3 
Anne x A (normative)  Determination of thermal stability  ................................ ................................ ...............  4 
A.1 Apparatus  ................................ ................................ ................................ ................................ ...............  4 
A.2 Procedure  ................................ ................................ ................................ ................................ ...............  4 
A.3 Results  ................................ ................................ ................................ ................................ ....................  4 
Annex B (normative)  Determination of pH  ................................ ................................ ................................ ...... 5 
B.1 Apparatus  ................................ ................................ ................................ ................................ ...............  5 
B.2 Reagents  ................................ ................................ ................................ ................................ ................  5 
B.3 Procedure  ................................ ................................ ................................ ................................ ...............  5 
Annex C (normative)  Determination of total fatty substance conten t ................................ .........................  6 
C.1 Outline of the method  ................................ ................................ ................................ ...........................  6 
C.2 Reagents  ................................ ................................ ................................ ................................ ................  6 
C.3 Procedure  ................................ ................................ ................................ ................................ ...............  6 
C.4 Calculation  ................................ ................................ ................................ ................................ .............  6 
Annex D (normative)  Determination of total viable count  ................................ ................................ .............  7 
D.1 Outline of the method  ................................ ................................ ................................ ...........................  7 
D.2 Apparatus  ................................ ................................ ................................ ................................ ...............  7 
D.3 Medial buffer  ................................ ................................ ................................ ................................ ..........  7 
D.4 Sterilization of apparatus  ................................ ................................ ................................ .....................  8 
D.5 Procedure  ................................ ................................ ................................ ................................ ...............  8 
D.6 Expression of results  ................................ ................................ ................................ ............................  8 
Annex E (normative)  Determination of Pseudomonas aeruginosa, Staphylococcus  aureus  and 
Candida albicans  in cosmetic products  ................................ ................................ ..............................  9 
E.1 Introduction  ................................ ................................ ................................ ................................ ............  9 
E.2 Principle of the method ................................ ................................ ................................ .........................  9 
E.3 Apparatus  ................................ ................................ ................................ ................................ ...............  9 
E.4 Reagents  ................................ ................................ ................................ ................................ ..............  10 
E.5 Micro -organisms  ................................ ................................ ................................ ................................ .. 11 
E.6 Product quality check  ................................ ................................ ................................ .........................  11 
E.7 Product preparation  ................................ ................................ ................................ ............................  11 
E.8 Bacterial inoculum preparation  ................................ ................................ ................................ .........  12 
E.9 Fungal inoculum preparation  ................................ ................................ ................................ .............  12 
E.10   Inoculum pools  ................................ ................................ ................................ ................................ ... 12 
E.11   Inoculation  ................................ ................................ ................................ ................................ ..........  12 
E.12   Sampling intervals  ................................ ................................ ................................ ..............................  13 
E.13   Aerobic Plate Count (APC)  ................................ ................................ ................................ ................  13 
E.14   Neutralization check ................................ ................................ ................................ ...........................  13 
E.15   Data analysis  ................................ ................................ ................................ ................................ ....... 13 EAS 786 : 2013 
iv © EAC 2013 – All rights reserved  
 Annex F (normative)  Test for lead using Atomic Absorption Spectrophotometer  (AAS)  .......................  15 
F.1 Principle of the method  ................................ ................................ ................................ ......................  15 
F.2 Reagents  ................................ ................................ ................................ ................................ ..............  15 
F.3 Procedure  ................................ ................................ ................................ ................................ ............  15 
F.4 Calculation  ................................ ................................ ................................ ................................ ...........  15 
Annex G (normative)  Test for arsenic using Atomic Absorption Spectrophotometer (AAS)  .................  16 
G.1 Scope  ................................ ................................ ................................ ................................ ...................  16 
G.2 Reagents  ................................ ................................ ................................ ................................ ..............  16 
G.3 Instrument conditions  ................................ ................................ ................................ ........................  16 
G.4 Sample preparation  ................................ ................................ ................................ ............................  17 
G.5 Analysis  ................................ ................................ ................................ ................................ ...............  17 
G.6 Calculation  ................................ ................................ ................................ ................................ ...........  17 
Annex H (normative)  Test for mercury using the Atomic Absorption Spectrophotometer (AAS)  .........  18 
H.1 Scope  ................................ ................................ ................................ ................................ ...................  18 
H.2 Reagents  ................................ ................................ ................................ ................................ ..............  18 
H.3 Instrument conditions  ................................ ................................ ................................ ........................  18 
H.4 Sample preparation  ................................ ................................ ................................ ............................  19 
H.5 Analysis  ................................ ................................ ................................ ................................ ...............  19 
H.6 Calculations  ................................ ................................ ................................ ................................ .........  19 
Bibliography  ................................ ................................ ................................ ................................ .....................  20 
 EAS 786 : 2013 
© EAC 2013 – All rights reserved  v 
 Foreword  
Development of the East African Standards has been necessitated by the need for harmonizing requirements 
governing quality of products and services i n 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 establ ished 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 im plementing .  
EAS 786 was prepared by Technical Committee EAS/TC 071, Cosmetics and cosmetic products.  
 EAS 786 : 2013 
vi © EAC 2013 – All rights reserved  
 Introduction  
Skin creams, lotions and gels are useful not only as skin beautifiers, but also guard the skin against adverse 
climatic conditions. The p erformance of any skin cream, lotion or gel depends on the type of skin on which it is 
applied, and the nature of ingredients added to effect the intended end use. This Uganda Standard covers the 
following preparations, vanishing creams and lotions, founda tion creams and lotions, cold creams and lotions, 
night creams, moisturizers, cleansers, hands creams and lotions, body creams and lotions, sun -protection 
preparations, toners, emollients, purifiers, nourishers and any other special treatment skin preparat ions e.g. 
AHA.  
 EAST AFRICAN STANDARD  EAS 786 :2013 
 
© EAC 2013 – All rights reserved  1 
 Skin care creams, lotions and gels  — Specification  
1 Scope  
This East African S tandard specifies  requirements  and methods of sampling  and test  for creams, lotions and 
gels for skin care.  
This East African  Standard do es not apply to  skin care products , for which therapeutic claims are made  and 
also does not apply to non -emulsified lotions and gels.  
2 Normative references  
The following referenced documents are indispensable for the application of this document. For date d 
references, only the edition cited applies. For undated references, the latest edition of the referenced 
document (including any amendments) applies.  
EAS 346 , Labelling of cosmetics — General requirements  
EAS 377 -1, Cosmetics and cosmetic products   Part 1 : List of substances prohibited in  cosmetic product s 
EAS 377 -2, Cosmetics and cosmetic products   Part 2 : List of substances which cosmetic  product s must not 
contain except subject to the restrictions laid down  
EAS 377 -3, Cosmetics and cosmetic p roducts   Part 3: List of colorants allowed in cosmetic products  
EAS 377 -4, Cosmetics and cosmetic products   Part 4: List of preservatives allowed in cosmetic products  
EAS 377 -5, Cosmetics and cosmetic products  Part 5: Use of UV filters in cosmetic products  
ISO 6887 -1, Microbiology of food and animal feeding stuffs — Preparation of test samples, initial suspension 
and decimal dilutions for microbiological examination — Part 1: General rules for preparation of the initial 
suspensions and decimal dilut ions 
ISO 11930, Cosmetics — Microbiology — Evaluation of the anti -microbial protection of a cosmetic product  
ISO 24153, Random sampling and randomization procedures  
3 Terms and definitions  
For the purposes of this Final East African Standard, the following  term and definition  shall apply  
special purpose product  
include cleansers, toners, purifiers, facial washes, nourishers, Alpha Hydroxy Acids (AHA) skin lightening 
preparations based on vitamins and acids, anti -wrinkle/aging preparations, facial scrubs, fa cial masks, facial 
wash, non -emulsified lotions and gels and such other products  EAS 786 : 2013 
2 © EAC 2013 – All rights reserved  
 4 Requirements  
4.1 General  requirements  
4.1.1 All ingredients used including dyes, pigments and colours shall  comply with all the parts of EAS 377.  
4.1.2 The preparation sha ll be clear or of uniform colour.  
4.1.3 The cream, lotion or gel shall be free from visible impurities.  
4.2 Specific  requirements  
4.2.1 All creams, lotions and gels for skin care shall comply with the chemical requirements given in Table 1 
when tested acco rding to the methods described therein.  
Table 1 — Chemical r equirements for creams, lotions and gels for skin care  
SL No.  Characteristic  Requirement  Method of test  
i Thermal stability  To pass test  Annex A  
ii pH range  
 Hand and body lotion, creams and 
gels (including baby products)  
 Special purpose products*  4.5 - 8.5 
3.5 - 8.5 Annex B  
iii Total fatty substance content, % by mass, min  5 Annex C  
* These include cleansers,  toners, purifiers, facial washes,  nourishers,  AHA’s skin lightening preparations based  on 
vitamins and acids, anti -wrinkle/aging preparations, facial scrubs, facial masks, facial wash, non -emulsified lotions and 
gels and such products  
 
4.2.2 All creams, lotions and gels for skin care shall comply with the microbiological limits given in T able 2  
when tested in accordance with the methods indicated therein . 
Table 2  Microbiological limits  
Micro -organisms  Limits, max.  
cfu/g  Method of test  
Total viable count  100 in 0.5 g1) Annex D 
100 in 0.1 g2) 
Pseudomonas aeruginosa3)  
Not detectable  Annex E  
Staphylococcus aureus3) 
Candida albicans3) 
1) For p roducts specifically intended for children under 3 years, eye area and mucous membranes  
2) For other products  
3) For Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans , the limits sha ll not be 
detectable in 0.5 g for products specifically intended for children under 3 years, eye area and mucous 
membranes and in 0.1 g for other products.  
 EAS 786 : 2013 
© EAC 2013 – All rights reserved  3 
 4.2.3 The products shall comply with the limits for heavy metal contaminants in accordance with T able 3.  
Table 3  Limits  for heavy metal contaminants  
S/N Characteristic  Limits t Method of test  
i.  Lead, ppm, max . 20 Annex F 
ii.  Arsenic, ppm, max . 2 Annex G 
iii.  Mercury, ppm, max . 2 Annex H 
NOTE  The total amount of heavy metals as lead, mercury and arsenic, in combination, in the finished 
product should not exceed 20 ppm.  
 
5 Packaging  and labelling  
5.1 Packaging  
The product shall be pack aged  in suitable well -sealed containers that shall protect the contents a nd shall not 
cause any contamination or react with  the product . 
5.2 Labelling  
5.2.1 The labelling shall be in English, Kiswahili or French or in combination as agreed between the 
manufacturer and supplier . 
5.2.2 The labelling shall comply with the requirements of EAS 346 . 
5.2.3 Special purpose products shall in addition bear the following warning on the label:  
a) These products shall not be used on children below the age of 12 years ; 
b) This product may cause irritation. If irritation persists  discontinue use ; and  
c) Precaution/warning shall be ‘'do patch test be fore use’’ . 
6 Sampling  
Random samples of the product shall be drawn for test in accordance with ISO 24153 from the market, factory 
or anywhere else  
. EAS 786 : 2013 
4 © EAC 2013 – All rights reserved  
 Annex A  
(normative)  
 
Determination of thermal stability  
A.1 Apparatus  
Thermostatically controlled oven , capable of maintaining a temperature of 37 °C ± 1 °C 
A.2 Procedure  
Place a fresh, unopened sample of the cream or lotion in its original container into a thermostatically 
controlled oven at 37 °C ± 1 °C for 48 h, making sure that the sample is securely seale d. If the product is 
packed in an opaque container (for example, a tube), remove 50 g of the sample and place into an effectively 
sealed glass tube or vial and test as above.  
A.3 Results  
The product shall be taken to have passed the test if, on removal fro m the oven, the following indications of 
instability are not observed:  
(i) change of colour;  
(ii) change of smell or odour;  
(iii) phase separation;  
(iv) formation of granules or crystal growth; and  
(v) shrinkage due to evaporation of water.  
 EAS 786 : 2013 
© EAC 2013 – All rights reserved  5 
 Annex B  
(normative)  
 
Determination o f pH 
B.1 Apparatus  
B.1.1 pH meter , equipped with a glass electrode  
B.1.2 Beaker , of 100 mL capacity   
B.2 Reagents  
B.2.1 pH 7.0 buffer solution  
B.2.2 pH 4.0 and pH 9.0 buffer solutions  
B.2.3 Deionised water  
B.3 Procedure  
B.3.1 Dip the pH meter into about 50  mL of pH 7.0 buffer solution. Ensure that the reading is 7.0.  
B.3.2 Rinse the meter with deionised water, and dip it into about 50 mL of pH 4.0 buffer solution.  Ensure 
that the reading is 4.0. Repeat using pH 9.0 buffer solution.  
B.3.3 For oil -in-water emulsion creams , weigh accurately 5 g ± 0.01 g of the cream in a 100-mL beaker. Add 
45 mL of water and disperse the cream in it. Determine the pH of the suspension at 25 °C using the pH meter.  
B.3.4 For water -in-oil emulsion creams , weigh 10 g of the cream  to the nearest 0.1 g. Add 90 mL of rectified 
spirit previously adjusted to pH 6.5  to 7.0. Warm, if necessary, to 45 °C  and stir thoroughly for 15 min. Filter 
the alcoholic layer through a filter paper and measure the pH of the filtrate at 25 °C  using the pH meter.  
The type of cream is determined by placing some of it on a spot tile and sprinkling with a mixed indicator 
consisting of an intimate mixture of an oil soluble dye of one colour, e.g. oil orange and a water -soluble dye of 
a different colour, for e xample, methylene blue. After a few minutes the predominant colour indicates whether 
the continuous phase is oil or water. In case of doubt this matter is confirmed by checking whether the product 
is capable of conducting electricity and, if so, then the c ream is deemed to be water -continuous.  EAS 786 : 2013 
6 © EAC 2013 – All rights reserved  
 Annex C  
(normative)  
 
Determination of total fatty substance content  
C.1 Outline of the method  
The emulsion is broken with dilute mineral acid and the fatty matter is extracted with petroleum ether. It is 
weighed after  removal of the solvent.  
C.2 Reagents  
C.2.1  Dilute hydrochloric acid 1:1 (v/v)  
C.2.2  Petroleum , B.P. 40 °C to 60 °C  
C.2.3  Methyl orange indicator solution  — Dissolve 0.1 g of methyl orange in 100 mL of water.  
C.2.4  Sodium sulphate , desiccated  
C.3 Procedure  
Weigh accurately about 2 g of the material into a conical flask; add 25 mL of dilute hydrochloric acid, fit a 
reflux condenser into the flask and boil the contents until the solution is perfectly clear. Pour the contents of 
the flask into a 300 mL separat ion funnel and allow it to cool to 20 °C. Rinse the conical flask with 50  mL of 
petroleum ether in portions of 10 mL. Pour the ether rinsings into the separation funnel shake the separation 
funnel well and leave until the layers separate. Separate out the aqueous phase and shake it out with 50  mL 
portions of ether twice. Combine all the ether extracts and wash them with water until free of acid (when 
tested with methyl orange indicator solution). Filter the ether extracts through a filter paper containing s odium 
sulphate into a conical flask which has been previously dried at a temperature of 60 °C ± 2 °C and then 
weighed. Wash the sodium sulphate on the filter with ether and combine the washings with the filtrate. Distil 
off the ether and dry the material r emaining in the flask at a temperature of 60 °C ± 2 °C  to constant mass.  
C.4 Calculation  
The total fatty substance shall be calculated as follows:  
Total fatty substance, percent by mass =
21 100
MM , 
where,  
M1 is the mass, in g rams , of the resi due, and  
M2 is the mass, in g rams , of the material taken for the test.  EAS 786 : 2013 
© EAC 2013 – All rights reserved  7 
 Annex D  
(normative)  
 
Determination of total viable count  
D.1 Outline of the method  
The test consists of plating a known dilution of the sample or any digest agar medium (soya bean casei n is 
recommended) suitable for the total count of bacteria and fungi after incubating them for a specified period to 
permit the development of visual colonies.  
IMPORTANT   Take precaution in ascertaining that only fresh samples, from carefully sealed conta iners 
that had not been opened before, are used for this test.  This is very necessary for getting accurate results.  
D.2 Apparatus  
D.2.1 Tubes , of resistant glass, provided with closely fitting metal caps  
D.2.2 Autoclaves , of sufficient size. They shall ke ep uniform temperature within the chamber up to and 
including the sterilizing temperature of 122 C. They shall be equipped with an accurate thermometer, located 
so as to register the minimum temperature within the sterilizing chamber, a pressure gauge and  properly 
adjusted safety valves.  
D.2.3 Petri dishes , of 100 mm diameter and 15 mm depth. The bottom of the dishes shall be free from 
bubbles and scratches and shall be flat so that the medium is of uniform thickness throughout the plate.  
D.2.4 Colony cou nter, an approved counting aid, such as Quebec colony counter. If such a counter is not 
available, counting may be done with a lens giving a magnification of 1.5 diameter. In order to ensure 
uniformity of conditions during counting, illumination equivalent  to that provided by Quebec colony counter 
shall be employed.  
D.2.5 Balance  
D.2.6 pH meter  
D.2.7 Water bath  
D.3 Medial buffer  
D.3.1  Soya bean casein digest agar medium  
Dissolve 15 g of pancreatic digest of casein 5 g of papaic digest of soya bean meal, a nd 5 g of sodium 
chloride in 100 mL of distilled water contained in a 2 -litre beaker by heating in a water bath. Add 15 g of 
powdered agar and continue boiling until the agar is completely digested. Adjust the pH to 7.5 with sodium 
hydroxide solution. Dist ribute in 20 -mL quantities, close the tubes with metal caps and autoclave at 123 C for 
20 min. After autoclaving, store the tubes in a cool place and use them within three weeks.  
D.3.2  Stock solution pH 7.2 phosphate buffer  
Dissolve 34 g of monobasic pot assium phosphate in about 100 mL of water contained in a 500 -mL volumetric 
flask. Adjust the pH 7.2 ± 0.1 by the addition of 4 % sodium hydroxide solution. Add water to volume and mix. 
Sterilize at 122 C for 20 min. Store under refrigeration.  EAS 786 : 2013 
8 © EAC 2013 – All rights reserved  
 D.3.3  Dilute  phosphate buffer solution pH 7.2  
Dilute 1 mL of stock solution with distilled water in the ratio of 1:800. Fill 50 mL in each of the conical flask of 
100 mL capacity. Plug the flasks with cotton and sterilize at 122 C for 20 min.  
D.4 Sterilization of ap paratus  
D.4.1  Tubes  
These shall be sterilized in the autoclave at 122 C temperature and 1.05 kg/cm2 pressure for 20 min or in a 
hot oven at 160 C for 1 h.  
D.4.2  Petri dishes   
These shall be packed in drums and autoclaved at 122 C temperature and 1.05 k g/cm2 pressure for 20 min or 
individually wrapped in kraft paper and sterilized  in a hot air oven at 160 C for 1 h.  
D.4.3  Pipettes  
These shall be placed in pipette cones (copper, stainless steel or aluminium) after plugging the breeder end 
with cotton and sterilized in the autoclave at 122 C temperature and 1.05 kg/cm2 pressure for 20 min, or at 
160 C for 1 h in the air oven.  
D.5 Procedure  
D.5.1 Melt a sufficient number of soya bean casein digest agar medium tube in a hot water bath and 
transfer while  hot in to a constant temperature water bath maintained at 48 C ± 20 C.  
D.5.2 Weigh and transfer aseptically 1 g of the sample to a conical flask containing sterile 50 mL or any 
suitable dilution factors of dilute phosphate buffer at pH 7.2. Shake well.  Pipette out in 1 mL portions into three 
sterile petri dishes. Pour melted and cooled (at 45 C) soya bean casein digest agar medium over it, and 
rotate to mix thoroughly. Incubate the plate at 32 C for 72 h in an inverted position.  
D.6 Expression of res ults 
Get the average number of colonies on soya bean casein digest agar medium plates and determine the 
number of micro -organisms per gram of the sample. If no colony is recovered from any of the plates micro -
organisms can be stated as being less than 50 p er gram. For calculations, refer to ISO 6887 -1. EAS 786 : 2013 
© EAC 2013 – All rights reserved  9 
 Annex E  
(normative)  
 
Determination of Pseudomonas aeruginosa, Staphylococcus  aureus  and Candida 
albicans  in cosmetic products  
E.1 Introduction  
A general aseptic and safety procedures should be followed.  Table  E.1 gives the results of the interlaboratory 
study supporting the acceptance of the method.  
Table E.1 — Inter -laboratory study results for determination of preservation of non -eye area water -
miscible cosmetic and toiletry formulations  
 
Incidence of false - negatives among total positive samplesa) Incidence of false -positives among total 
negative samplesb) 
Product name  Number  Percentage  Sensitivity 
rate Number  Percentage  Sensitivity 
rate 
Shampoo  2/49 4 96 0/53 0 100 
Conditioner  3/48 6 94 0/54 0 100 
Water -in-oil emulsion  0/52 0 100 1/50 2 98 
Oil-in-water emulsion  0/51 0 100 0/51 0 100 
All combined  5/200  2 98 1/208  0.5 99.5 
a) False -negative analysis indicates a sample is adequately preserved.  
b) False -positive analysis indicates a sample is not adequate ly preserved.  
 
E.2 Principle of the method  
Bacteria yeast and mould grown on laboratory media, harvested, calibrated, and inoculated into test products.  
Using serial dilutions and plate counts; the numbers of organisms surviving in the test products are 
determined over time.  Products meeting the specified criteria are considered adequately preserved for 
manufacture and consumer use.  Products not meeting criteria are considered inadequately preserved.  
E.3 Apparatus  
E.3.1 Jars, 2-4 oz wide -mouth, straight -side flint glass ointment jars with linerless metal, polypropylene or 
teflon lined scre w caps  
E.3.2 Disposable borosilicate glass culture tubes , 16 mm x 125 mm, with caps  
E.3.3 Disposable borosilicate glass culture tubes, 20 mm x 150 mm, with screw caps  
E.3.4 Petri plates Z , 100 mm x 15 mm  
E.3.5 Sterile 2.2 mL pipettes  
E.3.6 Steri le swabs  EAS 786 : 2013 
10 © EAC 2013 – All rights reserved  
 E.3.7 Glass beads  
E.3.8 Sterile gauze  
E.3.9 10 l -20 l inoculating loops  
E.3.10   Vortex mixer  
E.4 Reagents  
E.4.1 Preparation  
For convenience, dehydrated media of any br and equivalent in function may be used. Test each lot of medium 
for sterility and growth -promotion using suitable organisms.  
E.4.2 List of reagents  
E.4.2.1 Letheen agar, contains 5.0 g pancreatic digest of casein 1.0 g dextrose, 3.0 g beef extract, 1.0 g 
lecithin, 7.0 g polysorbate 80 g, and 15.0 g agar per L.  Prepare according to manufacturer’s directions.  
Dispense into suitable containers and sterilize by autoclaving at 121 C for 15 min. Final pH should be 7.0  
0.2 at 25 C . Place in 45 C water bath  until agar is 45 C  2 C.  Use for pour plates.  
E.4.2.2 D/E Neutralizing broth (Dey/Engley), contains 5.0 g pancreatic digest of casein, 2.5 g yeast 
extract, 10 g dextrose, 1.0 g sodium thiogycollate, 6.0g Na 2 S2O3 . 5H2O, 2.5 g NaHSO 3, 7.0 g lecithin, 5.0 g 
polysobate 80 g, and 0.02 g bromcresol purple per L.  
Prepare according to manufacturer’s directions. Dispense 9  mL or 9.9 mL aliquot into tubes and sterilize by 
autoclaving at 121 C for 15 min.  Final pH should be 7.6  0.2 at 25 C.  Use for aerobi c plate count, L, 
dilutions.  
E.4.2.3 Nutrient agar,  contains 5.0 g pancreatic digest of gelatin 3.0 g beef extract, and 15.0 g agar per 
L. Prepare according to manufacturer’s directions. Dispense into tubes and sterilize by autoclaving at 121 C 
for 15 min . Final pH should be 6.8  0.2 at 25 C. Cool in inclined position to form a slant. Use for bacterial 
culture maintenance and inoculum preparation.  
E.4.2.4 Y/M agar (yeast/malt extract),  contains 3.0 yeast extract, 3.0 g malt extract , 5.0 g peptone.10.0 
g dextrose  and 20.0 g agar per L. Prepare according to manufacturer’s directions.  Dispense into tubes and 
sterilize by autoclaving at 121 C for 15 min.  Final pH should be 6.2  0.2 at 25 C. Cool in slanted position.  
Use for yeast culture maintenance and  inoculum preparation.  
E.4.2.5 Potato dextrose agar (PDA), contains 200 g potato infusion, 20.0 g dextrose, and 15.0 g agar 
per L. Prepare according to manufacturer’s directions. Dispense into tubes and sterile by autoclaving at 121 
C for 15 min. Final pH  should be 5.6  0.2 at 25 C.  Cool in slanted position. Use for mould culture 
maintenance and inoculum preparation.  
E.4.2.6 0.85 % Saline, dissolve 8.50 g NaCI in water and dilute to 1 L.  Dispense into flasks or bottles 
and sterilize by autoclaving at 1 21 C for 15 min.  
E.4.2.7 0.85 % Saline with 0.05 % Polysorbate 80,  dissolve 8.5 g NaCI in water, mix in 0.50 g 
polysorbate 80 g, and dilute to 1  L.  Dispense into suitable containers and sterilize by autoclaving at 121 C 
for 15 min.  
E.4.2.8 Barium sulpha te standard No. 2   
E.4.2.8.1  Prepare a 1.0 % BaCI 2 solution by dissolving 1.0 g BaCI 2. 2H 2O in 100 mL water. Let this be 
referred to as solution 1.  EAS 786 : 2013 
© EAC 2013 – All rights reserved  11 
 E.4.2.8.2  Prepare a 1.0 % H 2SO 4  solution by mixing 1.0 mL H 2SO 4 in 100 mL water. Let this be referred to 
as solution 2.  
E.4.2.8.3 Mix 0.2 mL of solution (1) with9.8 mL solution (2), in scre w-capped test tube. Cap tightly  and store 
in the dark at room temperature.  
E.4.2.9 Barium sulphate standard No. 7, use solutions from E.4.2.8. Mix 0.7 mL of solution E.4.2.8.1, 
with 9.3 mL of solution E.4.2.8.2, in a screw -capped test tube. Cap tightly and store in the dark at room 
temperature.  
E.5 Micro -organisms  
E.5.1 Staphylococcus aureus   — ATCC 6538  
E.5.2 Staphycloccuss epidermidis  — ATCC12228  
E.5.3 Klebsiella pneumoniae   — ATCC10031  
E.5.4 Escherichia coli    — ATCC 8739  
E.5.5 Enterobacter gergoviae   — ATCC 33028  
E.5.6 Pseudomonas aeruginosa  — ATCC 9027  
E.5.7 Burkholderia cepacia   — ATCC25416  
E.5.8 Acinetobacter baumannii  — ATCC 19606  
E.5.9 Candida albicans    — ATCC10231  
E.5.10 Aspergillus niger    — ATCC 16404  
NOTE  Environmental micro organisms (s) likely to be contaminants of concern during product manufacture or use be 
included as a separate inoculum. Predominant environmental microbes isolated during manufacturing, equipm ent 
cleaning, and sanitizing, or from related deionized water systems are used as supplemental test inocula).  
E.6 Product quality check  
E.6.1 Weigh 1.0 g product into a screw -capped culture tube containing 9.0 mL sterile neutralizing broth to 
make a 1:10 d ilution.  If necessary to disperse product, add ten to twenty 3 -mm diameter glass beads to tube. 
Mix on Vortex mixer until homogeneous.  
E.6.2 Pipette 1.0 mL of the 1:10 dilution into each of four sterile petri plates. Pour 15 mL - 20 mL sterile 
molten Lethe en agar (45 C  2 C) into each plate. Mix by rotating plates to disperse the dilution thoroughly. 
Let solidify.  
E.6.3 Invert and incubate 2 plates at 35 C  2 C for 48 h and two plates at 25 C  2 C for 5 days.  
E.6.4 Count the number of colonies on a ll plates, add, and multiply by 2.5 to determine the number of 
colony forming units per gram (cfu./g) in the product.  
E.6.5 Save plates to be used for the neutralization validation by refrigerating.  
E.7 Product preparation  
E.7.1 Measure 20 mL sterile salin e into four sterile jars, E.3.1. Cap tightly and store at room temperature.  EAS 786 : 2013 
12 © EAC 2013 – All rights reserved  
 E.7.2 Weigh 20 g produc t into each of four sterile jars, E.3.1. Cap tightly and store at room temperature.  
E.8 Bacterial inocul um preparation  
E.8.1 Streak each bacteria culture, E.5.1 – E.5.10, onto a nutrient agar  slant , E.4.2.3.  Incubate for 48 h at 
35 C  2  C. Wash each slant with 5.0 mL sterile saline, loosening the culture from the agar surface. 
Transfer the suspension into a sterile tube. Repeat the wash with second 5.0 mL aliquot of saline. Combine 
washes and mix on Vortex mixer to disperse evenly.  
E.8.2 Adjust each wash with sterile saline to yield a suspension of ca 108 cfu/mL using Mc Farland BaSO 4 
standard No, 2,  E.4.2.8, direct microscopic count, turbidimetry, absorban ce, or other method correlated to  an 
aerobic plate count (APC), E.13. Perform an APC, E.13, on each suspension to confirm standardization.  
E.9 Fungal inoculum preparation  
E.9.1 Streak  C. albicans,  E.5.9, on 3 slants of Y/M agar, E.4.2.4. Incubate at 25 C  2 C for 48 h. Wash 
each slant sequentially with 5.0 mL aliquot of sterile saline.  Repeat with a second 5.0 mL aliquot of sterile 
saline. Combine washes to produce 10 mL suspension. Mix on Vortex mixer to disperse evenly.  
E.9.2 Adjust the wash with ster ile saline to yield a suspension of ca 107 cfu/ml using a Mc Farland Ba SO 4 
standard No. 7, E.4.2.8, direct microscopic count, turbidimetry, absorbance, or other method that has been 
correlated to an APC, E.13. Perform an APC, E.13, on the suspension to co nfirm standardization.  
E.9.3 Streak  A. Niger , E.5.10, on 5 slants of potato dextrose agar E.3.5. Incubate at 25 C  2 C for 10 
days. Dislodge mould spores by adding 5.0 mL sterile saline containing 0.05 % polysorbate 80 to each tube 
and vigorously rubbin g the surface of the agar slant with a sterile swab.  Repeat with a second 5.0 mL aliquot 
in each tube.  Combine the 10 washes to produce 50 mL suspension. Filter into a sterile container through 3 -5 
layers of sterile gauze supported in funnel.  Perform an  APC, E.13, using appropriate dilutions. Adjust mould 
suspension to ca 107 per ml using sterile saline.  Use immediately or refrigerate at 2 C - 5 C for up to 1 
month. Verify mould viability by an APC, E.13, before each use.  
E.10  Inoculum pools  
E.10.1 Pool equal parts of the S. aureus  and S . epidermidis   suspensions, E.8.2 in a sterile container to 
make incoculum pool 1: Gram -postive cocci.  
E.10.2 Pool equal parts of the K pneumoniae  , E . coli, and E . gergoviae  suspensios, E.8.2, in  a sterile  
contain er to make inoculum pool  2: Gram - negative fermentors.  
E10.3 Pool equal parts of the P. aeruginosa , B .cepacia  and A. baumanii suspensions, E.8.2, in a sterile 
container to make Inoculum Pool 3: Gram -negative nonfermentors.  
E.10.4 Pool equal parts of C. Albicans, E.9.2, and A. Niger, E.9.3, suspensions in a sterile container to make 
innoculum pool 4: Fungi.  
E.10.5 Use organism pools immediately or refrigerate them at 2 C - 5 C for more than 72 h.  
E.11  Inoculation  
E.11.1 Inoculate each of the four 20.0 m L aliquots of sterile saline, E.7.1, with 0.2 mL of its respective 
inoculum pool, E.10.1 - E.10.4. Mix thoroughly.  Use these suspensions to determine inoculums counts (see 
Ka). EAS 786 : 2013 
© EAC 2013 – All rights reserved  13 
 E.11.2 Inoculate each of the four 20 g products suspensions, with 2.0 mL of it s respective inoculums , E.10.1 
– E.10.4 . Mix thoroughly by shaking, Vortex mixing or stirring, so that each suspension contains 106 bacteria 
or 10 5 fungi per gram, evenly distribute throughout the product. Tightly close inoculated containers and store 
at ambient temperature (20 C - 25 C). 
E.12  Sampling intervals  
E.12.1 Sample each inoculated saline suspension, E.11.1, for APC, E.13, within 1 h after inoculation to 
obtain inoculum count.  
E.12.2 Test each inoculated product, E.11.2 for APC, E.13, at 7, 14  and 28 days after inoculation to obtain 
product interval count.  
E.13  Aerobic Plate Count (APC)  
E.13.1 Mix suspension thoroughly . Weigh 1.0 g product into screw -capped culture tube containing 9.0 mL 
sterile neutralizing broth for a 1:10 dilution. If neces sary to disperse product, add 10 - 20 sterile 3 mm 
diameter glass beads to the tube. Mix on Vortex mixer until homogeneous.  
E.13.2 Aseptically pipette 0.1 mL of the 1: 10 dilution into 9.9 mL tube of neutralizing broth to obtain a 1:1000 
dilution. Vortex m ix. Pipette 0.1 mL of the 1:1000 dilution into 9.9 mL neutralizing broth to obtain a 1: 100 000 
dilution.  
The number of dilutions may be decreased if previous counts microbial populations show reduction.  
E.13.3 Using a 2.2 mL pipette, aseptically pipette 1 .0 mL and 0.1 mL aliquots from the 1:10 dilution into 
duplicate petri dishes for the 1:10 and 1:100 plates. If necessary, transfer duplicate 1.0  mL and 0.1 mL 
aliquots from the 1: 1000 dilution for plates 1:1000 and 1:10 000, and from the 1:100 000 dilutio n for plates 
1:100 000 and 1:1000 000. Pour 15  mL - 20 mL sterile Letheen agar E.3.1, (45 C  2 C into each plate. Mix 
by rotating the plates to disperse the suspension thoroughly, and let solidify.  
E.13.4 Invert bacterial plates and incubate at 35 C  2 C.  Examine bacterial plates after 48 h - 72 h.  Count 
in suitable range (30 - 300 colonies). If no countable plates fall in that range, count the plate(s) nearest that 
range showing distinct colonies. Average duplicate plates counts and express results  as cfu/g of product.  
E.13.5 Invert and incubate fungal plates at 25 C  2 C.  Read fungal plates at 2 - 3 days and record results. 
Count plates in a suitable range (30 - 300 colonies). If no countable plates fall in that range, count the plate(s) 
neares t that range showing distinct colonies. Re -incubate plates for another 2 - 3 days. Read and record 
additional colonies. Add to previous results to obtain total counts. Average duplicate plate counts and record 
as cfu/g of products.   
E.14  Neutralization c heck  
Make a 1:10 000 dilution in steri le saline of pools 1, 2 and 3, E.10.1 - E.10.3, and  a 1:1000 dilution of pool 4, 
E.10.4. Streak each dilution for isolation with a 10 l loop on the plates s aved from E.6.5.  If plates are not 
usable due to either desi ccation or surface growth, repeat F .6, and streak freshly pr epared plates.  Incubate 
as in E.13.4 - E.13.5 
E.15  Data analysis  
E.15.1 Product quality check , E.6.4, should  be found to contain, <100 cfu/g to proceed with the challenge 
test. 
E.15.2 Inoculums counts , E.12.1, should be between 1 to 9.9 x 106 cfu /g product for bacteria and 1 to 9.9 x 
105 cfu/g product for fungi, or the test should be repeated with different dilutions.  EAS 786 : 2013 
14 © EAC 2013 – All rights reserved  
 E.15.3 Neutralization check , E.14, should  show significant growth of all pools  to confirm adequate 
neutralization.  A neutralizing broth other than D/E broth can be used.  If neutralization does not occur, the test 
is invalid.  Refer to references 4 -6 in AOAC  official method 998.10  for assistance.  
E.15.4 Calculate the percentage red uction:  
count Inoculum100 count interval product count Inoculum% Reduction,
 
 
E.15.5 The test product is considered adequately preserved if  
a) bacteria show at least 99.9 % (3 log) reduction within one week following challenge and remain at or 
below that level thereafter, and  
b) fungi show at least a 90 % (1 log) reduction within one week following challenge, and remain at or below 
that level thereafter. This criteria applies to freshly prepared formulations . EAS 786 : 2013 
© EAC 2013 – All rights reserved  15 
 Annex F 
(normative)  
 
Test for lead using Atomic Absorption Spectrophotometer  (AAS)  
F.1 Principle of the method  
The sample is passed through wet digestion using pressure decomposition. The amount of lead is then 
determined using AAS.  
F.2 Reagents  
The reagents used shall be of analytical reagent grade. Water shall be distilled or de -ionised.  
F.3 Proced ure 
F.3.1 Weigh accurately 0.4 g of sample and put this into a 50  mL decomposition pressure tube. Add 7.0 mL 
of concentrated nitric acid. Add 2.0 mL of water. Close the pressure vessel, and apply between 15 Nm - 20 Nm 
of pressure. Digest for about 2 h. Tran sfer to a 20 mL volumetric flask and make to the mark.  
F.3.2 Prepare the standard solutions for lead. Aspirate into the flame each of the standard solutions in 
ascending order of concentration. Take the absorbance reading for each concentration using the A AS. Plot a 
calibration graph of the concentration of lead in the standard solutions against the corresponding values of 
absorbances.  
F.3.3 Aspirate into the flame the sample solution. Take the absorbance reading from the AAS  and give it a 
value X. From the  graph, use the value of absorbance X to read the corresponding value of concentration. Let 
this value of concentration be C.  
F.4 Calculation  
The amount of lead shall be calculated as follows:  
Amount of lead = 
MDC  
where  
C is the c oncen tration of the sample solution;  
M is the  mass of the sample in grams; and  
D is the  Dilution factor.  EAS 786 : 2013 
16 © EAC 2013 – All rights reserved  
 Annex G  
(normative)  
 
Test for arsenic using Atomic Absorption Spectrophotometer (AAS)  
G.1 Scope  
This method describes the determination of arsenic in variou s cosmetic products.  
G.2 Reagents  
The reagents used should be of analytical reagent grade. Water shall be distilled or di -ionised.  
G.3 Instrument conditions  
G.3.1 Standard atomic absorption conditions for arsenic  
G.3.1.1  Recommended flame; air -acetylene,  reducing (rich, slightly yellow)  
G.3.1.2  Data obtained with a standard nebulizer and flow spoiler. Operation with a High Sensitivity 
nebulizer or impact bead will typically provide a 2 -3X sensitivity improvement.  
G.3.1.3  Characteristic concentration with a N 2O-C2H2 flame at 193.7 M: 1.4mg/L  
G.3.1.4  Table contains EDL data.  HCL sensitivity values are more than 25 % poorer.  
Table G.1 — HCl data EDL sensitivity values  for arsenic  
Wavelength ,  
nm Slit,  
nm Relative noise  Characteristic 
concentration ,  
mg/L  Characteristic 
concentration 
check ,  
mg/L  Linear range , 
mg/L  
193.7  0.7 1.0 1.0 45.0 100.0  
189.0  0.7 1.8 0.78 40.0 180.0  
197.2  0.7 0.95 2.0 90.0 250.0  
 
G.3.2  Stock standard solution  
Arsenic , 1000 mg/L . Dissolve 1.320 g of Arsenious oxide As 2O3, in 25  mL o f 20 % (w/v) KOH solution. 
Neutralize with 20  % (v/v) H 2SO 4 to a phenolphthalein endpoint. Dilute to one litre with 1  % (v/v) H 2SO 4. The 
standard solutions shall be of the following concentrations: 0 ppm, 2 ppm, 4 ppm, 6 ppm, 8 ppm and 10 ppm.  
G.3.3  Flame s 
The air -acetylene flame absorbs or scatters more than 60 % of the light source radiation at the 193.7 nm 
arsenic line. Flame absorption is reduced with the use of the nitrous oxide -acetylene flame, although 
sensitivity is also reduced. Use of background correction is recommended, as it will correct for flame 
absorption and thus improve the signal to noise ratio.  EAS 786 : 2013 
© EAC 2013 – All rights reserved  17 
 G.3.4  Light  sources  
Both HCL and EDL sources are available for arsenic. EDLS, which are more intense, provide better 
performance and longer life . 
G.3.5  Interferences  
Sample with high total salt content (greater than 1 %) can produce non -specific absorption at the 193.7 mn 
arsenic line, even when the metal is absent. It is therefore advisable to set background correction.  
G.4 Sample preparation  
As in F.4.  
G.5 Analysis  
As in F.6.  
G.6 Calculation  
As in F.7 EAS 786 : 2013 
18 © EAC 2013 – All rights reserved  
 Annex H  
(normative)  
 
Test for mercury using the Atomic Absorption Spectrophotometer (AAS)  
H.1 Scope  
This method describes the determination of mercury in various cosmetic products.  
H.2 Reagents  
The reagents used should be analytical reagent grade.  Water shall be distilled or de -ionised.  
H.3 Instrument conditions  
H.3.1 Standard atomic absorption conditions for mercury  
H.3.1.1  Recommended flame: air -acetylene, oxidizing (lean, blue)  
H.3.1.2  Data ob tained with a standard nebulizer and flow spoiler. Operation with a High Sensitivity 
nebulizer or impact bead will typically provide a 2 -3X sensitivity improvement.  
H.3.1.3  Characteristic Concentration with a N 2O-C2H2 flame at 253.7 nm: 12 mg/L  
H.3.1.4  Table contains EDL data.  HCL sensitivity values more than 25 % poorer  
Table H.1 — HCl data EDL sensitivity values  for mercury  
Wavelength ,  
nm Slit,  
nm Relative noise  Characteristic 
concentration ,  
mg/L  Characteristic 
concentration 
check ,  
mg/L  Linear range , 
mg/L  
253.7  0.7 1.0 4.2 200.0  300.0  
 
H.3.2 Standard flame emission conditions for mercury  
Table H.2 — Flame  emission conditions for mercury  
Wavelength (nm)  Slit (nm)  Flame  
253.7  0.2 Nitrous oxide - acetylene  
H.3.3 Stock standard solution  
Mercury , 1000 mg/L — Dissolve 1.080 g of mercury (II) oxide, HgO, in a minimum volume of (1 + 1) HCI. 
Dilute to one litre with deionized water. The standard solutions shall be of the following concentrations: 0  ppm, 
2 ppm, 4  ppm, 6  ppm, 8  ppm and 10 ppm.  EAS 786 : 2013 
© EAC 2013 – All rights reserved  19 
 H.3.4 Light sou rces 
Both Electrodeless Discharge Lamps (EDLS) and Hollow Cathode Lamps are available for Mercury. However, 
the light output of mercury Hollow Cathode Lamp is significantly poorer than with EDLs, and the sensitivity and 
detection limit achieved also are mu ch poorer. In addition, the life of Hollow Cathode Lamps is much shorter.  
H.3.5  Interferences  
Large concentrations of cobalt will absorb at the mercury 253.7 nm resonance line. A 1000 mg/L cobalt  
solution produces approximately 10 % absorption. Ascorbic a cid, stannous chloride, or other reducing agents 
may reduce the mercury present to Hg (I) or elemental mercury. These give higher sensitivities than Hg (II), 
and their presence can generate erroneously high results.  
H.4 Sample preparation  
As in F.4.  
H.5 Analysis  
As in F.6.  
H.6 Calculations  
As in F.7  
 EAS 786 : 2013 
20 © EAC 2013 – All rights reserved  
 Bibliography  
[1] IS 6608 : 1972 , Specification for skin creams;  
[2] IS 4011 : 1982, Methods  for dermatological tests for cosmetics  
[3] KS 03 -580: 1998, Specification for creams, lotions and gels for skin care  
[4] Official Journal of the European Communities N. L26217  
 
 
 
 
 
 
  
 EAS: 2013   
 
© EAC 2013 – All rights reserved