1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS ISO 13110:2012Cigarettes Determination ofmenthol in smoke condensates Gas-chromatographicmethodBS ISO 13110:2012 BRITISH STANDARDNational forewordThis British Standard is the
2、UK implementation of ISO 13110:2012. The UK participation in its preparation was entrusted to TechnicalCommittee AW/40, Tobacco and tobacco products.A list of organizations represented on this committee can be obtained on request to its secretary.This publication does not purport to include all the
3、necessary provisions of a contract. Users are responsible for its correct application. The British Standards Institution 2012. Published by BSI Standards Limited 2012ISBN 978 0 580 75142 4 ICS 65.160 Compliance with a British Standard cannot confer immunity from legal obligations.This British Standa
4、rd was published under the authority of the Standards Policy and Strategy Committee on 31 October 2012.Amendments issued since publicationDate T e x t a f f e c t e dBS ISO 13110:2012 ISO 2012Cigarettes Determination of menthol in smoke condensates Gas-chromatographic methodCigarettes Dosage du ment
5、hol dans les condensats de fume Mthode par chromatographie en phase gazeuseINTERNATIONAL STANDARDISO13110First edition2012-10-15Reference numberISO 13110:2012(E)BS ISO 13110:2012ISO 13110:2012(E)ii ISO 2012 All rights reservedCOPYRIGHT PROTECTED DOCUMENT ISO 2012All rights reserved. Unless otherwise
6、 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 of the requester.ISO copyright offi
7、ceCase postale 56 CH-1211 Geneva 20Tel. + 41 22 749 01 11Fax + 41 22 749 09 47E-mail copyrightiso.orgWeb www.iso.orgPublished in SwitzerlandBS ISO 13110:2012ISO 13110:2012(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO m
8、ember bodies). The work of preparing International Standards 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, governm
9、ental and non-governmental, in liaison with ISO, also take 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 Dire
10、ctives, Part 2.The main task of technical committees is to prepare International Standards. 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 the member bodi
11、es casting a vote.Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights.ISO 13110 was prepared by Technical Committee ISO/TC 126, Tobacco and tobacco produc
12、ts. ISO 2012 All rights reserved iiiBS ISO 13110:2012ISO 13110:2012(E)IntroductionNo machine smoking regimen can represent all human smoking behaviour: it is recommended that cigarettes also be tested under conditions of a different intensity of machine smoking than those specified in this Internati
13、onal Standard; machine smoking testing is useful to characterize cigarette emissions for design and regulatory purposes, but communication of machine measurements to smokers can result in misunderstandings about differences in exposure and risk across brands; smoke emission data from machine measure
14、ments may be used as inputs for product hazard assessment, but they are not intended to be nor are they valid as measures of human exposure or risks. Communicating differences between products in machine measurements as differences in exposure or risk is a misuse of testing using ISO standards.iv IS
15、O 2012 All rights reservedBS ISO 13110:2012Cigarettes Determination of menthol in smoke condensates Gas-chromatographic method1 ScopeThis International Standard specifies a method for the gas-chromatographic determination of menthol in the total particulate matter (TPM) of mentholated cigarette smok
16、e condensates. The smoking of cigarettes and the collection of mainstream smoke are carried out in accordance with ISO 4387 with some exceptions (see Annex A).Encapsulated menthol in specific products may lead to specific handling not described in this International Standard.2 Normative referencesTh
17、e 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.ISO 4387, Cigarettes Determination of total and nico
18、tine-free dry particulate matter using a routine analytical smoking machineISO 8243, Cigarettes Sampling3 PrincipleThe total particulate matter from the mainstream smoke is dissolved in a solvent containing an internal standard. The menthol content of an aliquot of the solution is determined by gas
19、chromatography, and the menthol content of the total particulate matter of the mainstream smoke is calculated.4 ReagentsUse only reagents of recognized analytical grade.4.1 Carrier gas: helium, nitrogen or hydrogen of high purity.4.2 Auxiliary gases: air and hydrogen of high purity for the flame ion
20、ization detector.4.3 Propan-2-ol, methanol or ethanol may also be used.4.4 Internal standard: n-heptadecane (of purity at least 99 %). Other appropriate internal standards (such as, but not limited to, carvone, anethole) may be used after assessment of their stability, purity and confirmation that t
21、he internal standard does not co-elute with other components of the smoke extract from the gas chromatographic column.The peak area of internal standard in the smoke extract of samples and standards should be consistent. In cases where inconsistencies are found, analysis of an extraction of a smoke
22、sample without the internal standard in the extraction solution should be performed to confirm the absence of a peak in the smoke extract eluting at the same time as the internal standard.INTERNATIONAL STANDARD ISO 13110:2012(E) ISO 2012 All rights reserved 1BS ISO 13110:2012ISO 13110:2012(E)4.5 Ext
23、raction solvent: propan-2-ol (4.3) containing an appropriate mass concentration of internal standard (4.4); this is normally in the range of 0,2 mg/ml to 0,5 mg/ml. Other appropriate solvents (such as, but not limited to, ethanol, methanol) may be used after assessment of their extraction efficiency
24、. Solvent not stored in a temperature-controlled laboratory shall be allowed to equilibrate to room temperature before use.NOTE Room temperature should be indicative of temperatures around 22 C. If the room temperature is substantially different from 22 C then all solvents and internal standards nee
25、d to be tested to prove their viability for use in the method under the temperature conditions of the laboratory.4.6 Reference substance: menthol, of known purity ( 99 %).NOTE It is recommended to store menthol in an air tight container not exposed to a heat source and light. Storage at a temperatur
26、e lower than 4 C is recommended.4.7 Calibration solutionsDissolve the menthol (4.6) in the extraction solvent (4.5) to produce a series of at least four calibration solutions whose mass concentrations cover the range expected to be found in the test portion (usually 0,02 mg/ml to 1,0 mg/ml). Solvent
27、 and solutions stored at low temperatures shall be allowed to equilibrate to room temperature before use.For the stock solution, store in a refrigerator and replace after a maximum of three months, or a time proven to show equivalent stability.For the working standards, store in a refrigerator and r
28、eplace after a maximum of one month or a time proven to show equivalent stabilityNOTE Room temperature should be indicative of temperatures around 22 C. If the room temperature is substantially different from 22 C then all solvents, internal standards, and calibration standards need to be tested to
29、prove their viability for use in the method under the temperature conditions of the laboratory.5 ApparatusUsual laboratory apparatus and, in particular, the following items.5.1 Gas-chromatograph, equipped with a split/splitless multimode injector, a flame ionization detector, and a computerized cont
30、rolled data acquisition and processing system.NOTE A recorder and integrator are acceptable if proven to be operational for intended purpose.5.2 Column, DB-WAX1), 1 m film thickness, 0,53 mm internal diameter and preferably 30 m length or any other type of column showing equivalent separation capabi
31、lity. An example chromatogram is given in Figure C.1.6 Procedure6.1 SamplingSampling is done in accordance with ISO 8243.NOTE For special precautions to be taken in sampling, see Annex B.1) DB-WAX is an example of a suitable product available commercially. This information is given for the convenien
32、ce of users of this document and does not constitute an endorsement by ISO of this product.2 ISO 2012 All rights reservedBS ISO 13110:2012ISO 13110:2012(E)6.2 Test portionPrepare the test portion by dissolving the total particulate matter obtained by the machine smoking of a known number of cigarett
33、es in a fixed volume of the extraction solvent (4.5) of 20 ml for 44 mm filter discs (pads), or 50 ml for 92 mm filtered discs (pads), ensuring that the disc (pad) is fully covered and shake the sample on an orbital shaker or shaker shown to be equivalent for minimum of 20 min. The volume may be adj
34、usted to give a concentration of menthol appropriate for the calibration curve (see 6.4) provided that there is adequate volume for effective extraction of the smoke condensate. For standard smoking, refer to ISO 4387 and Annex A of this International Standard (i.e. ISO 13110).6.3 Setting up the app
35、aratusSet up the apparatus and operate the gas chromatograph (5.1) in accordance with the manufacturers instructions. Ensure that the peaks for solvent, internal standard, menthol and other smoke component peaks are well resolved. It is recommended to equip the gas chromatograph with an autosampler
36、for sample injection.Suitable operating conditions may be as follows for the column described in 5.2: carrier gas: helium at a flow rate of about 12 ml/min; make up gas: helium at a flow rate of about 5 ml/min; injection temperature: 200 C; split ratio (approximately): 10/1; injection volume: 1 l; o
37、ven temperature 1: 100 C; time period 1 (initial): 2 min; temperature program 1: 10 C/min; oven temperature 2: 150 C; time period 2 (intermediate): 0,2 min; temperature program 2: 20 C/min; oven temperature 3: 200 C; time period 3 (final): 3 min; detector temperature: 250 C.Using the above condition
38、s, the analysis time is about 6 min to 8 min per sample.NOTE The column conditions are dependent on the column specified in 5.2, or an appropriate alternative that has been tested.6.4 Calibration of the gas chromatographInject an aliquot (such as 1 l) of each of the calibration solutions (4.7) into
39、the gas chromatograph. Record the peak areas of the menthol and internal standard (4.4).Calculate the ratio of the menthol peak to the internal standard peak from the peak area data for each of the calibration solutions. Generate a calibration curve by calculating a linear regression equation of the
40、 peak area ratios as a function of the concentration of menthol. The intercept of the regression line should be close to zero; if the intercept is not close to zero an investigation should occur to explain the situation, and the calibration should be repeated if necessary. ISO 2012 All rights reserv
41、ed 3BS ISO 13110:2012ISO 13110:2012(E)Perform this full calibration procedure daily before use. In addition, inject an aliquot of an intermediate concentration standard, which should be prepared from a separate stock, after every 20 sample determinations. If the calculated concentration for this sol
42、ution differs by more than 3 % from the original value, repeat the full calibration procedure.6.5 DeterminationInject aliquots (1 l) of the test portion (6.2) into the gas chromatograph. Calculate the ratio of the menthol peak/internal standard peak from the peak area data and obtain the concentrati
43、on of menthol in the solution by input of this ratio in the calibration curve.7 Expression of resultsFrom the concentration of menthol in the test portion, determine the amount of menthol in the total particulate matter. Express the test results in milligrams per cigarette, mM, for each channel to t
44、he nearest 0,01 mg, and the average per cigarette to the nearest 0,01 mg.8 Repeatability and reproducibilityAn international collaborative study involving 17 laboratories and three samples, conducted in 2010, showed that when cigarettes are smoked in accordance with ISO 4387 and the resulting smoke
45、solutions are analysed by this method, the following values for the repeatability limit (r) and the reproducibility limit (R) are obtained.Data analysis gave the estimates as summarized in Table 1.Table 1 Estimates given by data analysisBrand Mean value Repeatability limit Reproducibility limitmMr R
46、mg per cigarette mg per cigarette mg per cigaretteA 0,173 0,014 0,060B 0,079 0,018 0,045C 0,741 0,047 0,1609 Test reportThe test report shall state the yield of menthol per cigarette smoked and the method used, and shall include all conditions which may affect the result (e.g. test conditions, smoki
47、ng regime). It shall also give all details necessary for the identification of the cigarette sample smoked.4 ISO 2012 All rights reservedBS ISO 13110:2012ISO 13110:2012(E)Annex A (normative) Smoking procedure for the determination of menthol in smoke condensatesThe cigarettes are smoked according to
48、 the procedure described in ISO 4387. However, in the determination of the menthol content, the following exceptions concerning the preparation and conditioning of the cigarette samples apply. After sample receipt, samples shall be stored in such a way as to avoid loss of menthol, and the possible c
49、ontamination of non-mentholated products. Conditioning of the cigarettes cannot be carried out as usual due to the sublimation of menthol, therefore cigarettes have to be conditioned in their sealed pack. During sample preparation for smoking, samples shall be prepared as quickly as possible on the day of analysis in order to minimize loss of menthol. After sample preparation, including butt marking, is complete, the cigarettes shall be kept in a closed container until they are smoked. The maximum stor
