DEAS 996: 2019  
ICS 67.220.20  
 
© EAC 2019                                                                                                                               First Edition 2019   
 DRAFT EAST AFRICAN STANDARD  
Food grade aspartame  — Specification  
EAST AFRICAN COMMUNITY  DEAS 996: 2019  
ii © EAC  2019  – All rights reserved  
 Copyright notice  
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participants in the EAC standards development process is p ermitted without prior permission from EAC, 
neither this document nor any extract from it may be reproduced, stored or transmitted in any form for 
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© East African Community  2019  — All rights reserved  
East African Community   
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Arusha  
Tanzania  
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Reproduction for sales purposes may be subject to royalty payments or a licensing agreement.  
Violators may be prosecuted . DEAS 996: 2019  
© EAC 2019  – All rights reserved  iii 
 Contents  Page 
Foreword  ................................ ................................ ................................ ................................ ...................... iv 
1 Scope  ................................ ................................ ................................ ................................ ................ 1 
2 Normative references  ................................ ................................ ................................ ....................... 1 
3 Terms and definitions  ................................ ................................ ................................ ...................... 1 
4 Requirements  ................................ ................................ ................................ ................................ ...2 
4.1 General requirements ................................ ................................ ................................ ....................... 2 
4.2 Specific requirements  ................................ ................................ ................................ ...................... 2 
5 Hygiene  ................................ ................................ ................................ ................................ ............. 2 
6 Contaminants  ................................ ................................ ................................ ................................ ...2 
7 Packaging  ................................ ................................ ................................ ................................ ......... 3 
8 Weights and measures  ................................ ................................ ................................ ..................... 3 
9 Labelling  ................................ ................................ ................................ ................................ ........... 3 
10 Sampling  ................................ ................................ ................................ ................................ ........... 3 
Annex  A (normative)  Assay: Test for purity  ................................ ................................ ............................... 4 
Annex  B (normative)  Loss on drying  ................................ ................................ ................................ .......... 5 
Annex  C  (normative)   Determination of sulphated ash  ................................ ................................ ............. 6 
Annex  D (normative)  Determination of pH (Potentiometric method)  ................................ ......................... 7 
Annex  E (normative)  Test for 5 -Benzyl–3,6–dioxo–2–piperazineacetic acid  ................................ ............. 8 
Annex  F (normative)  Test for amine group  ................................ ................................ ...............................  10 
Annex  G (normative)  Test for ester group  ................................ ................................ ................................  11 
Bibliography  ................................ ................................ ................................ ................................ ................  12 
 DEAS 996: 2019  
iv © EAC  2019  – All rights reserved  
 Foreword  
Development of the East African Standards has been necessitated by the need fo r 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 wil l 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 wi th 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, privat e 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 
accord ance 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 Technical Committee EASC /TC 005 Food additives . 
Attention is drawn to the possibility that some of the elements of this document may be subjec t of patent 
rights. EAC shall not be held responsible for identifying any or all such patent rights.  DEAS 996: 2019  
© EAC 2019  – All rights reserved  v 
 Introduction  
Aspartame (3 -Amino -N-(α-carbomethoxy -phenethyl) -succinamic acid, N -L-α-aspartyl -L-phenylalanine -1-
methyl ester, C14H18N2O5) is a low calorie artificial sweetener, sugar substitute and flavour enhancer. It is 100 
– 200 times sweeter than sucrose. It is one  of the most popular artificial  sweeteners  and it is widely used in 
the preparation of beverages, desserts, sweets, dairy products, chewing gu ms, energy -reduced and weight 
control products, as a table -top sweetener and in the preparation of food for diabetics.  
Aspartame is made of two naturally occurring amino acids:  phenylalanine and aspartic acid . The 
phenylalanine  is slightly modified by add ing a methyl group  to give  the product its sweet taste. In the 
gastrointestinal tract, aspartame is rapidly and completely hydrolysed to phenylalanine, aspartic acid and 
methanol. The products of hydrolysis are absorbed and enter normal endogenous metaboli c pathways . 
In the Codex Alimentarius Commission International Numbering System, aspartame is assigned as INS 951 
and the  FAO/WHO Joint Experts Committee on Food Additives (JECFA) established the Acceptable Daily 
Intake (ADI) for aspartame at 0 mg/kg – 40 mg/kg body weight.  
The use of aspartame as a sugar replacer remains a controversial topic with some scientific studies validating 
its safety while others suggest it could have effects on the consumer.  This standard has therefore been 
developed to ensure th at the aspartame traded and/or used in food products  in the East African Community  is 
safe for human consumption . DEAS 996 : 2019    
 
© EAC 2019  – All rights reserved  
  
 Food grade aspartame — Specification  
1 Scope  
This Draft East African Standard specifies requirements, sampling and test methods for food grad e aspartame 
for the food industry.   
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.  
AOAC 999 .11, Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods, Atomic Absorption 
Spectrophotometry after Dry Ashing  
CODEX STAN 107 , General standard for the labelling of food additives when sold as such  
CAC/GL 50 , General guidelines on sampling  
AOAC 952.13 , Arsenic in food. S ilver diethyldithiocarbonate method  
EAS 39, Hygiene in the food and drink manufacturing industry — Code of practice  
CODEX STAN 193, General standard for contaminants and toxins in food and feed  
3 Terms and definitions  
For the purposes of this document, the following terms and definitions 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 
food grade material  
material, made of substances that  are safe and suitable for their intended use and which will not  impart any 
toxic substance or undesirable odour or flavour to the product  
3.2 
aspartame  
low calorie, artificial sweetener and flavour enha ncer derived from phenylalanine and aspartic acid  DEAS 996: 2019  
2 © EAC  2019  – All rights reserved  
  
3.3 
artificial sweetener  
synthetic substance used as a sugar subst itute to sweeten food and drink  
4 Requirements  
4.1 General  requirements  
Food grade aspartame shall be:  
 a white , odourless,  crystal line powder with a strong sweet taste ; and  
 Slightly soluble in water and practically insoluble or insoluble in ethanol . 
A satu rated aqueous solution of the product shall be acidic . 
4.2 Specific requirements  
Food grade aspartame shall comply with the specific requirements given in Table 1  when tested in 
accordance with the test methods  specified therein.  
Table 1 — Specific r equir ements for food grade aspartame  
S/N Characteristic  Requirement  Test method  
i) Purity as C14H18N2O5, % m/m (dry basis)  98 – 102 Annex A  
ii) Loss on drying,  %m/m, max.  4.5 Annex B  
iii) Sulphated ash, % m/m (dry basis) , max.  0.2 Annex C 
iv) pH (0.8  % so lution)  4.5 – 6.0  Annex D 
v) 5-Benzyl –3,6–dioxo –2–piperazineacetic acid, 
(diketopiperazine), % m/m, max.  1.5  Annex E 
vi) Test for amine group  To pass test  Annex F 
vii) Test for ester group  To pass test  Annex G 
5 Hygiene  
Food grade aspartame shall be manufactured and han dled in accordance with EAS 39.  
6 Contaminants  
Food grade aspartame  shall comply with the Maximum Levels of contaminants  given in Table 2 when tested 
in accordance with the test methods specified therein.   
 DEAS 996: 2019  
© EAC 2019  – All rights reserved  3 
 Table 2 — Maximum limits for metal contaminants in food grade aspartame  
S/N Contaminant  Limit  Test method  
i) Lead  (as Pb), mg/kg, m ax. 1 AOAC 9 99.11 
ii) Arsenic (as As), mg/kg, max.  3 AOAC 952.13  
7 Pack aging 
The product shall be securely packaged in containers made of food grade material s. The packages shall 
preserve the quality and safety of the product and preclude contamination  from the external environment.  
8 Weights and measures  
The product s shall comply with the Weights and Measures Regulations  of respective Partner States . 
9 Labelling  
In addition to the requirements of CODEX STAN 107,  the product packages shall be legibly and indelibly 
labelled with the following information:  
a) name of the product  as “Food grade aspartame”;  
b) name and physical address of the processor/packer/imp orter;  
c) date of manufacture;  
d) expiry date;  and 
e) net weight  of the product in metric  units. 
10 Sampling  
Representative samples of the product shal l be drawn in accordance with CAC/GL 50.  DEAS 996: 2019  
4 © EAC  2019  – All rights reserved  
 Annex  A 
(normative)  
 
Assay: Test for purity  
A.1 Apparatus  
A.1.1 Titration  vessel  
A.1.2  Aluminium foil  
A.2 Reagents  
A.2.1 Dimethylformamide  
A.2.2  Thymol Blue  
A.2.3 Lithium Methoxide/Sodium Methoxide  
A.3 Procedure  
Accurately weigh 150 mg of the sample, previously dried at 105 °C for 4 hours. Dissolve in 35 ml of 
dimethylformamide , add 5 drops of thymol blue, and titrate with a micro burette to a dark blue end -point with 
0.1N lithium methoxide or sodium methoxide. Perform a blank determination and make any necessary 
correction. Each ml of 0.1N lithium methoxide/sodium methoxide is equivalent to 29.43 mg of C 14H18N2O5. 
Caution : Protect the solution from absorption of carbon dioxide and moisture by covering the titration vessel with 
aluminium foil while dissolving the sample and during the titration.  DEAS 996: 2019  
© EAC 2019  – All rights reserved  5 
 Annex  B 
(normative)  
 
Loss on drying  
B.1 Apparatus  
B.1.1 Titration vessel  
B.1.2 Aluminium foil  
B.2 Reagents  
B.2.1 Dimethylformamide  
B.2.2 Thymol Blue  
B.2.3 Lithium Methoxide/Sodium Methoxide  
B.3  Procedure  
Place the bottle with its contents  in the drying chamber, removing the stopper and leaving it also in the 
chamber, and dry the sample at the 105 ºC for 2 hours. Upon opening the chamber, close the bottle promptly 
and allow it to come to room temperature in a desiccator. Weigh the cool bott le and its contents (M 3).  
Calculate the loss on drying from the following equation:  
Loss on drying (%w/w) = 
  
where  
 M1 is the mass of sample in grams;  
 M2 is the mass of sample and weighing bot tle in grams before drying; and  
 M3 is the mass of sample and weighing bottle in grams after dryi ng and cooling in a desiccator.  
If the sample melts at a temperature lower than 105 ºC, prepare the sample as described above, then place it 
in a vacuum desiccator containing sulfuric acid. Evacuate the desi ccator to 130 Pa (1 mm of mercur y), 
maintain this vacuum for 24 h, and then weigh the dried sample. Calculate the loss on drying using the same 
equation above.  DEAS 996: 2019  
6 © EAC  2019  – All rights reserved  
 Annex  C  
(normative)  
 
Determination of sulphated ash  
C.1 Apparatus  
C.1.1 Platinum dish  
C.1.2  Desiccator  
C.2 Reagents  
C.1.1 Dilute Sulphric acid  – 10 percent (m/v)  
C.2  Procedure  
Transfer about 2 g of the sample, accurately weighed, to a tared 50 -ml to 100 -ml platinum dish or other 
suitable container and add sufficient dilute sulphuric acid to moist en the entire sample. Heat gently, until the 
sample is dry and thoroughly charred, then continue heating until all the sample has been volatilized or nearly 
all of the carbon has been oxidized. Cool, moisten the residue with 0.1 ml of sulphuric acid, and h eat in the 
same manner until the remainder of the sample and any excess sulphuric acid have been volatilized. Finally 
ignite in a muffle furnace at 800 °C ± 25 °C for 15 minutes. Cool in a desiccator and weigh.  
C.3 Calculation  
Sulphated ash, percent by mas s = 
0X 101
MM  
where  
M1 is the mass, in grams, of residue after igniting, and  
M is the mass, in grams, of the sample tested.  DEAS 996: 2019  
© EAC 2019  – All rights reserved  7 
 Annex  D 
(normative)  
 
Determination of pH (Potentiometric method)  
D.1 Apparatus  
D.1.1 pH meter  
D.1.2 Distilled/deionised water  
D.1.3 Absorbent tissue  
D.1.4 Two standard buffer solutions (High pH and low pH, respectively)  
D.1.5 Weighing scale  
D.1.6 Erlenmeyer flask s 
D.2  Sample preparation  
Prepare a 1:125 solution.  
Measure 1 g of sample into a 250 -ml Erlenmeyer flask . Add 50 ml of distilled water and dissolve the sample. 
Add distilled water to the mixture till the 125ml mark and agitate the flask to obtain a uniform mixture.  
D.3 Procedure  
Use a calibrated pH meter and follow  the manufacturer's instructions. Each time the electrodes are used, rinse 
them with distilled or deionised water and carefully blot them dry with clean absorbent tissue. Form a fresh 
reference electrode liquid junction. Rinse the sample vessel three times  with each new solution to be 
introduced.  
Choose two standard buffers to bracket, if possible, the anticipated pH of the unknown. Warm or cool these 
standards as necessary to match within 2 °C the temperature of the unknown, and initially set the temperatu re 
compensator to that temperature.  
Immerse the electrodes in a portion of the first standard buffer, and following the manufacturer's instructions 
adjust the appropriate standardization control (knob, switch, or button) until the pH reading is that of th e buffer. 
Repeat this procedure with fresh portions of the first standard buffer until two successive readings are within ± 
0.02 pH unit without an adjustment of the standardization control.  
Rinse the electrodes, blot dry, and immerse them in a portion of the second standard buffer of lower pH. Do 
not change the setting of the standardization control. Following the manufacturer's instructions, adjust the 
slope control (thumbwheel switch, knob, or temperature compensator) until the exact buffer pH is display ed. 
Repeat the sequence of standardization with both buffers until the pH readings are within ± 0.02 pH unit for 
both buffers without any adjustment of either control (the amount of sample to be used in sample preparation 
is given where applicable in the i ndividual specification).  
The pH of the unknown solution may then be measured. The difference between the results of two pH 
determinations when carried out simultaneously on in rapid succession by the same analyst, under the same 
conditions, should not exc eed 0.05 pH unit.  
Note: Always re -standardize the instrument after even a short period during which the amplifier is turned off.  DEAS 996: 2019  
8 © EAC  2019  – All rights reserved  
 Annex  E 
(normative)  
 
Test for 5-Benzyl–3,6–dioxo–2–piperazineacetic acid  
E.1 Apparatus  
E.1.1 Gas Chromatograph  - of a suitable type, equipped with a hydrogen flame  ionization detector and 
designed for handling glass  colum ns with on -column injection (Micro -Tek 220 or  equivalent), containing a 1.83 
m x 4 mm (inside diameter) glass c olumn packed with 3 percent OV -1 on 80/100 -mesh supelcoport. Condition 
the column  overnight  at 250  °C before readjus tment and equilibration to  the operating condition. To preclude 
build -up of  silicon  oxide, clean the detector with acetone frequently.  
E.1.1.1 Operating conditions  - the operating param eters may vary depending on the particular instrument 
used, but a suitable chromatogram  may be obtained using the following conditions:  
a) Column tem perature   200 °C  
b) Inlet temperature    200 °C  
c) Detector tem perature    275 °C 
d) Carrier gas – Nitrogen, flowing at a rate of 75 ml per minute  
e) Hydrogen and air flow to burner – optimised  to give maximum sensitivity  
f) Recorder – 1 mV  full scale  
E.2 Reagents  
E.2.1 Silation reagent : Just before use, dilute 3 parts, by volu me, of N , O-bis-(trimethylsilyl) acetamide 
with 2 parts of dimethylformamide.    
E.2.2  Standard preparation: Transfer about 25 mg of 5 -Benzyl -3, 6-dioxo -2-piperazineacetic acid 
Refere nce Standard , accurately  weighed, into a 50 -ml volumetric flask, dissolve in metha nol, dilute to volume  
with methanol, and mix. Pipet 10 ml of this solution into a second 100 -ml volumetric flask, dilute to volume 
with methanol, and mix. Pipet 3 ml of the  second solution into a 2 -dram vial, with Teflon -lined cap, and 
evaporate to  dryness  on a steam bath. Add 1 ml of the Silylation reagent to the residue,  cap the vial tightly, 
shake and heat in an oven at 80  °C for 30 min utes. Remove the  vial from the oven, shake for 15 sec onds , and 
cool to room temperature.  
E.2.3  Sample preparation: Trans fer about 10 mg of the  sample , accurately weighed, into a 2 -dram vial, 
with Teflon -lined cap, add 1 ml of the Silylation reagent, cap tightly,  shake, and heat in an oven at 80  °C for 30 
minutes. Remove the vial from the oven,  shake for 15 sec onds , and cool  to room temperature.  
E.3 Procedure  
Inject a 3 µl portion of the standard preparation into the gas chromatograph and obtain the chromatogram. 
Measure the height of the peak produced by the 5 -benzyl -3, 6-dioxo -2-piperazineacetic acid, and record it as 
P. Under the stated conditions, the elution time is about 7 -9 min. Similarly, inject a 3 µl portion of the sample 
preparation, obtain the chromatogram and measure the height of the peak produced by any 5 -benzyl -3,6-
dioxo -2-piperazineacetic acid contained in the  sample, and record it as p. 
 DEAS 996: 2019  
© EAC 2019  – All rights reserved  9 
 The percentage of 5 -Benzyl –3, 6–dioxo –2–piperazineacetic acid in the sample can be computed as  
PmpM
 x  x 500 x  x 3
   
where,  
 M is the mass, in milligrams, of the reference standard taken  
 m is the mass in milligrams, of as partame analysed  
  p is the height of  peak produced by 5 -benzyl -3, 6-dioxo -2-piperazineacetic acid contained in sample;   
and 
 P height of peak produced by 5 -benzyl -3, 6-dioxo -2-piperazineacetic acid contained in standard.  DEAS 996: 2019  
10 © EAC  2019  – All rights reserved  
 Annex  F 
(normative)  
 
Test for amine group  
F.1 Apparatus  
F.1.1  Test tube  
F.2 Reagents  
F.2.1  Ninhydrin  
F.2.2  Dimethylsulfoxide  
F.2.3 Hydrindantin  
F.2.4 4M Lithium acetate  
F.2 Procedure  
Dissolve 2 g of ninhydrin in 75 ml of dimethylsulfoxide, add 62 mg of hydrindantin, dilute to 100 ml with 4M 
lithium acetate buffer solution (pH 9), and filter. Transfer about 10 mg of the sample to a test tube, add 2 ml of 
the reagent solution, and heat. A dark purple colour is formed.   DEAS 996: 2019  
© EAC 2019  – All rights reserved  11 
 Annex  G 
(normative)  
 
Test for ester group  
G.1 Reagents  
G.1.1 Methanol  
G.1.2 Methanol  saturated with hydroxylamine hydrochloride  
G.1.3 5N potassium hydroxide  
G.1.4 Hydrochloric acid  
G.1.5 Ferric chloride  
G.2 Procedure  
Dissolve abo ut 20 mg of sample in 1 ml of methanol, add 0.5 ml of methanol saturated with hydroxylamine 
hydrochloride, mix, and then add 0.3 ml of 5N potassium hydroxide in methanol. Heat the mixture to boiling, 
then cool, adjust the pH to between 1 and 1.5 with hydro chloric acid, and add 0.1 ml of ferric chloride. A 
burgundy/maroon colour is produced.  DEAS 996: 2019  
12 © EAC  2019  – All rights reserved  
 Bibliography  
[1] JECFA specification (2001):  Aspartame monograph   
[2] IS 13657 : 1993 (Reaffirmed in 2003) , Aspartame, food grade — Specification  
[3] Food chemicals cod ex (5th Edition)  
[4] EFSA Journal (2013) – Scientific opinion on a spartame  
[5] JECFA (2006), Analytical methods, test procedures and laboratory solutions used by and referenced in 
the food additive specifications, Volume 4.