ISO-11027-1993.pdf
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1、INTERNATIONAL STANDARD ISO 11027 First edition 1993-07-15 Pepper and pepper oleoresins - Determination of piperine content - Method using high-performance liquid chromatography Poivres, okorbines de poivres - Determination de Ia teneur en pipbine - Methode par Chromatographie en Phase liquide 2 haut
2、e Performance Reference number ISO 11027:1993(E) ISO 11027:1993(E) Foreword ISO (the International Organization for Standardization) is a worldwide federation of national Standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical
3、 committees. Esch member body interested in a subject for which a technical committee has been established 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. ISO collaborates closely with
4、 the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. Draft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of th
5、e member bodies casting a vote. International Standard ISO 11027 was prepared by Technical Committee lSO/TC 34, Agricultural food products, Sub-Committee SC 7, Spices and condimen ts. Annex A of this International Standard is for information only. 8 ISO 1993 All rights reserved. No patt of this publ
6、ication may be reproduced or utilized in any form or by any means, electronie or mechanical, including photocopying and microfilm, without per- mission in writing from the publisher. International Organization for Standardization Case Postale 56 l CH-l 211 Geneve 20 l Switzerland Printed in Switzerl
7、and ii -,-,- INTERNATIONAL STANDARD ISO 11027:1993(E) Pepper and pepper oleoresins - Determination of piperine content - Method using high-performance liquid chromatography 1 Scope This International Standard specifies a method for the determination, by high-performance liquid chro- matography, of t
8、he piperine content of peppers (Piper nigrum Linnaeus), whole or powdered, as weil as their extracts (oleoresins). This method enables the Separation and, if necessary, the determination of the other alkaloids of pepper (isochavicine, isopiperine and piperittin). 2 Normative references The following
9、 Standards contain provisions which, through reference in this text, constitute provisions of this International Standard. At the time of publi- cation, the editions indicated were valid. All Standards are subject to revision, and Parties to agreements based on this International Standard are encour
10、aged to investigate the possibility of applying the most re- cent editions of the Standards indicated below. Members of IEC and ISO maintain registers of cur- rently valid International Standards. ISO 28253 981, Spices and condiments - Prepa- ration of a ground Sample for analysis. ISO 5564:1982, Bl
11、ack pepper and white pepper, whole or ground - Determination of piperine con ten t - Spectrophotometric method. 3 Principle 3.1 Ground pepper Extraction with ethanol under reflux, then determi- nation of piperine by high-performance liquid chro- 2 matography (HPLC), in accordance with the procedure
12、described in this International Standard. 3.2 Whole pepper Preparation by grinding the Sample, then extraction of the powder obtained, followed by determination of piperine by HPLC, in accordance with the procedure described in this International Standard. 3.3 Oleoresins of pepper Dilution of the ol
13、eoresin in ethanol, then determi- nation of piperine by HPLC, in accordance with the procedure described in this International Standard. 4 Reagents Use only reagents of recognized analytical grade and distilled or demineralized water or water of equivalent purity. 4.1 Reference substance Piperine of
14、 at least 98 % purity, determined by the spectrometric method described in ISO 5564. 9 WARNING - This product should be handled with care as it is strongly irritating. 4.2 Solvents 4.2.1 Ethanol 4.2.2 Acetoni , 96 % (VW). trile WARNING - This product should be handled with care as it is lachrymatory
15、. 4.2.3 Acetic acid, 1 % (VW) aqueous Solution. -,-,- ISO 11027:1993(E) 4.3 Elution solvent Mix 52 volumes of acetic acid Solution (4.2.3) and 48 volumes of acetonitrile (4.2.2). 5 Apparatus Usual laboratory equipment and, in particular, the fol- lowing. 5.1 Pipettes, of 2 ml, 5 ml and 10 ml capacit
16、y. 5.2 Volumetric flasks, of 25 ml, 50 ml and 100 ml capacity (made of antiactinic glass or wrapped in alu- minium feil). 5.3 Filtration diaphragms, non-adsorbing. 54 . Separation System 5.4.1 Chromatograph, for high-performance liquid chromatography. 5.4.2 Pumping System, which enables a flow rate
17、which is constant or programmed at high pressure to be obtained and maintained. 5.4.3 Degassing Systems, for solvents. 5.4.4 Suitable detector System, adjustable to a wavelength of 343 nm. 5.5 Recorder or integrator, the Performance of which is compatible with the apparatus as a whole. 5.6 Column Ma
18、terial: stainless steel or glass Length: 10 cm to 25 cm Inside diameter: 0,4 cm to 0,5 cm Stationary Phase: grafted silica with derived octadecyl functional group, C,a, particle size 5 Pm max. 5.7 Injection System Injection valve with 10 1 loop or any other System giving the same injection accuracy.
19、 5.8 Test sieve, of aperture size 500 Pm. 6 Calibration method WARNING - Throughout the Operation, it is imperative that the solutions are not exposed to light (for instance wrap the flasks in aluminium feil) and are used as quickly as possible because of the instability of the piperine solutions. 6
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