《ISO-14939-2001.pdf》由会员分享，可在线阅读，更多相关《ISO-14939-2001.pdf（22页珍藏版）》请在三一文库上搜索。
1、Reference number ISO 14939:2001(E) ISO 2001 INTERNATIONAL STANDARD ISO 14939 First edition 2001-08-15 Animal feeding stuffs Determination of carbadox content Method using high- performance liquid chromatography Aliments des animaux Dtermination de la teneur en carbadox Mthode par chromatographie liq
2、uide haute performance Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 03:40:03 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 14939:2001(E) PDF di
3、sclaimer This PDF file may contain embedded typefaces. In accordance with Adobes licensing policy, this file may be printed or viewed but shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In downloading this file, partie
4、s accept therein the responsibility of not infringing Adobes licensing policy. The ISO Central Secretariat accepts no liability in this area. Adobe is a trademark of Adobe Systems Incorporated. Details of the software products used to create this PDF file can be found in the General Info relative to
5、 the file; the PDF-creation parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below. ISO 2001 All
6、 rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISOs member body in the country o
7、f the requester. ISO copyright office Case postale 56 ? CH-1211 Geneva 20 Tel. + 41 22 749 01 11 Fax + 41 22 749 09 47 E-mail copyrightiso.ch Web www.iso.ch Printed in Switzerland ii ISO 2001 All rights reserved Copyright International Organization for Standardization Provided by IHS under license w
8、ith ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 03:40:03 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 14939:2001(E) ISO 2001 All rights reservediii ContentsPage Foreword.iv 1Scope 1 2Normative reference1 3Principle1 4Reagents.2 5Appa
9、ratus.4 6Sampling.5 7Preparation of test sample5 8Procedure.5 8.1General5 8.2Preparation of spiked sample.5 8.3Extraction .6 8.4Column chromatography7 8.5HPLC analysis7 9Confirmation.8 9.1General8 9.2Co-chromatography.8 9.3Diode array detector9 9.4Post-column derivatization.10 10Calculation of resul
10、ts 10 10.1General10 10.2Feeding stuffs containing 0,1 mg/kg to 10 mg/kg of carbadox.10 10.3Feeding stuffs containing 10 mg/kg to 100 mg/kg of carbadox11 10.4Premixtures containing up to 10 % of carbadox11 11Precision.11 11.1Interlaboratory test11 11.2Repeatability.11 11.3Reproducibility.12 12Test re
11、port12 Annex A (informative) Flow chart13 Annex B (informative) Results of interlaboratory test.14 Bibliography16 Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 03:40:03 MDTNo r
12、eproduction or networking permitted without license from IHS -,-,- ISO 14939:2001(E) iv ISO 2001 All rights reserved Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Stand
13、ards is normally carried out through ISO technical committees. Each 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 ta
14、ke part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 3. Draft International Standards adopted b
15、y the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibility that some of the elements of this International Standard may be the subjec
16、t of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. InternationalStandardISO 14939waspreparedbyTechnicalCommitteeISO/TC 34,Foodproducts, Subcommittee SC 10, Animal feeding stuffs. Annexes A and B of this International Standard are for information only
17、. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 03:40:03 MDTNo reproduction or networking permitted without license from IHS -,-,- INTERNATIONAL STANDARDISO 14939:2001(E) ISO 2
18、001 All rights reserved1 Animal feeding stuffs Determination of carbadox content Method using high-performance liquid chromatography 1Scope This International Standard specifies a high-performance liquid chromatographic (HPLC) method for the determination of the carbadox content in premixtures and a
19、nimal feeding stuffs. The method is applicable to animal feeding stuffs with a mass fraction of carbadox of 0,5 mg/kg (limit of quantification) to 100 mg/kg, and to premixtures with a mass fraction of carbadox up to 10 %. The lower limit of detection is 0,1 mg/kg. NOTE 1For animal feeding stuffs the
20、 mass fraction of carbadox is expressed in milligrams per kilogram, and for premixtures as a percentage by mass. NOTE 2Carbadox is a chemotherapeuticum belonging to the quinoxaline group. Carbadox is used as a growth-promoting feed additive for piglets. 2Normative reference The following normative d
21、ocument contains provisions which, through reference in this text, constitute provisions of this International Standard. For dated references, subsequent amendments to, or revisions of, any of these publications do not apply. However, parties to agreements based on this International Standard are en
22、couraged to investigate the possibility of applying the most recent editions of the normative documents indicated below. For undated references, the latest edition of the normative document referred to applies. Members of IEC and ISO maintain registers of currently valid International Standards. ISO
23、 6498:1998, Animal feeding stuffs Preparation of test samples. 3Principle Carbadox is extracted from the sample with a mixture of acetonitrile and methanol. Animal feeds are prewetted with water. The extract of animal feeds is purified through a short aluminium oxide column. The extract of premixtur
24、es is directly diluted with a mixture of water, acetonitrile and methanol. The final extract is analysed by reverse-phase HPLC with UV detection at a wavelength of 365 nm (see references 1 to 3). The presence of dimetridazole, nitrofurazone or sulfadimidine sodium can interfere with the determinatio
25、n of carbadox. Alternatively, carbadox may be determined after post-column derivatization with sodium hydroxide with detection at a wavelength of 420 nm. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not
26、 for Resale, 04/19/2007 03:40:03 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 14939:2001(E) 2 ISO 2001 All rights reserved 4Reagents Use only reagents of recognized analytical grade. 4.1Water, demineralized or deionized, with resistivity of at least 10 M?cm, or water
27、 of at least equivalent purity. 4.2Extraction solvent: mixture of acetonitrile and methanol (1:1 by volume). Combine equal volumes of acetonitrile and methanol. Mix well and allow to adjust to room temperature before use. 4.3Dilution solvent: mixture of extraction solvent (4.2) and water (4.1) (70:3
28、0 by volume). Mix 70 ml of extraction solvent (4.2) with 30 ml of water (4.1). 4.4Acetic acid, volume fraction, w(CH3CO2H) = 10 %. Dilute 10 ml of glacial acetic acid to 100 ml with water. 4.5Sodium acetate solution, c(C2H3NaO2) = 0,01 mol/l, pH = 6,0. Weigh 0,82 g of water-free sodium acetate into
29、a 1 000 ml one-mark volumetric flask. Dissolve in 700 ml of water. Adjust the pH to pH = 6,0 with acetic acid (4.4). Dilute to the mark with water and mix. 4.6Mobile phase for HPLC. Combine 825 ml of sodium acetate solution (4.5) and 175 ml of acetonitrile and mix. Filter the eluent through a 0,22 m
30、 filter using a solvent filtration system (5.2), and degas for 10 min in an ultrasonic bath (5.3) before use. 4.7Carbadox standard material, 3-(2-quinoxalinyl methylene) carbazic acid methy ester N,?N-dioxide (CAS number 6804-07-5). WARNING Because of the sensitivity of carbadox to light, conduct al
31、l operations in the absence of daylight or artificial white light. Avoid inhalation of and exposure to the toxic carbadox standard material and solutions thereof. Work in a fume cupboard when handling the solvents and solutions. Wear safety glasses and protective clothing. 4.8Carbadox stock solution
32、 (approximately 100 g/ml). Weigh 10 mg ? 1 mg of carbadox (4.7), to the nearest 0,1 mg, into a 100 ml one-mark volumetric flask. Dissolve in extraction solvent (4. 2), dilute to the mark and mix. Calculate the concentration taking into account the purity of the standard material. Prepare fresh every
33、 month. Store in the dark at 0 C to 8 C. 4.9Carbadox working solutions (approximately 2 g/ml and 10 g/ml). Pipette 1,0 ml and 5,0 ml of the carbadox stock solution (4.8) into separate 50 ml one-mark volumetric flasks. Dilute to the mark with dilution solvent (4.3) and mix. Prepare fresh for each ser
34、ies of samples. 4.10Carbadox working solutions (approximately 0,4 g/ml and 2 g/ml). Pipette 1,0 ml of the carbadox stock solution (4.8) into a 50 ml one-mark volumetric flask, dilute to the mark with mobile phase (4.6) and mix. Pipette 10 ml of this solution (2 g/ml) into a 50 ml one-mark volumetric
35、 flask, dilute to the mark with mobile phase (4.6) and mix. Prepare fresh for each series of samples. 4.11Dimetridazole standard material, 1,2-dimethyl-5-nitro-1H-imidazole (CAS number 551-92-8). WARNING Because of the sensitivity of dimetridazole to light, conduct all operations in the absence of d
36、aylight or artificial white light. Avoid inhalation of and exposure to the toxic dimetridazole standard Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 03:40:03 MDTNo reproductio
37、n or networking permitted without license from IHS -,-,- ISO 14939:2001(E) ISO 2001 All rights reserved3 material and solutions thereof. Work in a fume cupboard when handling the solvents and solutions. Wear safety glasses and protective clothing. 4.12Dimetridazole stock solution (approximately 100
38、g/ml). Weigh 10 mg ? 1 mg of dimetridazole (4.11), to the nearest 0,1 mg, into a 100 ml one-mark volumetric flask. Dilute to the mark with methanoland mix. Calculate the concentration taking into account the purity of the standard material. Prepare fresh every month. Store in the dark at 0 C to 8 C.
39、 4.13Dimetridazole working solution (approximately 20 g/ml). Pipette 2,0 ml of the dimetridazole stock solution (4.12) into a 10 ml one-mark volumetric flask. Dilute to the mark with water and mix. Prepare fresh for each series of samples. 4.14Sulfadimidinestandardmaterial,sodiumsaltof4-amino-N-(4,6
40、-dimethyl-2-pyrimidinyl)benzene sulfonamide (CAS number 1981-58-4). WARNING Avoid inhalation of and exposure to the toxic sulfadimidine standard material and solutions thereof. Work in a fume cupboard when handling the solvents and solutions. Wear safety glasses and protective clothing. 4.15Sulfadim
41、idine stock solution (approximately 200 g/ml). Weigh 10 mg ? 1 mg of sulfadimidine standard material (4.14), to the nearest 0,1 mg, into a 50 ml one-mark volumetric flask. Dilute to the mark with methanol and mix. Calculate the concentration taking into account the purity of the standard material. P
42、repare fresh every month. Store in the dark at 0 C to 8 C. 4.16Sulfadimidine working solution (approximately 20 g/ml). Pipette 1,0 ml of sulfadimidine stock solution (4.15) into a 10 ml one-mark volumetric flask. Dilute to the mark with water and mix. Prepare fresh for each series of samples. 4.17Ni
43、trofurazone standard material, 5-nitro-2-furaldehyde semicarbazone (CAS number 59-87-0). WARNING Because of the sensitivity of nitrofurazone to light, conduct all operations in the absence of daylight or artificial white light. Avoid inhalation of and exposure to the toxic nitrofurazone standard mat
44、erial and solutions thereof. Work in a fume cupboard when handling the solvents and solutions. Wear safety glasses and protective clothing. 4.18Nitrofurazone stock solution (approximately 100 g/ml). Weigh 10 mg ? 1 mg of nitrofurazone (4.17), to the nearest 0,1 mg, into a 100 ml one-mark volumetric
45、flask. Dilute to the mark with methanol and mix. Calculate the concentration taking into account the purity of the standard material. Prepare fresh every month. Store in the dark at 0 C to 8 C. 4.19Nitrofurazone working solution (approximately 20 g/ml). Pipette 2,0 ml of nitrofurazone stock solution
46、 (4.18) into a 10 ml one-mark volumetric flask. Dilute to the mark with water and mix. Prepare fresh for each series of samples. 4.20Neutral aluminium oxide, activity 1. For total de-activation 0 % to 1 % of water is necessary. 4.21Sodium hydroxide solution, c(NaOH) = 0,5 mol/l. Weigh 20 g of sodium
47、 hydroxide into a 1 litre one-mark volumetric flask and dissolve in 10 ml of water. Dilute to the mark with water and mix. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 03:40:0
48、3 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 14939:2001(E) 4 ISO 2001 All rights reserved 5Apparatus Usual laboratory apparatus and, in particular, the following. 5.1pH-meter. 5.2Solvent filtration system, all glass apparatus suitable for 0,22 m filters. 5.3Ultrasonic bath. 5.4Rotary shaker, horizontal rotation, rotation frequency 250 min ?1 to 300 min ?1 . 5.5Glass microfibre filter, diameter 15 cm. 5.6Glass wool. 5.7Glass column for chromatography, length 30 cm, inter