DRAFT EAST AFRICAN STANDARD
 DEA S 831 :2014  
ICS 67.120.30  
 
© EAC 2014 First Edition 2014  
 
 
 DRAFT EAST AFRICAN STANDARD  
 
  
Frozen fish fillets — specification 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
EAST AFRICAN COMMUNITY  
 DRAFT EAST AFRICAN STANDARDDEAS 831:2014  
© EAC 2014 — All rights reserved ii Foreword  
 
Development of the East African Standards has been necessitated by the need for harmonizing 
requirements governing quality of products and services in East Africa. It is envisaged that through 
harmonized standardization, trade barriers which are encountered when goods and services are 
exchanged within the Community will be removed.  
 
In order to meet the above objectives, the EAC Partner States have enacted an East African 
Standardization, Quality Assurance, Metrology and Test Act, 2006 (EAC SQMT Act, 2006) to make 
provisions for ensuring standardization, quality assurance, metrology and testing of products 
produced or originating in a third country and traded in the Community in order to facilitate industrial 
development and trade as well as helping to protect the health and safety of society and the 
environment in the Community. 
 
East African Standards are formulated in accordance with the procedures established by the East 
African Standards Committee. The East African Standards Committee is established under the 
provisions of Article 4 of the EAC SQMT Act, 2006. The Committee is composed of representatives of 
the National Standards Bodies in Partner States, together with the representatives from the private 
sectors and consumer organizations. 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 accordance with the procedures of the 
Community.  
 
Article 15(1) of the EAC SQMT Act, 2006 provides that “Within six months of the declaration of an 
East African Standard, the Partner States shall adopt, without deviation from the approved text of the 
standard, the East African Standard as a national standard and withdraw any existing national 
standard with similar scope and purpose”.  
 
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.  
 
© East African Community 2010 — All rights reserved* 
East African Community  
P O Box 1096  
Arusha  
Tanzania  
Tel: 255 27 2504253/8 
Fax: 255- 27-2504481/2504255 
E-Mail: eac@eachq.org  
Web: www.each.int 
                                                           
* 2014 EAC — All rights of exploitation in any form and by any means reserved worldwide for EAC Partner States’ NSBs. 
 DRAFT EAST AFRICAN STANDARDDEAS 831:2014  
© EAC 2010 — All rights reserved   Frozen fish fillets — Specification  
 
1 Scope 
 
This East Africa draft   standard specifies requirements and methods of sampling and test for frozen 
fish fillets intended for human consumption without further processing. 
 
 
2 Normative references 
 
The following referenced documents are indispensable for the application 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. 
 
CODEX STAN 192 - CODEX STAN 192 -1995, General standard for food additives  
EAS 39, Hygiene in the food and drink manufacturing industry ― Code of practice 
CAC/RCP 52[CD/K/521:2010] , Code of practice for fish and fishery products  
 
EAS 12,  Drinking (potable water) — Specification  
 
EAS 38, Labelling of pre-packaged foods — Specification 
 
EAS 103, Schedule for permitted food additives  
 
ISO 4833, Microbiology of food and animal feeding stuffs — Horizontal method for the enumeration of 
microorganisms — Colony-count technique at 30 degrees C  
 
ISO 6579, Microbiology of food and animal feeding stuffs — Horizontal method for the detection of 
Salmonella spp.  
 
ISO 6888- 1, Microbiology of food and animal feeding stuffs — Horizontal method for the enumeration 
of coagulase-positive staphylococci (Staphylococcus aureus and other species) — Part 1: Technique 
using Baird-Parker agar medium  
 
ISO 6888- 2, Microbiology of food and animal feeding stuffs — Horizontal method for the enumeration 
of coagulase-positive staphylococci (Staphylococcus aureus and other species) — Part 2: Technique 
using rabbit plasma fibrinogen agar medium  
 
ISO 6888- 3, Microbiology of food and animal feeding stuffs — Horizontal method for the enumeration 
of coagulase-positive staphylococci (Staphylococcus aureus and other species) — Part 3: Detection 
and MPN technique for low numbers  
 
ISO 7251, Microbiology of food and animal feeding stuffs — Horizontal method for the detection and 
enumeration of presumptive Escherichia coli — Most probable number technique 
 
ISO 11290- 2, Microbiology of food and animal feeding stuffs — Horizontal method for the detection 
and enumeration of Listeria monocytogenes — Part 2: Enumeration method 
 
ISO 7937, Microbiology of food and animal feeding stuffs — Horizontal method for the enumeration of 
Clostridium perfringens — Colony-count technique 
 
ISO 21527- 1, Microbiology of food and animal feeding stuffs -- Horizontal method for the enumeration 
of yeasts and moulds -- Part 1: Colony count technique in products wit h water activity greater than 0. 9 
 DRAFT EAST AFRICAN STANDARD  DRAFT EAST AFRICAN STANDARDDEAS 831:2014  
© EAC 2010 — All rights reserved   ISO 27085, Animal feeding stuffs -- Determination of calcium, sodium, phosphorus, magnesium, 
potassium, iron, zinc, copper, manganese, cobalt, molybdenum, arsenic, lead and cadmium by ICP-
AES 
ISO 16050,  Foodstuffs — Determination of aflatoxin B 1, and the total content of aflatoxin B 1, B2, G 1 
and G 2 in cereals, nuts and derived products  — High performance liquid chromatographic method  
 
AOAC 990.04 Mercury (Methyl) in sea food by liquid chromatography-Atomic Absorption 
Spectroscopy (LC-AAS) 
 
AOAC 977.15 / 974.14 Mercury in fish by Flame Atomic Absorption Spectroscopy (FAA) 
 
3 Terms and definitions 
 
For the purpose of this standard the following terms and definitions shall apply;   
3.1  
fillet  
slice of fish of irregular size and shape removed from the fish flesh by cuts made parallel to the 
backbone. 
 
3.2  
frozen fillets  
the fillet that undergone freezing process 
 
3.3  
freezing   
process of heat removal from fish such that the temperature falls fairly rapidly to just below 0°C, the 
freezing point of water  
 
 
4 Requirements 
 
4.1 General requirements 
4.1.1 Frozen fish fillets shall be prepared from sound fish which are of a quality fit to be sold fresh for 
human consumption 
4.1.2  Frozen fish fillets shall be prepared from fish that conforms to DEAS 827:2014 
4.1.3  Frozen fish fillets shall be checked for foreign matter, colour, texture and odour of the fillet prior 
freezing  
4.1.4  Water used for washing the fish shall comply with EAS 12. 
4.1.5  The frozen fish fillets shall have a temperature of -18ºC 
 
4.2 Specific requirements 
 
4.2.1  Fillets may be presented as boneless, skin-on, glazed, provided that deboning has been 
completed including the removal of pin-bones. 
4.2.2  When tested as per annex A or any other validated method the level of histamine in species with 
high levels of histidine such as scromboids shall not exceed 10ppm 
 
5 Hygie ne 
 
The product covered by the provisions of this standard shall be prepared and handled in accordance 
with, EAS 39 and shall comply with microbiological limits given in table 1: 
 
 
 
 DRAFT EAST AFRICAN STANDARDDEAS 831:2014  
© EAC 2010 — All rights reserved    
 
 
 
 
Table 1 — Microbiological limits for frozen fish fillets 
 
S/No Micro -organisms  Max. limits  Method s of test  
i Salmonella  per 25 g  Absent  ISO 6579  
ii E. coli  per gram         absent  ISO 7251  
iii listeria monocytogenes  absent  ISO 11290 PART 1 &2  
iv Staphylococcus aureus  cfu per gram  100 ISO 6888  
v Clostridium perfringens  per gram  Absent  ISO 7937  
vi Vibrio Spp  per gram  Absent  ISO/TS 21872  
vii Total viable count per gram  105 ISO 4833  
 
 
 
6 Contaminant limits 
 
Frozen fish fillets shall comply with the contaminant limits given in table 2. 
 
Table 2 — Contaminant limits for frozen fish fillets 
 
 
S/No  Type of contaminant  Maximum limit  
(mg/kg)  Methods of test  
i Arsenic  0.1 ISO 27085  
ii Mercury  0.5 AOAC 977.15/ 
974.14  
iii Lead  0.3 ISO 27085  
iv Cadmium  0.3 ISO 27085  
v Methyl  mercury  0.5 AOAC 990.04  
 
 
7 Weight and measures 
 
The weight of the product shall comply with Weight and Measures regulations of Partner States or 
equivalent legislation. 
 
8 Packaging and labelling 
 
8.1 Packaging 
 
8.1.1  Frozen fish fillets shall be packaged in food grade containers which will safeguard the hygienic, 
nutritional, and organoleptic qualities of the product.  
8.1.2  The containers, including packaging material, shall be made of substances which are safe and 
suitable for their intended use. They shall not impart any toxic substance or undesirable odour or 
flavour to the product. DRAFT EAST AFRICAN STANDARDDEAS 831:2014  
© EAC 2010 — All rights reserved    
 
 
8.2 Labelling 
 
In addition to the requirements in EAS 38, the following specific labelling requirements shall apply and 
shall be legibly and indelibly marked: 
a) name of the product shall be "... frozen fish fillets 
b) storage and transportation conditions declaring the temperature to be -18C or lower 
c) Name and physical address of processor 
d) Net weight in grams or kilograms 
e) Date of  production 
f) Batch or code number 
g) Expiry date 
h) instruction for use 
i) Country of origin 
8.3 Labelling of Non-retail Containers 
 
Information on the above provisions shall be given either on the container or in accompanying 
documents, except that the name of the product, lot identification, and the name and address of the 
processor or packer as well as storage instructions, shall appear on the container. 
 
However, lot identification, and the name and address of the processor or packer may be replaced by 
an identification mark provided that such a mark is clearly identifiable with the accompanying 
documents. 
 
9. Method of sampling and test 
 
Sampling and tests shall be done as per test methods described in respective tables  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 DRAFT EAST AFRICAN STANDARDDEAS 831:2014  
© EAC 2010 — All rights reserved    
 
Annex A 
(normative) 
 
Determination of histamine 
 
A.1 Principle 
 
Sample is extracted with 75% (v/v) methanol. Extract is passed through ion exchange column. o –
Phthaldialdehyde solution is added to eluate to form fluorescent histamine derivatives. Fluorescent 
intensity of derivatives is measured using fluorometer and histamine is quantified using external 
standards. 
 
A.2 Apparatus  
 
Rinse all plastic and glass containers with HCl (1 + 3) and H 2O before use. 
 
(a) Chromatographic tube — 200 
 7 id mm polypropylene tube fitted with small plastic stopcocks 
and ca 45 cm Teflon tubing. Control flow rate at >3 ml/min by adjusting height of column relative 
to tubing outlet. Alternatively, use 2-way valve in place of tubing. 
 
(b) Photofluorometer  — equipped with medium pressure Hg lamp with excitation at 350 nm and 
measuring emission at 444 nm. 
 
(c) Repipets  — 1 and 5 m. 
 
A.3 Reagents 
 
(a) Ion-exchange resin  — Bio-Rad AG 1-X8, 50 –100 mesh or Dowex 1-X8, 50 –100 mesh. 
Convert to -OH form by adding ca 15 m 2M NaOH/g resin to beaker. Swirl mixture and let stand 
<30min. Decant liquid and repeat with additional base. Thoroughly wash resin with H 2O, slurry 
into fluted paper and wash again with H 2O. Prepare resin fresh weekly and store under H 2O. 
Place glass wool plug in base of tube, A .2(a), and slurry in enough resin to form 8 cm bed. 
Maintain H 2O level above top of resin bed at all times. Do not regenerate resin in packed 
column; rather, use batch regeneration in beaker when necessary. Wash column with about 10 
ml H 2O before applying each extract. 
 
(b) Phosphoric acid  — 3.57N. Dilute 121.8 ml 85% H 3PO 4 to 1 L. For other concentration H 3PO 4, 
volume required for 1 L1.19M acid = 17493/(density H 3PO 4 
percent H 3PO 4). Standardize 
5.00 ml by titration with 1.00M NaOH to phenolphthalein end point, and adjust concentration if 
necessary. 
 
(c) o-Phthaldialdehyde (OPT) solution  — 0.1% (w/v). Dissolve 100 mg OPT in 100 ml distilled- in-
glass methanol. Store in amber bottle in refrigerator. Prepare fresh weekly. 
 
(d) Histamine standard solutions  — Store in refrigerator.  
 
(1) Stock solution  — 1 mg/ml as free base. Accurately weigh ca 169.1 mg histamine 2HCl 
(98%) into 100 ml volumetric flask, and dissolve and dilute to volume with 0.1M HCl. 
Prepare fresh weekly.  
 
(2) Intermediate solution  — 10 µg/ml. Pipet 1 ml stock solution into 100 ml volumetric flask, 
and dilute to volume with 0.1M HCl. Prepare fresh weekly.  
 
(3) Working solutions  — 0.5, 1.0, and 1.5 µg/5 ml. Pipet 1, 2, and 3 ml intermediate 
solution into separate 100 ml volumetric flasks, and dilute each to volume with 0.1M HCl. 
Prepare fresh daily. DRAFT EAST AFRICAN STANDARDDEAS 831:2014  
© EAC 2010 — All rights reserved    
(e) Methanol  — 75% (v/v). Place 75 ml MeOH (distilled in glass) into 100 ml volumetric flask or 
stoppered graduated cylinder. Dilute to volume with H 2O. Swirl flask while adding H 2O. 
 
A.4 Preparation of standard curve 
 
Pipet duplicate 5 ml aliquots of each working standard solution into separate 50 ml glass or 
polypropylene Erlenmeyers. Pipet in 10 mL 0.1M HCl to each flask and mix. Pipet in 3 ml 1M NaOH 
and mix. Within 5 min, pipet in 1 ml OPT solution and mix immediately. After exactly 4 min, pipet in 3 
ml 3.57NH 3PO 4 and mix immediately. It is important to mix thoroughly after each addition and at least 
once during OPT reaction. (Run 6 – 10 OPT reactions simultaneously by adding reagents to 
Erlenmeyers in set order.) Prepare blank by substituting 5 ml 0.1M HCl for histamine solution. Within 
1.5 h, record fluorescence intensity ( I) of working standard solutions with H 2O in reference cell, using 
excitation wavelength of 350 nm and emission wavelength of 444 nm. Plot I (corrected for blank) 
against µg histamine/5 ml aliquot. 
 
A.5 Determination 
 
Extract prepared sample with 75% (v/v) methanol. Pass 4 –5 ml H 2O through column, A .2(a), and 
discard eluate. Pipet 1 ml extract onto column and add 4 –5 ml H 2O. Immediately initiate column flow 
into 50 ml volumetric flask containing 5.00 ml 1.00M HCl. When liquid level is ca 2 mm above resin, 
add ca 5 ml H 2O and let elute. Follow with H 2O in larger portions until ca 35 ml has eluted. Stop 
column flow, dilute to volume with H 2O, stopper, and mix. Refrigerate eluate. 
 
Pipet 5 ml eluate into 50 ml Erlenmeyer, and pipet in 10 ml 0.1M HCl. Proceed as in A.4, beginning 
"Pipet in 3 ml 1M NaOH….”  
 
If test sample contains >15 mg histamine/100 g fish, pipet 1 ml sample –OPT mixture into 10 ml 
beaker containing exactly 2 ml blank –OPT mixture, and mix thoroughly. Read fluorescence of new 
solution. Dilute and mix aliquots with blank –OPT mixture as needed to obtain measurable reading. 
This approximation indicates proper dilution of eluate required prior to second OPT reaction needed 
for reliable quantitation of test sample. Alternatively, use sensitivity range control of fluorometer (if 
instrument has one) to estimate dilution. Use these approximations to prepare appropriate dilution of 
aliquot of eluate with 0.1NHCl, and proceed as in A.4, beginning "Pipet in 3 ml 1M NaOH . . ." 
 
A.6 Calculations 
 
Plot of I (measured by meter deflection or recorder response and corrected for blank) against  µg 
histamine/5 ml test solution should be straight line passing through origin with  
slope =  m = [(Ia /1.5) + Ib + 2I c ]/3. 
 
mg Histamine/100 g fish = (10)( F)(1/m)(Is) 
 
µg Histamine/g fish = 10 
 (mg histamine/100 g fish) 
 
where Is, Ia, Ib, and Ic = fluorescence from test sample, 1.5, 1.0, and 0.5 µg histamine standards, 
respectively; and F = dilution factor = (ml eluate + ml 0.1M HCl)/ml eluate. F = 1 for undiluted eluate. 
 
If calibration plot is not linear, use standard curve directly for quantitation. Each subdivision on 
abscissa should be ≤0.1 µg histamine/5 ml test solution. Read all values from curve to nearest 0.05 
µg histamine/5 ml test solution. 
 
mg Histamine/100 g fish = (10)( F)(W) 
 
µg Histamine/g fish = 10× (mg histamine/100 g fish) 
 
where W = µg histamine/5 ml test solution as determined from standard curve. 
 
 DRAFT EAST AFRICAN STANDARDDEAS 831:2014  
© EAC 2010 — All rights reserved