EN 15829:2010
(Main)Foodstuffs - Determination of ochratoxin A in currants, raisins, sultanas, mixed dried fruit and dried figs - HPLC method with immunoaffinity column cleanup and fluorescence detection
Foodstuffs - Determination of ochratoxin A in currants, raisins, sultanas, mixed dried fruit and dried figs - HPLC method with immunoaffinity column cleanup and fluorescence detection
This European Standard specifies a method for the determination of ochratoxin A in currants, raisins, sultanas, mixed dried fruit and dried figs by high performance liquid chromatography (HPLC) with immunoaffinity cleanup and fluorescence detection. This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples ranging from 1,1 µg/kg to 11 µg/kg.
For further information on the validation, see Clause 9 and Annex B.
WARNING - The use of this standard can involve hazardous materials, operations and equipment. This standard does not purport to address all the safety problems associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
Lebensmittel - Bestimmung von Ochratoxin A in Korinthen, Rosinen, Sultaninen, gemischtem Trockenobst und getrockneten Feigen - HPLC-Verfahren mit Reinigung an einer Immunoaffinitätssäule und Fluoreszenzdetektion
Diese Europäische Norm legt ein Verfahren zur Bestimmung von Ochratoxin A in Johannisbeeren, Rosinen, Sultaninen, gemischtem Trockenobst und getrockneten Feigen durch Hochleistungsflüssigchromatographie (HPLC) mit Reinigung an einer Immunoaffinitätssäule und Fluoreszenzdetektion fest. Dieses Verfahren wurde in einem Ringversuch durch die Untersuchung sowohl von natürlich kontaminierten als auch von aufgestockten Proben mit Gehalten von 1,1 µg/kg bis 11 µg/kg validiert.
Weitere Informationen zur Validierung siehe Abschnitt 9 und Anhang B.
WARNUNG — Bei der Anwendung dieser Norm ist es möglich, dass gefährliche Substanzen und Geräte angewendet bzw. gefährliche Arbeitsgänge durchgeführt werden. Diese Norm erhebt nicht den Anspruch, dass alle mit ihrer Anwendung verbundenen Sicherheitsprobleme angesprochen werden. Es liegt in der Verantwortung des Anwenders dieser Norm, geeignete Vorkehrungen für den Arbeits- und Gesundheitsschutz zu treffen und vor der Anwendung die Anwendbarkeit einschränkender Vor¬schriften zu bestimmen.
Produits alimentaires - Dosage de l'ochratoxine A dans les raisins de Corinthe, les raisins secs, les raisins secs de Smyrne, les mélanges de fruits secs et les figues sèches - Méthode CLHP avec purification sur colonne d'immuno-affinité et détection par fluorescence
La présente Norme Européenne spécifie une méthode pour déterminer la teneur en ochratoxine A dans les raisins de Corinthe, raisins secs, raisins secs de Smyrne, mélanges de fruits secs et figues sèches par chromatographie liquide à haute performance (CLHP) avec purification sur colonne d’immuno-affinité et détection par fluorescence. Cette méthode a été validée lors d’une étude interlaboratoires par l’analyse d’échantillons naturellement contaminés et d’échantillons dopés, de 1,1 µg/kg à 11 µg/kg.
Pour plus d’informations sur la validation, voir l’Article 9 et l’Annexe B.
AVERTISSEMENT - L’utilisation de la présente norme peut impliquer l’utilisation de produits et la mise en œuvre de modes opératoires et d’appareillages à caractère dangereux. La présente norme n’a pas pour but d’aborder tous les problèmes de sécurité liés à son utilisation. Il incombe à l’utilisateur de la présente norme d’établir, avant de l’utiliser, des pratiques d’hygiène et de sécurité appropriées et de déterminer l’applicabilité des restrictions réglementaires.
Živila - Določevanje ohratoksina A v korintah, rozinah, sultaninah, mešanem sušenem sadju in sušenih figah - Metoda HPLC s čiščenjem z imunoafinitetno kolono in fluorescenčno detekcijo
Ta evropski standard določametodo za določevanje ohratoksina A v korintah, rozinah, sultaninah, mešanem sušenem sadju in sušenih figah z metodo HPLC s čiščenjem z imunoafinitetno kolono in fluorescenčno detekcijo. Ta metoda je bila potrjena v medlaboratorijski študiji preko analize tako naravno kontaminiranih kot vzorcev z internimi dodatki v razponu od 1,1 μg/kg do 11 μg/kg. Za nadaljnje informacije o potrjevanju glej Klavzulo 9 in dodatek B.
General Information
Overview
EN 15829:2010 is a CEN (European) standard that defines a validated laboratory method for the determination of ochratoxin A in currants, raisins, sultanas, mixed dried fruit and dried figs. The procedure uses high performance liquid chromatography (HPLC) combined with immunoaffinity column (IAC) cleanup and fluorescence detection. The method was validated in an interlaboratory study on naturally contaminated and spiked samples over the range 1.1 µg/kg to 11 µg/kg (see Clause 9 and Annex B for validation data). A safety warning in the standard notes that ochratoxin A is hazardous and that appropriate health, safety and regulatory precautions are the user’s responsibility.
Key topics and technical requirements
- Principle: Extract sample with methanol + phosphoric acid, filter, dilute with phosphate buffered saline (PBS), capture ochratoxin A on an immunoaffinity column, elute, concentrate and quantify by reverse‑phase HPLC with fluorescence detection.
- Validated range: 1.1 µg/kg to 11 µg/kg (interlaboratory study).
- Immunoaffinity column: IAC must contain antibodies specific for ochratoxin A with capacity ≥ 100 ng and recovery ≥ 70 % for a 5 ng spike (specified solvent matrix).
- Sample preparation: Prepare a fruit slurry (five parts fruit : four parts water); typical analytical portion uses 45 g slurry (equivalent to 25 g dried fruit) extracted with 50 ml methanol + 5 ml phosphoric acid.
- HPLC and detection:
- Reverse‑phase column (e.g. C18) recommended to resolve OTA from other peaks.
- Fluorescence detection set at 333 nm excitation / 477 nm emission (or 390 nm / 440 nm when a post‑column pH shift is used).
- Typical injection volumes and flow rates are specified to ensure reproducible quantification.
- Reagents and consumables: PBS, acetonitrile, methanol, toluene, injection/mobile phase recipes, and certified ochratoxin A standards are required. EN ISO 3696:1995 (water quality) is normative.
Applications and users
- Who uses it: Food testing laboratories, public health and regulatory authorities, quality control teams in dried fruit processing and trading companies, import/export inspection labs, and research groups studying mycotoxin contamination.
- Practical purposes:
- Routine monitoring of ochratoxin A in dried fruit to support food safety compliance.
- Verification of supplier batches and import controls.
- Data generation for exposure/risk assessment and product recalls.
- Method transfer and accreditation (laboratories seeking ISO/IEC 17025 evidence).
Related standards and references
- Normative reference: EN ISO 3696:1995 (water for analytical laboratory use).
- Validation details: see Clause 9 and Annex B of EN 15829:2010.
- Safety and toxicology references: International Agency for Research on Cancer (IARC) classification noted in the standard.
This standard is essential for reliable, reproducible HPLC‑based ochratoxin A analysis in dried fruit matrices and supports consistent regulatory testing across CEN member states.
Frequently Asked Questions
EN 15829:2010 is a standard published by the European Committee for Standardization (CEN). Its full title is "Foodstuffs - Determination of ochratoxin A in currants, raisins, sultanas, mixed dried fruit and dried figs - HPLC method with immunoaffinity column cleanup and fluorescence detection". This standard covers: This European Standard specifies a method for the determination of ochratoxin A in currants, raisins, sultanas, mixed dried fruit and dried figs by high performance liquid chromatography (HPLC) with immunoaffinity cleanup and fluorescence detection. This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples ranging from 1,1 µg/kg to 11 µg/kg. For further information on the validation, see Clause 9 and Annex B. WARNING - The use of this standard can involve hazardous materials, operations and equipment. This standard does not purport to address all the safety problems associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
This European Standard specifies a method for the determination of ochratoxin A in currants, raisins, sultanas, mixed dried fruit and dried figs by high performance liquid chromatography (HPLC) with immunoaffinity cleanup and fluorescence detection. This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples ranging from 1,1 µg/kg to 11 µg/kg. For further information on the validation, see Clause 9 and Annex B. WARNING - The use of this standard can involve hazardous materials, operations and equipment. This standard does not purport to address all the safety problems associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
EN 15829:2010 is classified under the following ICS (International Classification for Standards) categories: 67.050 - General methods of tests and analysis for food products; 67.080.10 - Fruits and derived products. The ICS classification helps identify the subject area and facilitates finding related standards.
EN 15829:2010 is associated with the following European legislation: Standardization Mandates: M/383. When a standard is cited in the Official Journal of the European Union, products manufactured in conformity with it benefit from a presumption of conformity with the essential requirements of the corresponding EU directive or regulation.
You can purchase EN 15829:2010 directly from iTeh Standards. The document is available in PDF format and is delivered instantly after payment. Add the standard to your cart and complete the secure checkout process. iTeh Standards is an authorized distributor of CEN standards.
Standards Content (Sample)
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.ILQLWHWQRLebensmittel - Bestimmung von Ochratoxin A in Johannisbeeren, Rosinen, Sultaninen, gemischten Trockenfrüchten und getrockneten Feigen - HPLC-Verfahren mit Reinigung an einer Immunoaffinitätssäule und FluoreszenzdetektionProduits alimentaires - Dosage de l'ochratoxine A dans les raisins de Corinthe, les raisins secs, les raisins secs de Smyrne, les mélanges de fruits secs et les figues sèches - Méthode CLHP avec purification sur colonne d'immuno-affinité et détection par fluorescenceFoodstuffs - Determination of ochratoxin A in currants, raisins, sultanas, mixed dried fruit and dried figs - HPLC method with immunoaffinity column cleanup and fluorescence detection67.080.10Sadje in sadni proizvodiFruits and derived products67.050Splošne preskusne in analizne metode za živilske proizvodeGeneral methods of tests and analysis for food productsICS:Ta slovenski standard je istoveten z:EN 15829:2010SIST EN 15829:2010en,fr,de01-april-2010SIST EN 15829:2010SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 15829
January 2010 ICS 67.050; 67.080.10 English Version
Foodstuffs - Determination of ochratoxin A in currants, raisins, sultanas, mixed dried fruit and dried figs - HPLC method with immunoaffinity column cleanup and fluorescence detection
Produits alimentaires - Dosage de l'ochratoxine A dans les raisins de Corinthe, les raisins secs, les raisins secs de Smyrne, les mélanges de fruits secs et les figues sèches - Méthode CLHP avec purification sur colonne d'immuno-affinité et détection par fluorescence
Lebensmittel -Bestimmung von Ochratoxin A in Korinthen, Rosinen, Sultaninen, gemischtem Trockenobst und getrockneten Feigen - HPLC-Verfahren mit Reinigung an einer Immunoaffinitätssäule und Fluoreszenzdetektion This European Standard was approved by CEN on 18 December 2009.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the CEN Management Centre or to any CEN member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by translation under the responsibility of a CEN member into its own language and notified to the CEN Management Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre:
Avenue Marnix 17,
B-1000 Brussels © 2010 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 15829:2010: ESIST EN 15829:2010
Typical chromatogram . 13Annex B (informative)
Precision data . 14Bibliography . 15
Horizontal methods”, the secretariat of which is held by DIN. This European Standard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by August 2010, and conflicting national standards shall be withdrawn at the latest by August 2010. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights. This document has been prepared under a mandate give to CEN by the European Commission and the European Free Trade Association. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. SIST EN 15829:2010
Specification and test methods (ISO 3696:1987) 3 Principle A test portion is extracted with a mixture of methanol and phosphoric acid. The extract is filtered, diluted with phosphate buffered saline, and applied to an immunoaffinity column containing antibodies specific for ochratoxin A. The ochratoxin A is isolated, purified and concentrated on the column then released with elution solvent. Ochratoxin A is quantified by reverse-phase high performance liquid chromatography (HPLC) with fluorescence detection. 4 Reagents 4.1 General Use only reagents of recognized analytical grade and water complying with grade 1 of EN ISO 3696:1995, unless otherwise specified. Solvents shall be of quality for HPLC analysis. Commercially available solutions with equivalent properties to those listed may be used. WARNING — Dispose of waste solvents according to applicable environmental rules and regulations. Decontamination procedures for laboratory wastes have been reported by the International Agency for Research on Cancer (IARC), see [1]. 4.2 Helium purified compressed gas 4.3 Disodium hydrogen phosphate, anhydrous or Na2HPO4·12 H2O 4.4 Potassium chloride 4.5 Potassium dihydrogen phosphate 4.6 Sodium chloride SIST EN 15829:2010
Ochratoxin A is a potent nephrotoxin with immunotoxic, teratogenic and potential genotoxic properties. The International Agency for Research on Cancer (IARC) has classified ochratoxin A as a possible human carcinogen (group 2B). Protective clothing, gloves and safety glasses should be worn at all times, and all standard and sample preparation stages should be carried out in a fume cupboard. Dissolve 1 mg of the ochratoxin A or the contents of 1 ampoule (if ochratoxin A has been obtained as a film) in solvent mixture (4.20) to give a solution containing approximately 20 µg/ml to 30 µg/ml of ochratoxin A. To determine the exact concentration, record the absorption curve between a wavelength of 300 nm and 370 nm in a 1 cm quartz cell with solvent mixture (4.20) as reference using the spectrometer (5.12). Identify the wavelength for maximum absorption. Calculate the mass concentration of ochratoxin A, ota, in micrograms per millilitre using Equation (1):
bMA×××=ερ100maxota (1) where Amax is the absorption determined at the maximum of the absorption curve (here: at 333 nm); M is the molar mass, in grams per mole, of ochratoxin A (M = 403,8 g/mol); 0 is the molar absorption coefficient, in square metres per mole, of ochratoxin A in the solvent mixture (4.20) (here: 544 m2/mol, see [2]); b
is the optical path length, in centimetres, of the quartz cell.
Store this solution in a freezer at approximately - 18 °C. Allow to reach room temperature before opening. A solution stored in this way is usually stable for 12 months. Confirm the concentration of the solution if it is older than six months. 4.25 Ochratoxin A spiking solution Transfer an aliquot of the stock solution (4.24) containing 12,5 µg of ochratoxin A to a 5 ml volumetric flask. Evaporate to dryness under nitrogen at no more than 50 °C. Redissolve immediately in methanol (4.16) and make up to volume. This solution contains 2,5 µg/ml ochratoxin A.
Store this solution in a freezer at approximately - 18 °C. Allow to reach room temperature before opening. A solution stored in this way is usually stable for 12 months. Confirm the concentration of the solution if it is older than six months. 5
Apparatus 5.1 General Usual laboratory glassware and equipment and, in particular the following. 5.2 Silanised glass vials (optional) Prepare the vials by filling them with the silanising reagent (4.22) and leave this reagent in the vial for 1 min. Rinse the vial first with a solvent of low polarity, for example toluene (4.17) then with methanol (4.16) and dry before use. WARNING — The use of silanised glassware may prevent ochratoxin A bin
...
EN 15829:2010 기준은 건포도, 건자두, 설탕에 절인 과일, 혼합 건조 과일 및 건무화과에서 아프라톡신 A의 검출 방법을 규정하고 있습니다. 이 유럽 기준은 고성능 액체 크로마토그래피(HPLC)를 이용한 면역Affinity 컬럼 정제 및 형광 검출 방식을 채택하고 있어 높은 정밀도와 신뢰성을 제공합니다. 이 기준의 중요한 장점 중 하나는 HPLC 방법이 인터랩 연구를 통해 검증되었으며, 자연 오염 및 인위적으로 오염된 샘플을 분석하여 1.1 µg/kg에서 11 µg/kg 범위의 아프라톡신 A를 정확하게 측정할 수 있다는 점입니다. 이는 다양한 식품에서의 오염 물질 검출에 있어 신뢰할 수 있는 절차를 제공하여, 소비자의 안전을 확보하는 데 기여합니다. 또한, 이 기준은 면역Affinity 기술을 사용함으로써 복잡한 샘플 매트릭스 속에서 아프라톡신 A를 분리하고 정량화하는 데 필요한 체계적이고 단순한 절차를 제시합니다. 이로 인해 분석법의 적용 가능성이 확대되어, 다양한 건조 과일의 품질 관리 및 안전성 검증에 유용하게 활용될 수 있습니다. 그러나 사용자는 이 기준을 사용할 때 관련된 위험 물질 및 장비의 안전 문제를 고려해야 하며, 적절한 안전 및 건강 관행을 수립하는 책임이 있습니다. 이 경고는 연구개발 및 산업 현장에서의 안정성을 보장하는 데 필수적입니다. 결론적으로, EN 15829:2010은 아프라톡신 A의 정량화에 필수적인 방법론을 제공하며, 다양한 식품의 안전성을 높이는 데 있어 매우 중요한 표준으로 자리 잡고 있습니다. 이 기준은 식품 산업뿐만 아니라 공공 보건 분야에서도 그 relevance와 필요성을 갖추고 있어, 향후에도 지속적으로 참조될 것으로 기대됩니다.
SIST EN 15829:2010の標準文書は、食品に含まれるオクラトキシンAの測定方法を明確に定義しており、特にカレンツ、レーズン、サルタナ、ミックスドライフルーツ、干しイチジクに焦点を当てています。この標準は、免疫親和性カラムのクリーンアップと蛍光検出を用いた高性能液体クロマトグラフィー（HPLC）法に基づいており、正確かつ信頼性の高い結果を提供します。高感度な分析手法として、特に食品安全において重要な役割を果たすオクラトキシンAを測定するために重要です。 この標準の強みは、その検証プロセスにあります。自然汚染およびスパイクサンプルの分析に基づいた相互試験によって、この方法は1.1 µg/kgから11 µg/kgの範囲での測定に成功しています。このような厳密な検証により、業界全体での信頼性と一貫性が高まります。また、ユーザーが安全性を確保し、関連する規制の制限を適用する責任があることを明示しているため、適切な使用を促す重要な情報を提供します。 この標準の適用性は、食品業界全体において果たす役割によって明らかであり、オクラトキシンAの管理が求められる様々なシナリオにおいて、確実な測定を可能にします。この標準は、オクラトキシン A に関する最新の技術を取り入れているため、業界のニーズに応じた重要なリソースとして位置づけられます。
The EN 15829:2010 standard provides a robust framework for the determination of ochratoxin A in various dried fruit products, specifically currants, raisins, sultanas, mixed dried fruit, and dried figs. It employs a high-performance liquid chromatography (HPLC) method enhanced by immunoaffinity column cleanup and fluorescence detection. The scope of this standard is particularly relevant for food safety authorities and producers looking to ensure the quality and safety of dried fruit products in a competitive market. One of the key strengths of EN 15829:2010 is its validation process, confirmed through an interlaboratory study involving a range of naturally contaminated and artificially spiked samples. The validated concentration range of ochratoxin A, from 1.1 µg/kg to 11 µg/kg, establishes the standard’s credibility and applicability in real-world scenarios. This comprehensive method permits accurate detection, which is critical for the protection of consumer health and compliance with food safety regulations. Moreover, the inclusion of specific methodologies for hazardous material handling highlights the standard's commitment to safety. While the standard does not cover all safety issues related to its use, it emphasizes the importance of user responsibility in establishing safety practices and adhering to regulatory limits. This aspect makes EN 15829:2010 not only a technical document but also a vital resource for risk management in food safety. Overall, EN 15829:2010 is a pertinent and authoritative standard in the field of food safety, providing an essential testing method that aligns with current food safety practices and regulatory requirements for monitoring ochratoxin A in dried fruits.
Questions, Comments and Discussion
Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.
Loading comments...