1、BRITISH STANDARD BS ISO 18054:2004 Oils of orris rhizome (Iris pallida Lam. or Iris germanica L.) Determination of irone content Method using gas chromatography on a capillary column ICS 71.100.60 BS ISO 18054:2004 This British Standard was published under the authority of the Standards Policy and S
2、trategy Committee on 1 June 2004 BSI 1 June 2004 ISBN 0 580 43837 6 National foreword This British Standard reproduces verbatim ISO 18054:2004 and implements it as the UK national standard. The UK participation in its preparation was entrusted to Technical Committee AW/54, Essential oils, which has
3、the responsibility to: A list of organizations represented on this committee can be obtained on request to its secretary. Cross-references The British Standards which implement international publications referred to in this document may be found in the BSI Catalogue under the section entitled “Inter
4、national Standards Correspondence Index”, or by using the “Search” facility of the BSI Electronic Catalogue or of British Standards Online. This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application. Compliance with a Br
5、itish Standard does not of itself confer immunity from legal obligations. aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international
6、and European developments and promulgate them in the UK. Summary of pages This document comprises a front cover, an inside front cover, the ISO title page, pages ii to iv, pages 1 to 8, an inside back cover and a back cover. The BSI copyright notice displayed in this document indicates when the docu
7、ment was last issued. Amendments issued since publication Amd. No. Date Comments Reference number ISO 18054:2004(E)INTERNATIONAL STANDARD ISO 18054 First edition 2004-05-15 Oils of orris rhizome (Iris pallida Lam. or Iris germanica L.) Determination of irone content Method using gas chromatography o
8、n a capillary column Huile essentielle de rhizomes diris (Iris pallida Lam. ou Iris germanica L.) Dtermination de la teneur en irones Mthode par chromatographie en phase gazeuse sur colonne capillaire BSISO18054:2004DPlcsid Fremia ihTs PDF file mya ctnoian emdebt dedyfepcaes. In ccacnadrow eitA hebo
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11、 neeb sah takne tsne oeru taht teh file is siutlbae fosu re yb ISO memdob rebeis. In tlnu ehikletneve y ttah lborp aem lertait gno it is f,dnuo plsaee ifnrom ttneC ehlar Secterirata ta teh serddaig sleb nevwo. ii BSISO18054:2004 iiiForeword ISO (the International Organization for Standardization) is
12、 a worldwide federation of national standards bodies (ISO member 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 represen
13、ted on that committee. International organizations, governmental 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 dr
14、afted in accordance with the rules given in the ISO/IEC Directives, 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 S
15、tandard 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 document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. ISO 18054 was prepare
16、d by Technical Committee ISO/TC 54, Essential oils. BSISO18054:2004iv Introduction Since the description of methods of analysis by gas chromatography is very long, it is considered useful to establish general methods giving detailed information on all the recurrent parameters, apparatus, products, m
17、ethods, formulae, etc. and then standards with short details on the determination of specific constituents in essential oils, giving only those operating conditions specific to the pertinent determination. This is the case with the present International Standard, which refers to the general standard
18、 ISO 7609 for the general paragraphs. BSISO18054:2004 1Oils of orris rhizome (Iris pallida Lam. or Iris germanica L.) Determination of irone content Method using gas chromatography on a capillary column 1 Scope This International Standard specifies a method for the determination of irone content in
19、oils of orris 1)rhizome (Iris pallida Lam. or Iris germanica L.) using gas chromatography on a capillary column. 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 referen
20、ces, the latest edition of the referenced document (including any amendments) applies. ISO 356, Essential oils Preparation of test samples ISO 7609, Essential oils Analysis by gas chromatography on capillary columns General method 3 Principle Gas phase chromatography on small quantities of orris ess
21、ential oils is carried out on a capillary column, under the specified analytical conditions. The irone content is determined by the internal standard method. 4 Reagents and products 4.1 Reference substance: synthetic irones of which the sum of the isomers is greater than 90 %, determined by chromato
22、graphy under the test conditions. 4.2 Internal standard Select either one of the following recently distilled substances, depending on whether a polar or apolar column is being used. The internal standard selected shall elute as near as possible to the compound(s) to be determined, and shall not ove
23、rlap any of the peaks corresponding to the constituents of the essential oil in question. 4.2.1 -Ionone, having a minimum purity of 98 %, determined by gas chromatography under the test conditions, for determination on a polar column. 4.2.2 trans-Anethole, having a minimum purity of 98 %, determined
24、 by gas chromatography under the test conditions, for determination on an apolar column. 1) Commercially, this is usually called “orris butter”. BSISO18054:20042 4.3 Dilution solvent: dichloromethane, having a minimum purity of 99 %, determined by gas chromatography under the test conditions, or any
25、 other suitable solvent, of identical analytical quality. WARNING Attention is drawn to the hazards of using dichloromethane, which is a toxic solvent. 4.4 Additive substance: linalool, having a minimum purity of 99 %, determined by chromatography under the test conditions. 5 Apparatus 5.1 Chromatog
26、raph, recorder and integrator. See ISO 7609. 5.2 Column. Capillary column having the characteristics given in 5.2.1 and 5.2.2. 5.2.1 Polar capillary column. Length 10 m to 100 m Internal diameter 0,2 mm to 0,5 mm Stationary phase poly(ethylene glycol) 20 000 (FFAP , Supelcowax ), for example 5.2.2 A
27、polar capillary column. Length 10 m to 100 m Internal diameter 0,2 mm to 0,5 mm Stationary phase methyl silicone, for example 5.3 Flame ionization detector. 6 Preparation of test sample See ISO 356. 7 Operating conditions 7.1 Temperatures 7.1.1 Oven temperature The oven temperature shall be chosen s
28、o that the irone isomers are well separated from the other essential oil constituents. The following temperatures are given by way of indication. a) Polar capillary column Linear temperature programming from 100 C to 220 C, at a rate of 2 C/min to 3 C/min. BSISO18054:2004 3b) Apolar capillary column
29、 Linear temperature programming from 100 C to 250 C, at a rate of 2 C/min to 3 C/min. 7.1.2 Injection temperature About 230 C. 7.1.3 Detection temperature a) Polar capillary column About 230 C. b) Apolar capillary column About 260 C. 7.2 Carrier gas and auxiliary gases flow rates See ISO 7609. 8 Col
30、umn performance 8.1 Chemical inertness test Carry out the test as specified in ISO 7609. 8.2 Column efficiency Determine the efficiency of the column as specified in ISO 7609. The efficiency shall be at least 50 000 theoretical plates, determined on the linalol peak, under the test conditions. The i
31、rone isomers shall be well separated. 9 Determination of retention indexes See ISO 7609. 10 Identification of the irone isomers See the typical chromatograms given in Annex A. 11 Determination by internal standard method 11.1 Determination of the response factor Determine the response factor in acco
32、rdance with ISO 7609, using the irone (4.1) as the reference substance and either a) or b) of the following standard mixtures, depending on whether the determination is conducted on a polar or an apolar capillary column. Prepare the standard mixtures by weighing, taking into account the analytical p
33、urity of the standards in the calculation of the masses of the weighed products. BSISO18054:20044 a) Standard mixture for polar capillary column -Ionone (4.2.1) 0,1 g Synthetic irones 0,1 g Linalool (4.4) 0,8 g Dichloromethane (4.3) 4 g approximately b) Standard mixture for apolar capillary column t
34、rans-Anethole (4.2.2) 0,1 g Synthetic irones 0,1 g Linalool (4.4) 0,8 g Dichloromethane (4.3) 4 g approximately NOTE 1 The purpose of adding linalool to the standard mixtures is to obtain irone concentrations close to those of the synthetic irones. NOTE 2 The weighing accuracy given in ISO 7609 is n
35、ot applicable to the dilution solvent dichloromethane (4.3). A relative weighing accuracy of 5 % suffices. NOTE 3 For the calculation of the response factor, K, A Ris the sum of the areas of the peaks of the reference irone isomers. It is assumed that the response factors of the isomers are identica
36、l. 11.2 Determination of the irones in the sample Carry out a determination of the irone content of the essential oil being analysed in accordance with the internal standard method specified in ISO 7609, using either a) or b) of the following test mixtures, depending on whether a polar or an apolar
37、capillary column is used. Prepare the mixtures by weighing. a) Test mixture for polar capillary column -Ionone (4.2.1) 0,1 g Synthetic irones 0,9 g Dichloromethane (4.3) 4 g approximately b) Test mixture for apolar capillary column trans-Anethole (4.2.2) 0,1 g Synthetic irones 0,9 g Dichloromethane
38、(4.3) 4 g approximately NOTE 1 The weighing accuracy given in ISO 7609 is not applicable to the dilution solvent dichloromethane (4.3). A relative weighing accuracy of 5 % suffices. NOTE 2 For the determination of the irone content of the synthetic irones, A xis the sum of the areas of the peaks of
39、the irone isomers. It is assumed that the response factors of the isomers are identical. BSISO18054:2004 512 Expression of results See ISO 7609. NOTE Typical chromatograms of the analysis are given in Annex A, for information. 13 Precision 13.1 Interlaboratory test Details of an interlaboratory test
40、 on the precision of the method are summarized in Annex B. 13.2 Repeatability The absolute difference between two independent single test results, obtained using the same method on identical test material in the same laboratory by the same operator using the same equipment within a short interval of
41、 time, should not be greater than 5 %. 13.3 Reproducibility The absolute difference between two single test results, obtained using the same method on identical test material in different laboratories with different operators using different equipment, should not be greater than 10 %. 14 Test report
42、 See ISO 7609. BSISO18054:20046 Annex A (informative) Typical chromatograms of the determination by gas chromatography of the irone content in the oil of orris rhizome (Iris pallida Lam. or Iris germanica L.), of Moroccan origin A.1 Identification of irone isomers and determination of the response f
43、actor on an apolar phase standard mixture 11.1 b) Peak identification Operating conditions 1 Dichloromethane 2 Linalool 3 trans-Anethole 4 trans- -Irone 5 cis- -Irone 6 cis- -Irone 7 -Irone Column: quartz capillary; length 50 m; internal diameter 0,32 mm Stationary phase: poly(dimethyl siloxane) (CP
44、Sil 5 CB ) Film thickness: 0,4 m Oven temperature: temperature programming from 100 C to 270 C at a rate of 2 C/min Injector temperature: 230 C Detector temperature: 250 C Detector: flame ionization type Carrier gas: helium Volume injected: 1 l Carrier gas flow rate: 1 ml/min Figure A.1 Typical chro
45、matogram taken on an apolar column BSISO18054:2004 7A.2 Identification of irones on an apolar phase test mixture 11.2 b) Peak identification Operating conditions 1 Dichloromethane 2 trans-Anethole 3 trans- -Irone 4 cis- -Irone 5 cis- -Irone 6 -Irone 7 Myristic acid Column: quartz capillary; length 5
46、0 m; internal diameter 0,32 mm Stationary phase: poly(dimethyl siloxane) (CPSil 5 CB ) Film thickness: 0,4 m Oven temperature: temperature programming from 100 C to 250 C at a rate of 2 C/min Injector temperature: 230 C Detector temperature: 250 C Detector: flame ionization type Carrier gas: helium
47、Volume injected: 1 l Carrier gas flow rate: 1 ml/min Figure A.2 Typical chromatogram taken on an apolar column BSISO18054:20048 Annex B (informative) Results of the interlaboratory tests Interlaboratory tests were carried out in 1995, with the collaboration of nine laboratories. Table B.1 Determinat
48、ion of the irone content in oils of orris rhizome Laboratories 1 3 4 5 6 7 8 9 7,89 8,42 8,15 7,75 8,89 7,42 7,82 8,17 7,92 8,26 8,12 7,77 8,81 7,47 7,98 7,94 7,67 8,54 8,12 8,07 8,82 7,58 7,92 7,90 7,75 8,50 8,36 8,09 8,81 7,63 7,69 8,83 7,63 8,15 7,55 8,07 8,08 8,10 Minimum per laboratory 7,67 8,2
49、6 8,12 7,75 8,07 7,42 7,82 7,55 Maximum per laboratory 7,92 8,54 8,36 8,09 8,89 7,63 7,98 8,17 Mean per laboratory 7,81 8,43 8,19 7,92 8,51 7,53 7,91 7,81 Deviation, % 1,76 2,02 2,11 2,15 5,13 1,40 1,10 4,56 Repeatability limit (r) 5,13 % Variance 0,013 9 0,015 3 0,013 4 0,034 3 0,149 9 0,009 4 0,006 5 0,053 8 Standard deviation 0,117 9 0,123 8 0,115 9 0,185 1 0,387 1 0,096 8 0,080 8 0,231 9 General mean
