DRAFT UGANDA STANDARD  
  DUS 1625  
First Edition  
2015 -mm-dd 
 
 
 Reference number  
DUS 1625 : 2015 
© UNBS 2015   
  
Acid —Based instant hand sanitizers — Specification   
 DUS 1625 : 2015  
ii © UNBS 2015  – All rights reserved  
 Compliance with this standard does not, of itself confer immunity from legal obligations  
A Uganda Standard does not purport to include all necessary provisions of a contract. Users are 
responsible for  its correct application  
© UNBS 20 15  
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The Executive Director  
Uganda National Bureau of Standards  
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Kampala  
Uganda  
Tel: 256 41 4 505 995  
Fax: 256 41 4 286 123  
E-mail: info@unbs .go.ug  
Web: www.unbs.go.ug  DUS 1625 :2015  
© UNBS 2015  – All rights reserved  iii 
 Contents  Page 
Foreword  ................................ ................................ ................................ ................................ ............................  iv 
1 Scop e ................................ ................................ ................................ ................................ ......................  1 
2 Normative references  ................................ ................................ ................................ ............................  1 
3 Terms and definitions  ................................ ................................ ................................ ...........................  1 
4 Require ments  ................................ ................................ ................................ ................................ .........  1 
4.1 General requirements  ................................ ................................ ................................ ...........................  1 
4.2 Specific quality requirements  ................................ ................................ ................................ ..............  2 
5 Packaging and labelling ................................ ................................ ................................ ........................  2 
5.1 Packaging  ................................ ................................ ................................ ................................ ...............  2 
Annex A  (normative)  Determination of disinfecting efficacy  ................................ ................................ ....... 4 
A.1 Outline of the method  ................................ ................................ ................................ ...........................  4 
A.2 Apparatus  ................................ ................................ ................................ ................................ ...............  4 
A.3 Media ................................ ................................ ................................ ................................ .......................  4 
A.3.1  Growth media for test organisms  ................................ ................................ ................................ ........  4 
A.3.2  Recovery medium  ................................ ................................ ................................ ................................ .. 5 
A.3.4  Yeast suspension  ................................ ................................ ................................ ................................ .. 5 
A.3.5  Ringers solution, 25 % (v/v)  ................................ ................................ ................................ .................  6 
A.4 Selection of the most resistant organism by the minimum inhibitory concentration tes t ............  6 
A.5 Preparation of inoculum  ................................ ................................ ................................ .......................  6 
A.6 Preparation of the sanitizer dilutions  ................................ ................................ ................................ .. 7 
A.7 Test procedure  ................................ ................................ ................................ ................................ ....... 7 
 DUS 1625 : 2015  
iv © UNBS 2015  – All rights reserved  
 Foreword  
Uganda National Bureau of Standards (UNBS) is a parastatal under the Ministry of Trade , Industry and 
Cooperatives established under Cap 327 , of the Laws of Uganda , as amended .  UNBS is mandated to co -
ordinate the elaboration of standards and is  
(a) a member of International Organisation for Standardisation (ISO) and  
(b) a contact point for the WHO/FAO Codex Alimentarius Commission on Food Standards, and  
(c) the National Enquiry Point on TB T Agreement  of the World Trade Organisation (WTO).  
The work of preparing Uganda Standards is carried out through Technical Committees. A Technical 
Committee is established to deliberate on standards in a given field or area and consists of representatives of 
consumers, traders, academicians, manufacturers, government and other stakeholders.  
Draft Uganda Standards adopted by the Technical Committee are widely circulated to stakeholders and the 
general public for comments. The committee reviews the comments b efore recommending the draft standards 
for approval and declaration as Uganda Standards by the National Standards Council.  
Responsible Technical C ommittee  
 
 DRAFT UGANDA STANDARD  DUS 1625 : 2015  
 
© UNBS 2015  – All rights reserved  1 
 Acid —Based instant hand sanitizers  — Specification  
1 Scope  
This Draft uganda  standard specifies  the requirements and methods of test for acid based instant hand 
sanitizers.  
2 Normative references  
The following referenced documents are indispensable for the application of this document. For dated 
references, only the edition cited applies. For undat ed references, the latest edition of the referenced 
document (including any amendments) applies.  
EAS 104, Alcoholic beverages – Method of sampling and test  
EAS 377 -1, Cosmetics and cosmetics products — Part 1: List of substances prohibited in cosmetic pr oducts  
EAS 377 -2, Cosmetics and cosmetics products — Part 2: List of substances which cosmetic products must 
not contain except subject to restrictions laid down  
EAS 377 -3, Cosmetics and cosmetics products — Part 3: List of colourants allowed in cosmetic  products  
EAS 377 -4, Cosmetics and cosmetics products — Part 4: List of preservatives allowed in cosmetic products  
EAS 377 -5, Cosmetics and cosmetics products — Part 5: Use of UV filters in cosmetic products  
3 Terms and definitions  
For the purposes of t his standard terms and definitions given in I SO 862 and the following apply.  
hand sanitizers  
antiseptic agents used to cleanse the hands when soap and water are unavailable. They are often used to 
protect and prevent the passage of bacteria, virus and oth er pathogens that can cause infections.  
4 Requirements  
4.1 General requirements  
4.1.1  The sanitizer shall be clear, homogenous and in the form of liquid or gel.  
4.1.2  The sanitizer shall not have any objectionable  odour or smell.  
4.1.4  The substances used in the sanitizers shall conform to all the parts of EAS 377  DUS 1625 : 2015  
2 © UNBS 2015  – All rights reserved  
 4.2 Specific quality requirements  
The hand sanitizer shall also comply with the specific quality requirements given in Table 1 when tested in 
accordance with the corresponding test method.  
Table 1 — Specific quality requirements for instant hand sanitizer  
Sl. No  Characteristic  Requirement  Test method  
1 Alcohol content (ethanol and/or isopropanol, n -
propanol), %, v/v, min.  60.0 EAS 104  
2 pH (neat)  2-5  
3 Bactericidal efficacy  to pass te st Annex A  
4 Virucidal efficacy  to pass test  ASTM E1838 -02 
 
5 Packaging and labelling  
5.1 Packaging  
5.1.1  The sanitizer shall be supplied in suitable well -closed containers/packages.  
5.1.2  The containers/packages (including the closures) shall not int eract chemically with the sanitizer and 
shall be strong enough to protect the sanitizer adequately during normal handling, transportation and storage.  
5.1.3  Only containers/packages of the same size and bearing the same batch identification shall be packe d 
together in a bulk container.  
5.2 Labeling  
The container/package shall be securely closed and marked legibly and indelibly with the following 
information:  
 
a) name of the product as ― Acid- based hand sanitizer   
b) manufacturer‘s name and physical address  
c) NOTE The name, physical address of the distributor/supplier and trade mark may be added as required  
d) batch or code number;  
e) net content;  
f) list of ingredients used;  
g) general instructions for use (be in either English, or in combination as agreed between the 
manufacturer and supplier);  
h) date of manufacture and expiry date;  
i) country of origin/manufacture;  
j) the following precautionary warnings:  
   i)―Do not allow the sanitizer  to come into contact with eyes ;  DUS 1625 : 2015  
© UNBS 2015  – All rights reserved  3 
   ii) ―Keep Out Reach of Children ;  
  iii) ―If swallowed contact a doctor ; and  
 iv) ―Highly flammabl e, keep away from fire or flame .  DUS 1625 : 2015  
4 © UNBS 2015  – All rights reserved  
 Annex A  
(normative)  
 
Determination of disinfecting efficacy   
A.1 Outline of the method  
A.1.1  The sanitizer is tested at the recommended ‗use‘ -dilution‘ and concurrently at 0.5 and 1.5 times that 
dilution. The test consists of challenging the diluted sanitizer with bacterial inoculum, withdrawing a sample 
after a given time and culturing the sample in a suitable recovery medium. After this sampling, the mixture is 
again challenged by a second inoculum and after a second interval, is again sampled for culturing. This 
process is then repeated to provide a third chall enge.  
A.1.2  The sample is considered to have passed or failed the test according to the extent of growth shown in 
the first two cultured samples.  
A.2 Apparatus  
A.2.1  Facility, for incubation at 37 °C ± 1 °C  
A.2.2  Facility, for incubation at 2 °C ± 1 °C   
A.2.3  Stop clock, indicating in seconds  
A.2.4  Facility, for refrigeration at 4 °C ± 1 °C  
A.2.5  Universal containers, made of glass and having metal tops with rubber liners. Plastic containers or 
glass containers with plastic tops shall not be used.  
A.2.6 Test tubes, 19 mm X 150 mm  
A.2.7  Filter paper, No. 4 Whatman (sterile) or equivalent  
A.2.8  Facility, for autoclaving at 121 °C ± 1 °C  
A.2.9  Pipette, capable of dispensing 0.02 mL ± 0.005 mL  
A.2.10  pH meter  
A.2.11  Facility, to sterilize by filtrati on  
A.2.12  150 μm test sieve  
A.2.13  Oven, capable of maintaining temperature at 100 °C ± 1 °C  
A.3 Media  
A.3.1  Growth media for test organisms  
A.3.1.1  The growth media for test organisms shall be Wright and Mundy Broth with Dextrose (WMBD).  DUS 1625 : 2015  
© UNBS 2015  – All rights reserved  5 
 A.3.1.2  Dispense 10 mL and 6 mL quantities of the Wright and Mundy Broth into universal bottles, and 
autoclave at 121 °C ± 1 °C for 12 min.  
A.3.1.3  Add to this medium, 10 % (m/V) dextrose solution sterilized by filtration, to give a final dextrose 
concentration of 0. 1 % (m/v), (that is, to 10 mL broth add 0.1 mL dextrose solution and to 6.0 mL broth add 
0.06 mL dextrose solution).  
A.3.2  Recovery medium  
A.3.2.1  Composition  
A nutrient broth prepared as follows:  
a) beef extract, 10 g;  
b) peptone, 10 g;  
c) sodium chloride, 5  g; and  
d) polyoxyethylene sorbitan mono -oleate, 30 g  
 
A.3.2.2  Preparation  
Add the ingredients to 1 000 mL of water. Mix well. Dispense 10 mL quantities into test tubes and autoclave at 
121 °C ± 1 °C for 15 min .  
A.3.3  Hard water  
Standard hard water with 342 mg/L (ppm) hardness prepared as follows: dissolve 0.304 g of anhydrous 
calcium chloride hexahydrate (MgCl 2.6H 20) in distilled water and make up the volume to one litre. Sterilize the 
standard hard water by autoclaving at 121 °C ± 1 °C for 15 min. Allow  this to reach room temperature before 
use.  
A.3.4  Yeast suspension  
A.3.4.1  Weigh to the nearest gram about 65 g of active dry yeast. Cream by the gradual addition of sterile 
hard water (A.3.3) using a heavy glass rod for stirring. Decant the creamed port ion into a flask, add more hard 
water to any lumpy residue remaining and repeat the creaming and decantation until no residue remains, and 
500 mL of hard water has been used.  
A.3.4.2  Shake the contents of the flask vigorously and strain -through a 150 μm sieve (A.2.12) breaking 
down any remaining lumps.  
A.3.4.3  Add 500 mL sterile hard water, shake vigorously.  
A.3.4.4  Transfer 50 mL or 100 mL portions into screw -capped bottles, screw the caps tightly and 
autoclave at 121 °C ± 1 °C for 15 min. Allow the  autoclave to cool without releasing the pressure. Store cold 
but not freezing.  
A.3.4.5  Dry two glass petri -dishes to constant mass. Into each of these dishes, pipette 25 mL of sterilized 
yeast suspension and dry to constant mass at 100 °C. Calculate the average solids content of the suspension.  
A.3.4.6  Before use, pipette 25 mL of the sterilized yeast suspension into a beaker. Determine the pH 
using a glass electrode, and determine the volume of 40 g/L sodium hydroxide solution needed to adjust the 
pH to  7.0 ± 0.1.  DUS 1625 : 2015  
6 © UNBS 2015  – All rights reserved  
 A.3.4.7  Immediately before use, add to each bottle of sterilized yeast suspension a volume of sterile hard 
water and a volume of 40 g/L sodium hydroxide calculated to adjust the concentration of dry yeast to 5 % 
(m/v) and the pH to 7.0 ± 0.1. D iscard prepared yeast, two weeks after preparation.  
A.3.5  Ringers solution, 25 % (v/v)  
Dissolve 9.00 g of sodium chloride, 0.42 g of potassium chloride, 0.24 g of anhydrous calcium chloride and 
0.20 g of sodium bicarbonate in water and dilute to 1 000 mL . Add one volume of this solution to three 
volumes of water to give a 25 % solution. Dispense into test tubes fitted with suitable closures and sterilized 
by auto -claving at 121 °C ± 1 °C for 15 min.  
A.4 Selection of the most resistant organism by the min imum inhibitory 
concentration test  
A.4.1  The following organisms shall be used for the test:  
a) Pseudomonas aeruginosa (NCTC 6749 or equivalent);  
b) Proteus vulgaris (NCTC 4635 or equivalent); and  
c) Staphyloccus aureus (NCTC 4163 or equivalent).   
These organis ms may be obtained as freeze dried cultures. Once sub -cultured, the organisms shall be 
maintained on agar slopes of suitable nutrient medium at 4 °C ± 1 °C.  
A.4.2  Subculture each organism daily into a universal bottle containing 6 mL of growth medium (see  A.3.1) 
and incubate for 24 h ± 2 h at 37 °C ± 1 °C.  
A.4.3  Dilute one part of freshly grown sub -culture of each organism, which is at least a fifth sub -culture and 
not more than a fourteenth, with ten parts of the growth medium (see A.3.1) before dilution , the P. aeruginosa , 
culture shall be filtered using a Whatman No.4 filter paper.  
A.4.4  Prepared three sets of ten, doubling dilutions of the sanitizer in universal containers (A.2.5). For this 
purpose dilute the neat sanitizer in the growth medium (see A .3.1) or the recovery medium (A.3.2) to give a 
final volume of 5 mL of the diluted sanitizer for each dilution.  
A.4.5  Inoculate each dilution in one set with 0.02 mL of a diluted culture of one organism (see A.4.3).  
A.4.6  Incubate all the three sets of i noculate dilutions at 37 °C ± 1 °C for 72 h, and examine to determine 
the organism most resistant to the sanitizer, that is the organism for which the minimum inhibitory 
concentration is highest.  
A.5 Preparation of inoculum  
A.5.1  Daily sub -cultures of the  test organism selected as in A.4.6 shall be grown in 6 mL quantities of the 
growth medium (A.3.1) and incubated at 37 °C ± 1 °C for 24 h ± 2 h.  
A.5.2  The day before the test, inoculate 10 mL of the growth medium (A.3.1) with the test organism from a 
daily sub -culture and not more than a fourteenth. Incubate the inoculated, broth at 37 °C ± 1 °C for 24 h ± 2 h.  
A.5.3  Add 6 mL of the test organism culture (A.5.1) and (A.5.2) to 4 mL of the yeast suspension (A.3.4) thus 
making a final concentration of 2 % ( m/v) of yeast in the yeast/organism suspension. If a culture of P. 
aeruginosa is used, it shall be filtered using a Whatman No.4 filter paper before addition.  
A.5.4  Shake the yeast/organism suspension for one minute with a few sterile glass beads. Immedia tely 
before the test, count the number of viable organisms in the inoculum by decimal dilutions in 25 % Ringers DUS 1625 : 2015  
© UNBS 2015  – All rights reserved  7 
 solution (see A.3.5) and by the drop plate method. The viable count shall be not less than 10 8 organisms/mL 
or more than 10 10 organisms/mL or th e test results are considered invalid.  
A.6 Preparation of the sanitizer dilutions  
Prepare three dilutions of t he sanitizer in hard water (A.3.3) based on the recommended ‗use dilution‘ of the 
sanitizer, as follows:  
 A = 0.5 times the recommended ‗use -dilution‘;  
 B = 1.0 times the recommended ‗use -dilution‘; and  
 C = 1.5 times the recommended ‗use -dilution‘.  
 
The sanitizer dilutions shall be prepared and tested on the same day.  
A.7 Test procedure  
A.7.1  The test shall be carried out at 27 °C ± 1 °C.  
A.7.2  Dispense 3 mL of each dilution of sanitizer (A6) into separate universal bottles labelled A, B, and C, 
then a llow to equilibrate to 27 °C ± 1°C.  
A.7.3  Add 1 mL of the inoculum to A, B and C at 0, 1 and 5 min respectively and mix by swirling gently.  
A.7.4  Eight minutes after the addition of the inoculum, remove a sample of the inoculum/sanitizer mixture 
and put 0.02 mL into each of the first group of five tubes of recovery broths. Return the remainder of the 
mixture in the pipette to the universal container.  
A.7.5  Ten minutes after the first addition of the inoculum, add another 1 mL of the inoculum to each of t he 
sanitizer dilutions and mix by swirling gently  
A.7.6  After 8 min, remove a sample of the mixture as put before (A.7.4) and put 0.02 mL into each of the 
second group of five tubes of recovery broths.  
A.7.7  Twenty minutes after the first addition of the  inoculum, add a further 1 mL of inoculum to each of the 
sanitizer dilutions and mix by swirling gently.  
A.7.8  After 8 min, remove a sample of the mixture as before and place 0.02 mL into each of the third group 
of five tubes of recovery broths.  
A.7.9  Swirl the recovery broths and incubate at 37 °C ± 1 °C for 48 h ± 2 h. Examine the growth and record 
the results.  
 
 DUS 1625 : 2015  
© UNBS 2015  – All rights reserved  9 
 Certification marking  
Products that conform to Uganda standards may be marked with Uganda Na tional Bureau of Standards 
(UNBS) Certification Mark shown in the figure below.  
The use of the UNBS Certification Mark is governed by the Standards Act, and the Regulations made 
thereunder. This mark can be used only by those licensed under the certificat ion mark scheme operated by 
the Uganda National Bureau of Standards and in conjunction with the relevant Uganda Standard. The 
presence of this mark on a product or in relation to a product is an assurance that the goods comply with the 
requirements of that  standard under a system of supervision, control and testing in accordance with the 
certification mark scheme of the Uganda National Bureau of Standards. UNBS marked products are 
continually checked by UNBS for conformity to that standard.  
Further particu lars of the terms and conditions of licensing may be obtained from the Director, Uganda 
National Bureau of Standards.  
 
 DUS 1625 : 2015    
 
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© UNBS 2015 – All rights reserved