UGANDA STANDARD
 
  US ISO 5564
      First Edition
         2009-09-04
 
    Reference numbe r
         US ISO 5564:1982
 © UNBS 2009
  
Black pepper and white pepper, whole or ground — 
Determination of piperine content — Spectrophotometric 
method 
 US ISO 5564:1982 
 
© UNBS  2009 – All rights reserved ii Compliance with this standard does not, of itself confer immunity from legal obligations 
A Uganda Standard does not purport to include all necessary provisions of a contract. Users are 
responsible for its correct application  
© UNBS 2009 
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilised in any 
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The Executive Director 
Uganda National Bureau of Standards 
P.O. Box 6329 
Kampala  
Uganda 
Tel: 256 41 505 995 
Fax: 256 41 286 123 
E-mail: unbs@infocom.co.ug
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       US ISO 5564:1982  
© UNBS  2009 - All rights rese rved                iii National foreword 
Uganda National Bureau of Standards (UNBS) is a parastatal under the Ministry  of Tourism, Trade and 
Industry established under Cap 327, of the Laws of U ganda. UNBS is mandated to co-ordinate the elaboration 
of standards and is  
(a) a member of International Organisation for Standardisation (ISO) and  
(b) a contact point for the WHO/FAO Codex A limentarius Commission on Food Standards, and  
(c) the National Enquiry Point on TBT/SPS Agr eements of the World Trade Organisation (WTO). 
The work of preparing Uganda Standards is carried out through Technical Committees. A Technical 
Committee is established to deliberate on standards in a gi ven field or area and consists of representatives of 
consumers, traders, academician s, manufacturers, governme nt and other stakeholders. 
Draft Uganda Standards adopted by the Technical Committ ee are widely circulated to stakeholders and the 
general public for comments. The committee reviews the comments before recommending the draft standards 
for approval and declaration as Uganda Standards by the National Standards Council. 
This Uganda Standard, US ISO 5564, Black pepper and white pepper, whole or ground — Determination
of piperine cont ent — Spectrophotometric method,  is identical with and has been reproduced from an
International Standard, ISO 5564:1982  Black pepper and white pepper, whole or ground — 
Determination of piperine conten t — Spectrophotometric method, and adopted as a Uganda Standard.
  
This standard was developed by the Food and agricu lture Standards Technical Committee (UNBS/TC 2).  
Wherever the words, "International Standard" a ppear, they should be replaced by "Uganda Standard."
  
 International Standard 5564 
INTERNATIONAL ORGANIZATION FOR STANDARDIZATIONWIE~YHAPOAHAR OPTAHHSALWIR fl0 CTAH~PTM3ALplM~RGANISATION INTERNATIONALE DE NORMALISATION 
Black pepper and white pepper, whole or ground - 
Determination of piperine content - Spectrophotometric 
method 
Pokre nok et pokre blanc, entier ou en poudre - DtSterrnination de Ia teneur en pipkine - Methode spectropho tomhique 
First edition - 1982-11-15 
iz UDC 633.841: 543.8 Ref. No. ISO 55644982 (E) - 
Descriptors : agricultural products, spices, pepper, tests, spectrophotometric analysis. 
Price based on 2 pages Forewoad 
ISO (the International Organization for Standardization) is a worldwide federation of 
national Standards institutes (lS0 member bodies). The work of developing Inter- 
national Standards is carried out through ISO technical committees. Every member 
body interested in a subject for which a technical committee has been set up has the 
right to be represented on that committee. International organizations, governmental 
and non-governmental, in liaison with ISO, also take part in the work. 
Draft International Standards adopted by the technical committees are circulated to 
the member bodies for approval before their acceptance as International Standards by 
the ISO Council. 
International Standard ISO 5564 was developed by Technical Committee ISO/TC 34, 
dlgricultu~alfoodpaoduct~, and was circulated to the member bodies in February 1981. 
lt has been approved by the member bodies of the following countries : 
Austria 
Canada 
Czechoslovakia 
Egypt, Arab Rep. of 
Ethiopia 
France 
Germany, F. R. 
Hungary 
lndia 
lreland Italy 
Kenya 
Korea, Dem. P. Rep. sf 
Korea, Rep. of 
Malaysia 
Netherlands 
New Zealand 
Peru 
Philippines 
Poland 
No member body expressed disapproval of the document. Portugal 
Romania 
South Africa, Rep. of 
Spain 
Sri Lanka 
Thailand 
Turkey 
United Kingdom 
USSR 
Y ugoslavia 
0 International Organkation for Standardkation, 1982 
Printed in Switzerland INTERNATIONAL STANDARD ISO 5564-1982 (E) 
Black pepper and white epper, whole’or gr,ound - 
Determination of pi erine content - Spectrophotometric 
method 
0 Introduction 
The method described in this international Standard is based 
on a number of international collaborative studies carried out 
over a Iong period of time. Necessarily, the method seeks to 
optimize a number of variables in an attempt to define pro- 
cedures and provide a common measure of the pungency of 
PePPer- 
1 Scope and field of application 
This International Standard specifies a spectrophotometric 
method for the determination of the piperine content of black 
or white pepper (Pipernigrum L.), in whole or in ground form. 
2 References 
ISO 939, Spices and condiments - Determination of moisture 
content - En trainmen t me thod. 
ISO 948, Spices and condiments - Sampling. 
ISO 2825, Spices and condiments - Reparation of a ground 
Sample for analysis. 
3 Definition 
3.1 piperine content : The quantity of pungent compounds 
(piperine and its analogues) in the product, determined by 
the procedure specified in this International Standard, and ex- 
pressed as a percentage by mass. 
4 Principle 
Extraction of the pungent compounds with ethanol and spec- 
trophotometric measurement at 343 nm. 5 Reagent 
5.1 Ethanol, 96 % WV. 
6 Apparatus 
Usual laboratory equipment, and in particular 
6.1 Extraction apparatus, comprising the following com- 
ponents joined together by means of ground-glass joints. 
6.1.1 Flask, round-bottomed, of capacity 100 ml. 
Kl.2 Reflux condenser. 
6.1.3 Anti-bumping granules : glass beads. 
6.2 Qne-mark wolumetric flasks, of capacities 25, 50 and 
100 ml, complying with the requirements of ISO 1042, and 
wrapped in aluminium foil or black tape (6.4). 
6.3 Filter Paper. 
6.4 Aluminium foil or black tape. 
6.5 Pipettes, one-mark, of capacity 5 ml, complying with 
the requirements of ISO 648. 
6.6 Spectrophotometer, capable of accurately measuring 
absorbances at 343 nm, and provided with a pair sf matched 
silica cells, or cells transparent to ultraviolet radiation, of Optical 
path length 1 cm. 
6.7 Analytical balance. 
1 ISO 55644982 (EI 
7 Sampling 
Sample the material by the method described in ISO 948. 
8 Procedure 
NOTE - It is essential during the operations described below, and 
whiist the piperine is in soiution, to avoid exposure to light. The 
isomerisation of piperine is catalysed by sunlight, and, therefore, 
during extraction and storage of the stock solution, iight should be 
excluded by wrapping the flasks with aluminium foil or black tape 
(sec 6.2). Measure the absorbance (Al of this Solution at 343 nm, by 
means sf the spectrophotometer (6.61, using, as the reference 
liquid, the ethanol (5.1). 
9 
The piperine content, expressed 
the dry basis, is equal to Expression of resuits 
A 50 25 100 100 -x-x-x 
J % x5 5 m 100~H 
lcm 
81 . Preparation of test Sample 
m 
Prepare the test Sample as described in ISO 2825. 
8.2 Test Portion 
Weigh, to the nearest 0,000 1 g, about 0,5 g of the’test Sample 
and transfer it to the round-bottomed flask (6.1 .l). 
8.3 Determination 
Transfer about 50 ml of the ethanol (5.1) to the flask (6.1.1) and 
add a few anti-bumping granules (6.1.3). Connect the con- 
denser (6.1.2) and, in a dark place (sec the introductory note to 
clause 81, boil under reflux for 3 h. Allow to cool and filter the 
Solution into a 160 ml volumetric flask (6.2). Wash the extrac- 
tion flask and filter Paper with successive 10 ml portions of the 
ethanol and use the washings to make up the volume in the 
volumetric flask to the mark. Transfer, by means of a pipette 
(6.51, a 5 ml aliquot Portion of this Solution to a 50 ml 
volumetric flask (6.2) and make up to the mark with ethanol. 
Transfer, by means of a pipette (6.51, 5 ml of this Solution into a 
25 ml volumetric flask (6.2) and again make up to the mark with 
ethanol. as a percentage bY mass 
is the mass, in grams, of the test Portion (8.2); on 
H is the moisture content, expressed as a percentage by 
mass, of the test Sample, determined by the method 
specified in ISO 939; 
A is the absorbance of the final test Solution; a 
A1 % 
lcm is the absorbance at 343 nm of a 1 % 
Solution of piperine in a cell of Optical path length 1 cm, 
which equals 1 238. 
10 Test report 
The test report shall Show the method used and the result ob- 
tained. lt shall also mention any operating conditions not 
specified in this International Standard, or regarded as op- 
tional, together with any circumstances that may have influenc- 
ed the result. 
The test report shall include all the information 
the complete identification of the Sample. necessary for 
2