DEAS 1119 -5: 2023  
ICS 59.080.30, 65.100  
 
© EAC 2023                                                                                                                             First Edition 2023   
 DRAFT EAST AFRICAN STANDARD  
Skin applied mosquito repellents  — Specification  — Part 5: Bracelets, 
wrist  bands and patches  
EAST AFRICAN COMMUNITY  DEAS 1119 -5: 2023  
ii © EAC  2023  – All rights  reserved  
 Copyright notice  
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© East African Community 2023  — All rights reserved  
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Reproduction for sales purposes may be subject to royalty payments or a licensing agreement.  
Violators may be prosecu ted. DEAS 1119 -5: 2023  
© EAC 2023  – All rights reserved  iii 
 Contents  Page 
Foreword  ................................ ................................ ................................ ................................ .............................  v 
1 Scope  ................................ ................................ ................................ ................................ ......................  1 
2 Normative references  ................................ ................................ ................................ ............................  1 
3 Terms and definitions  ................................ ................................ ................................ ...........................  1 
4 Active ingredients  ................................ ................................ ................................ ................................ . 2 
4.1 Natural repellents  ................................ ................................ ................................ ................................ .. 2 
4.2 Synthetic repellents  ................................ ................................ ................................ ..............................  3 
5 Requirements  ................................ ................................ ................................ ................................ .........  3 
5.1 General requirements  ................................ ................................ ................................ ...........................  3 
5.2 Specific requirements  ................................ ................................ ................................ ...........................  3 
5.3 Biological efficacy  ................................ ................................ ................................ ................................ . 5 
6 Packaging  ................................ ................................ ................................ ................................ ...............  5 
7 Labelling  ................................ ................................ ................................ ................................ .................  5 
Annex  A (normative)  Determination of DEET conten t ................................ ................................ ...................  7 
A.1 General  ................................ ................................ ................................ ................................ ...................  7 
A.2 Apparatus  ................................ ................................ ................................ ................................ ...............  7 
A.3 Preparation of calibration  curve  ................................ ................................ ................................ ..........  7 
A.4 Procedure  ................................ ................................ ................................ ................................ ...............  7 
A.5 Calculation  ................................ ................................ ................................ ................................ .............  8 
Annex  B (normative)  Determination of ethyl butylacetamidopropionate (IR3535)  ................................ ..... 9 
B.1 Outline of method  ................................ ................................ ................................ ................................ .. 9 
B.2 Reagents  ................................ ................................ ................................ ................................ ................  9 
B.3 Apparatus  ................................ ................................ ................................ ................................ ...............  9 
B.4 Procedure  ................................ ................................ ................................ ................................ ...............  9 
B.4.1  Chromatographic conditions (typical)  ................................ ................................ ................................  9 
B.5 Preparation of sample.  ................................ ................................ ................................ ........................  10 
B.6 System equilibration  ................................ ................................ ................................ ...........................  10 
B.7 Determination.  ................................ ................................ ................................ ................................ ..... 10 
B.8 Calculation  ................................ ................................ ................................ ................................ ...........  11 
Annex C (normative)  Determination of icaridin  ................................ ................................ ............................  12 
C.1 outline of method  ................................ ................................ ................................ ................................  12 
C.2 Reagents  ................................ ................................ ................................ ................................ ..............  12 
C.3 Apparatus  ................................ ................................ ................................ ................................ .............  12 
C.4 procedure  ................................ ................................ ................................ ................................ .............  12 
C.4.1  Chromatographic conditions (typical)  ................................ ................................ ..............................  12 
C.5 Preparation of sample  ................................ ................................ ................................ .........................  13 
C.6 Equilibration of the system  ................................ ................................ ................................ ................  13 
C.7 Determination  ................................ ................................ ................................ ................................ ...... 13 
C.8  Calculation  ................................ ................................ ................................ ................................ ..........  13 
Annex D  (normative)  Determination of total pyrethrins  ................................ ................................ .............  15 
D.1 General  ................................ ................................ ................................ ................................ .................  15 
D.2 Reagents  ................................ ................................ ................................ ................................ ..............  15 
D.3 Apparatus  ................................ ................................ ................................ ................................ .............  15 
D.4 Operating conditions  ................................ ................................ ................................ ..........................  15 
D.5 Preparation of the  standard  ................................ ................................ ................................ ...............  15 
D.6 Sample preparation  ................................ ................................ ................................ .............................  16 
D.7 Procedure  ................................ ................................ ................................ ................................ .............  16 DEAS 1119 -5: 2023  
iv © EAC  2023  – All rights  reserved  
 D.8 Calc ulation of the % total pyrethrins (active ingredient)  ................................ ................................  16 
Bibliography  ................................ ................................ ................................ ................................ .....................  17 
 DEAS 1119 -5: 2023  
© EAC 2023  – All rights reserved  v 
 Foreword  
Development of the East African Standards has been necessitated by the need f or 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 wi ll 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 w ith 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, priva te sector and other interested parties. 
Draft East African Standards are circulated to stakeholders through the National Standards Bodies in the 
Partner States. The comments received are discussed and incorporated before finalization of standards, in 
accor dance with the Principles and procedures for development of East African Standards. XXXXXX.  
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 th ey always have the latest versions of the standards 
they are implementing .  
The committee responsible  for this document is Technical Committee is Technical Committee EASC/TC 078, 
Healthcare and medical devices.  
Attention is drawn to the possibility that so me 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.  
WDEAS 1119 consists of the following parts, under the general title Skin applied mosquito repellents —
Spec ification:  
Part 1: Lotions, creams, gels and ointments  
Part 2: Sprays and roll -ons 
Part 3: Wipes  
Part 4: Bathing soaps  
Part 5: Bracelets, wristbands and patches  
Part 6: Jelly  
 DRAFT EAST AFRICAN STANDARD  DEAS 1119 -5:2023  
 
© EAC 2023  – All rights reserved  1 
 Skin applied mosquito repellents — Specification — Part 5: Bracelets, 
wrist ba nds and patches  
1 Scope  
This Draft East African Standard specifies the requirements, sampling and test methods for skin applied 
mosquito repellents prepared as bracelets, wristbands and patches.  
2 Normative references  
The following documents are re ferred 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 amen dments) applies.  
FDEAS 1120 -1, Mosquito repellent — Performance test guidelines — Part 1: Skin applied  
EAS 846, Glossary of terms relating to the cosmetic industry  
ISO 2589, Leather — Physical and mechanical tests — Determination of thickness  
ISO 22198, Textiles — Fabrics — Determination of width and length  
ISO 24153, Random sampling and randomisation procedures  
3 Terms and definitions   
For the purposes of this document, the te rms and definitions given in EAS 846 and the following apply.  ISO 
and IEC maintain  terminological databases for use in standardization at the following addresses:  — ISO 
Online browsing platform: available at http://www.iso.org/obp  
3.1 
bracelet  
ornament, band, or chain worn around the wrist  
3.2 
wristband  
small bracelet or band worn aroun d the wrist for identity purposes or to soak up sweat when playing sport  
3.3 
patch  
piece of cloth or other material used to mend or strengthen a torn or weak point  
3.4 
mosquito  
blood -sucking dipterous insect of the family Culicidae. Aedes , Anopheles,  Culex , Mansonia,  and Stegomyia  
which are genera containing most species involved in the transmission of protozoan and other disease  
producing parasites   
2 © EAC  2023  – All rights  reserved  
 3.5 
mosquito repellent  
substance applied to skin, clothing, or other surfaces which deters mosquito from appr oaching or settling  
3.6 
natural repellent  
repellent that contains plant -based compounds  
3.7 
synthetic repellent  
conventional repellent containing chemical compounds manufactured to imitate the natural compounds  
3.8 
DEET  
N, N-Diethyl -meta -toluamide or dieth yltoluamide  
3.9 
IR3535  
ethyl butylacetylaminopropionate  
3.10 
Picaridin/icaridin  
1-(1-methylpropoxycarbonyl) -2-(2-hydroxyethyl) piperidine or 2 -(2-hydroxyethyl) -1-piperidinecarboxylic acid 1 -
methylpropyl ester  
4 Active ingredients  
4.1 Natural repellents  
4.1.1 Active ingredients used in natural  repellents shall be plant -based compounds that are able to deter  
mosquitoes from approaching or settling on the surface. Such shall be essential oils or any other plant extract 
approved as mosquito repellents.  
4.1.2 The manufacturer shall provide adequate  data on the repellence of such ingredients.  
4.1.3 The manufacturer shall have adequate data justifying the proportion of ingredient(s) for which claims 
are made, used in the product.  
4.1.4 The essential oils and other plant extracts used in natural repell ents shall be, but not limited to:  
a) cedar wood oil;  
b) tea tree oil;  
c) geranium oil;  
d) rosemary oil;  
e) lemongrass oil;  
f) citronella oil;  
g) eucalyptus oil;  
h) cinnamon oil;  
i) neem oil ;  
© EAC 2023  – All rights reserved  3 
 j) peppermint oil; 
k) pine oil; 
l) garlic oil; 
m) amyris oil; 
n) Indian rose wood oil; and 
o) cajeput oil  
 
4.1.5 The proportion of single or blended active ingredient(s) in natural repellents shall be set by the  
manufacturer  in accordance with the relevant  standard and records shall be availed.  
4.1.6 Pyrethrum extracts such as pyrethrins when considered in natural repellents . 
4.2 Synthetic repellents  
4.2.1 Synthetic repellents shall contain chemical compounds that are able to deter mosquitoes from  
approaching or settling on the surface.  
4.2.2 If the  synthetic active ingredient is blended with other active ingredient(s), either natural or synthetic,  
the proportion shall be set by the manufacturer based on scientific research and records shall be availed.  
4.2.3 Synthetic repellents and their active ing redients shall be approved by relevant authority before being 
released to the market.  
5 Requirements  
5.1 General requirements  
5.1.1  The product shall constitute a mosquito repellent that is formulated as wristbands, bracelets or 
patches and shall be essentially a product which has active ingred ient(s) added to a certain level.  
5.1.2  The product shall be a pack of bands primarily composed of 100 % microfibre with no harmful 
chemicals used.  
5.1.3  Each band shall have an effective fragrance for repelling mosquitoes.  
5.2 Specific requirements  
5.2.1  Active ingredients and their content in mosquito repellents shall comply with the requirements given in 
Table 1 when tested in accordance with the test methods specified therein .  
4 © EAC  2023  – All rights  reserved  
 Table 1 — Requirements for a ctive ingredients content in skin applied  mosquito repellent bracelet, 
wristb and and patch  
S/N  Characteristic  Requirement  Test method  
i  DEETa, % w/w.  4 – 50 Annex A  
ii  Picaridina, % w/w.  5 – 20 Annex B  
iii  IR3535a, % w/w.  7.5 – 20.07  Annex C  
iv Pyrethrinsb, min % w/w  0.5 Annex D  
a applicable to synthetic mosquito repellen t bracelet, wristband and patch  
b  applicable to natural mosquito repellent bracelet, wristband and patch  
 
5.2.2  The product shall comply with the specific requirements given in Table 2 when tested in accordance 
with the test methods specified therein.   
© EAC 2023  – All rights reserved  5 
 Table 2 — Specific requirements for skin applied mosquito repellents in form of bracelets, wristbands 
and patches  
S/No  Parameter  Requirement  
mm Test method  
i) Length  150 - 250  
ISO 22198  ii) Width  12 - 25 
iii) Thickness  2 - 4 ISO 2589  
iv) Circumference  
 X-small  
 Small  
 Medium  
 Large  
 X-large   ISO 22198  
 165.1   
 177.8   
 190.5   
 203.2   
 215.9   
 
5.3 Biological efficacy  
When tested in accordance with DEAS 1120 -1, the product shall repel 100 % of the mosquitoes from 
appro aching or settling on that surface, within protection time indicated by the manufacturer.  
6 Packaging  
The product shall be packaged in suitable well -closed containers to protect the integrity of the product during 
handling, transportation, storage and sale . 
7 Labelling  
The package shall be legibly and indelibly labelled English and/or Kiswa hili/French  with the following 
information:  
a) name of product as “ mosquito repellent  bracelet ”, “mosquito repellent wristband” or “ mosquito 
repellent patch”;  
b) name and physical address of manufactu rer; 
c) name and physical address of the importer/distributor (if applicable);  
d) active ingredient(s) conte nt; 
e) net weight;  
f) pack size ; 
g) instruction for use, storage and disposal;  
h) country of origin;  
i) date of manufacture;  
j) expiry or best before date;   
6 © EAC  2023  – All rights  reserved  
 k) batch /code /lot /serial  number ; 
l) safety precaution; and  
m) special population whose exposure is prohibited (children and pregnant women  and lactating  
mothers ). 
8 Sampling  
Sampling shall be done in accordance with ISO 24153.   
© EAC 2023  – All rights reserved  7 
 Annex  A 
(normative)  
 
Determinat ion of DEET content  
A.1 General  
The sample is dissolved i n carbon disulfide and the difference in absorbance at 14.18 μm and at 14.48 μm is 
determined. The quantity of meta -isomer is obtained from this value by means of a calibration curve prepared 
by the use of a reference standard.  
A.2 Apparatus  
A.2.1  Double -beam infrared spectrophotometer, Perkin -Elmer model 21 or equivalent  
A.2.2  Two equivalent infrared absorption cells, with sodium chloride windows and a path length of 
approximately 0.4 mm  
A.3 Preparation of calibration curve  
A.3.1  Weigh (to the nearest 0.1 mg) into four volumetric flasks sufficient amounts of the reference DEET 
standard of known purity to give concentrations of approximately 20 g/l, 40 g/l, 60 g/l and 80 g/l when 
dissolved in carb on disulfide.  
A.3.2  Fill the reference cell with carbon disulf ide and the sample cell with each of the standard solutions in 
turn, and record the spectra. The spectrum may be scanned rapidly, except for the region 12 μm to 15 μm, 
where a normal speed should be used. Carry out a blank measurement with carbon disulfide  to correct for any 
inequality in the paired cells and to determine whether a cell correction is required.  
A.3.3  Measure the absorbance at 14.18 μm and at 14.48 μm and calculate the difference between these 
values, ΔA, for each of the solutions. Plot the values of ΔA against the concentration (g/L) of the meta -isomer.  
A.3.4  If a cell correction is required, the valu e of ΔA is determined from the formula:  
ΔA = (A14.18 – A14.48)ref. – (A14.48)blank  
where  
ref. is the determination with reference standard;  
blank  is the determination on carbon disulfide (CS2) blank.  
A.4 Procedure  
Weigh (to the nearest 0.1 mg) about 0.5 g of the sample, transfer quantitatively to a 10 -mL volumetric flask, 
and make up to the mark with carbon disulfide. Measure the infrared absorption at 14.18 μm and 14.48 μm 
using the same conditions as described in A.3. Determine the concentration of meta -isomer by comparing this 
value with the calibration curve. A standard sample should be run each day to check the calibration of the 
instrument.   
8 © EAC  2023  – All rights  reserved  
 A.5 Calculation  
DEET conte nt, expressed in grams per kilogram, shall be calculated using the formula below.  
where  
C1 is the concentration, in grams per litre (g/l), of standard DEET found from calibration curve;  
P is the purity, in grams per kilogram (g/kg), of the reference standard;  
C2 is the concentration, in grams per litre (g/l), o f sample taken.  
 
© EAC 2023  – All rights reserved  9 
 Annex  B 
(normative)  
 
Determination of ethyl butylacetamidopropionate (IR3535)  
B.1 Outline of method  
B.1.1  Ethyl butylacetamidopropionate is determined by capillary gas chromatography using flame ionisation 
detection and internal standardisation.  
B.1.2  The retention time of the ethyl butylacetamidopropionate peak of the sample solution should not 
deviate by more than 2 % from that of the calibration solution.  
B.2 Reagents  
B.2.1  Acetonitrile  
B.2.2 Ethyl butylacetamidopropionate, standard of known purity  
B.2.3  Methyl tridecanoate, internal standard  
B.2.4  Calibration solution.  
B.2.4.1  Weigh (to the nearest 0.1 mg in duplicate) into volumetric flasks (10 ml) about 100 mg ethyl 
butylacetamidopropio nate standard (s mg) and about 100 mg methyl tridecanoate (r mg).  
B.2.4.2  Dissolve in acetonitrile and fill to the mark with acetonitrile (solutions C1 and C2). The solutions 
are stable for one week at room temperature.  
B.3 Apparatus  
B.3.1  Gas chromatogra ph equipped with a split/splitless injection and a flame ionisation detector  
B.3.2  Capillary column fused silica, 25 m × 0.32 (i.d.) mm, coated with CP -Sil 5 CB, film thickness: 1.2 µm  
B.3.3  Electric integrator or data system  
B.4 Procedure  
B.4.1  Chromatogr aphic conditions (typical)  
B.4.1.1  Column  fused  silica,  25 m × 0.32 mm (i.d.),  film thickness: 1.2 µm, coated with 
CP-Sil 5 CB  
B.4.1.2  Injection system  
B.4.1.2.1  Injector: split injection  
B.4.1.2.2  Split ratio:1:50   
10 © EAC  2023  – All rights  reserved  
 B.4.1.2  Detector: Flame ionisation  
B.4.1. 3 Temperatures  
B.4.1.3.1  Injection port:300 °C  
B.4.1.3.2  Detector:310 °C  
B.4.1.3.3  Oven program  
B.4.1.3.3.1  initial: 120 °C  
B.4.1.3.3.2  Program rate: 10 °C/min  
B.4.1.3.3.3  Final: 260 °C  
B.4.1.4  Injection volume: 5 µl  
B.4.1.5  Gas flow rates  
B.4.1.5.1  Heliu m 1.1 ml/min  
B.4.1.5.2  Helium (make up)  45 ml/min  
B.4.1.5.3  Hydrogen  40 ml/min  
B.4.1.6  Retention times  
B.4.1.6.1  ethyl butylacetamidopropionate: about: 10.4 min  
B.4.1.6.2  methyl tridecanoate: about: 10.9 min  
B.5 Preparation of sample  
B.5.1  Weigh in dupl icate (to the nearest 0.1 mg) into volumetric flasks (10 ml) sufficient sample to contain 
about 100 mg ethyl butylacetamidopropionate (w mg) and about 100 mg methyl tridecanoate (q mg).  
B.5.2  Dissolve in acetonitrile and fill to the mark with acetonitrile  (solutions S1 and S2). The solutions are 
stable for one week at room temperature . 
B.6 System equilibration  
Inject 0.1 µl portions of the calibration solution and repeat the injections until retention times and calibration 
factors vary by less than 0.1 % o f the mean of three successive injections.  
B.7 Determination.  
B.7.1  Inject 0.1 µl portions of the calibration solution and sample solutions in the following sequence: C 1, S1, 
C2, S2,…etc.  
B.7.2  Determine the peak areas and calculate the mean response facto r (f) of the calibration solution 
injections bracketing the injections of the sample solutions .  
© EAC 2023  – All rights reserved  11 
 B.8 Calculation  
B.8.1  Individual response factor 
  is calculated as follows:  
 
B.8.2  Ethyl butylacetamidopropionate shall be calculated as follows : 
 
where:  
fi individual  response  factor  
f mean  response  factor  
Hs peak  area of ethyl butylacetamidopropionate  in the calibration  solution  
Hw peak  area of ethyl butylacetamidopropionate  in the sample  solution  
Ir peak  area of the internal standard  in the calibration  solution  
Iq peak  area of  the internal standard  in the sample  solution  
s mass  of ethyl  butylacetamidopropionate  standard  in the calibration  solution  (mg) 
r mass  of internal  standard  in the calibration  solution  (mg) 
q mass  of internal  standard  in the sample  solution  (mg) 
w mass  of sample  taken  (mg) 
P purity  of ethyl  butylacetamidopropionate  standard  (g/kg)  
Repeatability  r 18 g/kg at 1002  g/kg active  ingredient  content  
Reproducibility  R 18 g/kg at 1002  g/kg active  ingredient  content  
  
12 © EAC  2023  – All rights  reserved  
 Annex C  
(normati ve) 
 
Determination of icaridin  
C.1 outline of method  
Icaridin is determined by capillary gas chromatograph using internal standardisation and flame ionisation 
detection.  
C.2 Reagents  
C.2.1  Icaridin reference standard with known content  
C.2.2  Dimethyl Phtha late internal standard  
C.2.3  Propan -2-ol 
C.2.4  Calibration solution.  
Weigh (to the nearest 0.1 mg) about 100mg icaridin reference substance ( s mg) and 100 mg dimethyl 
phthalate ( r mg) into a volumetric flask (20 ml). Fill to the mark with propan -2-ol and h omogenise.  
C.3 Apparatus  
C.3.1 Gas chromatograph capable of operating in the range 150 °C to 330 °C, fitted with a flame ionisation 
detector, a split injector and an autosampler  
C.3.2 Column quartz,30 m ×0.25 mm (i.d), coated with dimethyl polysiloxane/ di phenyl polysiloxane 95/5% 
(e.g. DB5), film thickness 0.25 µm 
C.3.3 Electronic integrator or data system  
C.4 procedure  
C.4.1 Chromatographic conditions (typical)  
C.4.1.1  Column: Quartz,30 m × 0.25  mm (i.d.),  coated with dimethyl polysiloxane/ diphenyl polys iloxane 
95/5% (e.g. DB5), film thickness 0.25 µm  
C.4.1.2  Injection system  
C.4.1.2.1  Injector: split injection  
C.4.1.2.2  Split flow:40ml/min  
C.4.1.2  Detector: Flame ionisation  
C.4.1.3  Temperatures   
© EAC 2023  – All rights reserved  13 
 C.4.1.3.1  Injector :240 °C 
C.4.1.3.2  Detector:3 30 °C 
C.4.1.3. 3 Oven program : 150 °C hold for 2 min, gradient: 10 °C/ min to 330 °C, hold for 3 min.  
C.4.1.4  Injection volume: 1 µl  
C.4.1.5  Gas flow rates  
C.4.1.5.1  Helium (carrier)  1.5 ml/min (100kpa)  
C.4.1.5.2  Hydrogen: about 30ml/min  
C.4.1.5.3  Air: about 3 00ml/min  
C.4.1.5.4  Nitrogen (makeup):  about 25ml/min  
C.4.1.6  Run time: about 25 min  
C.4.1.7 Retention times  
C.4.1.6.1  Dimethyl Phthalate:  about 3 min  
C.4.1.6.2  Icaridin : about 4.5 min  
C.5 Preparation of sample  
Weigh (to the nearest 0.1 mg) into a volumetric flask (2 0 ml) sufficient sample to contain about 100 mg icaridin 
(w mg) and about 100 mg dimethyl phthalate ( q mg). Fill to the mark with propan -2-ol and homogenise.  
C.6 Equilibration of the system  
Inject 1 µl portion of calibration solution and repeat the injecti ons until retention times and the icaridin to the 
internal standard peak area ration vary by less than 0.5% of the mean for successive injections.  
C.7 Determination  
C.7.1 Inject in duplicate 1 µ portions of the calibration solution (C 1 and C 2) and of the s ample solution (S 1, 
S2, etc.) in the following sequence:C 1, S1, S2, ...C 2).  
C.7.2 Determine the peak areas and calculate the response factor ( f) from the calibration solutions 
bracketing the injections of the sample solutions. Calculate the content of the  sample solutions.  
C.8  Calculation  
C.8.1  Response factor f is calculated as follows:  
 
C.8.2  Icaridin content shall be calculated in g/kg as follows:   
14 © EAC  2023  – All rights  reserved  
 
 
where:  
f mean  response  factor  
Hs peak  area of icaridin  in the calibration  solution  
Hw  peak area of i caridin in the sample solution  
Ir peak  area of dimethyl phthalate in  the calibration  solution  
Iq peak  area of  dimethyl phthalate in  the sample  solution  
s mass  of icaridin  in the calibration  solution  (mg) 
r mass  of dimethyl phthalate in  the calibration  solution (mg)  
q mass  of dimethyl phthalate  in the sample  solution  (mg)  
w mass  of sample  taken  (mg) 
P purity  of icaridin  standard  (g/kg)  
Repeatability r  12 g/kg to 14 g/kg at  986 g/kg active  ingredient  content  
Reproducibility R  14 g/kg at 986 g/kg active  ingred ient content  
  
© EAC 2023  – All rights reserved  15 
 Annex D  
(normative)  
 
Determination of total pyrethrins  
D.1 General  
The active ingredients in pyrethrum extract may be determined using a HPLC system first by injecting a 
solution of the analyte into the chromatograph, followed by the separat ion and comparison of peaks areas of 
the analytes in the sample with that of an external standard containing a known amount of the analytes. The 
peaks are eluted in the following order: Cinerin II, Pyrethrin II, Jasmolin II (total Pyrethrins II) and Cineri n I, 
Pyrethrin I, Jasmolin I (total Pyrethrins I).  
D.2 Reagents  
D.2.1  World pyrethrum standard, 50 %  
D.2.2  Acetonitrile, HPLC grade  
D.2.3  Water, HPLC grade  
D.3 Apparatus  
A liquid chromatography System equipped with an auto -sampler, a Variable Wavelength De tector (or 
equivalent) and a Column {Phenomenex, 250 x 4.6 mm Luna Phenyl -Hexyl 5μ Reverse Phase (or equivalent).  
D.4 Operating conditions  
D.4.1 Flow rate: 1.5 ml/min  
D.4.2 Composition: 40:60 (%, v/v water/acetonitrile)  
D.4.3 Elution: Isocratic  
D.4.4 Oven temperature: 40 ºC  
D.4.5 Wavelength: 240 nm  
D.4.6 Injection Volume: 15 μl  
D.4.7 Stop time: 22 min  
D.4.8 Post time: 1 min  
D.5 Preparation of the standard  
Weigh 20 mg of the pyrethrum standard to the nearest 0.0001 g in a 100 mL volumetric flask and dilute t o 
volume with acetonitrile and label it. Transfer a small portion to a sample vial and label it accordingly.   
16 © EAC  2023  – All rights  reserved  
 D.6 Sample preparation  
In a 100 ml volumetric flask, weigh 20 mg to the nearest 0.0001 g of the sample to be analyzed and dilute to 
volume with Acetonitrile. Sample this solut ion using a vial and label it accordingly.  
D.7 Procedur e 
After the chromatograph is stable, make a minimum of three injections for the standard solution as well as for 
the analyte and average the area counts. The relative standard deviation between injections should be within 
2 %.  
D.8 Calculation of the % tot al pyrethrins (active ingredient)  
The total p yrethrins , expressed as percent  shall be calculated as follows;  
=
  
  
© EAC 2023  – All rights reserved  17 
 Bibliography  
[1] RS 392 -5: 2018, Skin applied mosquito repellents — Specification — Part 5: Bracelets, wristbands and 
patch es 
[2] US 2296 -5:2022, Skin applied mosquito repellents - Specification - Part 5: Bracelets, wristbands and 
patches  
 DEAS 1119 -5: 2023  
© EAC 2023  – All rights reserved