DIN 10234-2003 Analysis of spices and condiments - Determination of capsaicinoid content of powdered paprika paprika oleoresins powdered chillies and chillie oleoresins - Method us).pdf
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1、ICS 67.220.10Untersuchung von Gewrzen und wrzenden Zutaten Bestimmung des Capsaicinoidgehaltes inPaprikapulvern, Paprikaoleoresinen, Chilipulvern und Chilioleoresinen Verfahren mitHochleistungsflssigchromatographie (HPLC)In keeping with current practice in standards published by the International Or
2、ganization for Standardization(ISO), a comma has been used throughout as the decimal marker.Ref. No. DIN 10234 : 2003-02English price group 08 Sales No. 010810.03DEUTSCHE NORM February 200310234Continued on pages 2 to 5. No part of this translation may be reproduced without the prior permission ofDI
3、N Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).Translation by DIN-Sprachendienst.In case of doubt, the German-language original should be consulted as the authoritative text.Determining the cap
4、saicinoid content ofpowdered paprika and chilli peppers by highperformance liquid chromatography (HPLC)ForewordThis standard has been prepared by Technical Committee Gewrze und wrzende Zutaten of theNormenausschuss Lebensmittel und landwirtschaftliche Produkte (Foodstuffs and Agricultural ProductsSt
5、andards Committee).1 ScopeThis standard specifies a routine method of determining capsaicinoid contents of greater than 10 mg/kg inpowdered paprika and paprika oleoresins by high performance liquid chromatography. If the extract issuitably diluted, it can also be used for examining chilli powders an
6、d chilli oleoresins.2 Normative referencesThis standard incorporates, by dated or undated reference, provisions from other publications. Thesenormative references are cited at the appropriate places in the text, and the titles of the publications arelisted below. For dated references, subsequent ame
7、ndments to or revisions of any of these publicationsapply to this standard only when incorporated in it by amendment or revision. For undated references, thelatest edition of the publication referred to applies.DIN 10230 Preparation of ground samples of spices and condiments for analysisISO 5725-1 :
8、 1994 Accuracy (trueness and precision) of measurement methods and results Part 1: Gen-eral principles and definitionsISO 5725-2 : 1994 Accuracy (trueness and precision) of measurement methods and results Part 2: Basicmethods for the determination of repeatability and reproducibility of a standard m
9、eas-urement method3 ConceptCapsaicinoid contentThe sum of the capsaicin, dihydrocapsaicin and nordihydrocapsaicin contents, in mg/kg, determined by themethod described in this standard.Page 2DIN 10234 : 2003-024 PrincipleThe sample is extracted with an acetonitrile/water mixture under reflux and an
10、aliquot of the solution obtainedis analysed by high performance liquid chromatography using fluorescence detection.5 Reagents5.1 GeneralUnless otherwise specified, analytical grade reagents shall be used and the water used shall be distilled or ofequivalent purity.The following reagents shall be use
11、d.5.2 Nonivamid (pelargonic acid vanillylamide, PAVA), C17H27NO3, having a molar mass of 293,4 g/mol and apurity of greater than 97 % (m/m), determined by HPLC, used as standard substance.CAUTION. This substance is a strong irritant and must be handled with caution.5.3 Natural capsaicinoid mixture (
12、capsaicin, dihydrocapsaicin and nordihydrocapsaicin) as reference sub-stance to aid in peak assignment.5.4 Solvents5.4.1 Acetonitrile.5.4.2 Water.5.4.3 99 % to 100 % (m/m) acetic acid.5.5 Eluent, prepared by mixing 50 parts by volume of acetonitrile, 50 parts by volume of water and 0,5 partsby volum
13、e of acetic acid and degassing.5.6 Solvent mixture, for standard solutions and sample extraction, prepared by mixing 50 parts by volumeof acetonitrile and 50 parts by volume of water.5.7 Standard solutions5.7.1 Stock solution, prepared by dissolving (70 t 0,1) mg of PAVA in 100 ml of solvent mixture
14、.5.7.2 Standard solution, prepared by diluting 5 ml of stock solution to 250 ml using the solvent mixture.5.7.3 Dilution series, prepared from standard solution for calibration purposes. Table 1 shows an exampleof a suitable dilution series. The linear calibration range shall encompass at least five
15、 dilution steps.Table 1: Dilution series6 ApparatusIn addition to standard laboratory equipment, the following shall be used.6.1 One-mark bulb pipettes, one having a nominal capacity of 5 ml and the other being of larger capacityfor collecting the sample extract when extracting oleoresins as describ
16、ed in subclause 7.3.2.6.2 Volumetric flasks, of nominal capacities 20 ml, 50 ml, 100 ml, 250 ml and 500 ml.6.3 Boiling aid (boiling chips or beads).6.4 Cellulose fluted filter, having a diameter of 18,5 cm, a mass of about 75 g/m2and a thickness of about0,20 mm.Page 3DIN 10234 : 2003-026.5 Separatin
17、g system6.5.1 High performance liquid chromatograph.6.5.2 Fluorescence detector, having an excitation wavelength of 280 nm and an emission wavelength of320 nm.6.5.3 Solvent degassing system.6.6 Columns6.6.1 Analytical column, having an internal diameter of 4 mm to 4,6 mm and a length of 250 mm, pack
18、ed withoctadecylsilane-modified silica gel (RP-18, end-capped) of particle size 5 m, capable of separating the baselines of nordihydrocapsaicin and capsaicin.6.6.2 Precolumn, suitable for the analytical column, having an internal diameter of 4 mm to 4,6 mm and alength of 4 mm, packed with the same s
19、tationary phase as in subclause 6.6.1, suitable for a flow rate of1 ml/min.6.7 Recorder or integrator.6.8 Injection valve, with 20 l sample loop or any other system operating with the same accuracy.7 Procedure7.1 CalibrationInject 20 l of each of the PAVA solutions in the dilution series spaced out
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