KENYA STANDARD                                                KS 1 529: 2012 
                                    ICS 71.100.70 
 
 
 
 
 
 
 
 
 
 
 
 
 
Pomades and solid brilliantines — 
Specification 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
© KEBS 2012        Third Edition 2012 KS 1529: 2012 
  © KEBS 2012 — All rights reserved  ii  
TECHNICAL COMMITTEE REPRESENTATION 
 
The following organizations were represented on the  Technical Committee: 
 
 
Consumer Information Network 
Kenyatta National Hospital — Dermatology Department  
Government Chemist 
Haco Industries Ltd. 
Rozeco Chemical Industries Ltd. 
Interconsumer Products Ltd. 
Kenya Industrial Research and Development Institute  (KIRDI) 
Beiersdorf (EA) Ltd. 
Pz Cussons E.A. Ltd 
Unilever Kenya Ltd. 
Henkel Chemicals (EA) Ltd. 
Intertek International 
Jomo Kenyatta University of Agriculture and Technol ogy (JKUAT)   
Ministry of Medical Services 
Kenya Association of Manufacturers 
Ministry of Public Health and Sanitation 
Kenya Bureau of Standards — Secretariat 
 
 
 
 
 
 
 
 
REVISION OF KENYA STANDARDS  
 
In order to keep abreast of progress in industry, K enya Standards shall be regularly 
reviewed. Suggestions for improvements to published  standards, addressed to the 
Managing Director, Kenya Bureau of Standards, are w elcome. 
 
 
 
 
 
 
© Kenya Bureau of Standards, 2012 
 
Copyright. Users are reminded that by virtue of Sec tion 25 of the Copyright Act, Cap. 12 of 2001  of th e Laws of Kenya, copyright 
subsists in all Kenya Standards and except as provide d under Section 26 of this Act, no Kenya Standard pro duced by Kenya Bureau of 
Standards may be reproduced, stored in a retrieval system in any form or transmitted by any means with out prior permission in writing 
from the Managing Director. 
 
 
 
 
ISBN 9966-23-478-0 
 
             KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                iii              ICS 71.100.70  
 
 
 
 
 
 
 
Pomades and solid brilliantines — 
Specification  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
KENYA BUREAU OF STANDARDS (KEBS) 
 
Head  Office:  P.O. Box 54974, Nairobi-00200, Tel.: (+254  020) 60 5490, 602350, Fax: (+254 020)  604031 
E-Mail: info@kebs.org,  Web:http://www.kebs.org   
 
Coast Region    Lake Region         North Rift Regi on  
P.O. Box 99376, Mombasa-80100  P.O. Box 2949, Kisum u-40100       P.O. Box 2138, Nakuru-20100 
Tel.: (+254  041) 229563, 230939/40 Tel.: (+254 057 ) 23549, 22396       Tel.: (+254 051) 210553, 21055 5  
Fax: (+254  041) 229448   Fax: (+254 057) 21814              
 KS 1529: 2012 
  © KEBS 2012 — All rights reserved  iv   
Contents           Page  
 
Foreword .......................................... ................................................... ................................... iv 
1 Scope .............................................. ................................................... ............................. 1 
2 Normative references ............................... ................................................... .................... 1 
3 Definitions ....................................... ................................................... .............................. 1 
4 Types ............................................. ................................................... ............................... 2 
5 Requirements ...................................... ................................................... ......................... 2 
Table 1 – Sulphur and sulphide limits ............. ................................................... ............ 3 
Table 2 – Chemical requirements  ................... ................................................... ........... 3 
6 Packaging ......................................... ................................................... ............................ 4 
7 Marking and labelling  ............................ ................................................... ...................... 4 
8 Sampling  ......................................... ................................................... ............................ 4 
9 Methods of test .................................... ................................................... ......................... 4 
Annex A (normative) Testing of total sulphur content  . ................................................... ...... 5 
Annex B (normative) Test for sulphur and sulphides  ..... ................................................... ... 6 
Annex C (normative) Determination of sulphated ash  . ................................................... ...... 7 
Annex D (normative) Determination of lead content in  pomades and solid brilliantines  
by graphite furnace atomic absorption spectrophotomet er (AAS) ........................... 8 
Annex E (normative) Test for arsenic using atomic absor ption spectrophotometer  
(AAS)   ........................................... ................................................... ........................ 12 
Annex F (normative) Test for mercury using atomic absorp tion spectrophotometer  
(AAS)   ........................................... ................................................... ..................................... 14 
Annex G (normative) Total amount of heavy metals  .... ................................................... ..... 17 
Annex H (normative) Determination of melting point  ................................................... ......... 18 
Annex J (normative) Determination of consistency  ..... ................................................... ...... 19 
Annex K (normative) Test for rancidity  .............. ................................................... ................ 21 
Annex L (normative) Bleed number  .................. ................................................... ................. 22 
Annex M (normative) Test for stability  .............. ................................................... ................. 23 
Annex N (normative) Determination of yeasts and mould s in pomades and solid  
brilliantines ..................................... ................................................... ........................ 24 
Annex P (informative) List of raw materials conventi onally used in the formulation of  
pomades and solid brilliantines  .................... ................................................... ........ 30 
Annex Q (normative) Antibacterial test  .............. ................................................... ................ 31  
Annex R (informative) Available Kenya standards on fats,  oils and waxes  ......................... 32  
 
 
 
 
 
             KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                v   
 
Foreword 
 
This third Edition of this Kenya Standard was prepare d by the Technical Committee on the Cosmetics and 
related products, under the guidance of Standards Pro jects Committee and it is in accordance with the 
procedures of the Bureau. 
 
Pomades and brilliantines are products with a wide ap plication area, though they are mainly used for hair  
and skin care.  A limit for heavy metals is therefor e given for safety purposes.  Other requirements spec ified 
include melting point, sulphated ash, penetration con e, rancidity, bleed number, stability, and microbiolo gical 
examination. 
 
This standard only covers pomades and solid brillianti nes. Liquid brilliantines are covered in the specificati on 
for hair oils. 
 
This standard was first issued in the year 2000. In this second edition of 2004, a provision was made for 
products containing sulphur for control of itchy scalp. Also, methods of test for lead arsenic and mercury 
were replaced with current ones. In addition, testing of specific pathogenic micro-organisms was introduced,  
and the marking clause was altered. Also, the scope w as widened to cover pomades or solid brilliantines 
containing insect repellents.  
 
During the preparation of this standard, reference was made to the following documents: 
 
IS 9339-1988: Specification for pomades and brilliant ines. 
 
Regulation (EC) No. 1223/2009 of the European Parli ament and of the Council of the European Union. 
(2009): Cosmetic Products . Brussels. 
 
Acknowledgement is hereby made for the assistance derive d from these sources. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
          KENYA STANDARD            KS 1529: 20412 
© KEBS 2012 — All rights reserved  1Pomades and solid brilliantines — Specification 
 
 
1 Scope 
 
1.1  This Kenya Standard prescribes requirements and method s of test for pomades and solid 
brilliantines. 
 
1.2  This standard applies to: 
 
i) pomades and solid brilliantines based on vegetable oils, mineral oils, animal fats, waxes, or 
fatty acids but excludes oil emulsions. 
ii) pomades and solid brilliantines to which sulphur h as been added as an intrinsic part of the 
formulation. 
iii) insect repellents manufactured in the form of pomades or solid brilliantines. 
 
1.3 This standard does not cover: 
 
i) liquid brilliantines. These are covered in KS 03-60 2 ∗. 
ii) pomades and brilliantines which contain ingredient s that affect the physiological functions of the 
body, scalp or hair, or for which therapeutic claims are made. such products shall be registered by 
the Ministry responsible for registration of medicated  products  
 
2 Normative references 
 
The following standards are necessary adjuncts to thi s standard. They contain provisions which through 
reference in this text constitute provisions of this st andard. 
 
       
KS 1474     Classification of cosmetic raw materials and adjuncts.  
KS 1707  Labelling of cosmetic products . 
KS 1668   Methods of sampling cosmetics . 
 
3 Definitions 
 
For the purposes of this standard, the following defi nitions shall apply: 
 
3.1         
solid brilliantine              
a hair dressing preparation based on petroleum jelly.  May contain dyes and perfumes 
 
3.2  
liquid brilliantine    
liquid mineral, vegetable or animal oil for body and  hair care 
 
3.3         
pomade                             
an ointment used for cosmetic purposes.  May be greas y, soft or hard, perfumed or non-perfumed.  May 
contain mineral oil, vegetable oil, animal fat, or a  combination of the three 
 
3.4  
petroleum jelly   
a mixture of petroleum hydrocarbons used as an emollie nt. Usually consists of mineral oil and waxes like 
microcrystalline wax 
 
                                                 
∗ Specification for hair oils. KS 1529: 2012 
© KEBS 2012 — All rights reserved  2  
 
4 Requirements 
 
4.1       Visual appearance    
 
The pomade and solid brilliantine shall be in the fo rm of a soft, homogenous unctuous mass. 
 
4.2        Ingredients  
 
4.2.1  All raw materials used in the manufacture of pomades  and solid brilliantines shall comply with all 
parts of KS 1474. In addition, the mineral oils, vege table oils, animal fats and waxes shall comply with th e 
relevant Kenya standards, wherever available (see An nex P). 
 
4.2.2  All products containing ingredients that affect the p hysiological functions of the body, scalp or hair, o r 
medicinal products for which therapeutic or medical cl aims are made shall be registered with the Ministry of  
Health. 
 
4.2.3  All essential oils/herbs used shall comply with the a pproved standards where such exist. 
 
4.2.4  All products containing herbal extracts known to have antibacterial activity e.g. neem oil/extract, and 
aloe oil/extract, shall pass the test for antibacteria l and antifungal activity outlined in Annex Q. Furth er details 
may also be obtained from Annex N. All products claim ing antibacterial effects shall be also be tested for 
antibacterial activity.   
 
4.2.5  All insect repellents manufactured in the form of pomades or solid brilliantines shall be registered 
with the Pest Control Products Board, which shall dete rmine their efficacy before issuing a licence.   
 
4.2.6  All ingredients used shall be in sufficient quantiti es to effect the claims made. 
 
4.2.7  The raw materials used shall be free from any harmfu l effect on the skin, scalp or hair and the               
interaction of the raw materials used in the finished  pomade or brilliantine shall be free from toxic or 
sensitizing effects. 
 
4.2.8   A list of ingredients conventionally used in the f ormulation of pomades and brilliantines of various 
types is given for information only in Annex P. 
 
4.2.9 Use of sulphur as an ingredient 
 
4.2.9.1   Sulphur may be added to products as an ingredient to form an intrinsic part of the formulation. 
 
4.2.9.2 The main purpose of the product shall be to sooth i tchy scalp. 
 
4.2.9.3  Such a product shall fulfil the following conditions : 
 
i) shall contain a maximum of 3 % sulphur (w/w) when te sted in accordance with Annex A  
 (see Table 1); 
 
ii) shall be clearly labelled as per 7.1 and 7.2; 
 
4.2.9.4  All other products in which sulphur is not added  as an intrinsic part of the formulation shall not         
contain sulphur or sulphides when tested in accordance wi th Annex B (see Table 1). 
 
4.3  Other requirements 
 
 All pomades and brilliantines shall also comply with the requirements given in Tables 2, 3 and 4 when te sted 
according to the methods prescribed therein. 
 
 
Table 1 — Sulphur and sulphide limits 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 3  SL  
No.  
 
Characteristic 
 Requirement  
 Test method  Products containing sulphur 
as an ingredient  Products without 
sulphur 
i) 
 Total sulphur content  3 % max. (m/m) N/A Annex A 
ii) 
 Sulphur and sulphides  N/A To pass the test Annex B 
 
 
Table 2 — Contaminants 
 
 
SL No. 
 Characteristic Limits Test method  
i) Sulphated ash, % by mass, max a). 
 0.10 Annex  C 
ii) Lead, parts per million, max. 
 20 Annex  D 
iii) Arsenic, parts per million, max. 
 2 Annex  E 
iv) Mercury, parts per million, max. 
 2 Annex  F 
v) 
 Total amount of heavy metals, ppm, 
max. 20 Annex  G 
a)  For products containing sulphur for soothing of the sca lp, refer to 4.2.5. 
 
 
   Table 3 —  Requirements for pomades and solid br illiantines 
 
SL No.  
 Characteristic Requirements Test method  
i) Melting point, ºC 
 35 to 60 Annex  H 
ii) Consistency, 1/10 mm 
 100 to 275 Annex  J 
iii) Test for rancidity Shall be free from 
rancidity Annex  K 
iv) Bleed number 
 5 to 15 Annex  L 
v) Stability To pass the test Annex M 
 
 
Table 4 — Microbiological examination 
 
SL No.  
 Char acteristic  Limits  Test method  
i) Microbiological examination   
 
Total viable count for aerobic 
mesophyllic micro-organisms per g, 
max.  
 
100  
 
Annex N 
ii) Psedomonas aeruginosa     ) 
Staphylococcus aureus       ) 
Candida albicans                 ) Not detectable in 0.5 g 
of cosmetic product Annex N 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  4 5 Packaging 
 
The product shall be packed in suitable, airtight cont ainers. 
 
6 Marking and labelling 
 
6.1  Labelling shall be as per the requirements outlined in KS 1707 1. 
 
 
6.2  In addition to the above, all products containing sul phur as part of the formulation (see 4.2.5) shall be 
marked with the following warnings: 
 
“Contains Sulphur ” 
 
“If irritation develops, discontinue use ” 
 
7 Sampling 
 
Representative unopened samples shall be drawn for tes t from the market, factory or anywhere else 
following the procedures outlined in KS 1668. 
 
The samples shall be declared as complying with the speci fication if they satisfy all the specified 
requirements. 
 
8 Methods of test 
 
Unless otherwise stated, analytical grade chemicals an d distilled or de-ionized water shall be used in test s. 
All calculations shall be to 3 decimal places. 
 
 
                                                 
1 KS 1707: labeling of cosmetic products                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 5  Annex A 
(normative) 
 
Testing of total sulphur content in pomades 
 
A.1 Procedure 
 
Transfer about 0.5 g of sulphur ointment, accurately we ighed, to a suitable conical flask. Add 5 mL of nitr ic 
acid and 3 mL of bromine. Warm slightly and allow to stand overnight. Heat gently on a steam bath until t he 
excess bromine has been dissipated.  Add 30 mL of wate r and then 30 mL of ether, and swirl to dissolve 
most of the ointment base.  Transfer the mixture to a separator, with the aid of a 20-mL and a 10-mL port ion 
of ether, followed by two 10-mL portions of water. S hake the mixture, and draw off the water layer into a  
suitable beaker or flask. Extract the ether layer with two 40-mL portions of water each containing 1 mL of 
hydrochloric acid.   Heat the combined water extracts on  a steam bath to remove all traces of ether. Dilute 
the solution with water to about 200 mL and heat to  boiling.  Add slowly, with constant stirring, about 2 0 mL 
of hot barium chloride TS.  Heat on a steam bath for  1 hour, and then collect the precipitate on a filter . Wash 
it well with hot water, dry, and ignite to constant w eight. 
 
A.2       Results  
  
The weight of the barium sulphate so obtained, multip lied by 0.1374, represents the weight of sulphur.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  6 Annex B 
(normative) 
 
Test for sulphur and sulphides 
 
B.1  Apparatus 
 
B.1.1  Copper strips , 1 cm in width, and freshly polished. 
 
B.2  Procedure 
 
Melt in a beaker about 100 g of the sample and keep in a water bath at a temperature of 95 ºC.  Then place a 
strip of copper in the melted sample so that it is par tially immersed in it and allow to remain for 10 mi nutes. 
 
B.3  Results 
 
The material shall be taken to have passed the test if the copper strip used in the test shows no tarnishin g 
when compared with another freshly polished copper strip. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 7   
Annex C 
 
(normative) 
 
Determination of sulphated ash 
 
C.1  Reagents 
 
C.1.1  Dilute sulphuric acid , approximately 5N. 
 
C.2  Procedure 
 
Heat a porcelain or silica dish of 50 mL to 100 mL capa city to redness; cool in a desiccator and weigh.  Place 
about 20 g of the sample, accurately weighed, in the dish.  Heat the dish gently by means of a bunsen burner  
until the oil can be ignited at the surface.  Remove the burner and allow the oil to burn completely, taki ng 
care that all the free carbon on the sides of the dish is completely burnt.  Heat the residue with a strong fl ame 
or in a muffle furnace until all the carbonaceous ma tter has disappeared.  Cool the dish; add a few drop s of 
dilute sulphuric acid; heat gently to drive off the a cid and then heat strongly.  Cool the dish again in the 
desiccator and weigh it.  Repeat the heating, coolin g and weighing until constant mass is obtained. 
 
C.3  Calculation 
 
Sulphated ash, %, by mass    = 
21100  x 
MM 
  
where 
 M1   = mass in g of the residue; and 
 M2  = mass in g of the sample taken for the test. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  8  
Annex  D 
 
(normative) 
 
Determination of lead content in pomades and solid brilliantines by 
graphite furnace atomic absorption spectrophotomete r (AAS) 
 
 
D.1  Scope 
 
This test method specifies an electrothermal atomiz ation technique by using graphite furnace AAS metho d 
for the determination of lead content of pomades an d solid brilliantines. 
 
D.2  Warning and safety 
 
The acids used in the test are highly corrosive and  shall be handled with maximum care and where 
appropriate, use a fume hood during preparation of standards.  Lead is very toxic/carcinogenic and mus t be 
handled with maximum care avoiding physical contact . 
 
If spillage occurs, use adequate amounts of water a nd soap to wash off the spill. 
 
D.3  Principle 
 
Injecting of the prepared solution into a graphite furnace. Spectrometric measurements of the atomic 
absorption of the 228.8 spectral line emitted by le ad hollow cathode lamp. 
 
D.4  Materials 
 
D.4.1  Reagents, chemicals and standards 
 
D.4.1.1 Nitric acid, ρ about 1.4 g/mL. 
 
D.4.1.2 Nitric acid (1+1) v/v  
 
Mix 1 volume of conc. HN0 3 with 1 volume of  distilled water.  
 
D.4.1.3 Nitric acid (0.1M)  
 
Place 17 mL of concentrated acid in 100 mL volumetr ic flask then top to the mark with distilled water and mix. 
 
D.4.1.4 Lead standard solution , 1 000 ppm 
 
In 1 litre volumetric flask, dissolve 1.598 g of Pb  (N0 3)2 in minimum volume of 1% v/v HN0 3 and finally top the 
mark using 1% HN0 3.   
 
NOTE   Commercial grade standards can also be used when available. 
 
D.4.1.5 Lead standard solution ,  100 ppb 
 
This shall be prepared freshly by serial dilution o f the lead solution (D.4.1.4).  
 
D.4.1.6 Purge gas , argon 
 
Sufficiently pure, free from water and oil and free  from lead. 
 
 
 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 9   
D.4.2  Apparatus and equipment 
 
D.4.2.1 Atomic absorption spectrometer fitted with graphite furnace 
 
The atomic absorption spectrometer used will be sat isfactory if after optimization according to the 
manufacturer’s instructions and when in reasonable agreement with the values given by the manufacturer  
and it meets the performance criteria as set out in  the manual. 
 
D.4.2.2 Lead hollow cathode lamp 
 
D.4.2.3 Ordinary laboratory apparatus 
 
NOTE  All glassware shall first be washed in hydroc hloric acid ( ρ about 1.19 g/mL, diluted). 
 
D.5  Performance 
 
D.5.1  Sample preparation 
 
Ignite 1 g of sample at 500 ± 2 °C to ash. Extract the lead from the ash with 20 mL of 2 N HNO 3 , and repeat 
with 10 mL of 2 N HNO 3. Combine the extracts and dilute to 50 mL with 0.5  N HNO 3. 
 
D.5.2  Calibration 
 
D.5.2.1 Preparation of calibration curve 
 
D.5.2.1.1     Dilute the stock 100 ppb solution with 0.1 M   HNO 3 to obtain solutions with 10 ppb, 20 ppb, 40 
ppb, 60 ppb, 80 ppb and 90 ppb of lead. 
 
D.5.2.1.2     Inject 20 microlitres each of the six solutions  in turns at the same rate starting from the lowest  
concentrated solution to the highest concentrated s olution. 
 
             D.5.2.1.3  Record the corresponding absorbance values and plot  calibration curve. 
 
D.5.3  Quality control checks 
 
D.5.3.1 Duplicates 
 
D.5.3.1.1   All samples will be analyzed in duplicates and th e stated acceptance criteria shall apply:  The 
absolute difference between two independent test re sults obtained using the same procedure shall be no t 
greater than 10 % of the arithmetic mean of the two  results. 
 
D.5.3.1.2  Spike distilled water with 10.0 ppb of lead and obt ain the recovery percentage. 
 
D.5.3.1.3  Recovery %  ≥96.  
 
D.6  Procedure 
 
D.6.1     Test portion 
 
Use sample as prepared in (D.5.1). 
 
D.6.2       Blank test 
 
D.6.2.1   Run a parallel reagent blank determination replacin g the test solution with distilled water. 
 
D.6.2.2    Reagent blank shall be ≤ 0.000 1 ppb of lead. 
 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  10 D.6.3  Instrumentation 
 
D.6.3.1     Follow the manufacturer’s instructions for preparin g the instrument for use. 
 
D.6.3.2   Install the appropriate lamp and adjust the curre nt to the recommended value. 
 
D.6.3.3   Ensure that the autosampler pipette is correctly aligned and that the drain is available. 
 
D.6.3.4 Select the sample tray type installed. 
 
D.6.3.5  Ensure that the graphite tube is in good condition  and correctly aligned. 
 
D.6.3.6  Switch on the cooling system, turn on the purge ga s and finally start the instrument software. 
 
D.6.3.7  Select the relevant method and then condition tube .   
 
D.6.4     Instrument conditions  
 
The following conditions shall be used for the furn ace during analysis of lead: 
 
D.6.4.1 Wavelength:                283.3 
 
D.6.4.2  Slit:                               0.7 
 
D.6.4.3 Atomization site:  pyro/platform 
 
D.6.4.3  The furnace conditions for lead are given in Table  D.1. 
 
Table D.1 — Furnace conditions for lead 
 
 
Step  
Temperature  0C  
Ramp time 
(seconds)  
Hold time 
(seconds)  
Internal 
gas flow 
(L/min.)  
Gas type 
1 (drying) 120 10 60 250 Normal 
2 (pretreatment) 
 500 1 30 250 ,, 
3 (cooling) 
   20 1 15 250 ,, 
4 (atomization) 
 1 800 0 5   0 ,, 
5 (cleanout) 
 2 600 1 5 250 ,, 
 
D.6.5  Spectrometric measurements 
 
D.6.5.1  Inject 20 µl of the calibration standards, the blank solution and the test solution into the graphite 
furnace.  
 
D.6.5.2  Record the absorbance reading. 
 
D.6.5.3  If the absorbance of the sample in greater than th e highest calibration standard, dilute the test 
solution appropriately using 0.1 M HN0 3 for lead then measure the absorbance. 
 
D.6.5.4 Inject the calibration solutions in ascending order . 
 
NOTE 1 The calibration curve shall only be acceptab le for analysis when the correlation coefficient  (  r) ≥ 0.99. 
 
 
 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 11   
 
D.7   Expression of results 
 
D.7.1  Method of calculation 
 
The lead content of the sample expressed in mg/L of  product is equal to: 
 
[ (C1   - C2) * V ] / M0 
 
where  
C1      =  lead content of test solution expressed in  mg/L read from calibration curve; 
 
C2      =  lead content of blank solution expressed i n mg/L read from calibration curve; 
M0  =  grams of sample taken for analysis (5 g); 
 
V  =  Volume of sample taken for analysis (100 mL). 
 
NOTE  If the test solution was diluted, then the di lution factor shall be taken into consideration in calculation. 
 
D.7.2  Expression of results 
 
D.7.2.1 Report results of manganese content in mg/L as Pb i n the sample into two decimal points. 
 
D.8  Method validation 
 
D.8.1  The method validation data shall be as shown in Tab le D.2.  
 
   Table D.2 — M ethod validation data  
 
Element  
 Linearity  LOQ ppb  LOD ppb  
Pb 
 r ≥ 0.99 32.356 3.804 
 
D.8.2  Precision:  Repeatability  
 
The absolute difference between two independent tes t results obtained using the same method on identic al 
test material in the same laboratory by the same op erator using the same equipment within a short inte rval of 
time shall not be greater than 10 % of the arithmet ic mean of the two results. 
 
D.8.3  Working range 
 
Pb      10-100 ppb. 
 
D.8.4  Reporting limits 
 
Pb   30 ppb. 
   
 
 
 
 
 
 
 
 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  12  
Annex E 
 
(normative) 
 
Test for arsenic using atomic absorption spectropho tometer (AAS) 
 
E.1 Scope 
 
This method prescribes the determination of arsenic  in pomades and solid brilliantines. 
 
E.2  Reagents 
 
E.2.1  0.15 mol/L ( ≅≅ ≅≅ 1.5 % V/V) hydrochloric acid  
 
Carefully add 15 mL conc. HCl to deionized water an d make up to 1 L. 
 
E.2.2  0.25 mol/L ( ≅≅ ≅≅ 1 % W/V) NaOH solution  
 
Carefully dissolve 10 g sodium hydroxide flakes in deionized water and make up to 1 L. 
 
E.2.3  0.8 mol/L ( ≅≅ ≅≅ 3 % W/V) NaBH 4 solution  
 
Dissolve 3 g sodium tetrahydroborate in 1 % NaOH so lution and make up to 100 mL with 1 % NaOH solution . 
 
E.3  Stock solution 
 
The stock solution contains 1 000 mg/L As.  The use  of commercially available concentrated calibration  
solutions for AAS is recommended. 
 
WARNING:  Arsenic solutions are toxic.  
 
E.4  Calibration solution   
 
1 mg As/L (in 1.5 % HCl). 
 
E.4.1  Aliquots for calibration:  10, 25, 50 µL 
 
E.4.2  Corresponding to:  10, 25, 50 ng As 
 
E.4.3 Diluent:    1.5 % (V/V) hydrochloric acid 
 
E.4.4  Calibration volume:  10 mL 
 
E.5  Reductant solution   3 % NaBH 4 in 1 % NaOH solution 
 
E.6  Oxidation state 
 
The hydride is generated much more slowly from As(V ) than from As(III).  To prevent interferences, As( V) 
must be prereduced to As(III) prior to the determin ation. 
 
Prereduction can be performed with KI in semi conce ntrated (5 mol/L) HCl solution or, preferably, with  L-
cysteine 
 
 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 13   
 
 
E.7 Pre-reduction 
 
E.7.1  KI solution   
 
Dissolve 3 g KI and 5 g L (+)- ascorbic acid in 100  mL water.  Prepare fresh daily.  Add 1 mL of the K I 
solution per 10 mL of the sample solution in 5 mol/ L HCl and stand for 30 min. 
    
Or, 
 
E.7.2  L-cysteine solution: Dissolve 5 g L-cysteine in 10 0 mL 0.5 mol/L HCl.  This solution is stable for at  
least a month.  Add 2 mL of the L-cysteine solution  per 10 mL of the sample solution and stand for 30 min. 
 
E.8  Instrument conditions  
 
E.8.1 Analytical wavelength:   193.7 nm 
 
E.8.2 Slit width and height:  0.7 nm low 
 
E.8.3 Radiation source:   Electrodeless discharge lamp for As  
 
E.8.4 QTA heating:    Heat the QTA in a lean, blue air-acetylene flame 
 
E.8.5 Prepared measurement volume:  10 mL minimum to 50 mL maximum 
 
E.8.6 Pre-reaction purge time:   Approx. 50 s 
 
E.8.7 Post-reaction purge time:  Approx. 40 s 
 
E.8.8    Characteristic mass:   0.95 ng As for 1 % absorption (A = 0.0044)  
 
E.8.9 Characteristic concentration:       0.095 µg/L 1 % absorption for 10 mL calibration volume 
 
E.8.10  Characteristic concentration check:50 µL of the 1 000 mg/L As stock solution (50 ng) give an 
absorbance of approx. A = 0.2. 
 
E.8.11  Alternate analytical wavelengths are given in Tabl e E.1: 
 
Table E.1 — Alternate analytical wavelengths 
 
Wavelength  
nm Slit width  
nm Sensitivity relative to main  
analytical wavelength 
189.0 
197.2 0.7 
0.7 0.8 
2.0 
 
NOTE    Condition the QTA in cold hydrofluoric acid if ther e is a decrease in sensitivity(and other causes are  excluded). 
 
 
 
 
 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  14  
Annex F 
 
(normative) 
 
Test for mercury using atomic absorrption spectroph otometer (AAS) 
 
F.1        Method 1 — Using sodium tetrahydroborate (Na BH 4) as reductant 
 
F.1.1 Scope 
  
This method prescribes the determination of mercury  in pomades and solid brilliantines using sodium 
tetrahydroborate (NaBH 4) as reductant. 
 
F.1.2 Reagents 
 
F.1.2.1 0.15 mol/L ( ≅≅ ≅≅ 1.5 % V/V) hydrochloric acid  
 
Carefully add 15 mL conc. HCl to deionized water an d make up to 1 L. 
 
F.1.2.2 0.22 mol/L ( ≅≅ ≅≅ 1.5 % V/V) nitric acid  
 
Carefully add 15 mL conc. HNO 3 to deionized water and make up to 1 L.  
 
F.1.2.3 5 % (W/V) KMnO 4 solution 
 
Dissolve 5 g potassium permanganate in deionized wa ter and make up to 100 mL. 
 
F.1.2.4 0.25 mol/L ( ≅≅ ≅≅ 1 % W/V) NaOH solution  
 
Carefully dissolve 10 g sodium hydroxide flakes in deionized water and make up to 1 L. 
 
F.1.2.5 0.8 mol/L ( ≅≅ ≅≅ 3 % W/V) NaBH 4 solution 
 
Dissolve 3 g sodium tetrahydroborate in 1% NaOH sol ution and make up to 100 mL with 1% NaOH solution. 
 
F.1.3  Stock solution 
 
The stock solution contains 1 000 mg/L Hg. The use of commercially available concentrated calibration 
solutions for AAS is recommended. 
 
WARNING   Mercury solutions are toxic.  
 
F.1.4  Calibration solution 
 
1 mg Hg/L (in 1.5 % HNO 3 stabilized by the addition of a few drops of  5 % KMnO 4 solution). 
 
F.1.4.1  Aliquots for calibration:  100, 200, 500 µL 
 
F.1.4.2 Corresponding to:  100, 200, 500 ng Hg 
 
F.1.4.3 Diluent:    1.5 % (V/V) nitric acid or 1.5 % (V/V) hydrochloric acid 
 
F.1.4.4 Calibration volume:  10 mL 
 
F.1.5  Reductant solution:  3 % NaBH 4 in 1 % Na0H solution 
 
 
 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 15   
 
 
F.1.6  Instrument conditions  
 
F.1.6.1 Analytical wavelength:  253.6 nm 
 
F.1.6.2 Slit width and height:  0.7 nm low 
 
F.1.6.3   Radiation source: Electrodeless discharge lamp or h ollow cathode lamp for Hg 
 
F.1.6.4   QTA heating:  No flame required. If condensation in  the QTA is a problem, heat the 
QTA gently by mounting an infrared lamp above it 
 
F.1.6.5  Prepared measurement volume: 10 mL minimum to 50 mL  maximum 
 
F.1.6.6 Pre-reaction purge time:  Approx. 5 s 
 
F.1.6.7 Post-reaction purge time: Approx. 50 s 
 
F.1.6.8 Characteristic mass:  4.68 ng Hg for 1 % absorption  (A = 0.0044) 
 
F.1.6.9  Characteristic concentration:       0.468 µg/L 1 % absorption for 10 mL calibration volume 
 
F.1.6.10 Characteristic concentration check:  250 µL of the 1000 mg/L Hg stock solution (250 ng) give an  
           absorbance of approx. A = 0.2. 
 
F.1.7 Notes 
 
F.1.7.1  Stabilize stock and calibration solutions by addin g KMnO 4 or KI solution. 
 
F.1.7.2 Stabilize all solutions in the reaction flask by ad ding 1 drop of 5 % (w/v) KMnO 4 solution before 
starting the determination. 
 
F.2  Method 2 — Using tin (II) chloride (SnCl 2) as reductant 
 
F.2.1  Scope 
  
This method describes the determination of mercury in pomades and solid brilliantines using tin (II) c hloride 
(SnCI 2) as reductant. 
 
F.2.2  Reagents 
 
F.2.2.1  0.15 mol/L ( ≅≅ ≅≅ 1.5 % V/V) hydrochloric acid  
 
Carefully add 15 mL conc. HCl to deionized water an d make up to 1 L. 
 
F.2.2.2 1 mol/L ( ≅≅ ≅≅ 10 % V/V) hydrochloric acid   
 
Carefully add 100 mL conc. HCl to deionized water a nd make up to 1 L. 
 
F.2.2.3 0.22 mol/L ( ≅≅ ≅≅ 1.5 % V/V) nitric acid  
 
Carefully add 15 mL conc. HNO 3 to deionized water and make up to 1 L. 
 
F.2.2.4  5% (W/V) KMn0 4 solution  
 
Dissolve 5 g potassium permanganate in deionized wa ter and make up to 100 mL. 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  16  
F.2.2.5       5 % (W/V) SnCl 2 
 
Dissolve 50 g tin (II) chlroride dihydrate (SnCl 2 2H 20) in 10 % HCl solution and make up to 1 L with 10 % HCl 
solution. 
 
F.2.3  Stock solution 
 
The stock solution contains 1 000 mg/L Hg.  The use  of commercially available concentrated calibration  
solutions for AAS recommended. 
 
WARNING:  Mercury solutions are toxic.  
 
F.2.4  Calibration solution 
 
1 mg Hg/L (in 1.5 % HNO 3 stabilized by the addition of a few drops of 5 % K MnO 4 solution). 
 
F.2.4.1  Aliquots for calibration: 100, 200, 500 µL 
 
F.2.4.2  Corresponding to:  100, 200, 500 ng Hg  
 
F.2.4.3    Diluent:     5 % (V/V) nitric acid or 1.5% (V/V)  hydrochloric acid 
 
F.2.4.4  Calibration volume:  10 mL 
 
F.2.5  Reductant solution  5 % SnCl 2 2H 2O in 10 % HCl solution. 
 
F.2.6  Instrument Conditions 
 
F.2.6.1 Analytical wavelength:  253.6 nm 
 
F2.6.2 Slit width and height:  0.7 nm low 
 
F2.6.3 Radiation source:  electrodeless discharge lamp or hollow cathode    
     lamp for Hg 
 
F.2.6.4   QTA heating: no flame required.  If condensation in  the QTA is a problem, heat 
the QTA gently by mounting aninfrared lamp above it . 
  
F.2.6.5 Prepared measurement volume:  10 mL minimum to 50 m L maximum. 
 
F2.6.6 Pre-reaction purge time:   Approx. 5 s 
 
F.2.6.7 Post-reaction purge time:  Approx. 50 s 
 
F.2.6.8 Characteristic mass :  4.68 ng Hg for 1 % absorptio n (A = 0.0044). 
 
F.2.6.9   Characteristic concentration:  0.468 µg / L / 1 % absorption for 10 mL calibration volume . 
 
F.2.6.10  Characteristic concentration check:   250 µL of the 1 000 mg/L Hg stock solution (250 ng) give  
       an absorbance of approx. A = 0.2 
 
F.2.7      Notes   
 
F.2.7.1 Stabilize stock and calibration solutions by adding  KMnO 4 solution.  Do not use KI solution since 
iodide interferes in the release of mercury.   
 
F.2.7.2 Stabilize all solutions in the reaction flask by ad ding 1 drop of 5 % (W/V) KMn0 4 solution before 
starting the determination. 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 17   
Annex G 
(normative) 
 
Total amount of heavy metals 
 
The total amount of heavy metals shall be calculate d by adding up the values obtained for lead (Annex D), 
arsenic (Annex E), and mercury (Annex F). The repor t shall be given in ppm. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  18  
Annex   H 
(normative) 
 
Determination of melting point 
 
H.1  Procedure 
 
H.1.1  Heat a quantity of the sample on a water bath whil e stirring until it reaches a temperature of 90 ºC to 
92 ºC.  Cool the molten sample to a temperature of 8 ºC to 10 ºC above the expected melting point.  
Chill the bulb of a thermometer (range 1 ºC to 100 ºC) to 5 ºC, wipe it dry and while it is still cold, dip it 
into the molten sample so that approximately half o f the bulb is submerged.  Withdraw it immediately 
and hold it vertically away from heat until the wax  surface dulls, then dip it for five minutes into a  
water bath having a temperature not higher than 16 ºC. 
 
H.1.2  Fix the thermometer prepared securely in a test tu be so that its lowest point is about 15 mm above 
the bottom of the test tube.  Suspend the test tube  in a water bath adjusted to 16 ºC and raise the 
temperature of the bath at a rate of 2 ºC per minute up to 30 ºC, then change the rate of rise to 1 ºC 
per minute and note the temperature at which the fi rst drop of the melted sample leaves the 
thermometer.  Repeat the determination twice on a f reshly melted portion of the sample.  If the 
variation in three determinations is less than 1 ºC take the average of the three as the melting poin t.  
If the variation in three determinations is more th an 1 ºC, make two additional determinations and 
take the average of five. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 19   
Annex  J 
(normative) 
 
Determination of consistency  
 
J.1  Determination of consistency of the material is ma de by measuring the penetration of a standard 
cone at 25.0 ± 0.5 ºC.  
 
J.1. 2  Apparatus 
 
J.1.2.1  Penetrometer   
 
Any suitable penetro-meter which permits the specif ied cone to drop vertically without appreciable fri ction for 
at least 40 mm and which indicates accurately the d epth of penetration to the nearest 0.1 mm.  The 
instrument shall have a table to carry the test sam ple which may be adjusted to the horizontal before 
conducting the test.  A mechanism for releasing and  clamping the loaded cone shall be provided. 
 
J.1.2.2  Cone   
 
Consisting of a conical body of brass or corrosion resistant steel with detachable hardened steel tip,  
constructed to conform to the dimensions and tolera nces shown in Figure J.1.  The total moving mass, 
namely, that of the cone and its movable attachment s shall be  150.0 ± 0.1 g.  The attachments consist of a 
rigid shaft having a suitable device at its lower e nd for engaging the cone.  The outer surface shall be 
polished to a very smooth finish. 
 
All dimensions in millimetres 
Figure J.1— Penetrometer cone 
KS 1529: 2012 
© KEBS 2012 — All rights reserved  20  
J.1.2.3 Constant temperature bath    
 
A water bath capable of regulating the temperature at 25.0 ºC and of suitable design for conveniently bringing 
the sample container to the test temperature.  The bath shall be provided with a cover to maintain the  
temperature of the air above the sample at 25 ºC. 
 
J.1.2.4 Timing device   
 
A stop-watch or any other suitable instrument capab le of  measuring an interval of 5 seconds to an acc uracy 
of 0.2 seconds. 
 
J.1.2.5 Sample container   
 
Flat-bottomed; metal or glass cylinders that are 10 0 ±  5 mm in diameter and not less than 60 mm in heigh t. 
 
J.1.3  Procedure 
 
J.1.3.1  Melt a quantity of the sample at 82.0  ±  2.5 ºC, pour into one or more of the sample containers, filling 
to within 6 mm of the brim.  Cool at 25.0 ±  0.5 ºC over a period of not less than 16 hours, protecti ng from 
draughts.  Two hours before the test; place the con tainers in a water bath at 25.0  ±  0.5 ºC.  If the room 
temperature is below 23.5 ºC or above 26.5 ºC, adjust the temperature of the cone to 25.0  ± 0.5 ºC by 
placing it in a water bath. 
 
J.1.3.2  Without disturbing the surface of the sample, plac e the container on the penetrometer table, and 
lower the cone until the tip just touches the top s urface of the sample at a spot 25 mm to 39 mm from the 
edge of the container.  Adjust the zero setting and  quickly release the plunger, then hold it free for  5 seconds.  
Secure the plunger, and read the total penetration from the scale.  Make three or more trails each so spaced 
that there is no overlapping of the areas of penetr ation.  Where the penetration exceeds 20 mm, use a 
separate container of the sample for each trial.  R ead the penetration to the nearest 0.1 mm.  Calcula te the 
average of three or more readings; and conduct furt her trials to a total of 10 if the individual resul ts differ from 
the average by more than ± 3 %. 
 
 
 
J.I.4  Calculation 
 
Cosistency = 10 A 
 
where 
 A = mean of all the values of penetration in mm.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 21   
 
Annex  K 
(normative) 
 
Test for rancidity  
 
K.1  Reagents 
 
K.1.1 Concentrated hydrochloric acid,   Analytical reagent grade.  
 
K.1.2  Phloroglucinol solution    
 
Dissolve 0.1 g of phloroglucinol in 100 mL of dieth yl ether. 
 
K.2  Procedure   
 
Shake 10 mL of the material, melt if necessary, wit h 10 mL of concentrated hydrochloric acid and 10 mL  of 
phloroglucinol solution:  Shake for 1 minute. 
 
J.3  Results 
 
The material shall  be taken to have passed the tes t if no pink colour develops. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  22  
Annex  L 
(normative) 
 
Bleed number  
 
L.1  Procedure   
 
Heat the sample to 95 ºC.  Then allow to cool to  10 ºC above its melting point.  Dip a glass tube (or in ternal 
diameter 4 mm and wall thickness 1 mm) into the sam ple so that when it is removed with the upper end 
closed with a finger, it contains approximately 25 mm column of molten sample.  From approximately 12 mm 
above the filter paper (Whatman No. 1 or equivalent ), allow 5 evenly spaced drops of the sample to fal l 
separately on the paper.  The droplets shall have a  diameter of 6 mm to 8 mm.  When the droplets solid ify, 
place the paper on a watch glass and insert in an o ven kept at 30 ºC for 24 hours.  After 24 hours, determine 
the diameter of each droplet plus the oil ring whic h surrounds it.  Subtract the diameter of the dropl et from the 
oil ring and record the result in mm.  The average of these result in mm.     
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 23    
 
Annex  M 
(normative) 
 
Test for stability 
 
M.1  Apparatus  
 
M.1.1  Ultra violet lamp ,  with emission at 360 mm. 
 
M.2  Procedure  
 
Place 50 mL of the material in a 100 mL glass beake r.  Turn on the ultra violet lamp and expose the sa mples 
at a distance of 12 cm to 14 cm below the lamp for 6 hours.  After the specified time, remove the samp le, 
cool to room temperature and compare for any change  in odour or colour.  The same volume of material s hall 
be employed for all tests so that comparison is ens ured on a reproducible basis. 
 
NOTE   The output of the ultra violet lamp diminishes  wit h time in service.  A log of number of hours of the lamp in use shall be  
maintained.  The lamp is  to be replaced after the specified hours of service , as recommended by the lamp manufacturer. 
 
M.3  Evaluation   
 
Evaluation is done by comparing the test material a gainst an unexposed specimen from the same sample. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  24  
Annex N 
(normative) 
 
Determination of yeasts and moulds in pomades and s olid  brilliantines 
 
N.1  Introduction 
 
N.1.1  This method is obtained from the AOAC official meth od 998.10 entitled   
 
“Efficacy of preservation of non-eye area water-mis cible cosmetic and toiletry formulations”. 
 
N.1.2  Acknowledge of microbiological techniques is requi red for these procedures.  A general aseptic and 
safety procedures shall be followed. 
 
N.1.3  Table N.1 gives the results of the interlaboratory  study supporting the acceptance of the method. 
 
N. 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 pro ducts are 
determined over time.  Products meeting the specifi ed criteria are considered adequately preserved for  
manufacture and consumer use.  Products not meeting  criteria are considered inadequately preserved. 
 
N.3  Apparatus 
 
N.3.1  Jars  –2-4 oz wide-mouth, straight-side flint glass oint ment jars with linerless metal, polypropylene or 
teflon lined screw caps. 
 
N.3.2  Disposable borosilicate glass culture tubes , 16 x 125 mm, with caps. 
 
N.3.3  Disposable borosilicate glass culture tubes , 20 mm x 150 mm, with screw caps. 
 
N.3.4  Petri plates , 100 mm x 15 mm. 
 
N.3.5  Sterile  2.2 mL pipettes. 
 
N.3.6  Sterile swabs . 
 
N.3.7  Glass beads . 
 
N.3.8  Sterile gauze .  
 
N.3.9  µµ µµL 10 to 20 µµ µµL inoculating loops . 
 
N.3.10   Vortex mixer . 
   
 
  
 
 
 
 
 
 
 
 
 
 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 25   
Table N.1 — Interlaboratory study results for deter mination of preservation of non-eye area water-
miscible cosmetic and toiletry formulations 
 
                                    Incidence of false -negatives                       
among total positive samples a)  Incidence of false -positives among total 
negative samples b)   
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 adequ ately preserved. 
 
b)  False-positive analysis indicates a sample is not a dequately preserved. 
 
 
 
N.4  Reagents 
 
For convenience, dehydrated media of any brand equi valent in function may be used.  Test each lot of 
medium for sterility and growth-promotion using sui table organisms. 
 
N.4.1  Letheen agar   
 
Contains 5.0 g pancreatic digest of casein 1.0 g de xtrose, 3.0 g beef extract, 1.0 g lecithin, 7.0 g p olysorbate 
80, and 15.0 g agar per L.  Prepare according to ma nufacturer’s directions.  Dispense into suitable co ntainers 
and sterilize by autoclaving at 121 ºC for 15 min.  Final pH shall be 7.0  ± 0.2 at 25 ºC .  Place in 45 ºC water 
bath until agar is 45 ± 2 °C.  Use for pour plates. 
 
N.4.2 D/E neutralizing broth (Dey/Engley)  
 
Contains 5.0 g pancreatic digest of casein, 2.5 g y east extract, 10 g dextrose,1.0 g sodium thiogycoll ate, 6.0g 
Na 2 S2O3 . 5H 2O, 2.5 g NaHSO 3, 7.0 g lecithin, 5.0 g polysobate 80, and 0.02 g b romcresol purple per L.  
Prepare according to manufacturer’s directions.  Di spense 9 or 9.9 mL aliquot into tubes and sterilize  by 
autoclaving at 121 ºC for 15 min.  Final pH shall be 7.6 ± 0.2 at 25 ºC.  Use for aerobic plate count, L, 
dilutions. 
 
N.4.3  Nutrient agar    
 
Contains 5.0 g pancreatic digest of gelatin 3.0 g b eef extract and 15.0 g agar per L.  Prepare accordi ng to 
manufacturer’s directions.  Dispense into tubes and  sterilize by autoclaving at 121 ºC for 15 min.  Final pH 
shall be 6.8 ± 0.2 at 25 ºC .  Cool in inclined position to form a slant.  Us e for bacterial culture maintenance 
and inoculum preparation. 
 
N.4.4  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 i nto tubes and sterilize by autoclaving at 121 °C for 15 min.  
Final pH shall be 6.2 ± 0.2 at 25 ºC.  Cool in slanted position.  Use for yeast cultur e maintenance and 
inoculum preparation. 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  26  
N.4.5  Potato dextrose agar (PDA)  
 
Contains 200 g potato infusion, 20.0 g dextrose, an d 15.0 g agar per L.  Prepare according to manufact urer’s 
directions.  Dispense into tubes and sterile by aut oclaving at 121 ºC for 15 min. Final pH shall be 5.6 ± 0.2 at 
25 ºC.  Cool in slanted position.  Use for mould cultur e maintenance and inoculum preparation. 
 
N.4.6  0.85 % Saline   
 
Dissolve 8.50 g NaCI in water and dilute to 1 L.  D ispense into flasks or bottles and sterilize by aut oclaving at 
121 ºC for 15 min. 
 
N.4.7      0.85 % saline with 0.05 % polysorbate 80      
 
Dissolve 8.5 g NaCI in water , mix in 0.50 g polyso rbate 80, and dilute to 1 L.  Dispense into suitabl e 
containers and sterilize by autoclaving at 121 ºC for 15 min. 
 
N.4.8  Barium sulphate standard No. 2   
 
N.4.8.1 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.  
 
N.4.8.2 Prepare a 1.0 % H 2SO 4 solution by mixing 1.0 mL H 2SO 4 in 100 mL water. Let this be refered to as 
solution 2.  
 
N.4.8.3 Mix 0.2mL of solution 1 with 9.8 mL solution 2, in screw-capped test tube.  Cap tightly  and store in 
the dark at room temperature. 
 
N.4.9  Barium sulphate standard No. 7  
 
Use solutions from N.4.8.  Mix 0.7 mL of solution N .4.8.1, with 9.3 mL of solution N.4.8.2, in a screw -capped 
test tube. Cap tightly and store in the dark at roo m temperature. 
 
N.5  Micro-organisms 
 
N.5.1  Staphylococcus aureus   — ATCC 6538 
 
N.5.2 Staphyloccuss epidermidis — ATCC 12228 
 
N.5.3 Klebsiella pneumoniae   — ATCC 10031 
 
N.5.4 Escherichia coli    — ATCC 8739 
 
N.5.5 Enterobacter gergoviae   — ATCC 33028 
 
N.5.6 Pseudomonas aeruginosa  — ATCC 9027 
 
N.5.7 Burkholderia cepacia   — ATCC 25416 
 
N.5.8 Acinetobacter baumanii   — ATCC 19606 
 
N.5.9 Candida albicans   — ATCC10231 
 
N.5.10 Aspergillus niger   — ATCC 16404 
 
NOTE   Environmental micro organisms (s) likely to be conta minants of concern during product manufacture or us e be included as a 
separate inoculum.  Predominant environmental microb es isolated during manufacturing, equipment cleanin g, and sanitizing, or from 
related deionised water systems are used as supplem ental test inocula). 
 
 
 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 27   
 
N.6  Product quality check 
 
N.6.1  Weigh 1.0 g product into a screw-capped culture tu be containing 9.0 mL sterile neutralizing broth to 
make a 1:10 dilution.  If necessary to disperse pro duct, add ten to twenty 3 mm diameter glass  beads to 
tube. Mix on vortex mixer until homogeneous. 
 
N.6.2 Pipet 1.0 mL of the 1:10 dilution into each of 4 st erile petri plates.  Pour 15-20 mL sterile molten 
Letheen agar (45 ± 2 ºC ) into each plate .  Mix by rotating plates to di sperse the dilution thoroughly. Let it 
solidify. 
 
N.6.3 Invert and incubate 2 plates at 35 ± 2 ºC for 48 h and 2 plates at 25 ± 2 ºC for 5 days. 
 
N.6.4 Count the number of colonies on all plates, add, an d multiply by 2.5 to determine the number of 
colony forming units per gram (cfu./g) in the produ ct. 
 
N.6.5 Save plates to be used for the neutralization valid ation in N.14 by refrigerating. 
 
N.7  Product preparation 
 
N.7.1  Measure 20 mL sterile saline into 4 sterile jars , N.3.1. Cap tightly and store at room temperature. 
 
N.7.2 Weigh 20 g product into each of 4 sterile jars, N.3 .1. Cap tightly and store at room temperature. 
 
N.8   Bacterial inocula preparation 
 
N.8.1  Streak each bacteria culture, N.5.1 – N.5.10 onto a nutrient agar,  N.4.3, slant. Incubate for 48 h at 35 
± 2 °C.  Wash each slant with 5.0 mL sterile saline, loo sening the culture from the agar surface.  Transfer  the 
suspension into a sterile tube.  Repeat the wash wi th a second 5.0 mL aliquot of saline.  Combine wash es 
and mix on vortex mixer to disperse evenly. 
 
N.8.2 Adjust each wash with sterile saline to yield a sus pension of ca 10 8 cfu/mL using Mc-Farland BaSO 4 
standard No, 2, N 4.8, direct microscopic count, tu rbidimetry, absorbance, or other method correlated to an 
aerobic plate count (APC), N.13. Perform an APC, N. 13, on each suspension to confirm standardization. 
 
N.9       Fungal inoculum preparation 
  
N.9.1  Steak C. albicans , N.5.9, on 3 slants of Y/M agar,  N.4.4. Incubate at 25 ± 2 ºC for 48 h.  Wash each 
slant sequentially with 5.0 mL aliquot of sterile s aline.  Repeat with a second 5.0 mL aliquot of ster ile saline.  
Combine washes to produce 10 mL suspension.  Mix on  vortex mixer to disperse evenly. 
 
N.9.2 Adjust the wash with sterile saline to yield a susp ension of ca 10 7 cfu/mL using a Mc-farland BaSO 4 
standard No. 7, N.4.9, direct microscopic count, tu rbidimetry, absorbance, or other method that has be en 
correlated to an APC,  N.13. Perform an APC, N.13, on the suspension to co nfirm standardization. 
 
N.9.3 Streak A.Niger , N.5.10, on 5 slants of potato dextrose agar  N.4.5. Incubate at 25 ± 2 ºC for 10 days.  
Dislodge mould spores by adding 5.0 mL sterile sali ne containing 0.05 % polysorbate 80 to each tube an d 
vigorously rubbing the surface of the agar slant wi th a sterile swab.  Repeat with a second 5.0 mL ali quot in 
each tube.  Combine the 10 washes to produce 50 mL suspension.  Filter into a sterile container throug h 3-5 
layers of sterile gauze supported in funnel.  Perfo rm an APC, N.13, using appropriate dilutions. Adjus t mould 
suspension to ca 10 7 per mL using sterile saline.  Use immediately or r efrigerate at 2 – 5 ºC for up to 1 
month. Verify mould viability by an APC,  N.13, before each use. 
 
N.10    Inoculum pools 
 
N.10.1  Pool equal parts of the S. aureus  and S. epidermidis  suspensions, N.8.2  in a sterile container to 
make incoculum pool 1: gram-postive cocci. 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  28  
N.10.2     Pool equal parts of the K.  pneumoniae  , E . coli, and E . gergoviae  suspensios, N.8.2, in a sterile  
container to make inoculum pool . 2: gram — negativ e fermentors. 
 
N.10.3   Pool equal parts of the P. aeruginosa , B .cepacia  and A. baumanii suspensions, N.8.2, in a sterile 
container to make inoculum pool 3: gram — negative nonfermentors. 
 
N.10.4   Pool equal parts of C. Albicans, N.9.2, and A. Niger, N.9.3, suspensions in a sterile container to 
make innoculum pool 4: fungi. 
 
N.10.5   Use organism pools immediately or refrigerate them at 2 to 5 ºC for more than 72 h. 
 
N.11   Inoculation 
 
N.11.1  Inoculate each of the four 20.0 mL aliquots of ster ile saline, N.7.1,  with 0.2 mL of its respective 
inoculum pool, N.10.1  – N.10.4. Mix thoroughly.  Use these suspensions to d etermine inoculums counts (see 
K (a). 
 
N.11.2  Inoculate each of the four 20 g products suspension s, F (b) with 2.0 mL of its respective inoculums 
pool I(a) –d).  Mix thoroughly by shaking, vortex m ixing or stirring, so that each suspension contains  10 6 
bacteria or 10 5 fungi per gram, evenly distribute throughout the p roduct.  Tightly close inoculated containers 
and store at ambient temperature (20 to 25 ºC). 
 
N.12  Sampling intervals 
 
N.12.1  Sample each inoculated saline suspension, N.11.1, for APC,  N.13, within 1 h after inoculation to 
obtain inoculum count. 
 
N.12.2 Test each inoculated product,  N.11.2   for APC, N.13 at 7, 14 and 28 days after inoculatio n to obtain 
product interval count. 
 
N.13   Aerobic plate count (APC) 
 
N.13.1  Mix suspension thoroughly, weigh 1.0 g product int o screw-capped culture tube containing 9.0 mL 
sterile neutralizing broth for a 1: 10 dilution.  I f necessary to disperse product, add 10 – 20 steril e 3 mm 
diameter glass beads to the tube.  Mix on vortex mi xer until homogeneous. 
 
N.13.2 Aseptically pipette 0.1 mL of the 1: 10 dilution in to 9.9 mL tube of neutralizing broth to obtain a 1:  1 
000 dilution. Vortex mix. Pipette 0.1 mL of the 1:1  000 dilution into 9.9 mL neutralizing broth to obt ain a 1: 
100 000 dilution.  The number of dilutions may be d ecreased if previous counts microbial populations s how 
reduction. 
N.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 plate s. If necessary, transfer duplicate 1.0 and 0.1 mL aliquots 
from the 1: 1 000 dilution for plates 1:1 000 and 1 : 10 000, and from the 1: 100 000 dilution for plat es 1: 100 
000 and 1: 1 000 000.  Pour 15 – 20 mL sterile leth een agar N.4.1, at 45 ± 2 ºC into each plate. Mix by 
rotating the plates to disperse the suspension thor oughly, and let solidify. 
N.13.4 Invert bacterial plates and incubate at 35 ± 2 ºC.  Examine bacterial plates after 48 – 72 h.  Coun t in 
suitable range (30 – 300 colonies).  If no countabl e plates fall in that range, count the plates(s) ne arest that 
range showing distinct colonies.  Average duplicate  plates counts and express results as cfu/g of prod uct. 
N.13.5 Invert and incubate fungal plates at 25 ± 2 ºC.  Read fungal plates at 2 – 3 days and record res ults.  
Count plates in a suitable range  (30 – 300 colonie s).  If no countable plates fall in that range, cou nt the 
plate(s) nearest that range showing distinct coloni es.  Reincubate plates for another 2 – 3 days.  Rea d and 
record additional colonies.  Add to previous result s to obtain total counts.  Average duplicate plate counts and 
record as cfu/g of products.   
 
                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 29   
N.14  Neutralization check 
Make a 1:10 000 dilution in sterile saline of pools  1, 2 and 3, N.10.1 – N.10.3, and a 1: 1 000 diluti on of pool 
4, N.10.4.  Steak each dilution for isolation with a 10 µL loop on the plates saved from N.6.5.  If plates a re not 
usable due to either desiccation or surface growth,  repeat N.6, and steak freshly prepared plates.  In cubate 
as in N.13.4 and N.13.5.  
N.15  Data analysis 
N.15.1   Product quality check, N.6.4, must be found to co ntain, <100 cfu/g to proceed with the challenge tes t. 
N.15.2  Inoculums counts, N.12.1, shall be between 1 to 9.9  x 10 6 cfu/g product for bacteria and 1 to 9.9 x 
10 5 cfu/g product for fungi, or the test shall be repe ated with different dilutions. 
N.15.3 Neutralization check, N.14,  must show significant growth of all pools to confir m adequate 
nutralization.  A neutralizing broth other than D/E  broth can be used.  If neutralization does not occ ur, the test 
is invalid.   
N.15.4 Calculate the percentage reduction: 
Reduction, % =    100 x count  Inoculum count  interval product -  count  Inoculum  
N.15.5  The test product is considered adequately preserved  if:  
a) bacteria show at least 99.9 % (3 log) reduction within 1 week following challenge and remain at or 
 below that level thereafter; and  
b) fungi show at least a 90 % (1 log) reduction wit hin 1 week following challenge, and remain at or 
below that level thereafter.  These criteria apply to freshly prepared formulations.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  30  
Annex P 
(informative) 
 
List of raw materials conventionally used in the fo rmulation of pomades and 
brilliantines  
 
P.1  Type 1   
 
Based on mineral oils and waxes. 
 
a) paraffin wax; 
 
b) microcrystalline wax; 
 
c) petroleum jelly; 
 
d) mineral oil; 
 
e) perfume and colour. 
 
P.2 Type 2   
 
Based on vegetable oils and waxes. 
 
a) castor oil; 
b) beeswax;  
c) paraffin wax; 
d) coconut oil; 
e) perfume and colour. 
P.3  Type 3  
 
Based on mineral oils and fatty acids. 
 
a) stearic acid; 
b) mineral oil; 
c) perfume and colour. 
 
P.4  Type 4  
 
Based on mixture of mineral and vegetable oils, ani mal fats and waxes. 
 
a) coconut oil; 
b) mineral oil; 
c) beeswax; 
d) petroleum jelly; 
e) lanolin; 
f) perfume and colour                       KS 1529: 2012 
© KEBS 2012 — All rights reserved                                                                                                                 31   
 
Annex  Q 
(normative) 
 
Antibacterial test 
 
Q.1 Procedure 
 
Prepare nutrient agar for bacterial growth by dispe rsing 28 g of nutrient agar powder in 1 litre of de ionized 
water. Allow to soak for 10 minutes, swirl to mix a nd then heat gently with stirring to ensure uniform ity. 
Sterilize by autoclaving for 15 minutes at 121 ºC, cool at 47 ºC, mix well and then pour to sterilized petri 
dishes. Leave it  to solidify undisturbed. Plant  Staphylococcus aureus,  pseudomonas aeruginosa and 
candida albicans  on the so prepared nutrient agar in petri dishes. 
 
Meanwhile, prepare filter paper discs and sterilize  them by autoclaving. Dip in various samples. Place  in the 
petri dishes containing bacteria culture agar mixtu re. Incubate the petri dishes  at 35 ºC for 48 hours. 
Determine bacteria growth inhibition zones. 
 
Q.2 Antifungal test 
 
Dissolve potato dextrose agar (39 g) in 1 dm 3 of distilled water. Use the same procedure as for b acterial test  
(Q.1 ) above.  Test the cream against fusarium fung i . Obtain the results after 4 days and the tempera ture of 
incubation shall be 25 ºC. 
 
Q.3 Results 
 
The inhibition zone shall be at least 0.2 mm in dia meter. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 KS 1529: 2012 
© KEBS 2012 — All rights reserved  32  
 
 
Annex  R 
(informative) 
 
Available Kenya Standards on fats, oils and waxes  
 
R.1  Mineral oils and waxes 
 
a) mineral oil  - KS 03-796:1989 1)  
b) paraffin wax  - KS 03-1426:1997 2)  
c) microcrystalline wax - KS 03-1427:1997 3)  
d) petroleum jelly  - KS 03-581:1998 4)  
 R.2  Vegetable oils 
 
a)  sesame oil  - KS 03 797:1988 5) 
b) groundnut oil  - KS 03-798:1989 6)  
c) castor oil  - KS 03-799:1989 7) 
d) coconut oil  - KS 03-800:1989 8)  
e) mustard oil  - expressed raw type 
f)           avocado oil                    -            KS 1765 9)  
R.3  Animal fats 
 
a) lanolin   - KS 03-915:1989 10) 
 
 
 
 
 
                                                 
1)  Specification for mineral oil for cosmetic industr y 
2)  Specification for paraffin wax 
3)  Specification for microcrystalline wax of petroleu m origin  
4)  Specification for pure petroleum jelly for human u se 
5)  Specification for sesame oil for petroleum industr y 
6)  Specification for groundnut/peanut oil for cosmeti c industry 
7)  Specification for castor oil for cosmetic industry  
8)  Specification for coconut oil for cosmetic industr y 
9)  Specification for avocado oil for cosmetic industr y 
10)  Specification for anhydrous lanolin for cosmetic i ndustry