DEAS  1205 -1: 2024 
ICS 71.100.70 
 
© EAC 2024                                                                                                                          First Edition 2024 
 DRAFT EAST AFRICAN STANDARD  
Skincare special purpose product — Specification  — Part 1: anti-aging  
 
EAST AFRICAN COMMUNITY   
  
 DEAS 1205 -1: 2024 
ii © EAC  2024 – All rights reserved  
 Copyright notice  
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Reproduction for sales purposes may be subject to royalty payments or a licensing agreement.  
Violators may be prosecuted . DEAS 1205 -1: 2024  
© EAC 2024  – All rights r eserved  iii 
 Contents  Page 
Foreword  ................................ ................................ ................................ ................................ ............................  iv 
1 Scope  ................................ ................................ ................................ ................................ ......................  1 
2 Normat ive references  ................................ ................................ ................................ ............................  1 
3 Terms and definitions  ................................ ................................ ................................ ...........................  1 
4 Requirements  ................................ ................................ ................................ ................................ .........  2 
4.1 Ingredients  ................................ ................................ ................................ ................................ .............  2 
4.2 General requirements  ................................ ................................ ................................ ...........................  2 
4.3 Specific requirements  ................................ ................................ ................................ ...........................  2 
4.4 Microbiological limits  ................................ ................................ ................................ ............................  3 
4.5 Heavy metal contaminants  ................................ ................................ ................................ ...................  3 
5 Packaging  ................................ ................................ ................................ ................................ ...............  4 
6 Labelling  ................................ ................................ ................................ ................................ .................  4 
7 Sampling  ................................ ................................ ................................ ................................ ................  4 
Annex A (normative)  Determination of total fatty substance content  ................................ .........................  5 
A.1 Outline of the method  ................................ ................................ ................................ ...........................  5 
A.2 Reagents  ................................ ................................ ................................ ................................ ................  5 
A.3 Procedure  ................................ ................................ ................................ ................................ ...............  5 
A.4 Calculation  ................................ ................................ ................................ ................................ .............  5 
Annex B (normative)  Determination of hydroquinone content  ................................ ................................ .... 6 
B.1 Principle  ................................ ................................ ................................ ................................ .................  6 
B.2 Reagents  ................................ ................................ ................................ ................................ ................  6 
B.3 Apparatus  ................................ ................................ ................................ ................................ ...............  6 
B.4 Sample preparation  ................................ ................................ ................................ ...............................  7 
B.4.1  Preparation of the solvent  mixture  ................................ ................................ ................................ ...... 7 
B.4.2  Preparation of calibration standard  solutions  ................................ ................................ ...................  7 
B.4.3  Sample  preparation  ................................ ................................ ................................ ...............................  9 
B.5 Environmental  control  ................................ ................................ ................................ ..........................  9 
B.6 Quality  control  ................................ ................................ ................................ ................................ ....... 9 
B.7 Procedure: High Performance Liquid Chromatography (HPLC) Setup  ................................ ...........  9 
B.8 HPLC calibration  curve  ................................ ................................ ................................ .......................  10 
B.9 HPLC  determination  ................................ ................................ ................................ ............................  10 
B.10  Results  ................................ ................................ ................................ ................................ ..................  10 
B.10.1  Determination of hydroquinone concentration  ................................ ................................ ................  10 
B.10.2  Expression of results  ................................ ................................ ................................ ..........................  10 
Annex C (normative)  Determination of antioxidant activity  ................................ ................................ ........  11 
C.1 General  ................................ ................................ ................................ ................................ .................  11 
C.2 Procedure  ................................ ................................ ................................ ................................ .............  11 
C.3 Calculation  ................................ ................................ ................................ ................................ ...........  11 
Bibliography  ................................ ................................ ................................ ................................ ......................  12 
 DEAS 1205 -1: 2024 
iv © EAC  2024 – All rights reserved  
 Foreword  
Development of the East African Standards has been necessitated by the need for harmonizing requirements 
governing quality of products and services in the East African Community. It is envisaged  that through 
harmonized standardization, trade barriers that are encountered when goods and services are exchanged 
within  the Community will be removed.  
The Community has established an East African Standards Committee  (EASC)  mandated to develop and 
issue  East African Standards  (EAS) . The Committee is composed of representatives of the National 
Standards Bodies in Partner States, together with the representatives from the public and private sector 
organizations  in the community . 
East African Standards are developed through  Technical Committees that are representative of key 
stakeholders including government, academia, consumer groups, private sector and other interested parties. 
Draft East African Standards are circulated to stakeholders through the Nationa l Standards Bodies in the 
Partner States. The comments received are discussed and incorporated before finalization of standards, in 
accordance with the Principles and procedures for develop ment of East African Standards.  
East African Standards are subject to review, to keep pace with technological advances. Users of the East 
African Standards are therefore expected to ensure that they always have the latest versions of the standards 
they are implementing . 
The committee responsible for this document is Techn ical Committee EASC/TC 071, Cosmetics and related 
products.  
Attention is drawn to the possibility that some of the elements of this document may be subject of patent 
rights. EAC shall not be held responsible for identifying any or all such patent rights.  
EAS 1205  consists of the following parts, under the general title Skincare special purpose product — 
Specification : 
Part 1: Anti-aging  
Part 2: Sunscreen  
 
 DRAFT EAST AFRICAN STANDARD  DEAS 1205 -1: 2024  
 
© EAC 2024 – All rights reserved  1 
 Skincare special purpose product — Specification — Part 1: anti -aging  
1 Scope  
This Draft East African Standard  specifies requirements , sampling and test methods for anti-aging / anti -
wrinkle products . 
This standard  does not apply to skincare products covered by EAS 786, aromatherapy substances , sun 
protection products  and hair creams, lotions and gels . 
2 Normative references  
The following documents are referred to in the text in such a way that some or all of their content constitutes 
requirements of this document.  For dated references, only the edition cited applies. For undated references, 
the latest edition of the referenced document (including any amendments) applies.  
EAS 346, Labelling of cosmetics — General req uirements  
EAS 377 (all parts), Cosmetics and cosmetic products  
EAS 846 , Glossary of terms relating to the cosmetic industry   
EAS 847 -1, Cosmetics — Analytical methods  — Part 1: Glossary of terms  
EAS 847 -16, Cosmetics — Analytical methods — Part 16: Determi nation of lead, mercury and arsenic content  
EAS 847 -17, Cosmetics — Analytical methods — Part 17: Determination of pH  
EAS 847 -18, Cosmetics — Analytical methods — Part 18: Determination of thermal stability  
ISO 18416, Cosmetics — Microbiology — Detection o f Candida albicans  
ISO 21149, Cosmetics — Microbiology — Enumeration and detection of aerobic mesophilic bacteria   
ISO 21150 , Cosmetics — Microbiology — Detection of Escherichia coli  
ISO 22717, Cosmetics — Microbiology — Detection of Pseudomonas aeruginosa  
ISO 22718, Cosmetics — Microbiology —Detection of Staphylococcus aureus  
ISO 24153, Random sampling and randomization procedures  
3 Terms and definitions   
For the purposes of this document, the terms and definitions given in EAS 846  and EAS 847 -1 and the 
following apply.  
ISO and IEC maintain terminological databases for use in standardization at the following addresses:  DEAS 1205 -1: 2024  
2 © EAC  2024 – All rights reserved  
 — ISO Online browsing platform: available at http://www.iso.org/obp  
3.1 
cream  
semi -solid emulsion w hich contain mixtures of oil and water  as a cosmetic preparation applied to the skin . 
Examples include vanishing creams, foundation creams, cold creams, night creams, moisturizer  creams , hand  
creams, face creams, body creams, toning creams, emollient  cream s, purifier  cream s, nourisher  cream s, 
facial scrub  cream s, facial mask  cream s, facial wash cream s and such products among others.  
3.2 
lotion  
thick, smooth liquid preparation designed to be applied to the skin for cosmetic purposes. Examples include 
vanishi ng lotions, foundation lotions, cold lotions, night lotions, moisturizers, cleanser  lotion s, hand lotions, 
face lotions, body lotions, toner  lotion s, emollients, purifiers, and nourishers and such products among others . 
3.3 
gel 
semi -solid colloidal suspens ion of a solid dispersed in a liquid  to be applied to the skin for cosmetic purposes . 
Examples include serums and such products among others”.  
3.4 
anti-aging  skincare product  
cosmetic formulation designed to help reduce or mitigate the signs of aging on th e skin  such as but not limited 
to; fine lines, hyperpigmentation, age spots , dark eye circle and crow feet  
3.5 
anti-wrinkle  skincare product  
cosmetic formulation  intended to reduce the appearance of wrinkles in the skin  
3.6 
gel-based serum  
water or oil bas ed preparation containing plant extracts, carrier oils, essential oils, and sometimes hydrosols 
or other materi als with anti ageing properties  
4 Requirements  
4.1 Ingredients   
4.1.1  Skincare special purpose products shall contain antioxidants, exfoliants, m oisturizers and cell 
regulators such as retinols, peptides and growth factors. It may contain sunscreens.  
4.1.2  All ingredients used including dyes, pigments and colours shall comply with all parts of EAS 377 . 
4.2 General requirements  
Skincare special purp ose products shall be : 
a) clear or homogeneous and  
b) free from visible impurities.  
4.3 Specific requirements  
4.3.1  Anti-aging products  shall comply with the specific requirements given in Table 1 when tested in 
accordance with the test methods specified therei n. DEAS 1205 -1: 2024  
© EAC 2024  – All rights r eserved  3 
 Table 1 — Specific requirements for anti-aging products  
S/N Characteristic  Requirement  Test method  
i.  Thermal stability  To pass test  EAS 847 -18 
ii.  pH range  3.5 – 8.5 EAS 847 -17 
iii.  Total fatty substance 
contenta, % by mass, min . 5 Annex A  
iv.  Hydroquinone c ontent  Not detected  Annex B 
v.  Antioxidant activity,% 
Radical scavenging 
activity, min  10 Annex  C 
a applicable to products in form of creams and lotions  
 
4.3.2  Non-emulsified and gels anti-aging product shall comply with the specific requirements given in  
Table 2 when tested in accordance with the test methods specified therein.  
Table 2 — Specific requirements for Non-emulsified and gels anti -aging product  
S/N Characteristic  Requirement  Test method  
i.  pH range  Non skin lightening  products   4.5 - 8.5 EAS 84 7-17 
Skin lightening products  3.5 - 8.5 
ii.  Hydroquinone content  Not detected  Annex B  
 
4.4 Microbiological limits  
Anti-aging product s shall comply with the microbiological limits given in Table 3 when tested in accordance 
with the test methods specified  therein . 
Table 3 — Microbiological limits  for Anti-aging products  
S/N Micro -organism  Limit  Test method  
i.  Total viable count for aerobic mesophyllic 
microorganisms CFU/g or CFU/ ml, max.  100 ISO 21149  
ii.  Pseudomonas aeruginosa  
Not detect ed in 1 ml or 1  g 
of cosmetic product  ISO 22717  
iii.  Staphylococcus aureus  ISO 22718  
iv.  Candida albicans  ISO 18416  
v.  Escherichia coli Not detected in 1 g of 
cosmetic product  ISO 21150  
 
4.5 Heavy metal contaminants  
Anti-aging product s shall comply with the heavy metal limits giv en in Table 4 when tested in accordance with 
the test methods specified therein.  DEAS 1205 -1: 2024  
4 © EAC  2024 – All rights reserved  
 Table 4 — Heavy metal limits  for skincare special purpose creams, lotions and gels  
S/N Heavy metal  Maximum l imita 
mg/kg  Test method  
i.  Lead  10 
EAS 847-16 ii.  Arsenic  2 
iii.  Mercur y 2 
a The total amount of heavy metals as lead, mercury and arsenic, in combination in the finished product , shall not 
exceed 10 mg/kg.  
 
5 Packaging  
The product shall be packaged in suitable well -sealed containers that shall protect the contents and sha ll not  
cause any contamination or react with the product.  
6 Labelling  
In addition to the labelling requirements given in EAS 346, each package shall be legibly and indelibly labell ed 
with the following : 
a) product name as “ Cream ”, “Oil” and “ Jelly”, “Lotion” , “Serum ” or “Gel”, with an indication of where the 
product is to be applied ; and;  
b) an indication on whether the product is  “anti-aging” or “anti -wrinkle”.  
7 Sampling  
Sampling shall be carried out in accordance with ISO 24153 . DEAS 1205 -1: 2024  
© EAC 2024  – All rights r eserved  5 
 Annex A 
(normative)  
 
Determinat ion of total fatty substance content  
A.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.  
A.2 Reagents  
A.2.1 Dilute hydrochloric acid , 1:1 (v/v)  
A.2.2 Petroleum  ether , B.P. 40 °C to 60 °C  
A.2.3 Methyl orange indicator solution , dissolve 0.1 g of methyl orange in 100 ml of water.  
A.2.4 Sodium sulphate,  dehydrated  
A.3 Procedure   
Weigh accurately about 2 g of the material into a conical flas k; 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 separation funnel and allow it to cool to 20 °C. Rinse the conical fla sk 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 tw ice. 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 sodium  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 remaining in the flask at a temperature of 60 °C ± 2 °C to cons tant mass.  
A.4 Calculation  
The total fatty substance , expressed as percent by mass, shall be calculated as follows : 
 
where  
M1 is the mass, in grams, of the residue;  and 
M2  is the mass, in grams, of the material taken for the test.  
 DEAS 1205 -1: 2024  
6 © EAC  2024 – All rights reserved  
 Annex B 
(normative)  
 
Determination of hydroquinone  content  
B.1 Principle  
Hydroquinone is extracted from the cosmetic product using a mixture of water and methanol in the ratio of 
20:80 in an ultrasonic bath for 20 min. Determination of the analyte in the resulting solution is p erformed by 
reverse phase HPLC equipped with UV/VIS detector at a wavelength of 295  nm. 
B.2 Reagents  
B.2.1  HPLC grade  water  
B.2.2  HPLC grade  methanol  
B.2.3  Hydroquinone standard,  purity  99.9+  % 
B.2.4  HPLC grade  acetonitrile  
B.2.5  Mobile phase methanol/wate r (80/20)  
B.3 Apparatus  
B.3.1  HPLC equipped with UV/VIS  detector  
B.3.2  Analytical balance with accuracy of ±  0.000  1 g 
B.3.3  Ultrasonic  bath  
B.3.4  Amber volumetric flasks,  10-ml, 1 000-ml. 
B.3.5  10-ml and 100-ml beaker s 
B.3.6  250-ml measuring  cylinder  
B.3.7 0.45-µm Teflon syringe  filters  
B.3.8  Aluminium foil  paper  
B.3.9  10-ml glass  syringe  
B.3.10  Stainless steel column,  length 150 mm, internal diameter 4.6 mm, C18, particles size of 5 µm or 
equivalent.  
B.3.11  Wash -bottle  DEAS 1205 -1: 2024  
© EAC 2024  – All rights r eserved  7 
 B.4 Sample preparation  
B.4.1  Prepara tion of the solvent  mixture  
Prepare the solvent mixture by measuring about 200 ml of HPLC grade water using 250-ml measuring 
cylinder and transfer into a 1  000-ml volumetric flask. Fill the 1  000-ml volumetric flask to the mark with HPLC 
grade methanol usi ng a wash bottle. Cap the flask and shake well to ensure a homogeneous solution is 
obtained. Label the flask appropriately ( methanol/water 80/20) and allow the resultant solution to settle.  
B.4.2  Preparation of calibration standard  solutions  
Calibration st andard solutions are prepared as detailed in B.4.2.1 to B.4.2.8 . 
B.4.2.1   Principle  
Calibration standards are gravimetrically prepared by weighing known masses of standards into a known 
mass of solvent to obtain the desired concentration level of the stand ards. A stock solution of about 1  000 
mg/kg is prepared and is then serially diluted to the desired concentration level for working standards. 
Calibration standards are directly injected (neat) into the HPLC and their resultant peak areas are used to 
devel op a calibration curve for quantification of the unknown concentration of samples.  
B.4.2.2  Reagents  
All reagents shall be of HPLC grade or better. The purity shall be declared.  
B.4.2.3  Apparatus  
B.4.2.3.1  Analytical balance,  with an accuracy of  0.000  1 g 
B.4.2.3.2  Gas-tight syringes,  1-ml, 2.5-ml, and 5.0 -ml 
B.4.2.3.3  Amber volumetric flask,  10-ml 
B.4.2.3.4  Amber autosampler vials,  1.8-ml 
B.4.2.3.5  Beaker, 10-ml 
B.4.2.3.6  Ultrasonic bath  
B.4.2.3.7  Lint-free paper towel  
B.4.2.4  Procedure  
B.4.2.4.1  Preparat ion of stock solution , 1 000 mg/ kg 
B.4.2.4.1.1  Accurately weigh a labelled volumetric flask with cap on and record mass in grams as M1. 
B.4.2.4.1.2  Weigh empty 10-ml beaker and record mass in grams as M2. 
B.4.2.4.1.3  Add approximately 0.01 g of the standar d on the pre -weighed beaker and record mass in grams 
as M3. 
B.4.2.4.1.4  Add about 5 ml of solvent to the beaker and sonicate on a n ultrasonic bath.  
B.4.2.4.1.5  Transfer the standard solution into the pre -weighed volumetric flask, and thereafter wash the 
beaker twice with 2 ml of the solvent adding them to the standard solution in the volumetric flask . DEAS 1205 -1: 2024  
8 © EAC  2024 – All rights reserved  
 B.4.2.4.1.6  Top the volumetric flask to the mark with the solvent, wipe with lint -free paper towel and weigh, 
record mass in grams as M4. 
B.4.2.5  Preparation of working solutions  
B.4.2.5.1  Label five capped volumetric flasks as Level 1 to Level 5 and weigh them recording the masses 
in grams as VE1 to VE5. 
B.4.2.5.2  Aliquot 1  000 µl of the stock solution using a clean 1 -ml gas-tight syringe, wipe with lint -free 
paper towel and weigh on the analytical balance. Record mass in grams as WS I. 
B.4.2.5.3  Transfer 50 µl of the stock solution into the pre -weighed volumetric flasks, and top to the mark 
with solvent. Cap the flask and weigh and record the mass in grams as VF. 
B.4.2.5.4  Weigh the gas -tight syringe and record mass in grams as WS F. 
B.4.2.5.5  Repeat steps B.4.2.5.3 to B.4.2.5.4  for Level 2 to Level 5, transferring 100 µl, 150 µl, 200 µl and 
500 µl respectively, record masses in grams as WS F2 to WS F5. 
B.4.2.5. 6 The target concentrations (mg/kg) are 5, 10, 15, 20 and 50.  
B.4.2.6  Documentation and data control  
Readings of masses shall be recorded in the analyst’s workbook.  
B.4.2.7  Calculation and expression of results  
B.4.2.7.1  Concentration of calibration standards  
B.4.2.7.1.1  The concentration of stock solution , expressed as milligrams per kilogram,  is calculated using 
the following formula:  
 
where  
M1 is the mass , in grams,  of the  volumetric flask with cap on ; 
M2 is the mass , in grams,  of the  empty beaker;  
M3 is the mass , in grams,  of the  standard and the pre -weighed beaker ; and  
M4 is the mass , in grams,  of the volumetric flask topped to the mark with solvent and wiped with lint -free 
paper towel . 
B.4.2.7.1.2  The concentration of working standards is calculated usin g the following formula : 
 
where  
VF is the mass , in grams,  of the  pre-weighed volumetric flask  and solvent topped to the mark;  
 DEAS 1205 -1: 2024  
© EAC 2024  – All rights r eserved  9 
 VE   is the mass , in grams , of the  empty volumetric flask  with the cap  
WS 1 mass, in grams, of the gas-tight syringe with 1 000 µl of the stock solution and  wiped with lint -
free paper towel ; and  
WS F  is the mass , in grams,  of the  empty gas-tight syringe . 
B.4.2.8  Expression of r esults  
Levels  5 mg/ kg but < 50 mg/ kg to the nearest three significant figures . 
Levels  50 mg/ kg to the n earest whole number . 
Levels > 1 mg/ kg < 5 mg/ kg to the nearest two significant figures.  
B.4.3  Sample  preparation  
Weigh a100 -ml beaker and record the mass ( S1). Weigh approximately 0.5 g of sample into the pre -weighed 
beaker and record the mass ( S2). Disper se the sample in about 8 ml solvent mixture ( B.4.1). Cover the beaker 
with aluminium foil paper ( B.3.8) and place it in a n ultrasonic bath ( B.3.3) for 10 min. Weigh a 10-ml amber 
volumetric flask and record the mass ( S3). Transfer the sample solution into the volumetric flask and top to the 
mark with the solvent mixture. Weigh the volumetric flask containing sample mixture and record the mass ( S4). 
Transfer  the sample into appropriately labelled amber autosampler vials for HPLC determination. Test 
samples w ithin 24 h of  preparation.  
B.5 Environmental  control  
The analysis should be carried out in a well -ventilated air -conditioned room maintained at 20 ºC ± 2 ºC since 
the retention time of hydroquinone fluctuates with temperature changes.  
B.6 Quality  control  
The stability of the retention time shall be ensured when the Relative Standard Deviation (RSD) of successive 
injections do not vary one from the other by more than 2  %. If the linearity is below the acceptable limit, repeat 
the calibration exercise.  
The li nearity of the calibration curve should be above 0.999XX. If the linearity is below the acceptable limit, 
repeat the calibration exercise.  
B.7 Procedure: High Performance Liquid Chromatography (HPLC) Setup  
B.7.1  Degas mobile phase solvents in a sonicator f or 10 min and then connect solvent reservoirs to the 
instrument. Turn on the instrument and load the method of test in the software.  
B.7.2  Download the method into the instrument. Allow software to connect to the instrument and then open 
the pump valve.  
B.7.3 Press the purge button to prime the pump before use. Once purging is complete, close the valve and 
check if pressure is building up. Check for any air bubbles in the plumbing and purge the system if necessary. 
Install the appropriate column and check f or any leaks. If leaks are detected, tighten the  connectors.  
B.7.4  Allow the equipment to run for 20 min to equilibrate, with a different solvent other than the mobile 
phase flowing through  (in this case , the solvent is Acetonitrile). After the equipment h as stabilized, switch off 
the pump and switch to the mobile phase and allow the equipment to run for about 10 min. Ensure that the DEAS 1205 -1: 2024  
10 © EAC  2024 – All rights reserved  
 oven temperature of 40°C has been attained and any signals arising from the noise and drift of the equipment 
has been  reduced . 
B.7.5  Adjust the flow rate of the mobile phase to 1.5 ml/min, set runtime at 5 min, the pump mode to 
isocratic mode, the lamp to D2, and detector wavelength set at 295  nm. 
B.8 HPLC calibration  curve  
To obtain a calibration curve, inject 1 µ l of calibrati on standard solutions starting with the blank and then 
standards starting with the lowest concentration. Record the peak areas/heights for each corresponding 
concentration level. Ensure at least five replicate injections are made for each calibration stand ard. Prepare 
the calibration curve by plotting the peak areas/heights against hydroquinone concentration, in milligrams per 
kilogram ( mg/kg). Ensure that the results meet the acceptance criteria stipulated in B.6.  
B.9 HPLC  determination  
Inject 1 µ l of the samples and record the peak area/heights. Using the peak areas/heights of the sample , 
calculate the concentration of the sample using the Microsoft® Excel spreadsheet calibration curve. Ensure 
that at least three injections are made and the average is repo rted as the result if it passes the QC checks 
stipulated in  B.6. 
B.10  Results  
B.10.1  Determination of hydroquinone concentration  
Hydroquinone concentration , expressed as milligrams per kilogram, shall be calculated  using the following  
formula :  
 
where  
S1 is the mass , in grams , of empty beaker ; 
S2  is the  mass , in grams , of beaker with sample ; 
S3  is the mass , in grams , of the empty volumetric flask ; and  
S4  is the mass , in grams , of volumetric flask with sample solution . 
B.10.2  Expression of results  
Levels  ≥ 10 mg/ kg but < 100 mg/ kg to the nearest three significant figures  
Levels ≥ 100 mg/ kg to the nearest whole number  
Levels > 0.1 mg/ kg < 10 mg/kg to the nearest two significant figures  DEAS 1205 -1: 2024  
© EAC 2024  – All rights r eserved  11 
 Annex C  
(normative)  
 
Determination of antioxidant activity  
C.1 General  
The free RSA is tested using a 1, 1 - diphenyl -2-picryl hydrazyl (DPPH) technique.  
C.2 materials  
C.2.1 Analytical Reagent Methanol. Absolute ethanol can be used in place of Methanol (AR) 
C.2.2 UV/Vis spectrophotometer (able to read absorbance at 517 nm ) 
C.2.3 Amb er yellow glass test tubes  
C.2.4 Analytical balance 0.0001g  
Procedure  
C.2.1  Dissolve 24 m g of DPPH in 100 mL of methanol for making the stock solution. Filtration of DPPH 
stock  solution using methanol yields  a usable mixture with  an absorbance of around 0.973 at  517 nm. If not 
achieved, dilute the solution with the solvent until absorbance is below one. 
C.2.2  In a test tube, combine  3 mL DPPH workable solutions with 100 µL of  sample .  
C.2.3  Three millilit res of solution containing DPPH in 100 µL of methanol  kept in complet e darkness for 30 
minutes shall be used as is often given as a standard.  
C.2.4 keep the tubes in c omplete darkness for 30 min and d etermine the absorbance at 517 nm.  
C.3 Calculation  
The antioxidant activity  expressed as percent, shall be calculated using the formula below:  
 
Where  
Ac Control reaction absorbance; and  
As Testing specimen absorbance.  DEAS 1205 -1: 2024  
12 © EAC  2024 – All rights reserved  
 Bibliography  
[1] Bhavesh, J., Piyush, A., Deepak, S., & Ashok, D. (2013). Anti -aging Cosm etics. Journal of Drug 
Delivery & Therapeutics , 3(3), 158 -62. 
[2] EAS 786: 2013, Skin care creams, lotions and gels — Specification   
[3] Li, X. (2015, June). Anti -aging  cosmetics and its efficacy assessment methods. In IOP Conference 
Series: Materials Science and Engineering (Vol. 87, No. 1, p. 012043). IOP Publishing.  
[4] Official Journal of the European Communities N. L26217  
[5] Lee, K. T., Lee, S. Y., Jeong, J. H., &  Jo, B. K. (2002). New anti -wrinkle cosmetics. Journal of the 
Society of Cosmetic Scientists of Korea, 28(1), 71 -79. 
 
 DEAS  071-1: 2024    
 
© EAC 2022  – All rights reserved