EN 15054:2006

Non fatty foods - Determination of chlormequat and mepiquat - LC-MS method

This draft European Standard specifies a method using high performance liquid chromatography/mass spectrometry (LC-MS) for the determination of the growth regulators chlormequat and mepiquat in non fatty foods as chlormequat and mepiquat cation, respectively.
The method is applicable to all kinds of fruits, vegetables and cereal products. It has been collaboratively studied on mushrooms, pears, wheat flour and fruit puree, see [1].

Fettarme Lebensmittel - Bestimmung von Chlormequat und Mepiquat - LC-MS-Verfahren

Aliments non gras - Détermination du chlorméquat et du mépiquat - Méthode CL-SM

La présente Norme européenne spécifie une méthode utilisant la Chromatographie liquide à haute
performance/Spectrométrie de masse (CL-SM) pour la détermination des substances ou régulateurs de
croissance chlorméquat et mépiquat dans des aliments non gras respectivement sous la forme de cation
chlorméquat et de cation mépiquat.
La méthode s'applique à toutes les sortes de fruits, légumes et produits céréaliers. Elle a fait l'objet d'études
réalisées en collaboration sur des champignons, des poires, de la farine de blé et de la purée de fruit (voir [1]).

Nemastna živila – Določevanje klormekvata in mepikvata – Metoda LC-MS

General Information

• Status: Published
• Publication Date: 16-May-2006
• Withdrawal Date: 29-Nov-2006
• ICS: 67.060 - Cereals, pulses and derived products
• ICS: 67.080.01 - Fruits, vegetables and derived products in general
• Technical Committee: CEN/TC 275 - Food analysis - Horizontal methods
• Drafting Committee: CEN/TC 275/WG 3 - Pesticides and PCB's in foods of animal origin
• Current Stage: 9093 - Decision to confirm - Review Enquiry
• Start Date: 05-Jun-2023
• Completion Date: 14-Apr-2025
• Ref Project: SIST EN 15054:2006 - Non fatty foods - Determination of chlormequat and mepiquat - LC-MS method

Overview

EN 15054:2006 (CEN) specifies an HPLC–mass spectrometry (LC‑MS) method for the determination of the plant growth regulators chlormequat and mepiquat (reported as their respective cations) in non‑fatty foods. The method is designed for fruits, vegetables and cereal products and was collaboratively studied on mushrooms, pears, wheat flour and fruit puree. Keywords: EN 15054:2006, chlormequat, mepiquat, LC‑MS, non fatty foods, food analysis, pesticide residues.

Key topics and requirements

• Analytical principle: extraction with water/methanol, addition of deuterated internal standards, centrifugation, filtration and analysis by HPLC with positive electrospray ionisation (ESI) MS.
• Internal standards: use of isotopically labelled standards (d4‑chlormequat and d3‑mepiquat) to correct for matrix effects and improve quantification.
• Calibration and quantification: six calibration solutions covering a range that corresponds to 0.01–3.00 mg/kg for a 20 g test portion; calibration based on response ratios (analyte/internal standard).
• Sample handling: guidance on representative sampling, storage (0–5 °C up to 3 days or –20 °C for longer), and recommended test portion masses (typically 20 g; 10 g for dry matrices).
• Apparatus and reagents: common lab equipment plus LC‑MS with electrospray interface; specified solvents, ammonium acetate buffer and reference stock solutions.
• Validation and performance: inter‑laboratory precision data (repeatability and reproducibility) provided in Annex B for studied matrices; criteria for ion ratio confirmation (±30% tolerance) and procedures for confirmatory testing (e.g., alternate LC column).
• Reporting: mandatory elements for the test report include sample ID, method reference, results and units, sampling and testing dates, deviations and optional operations.

Applications and users

• Who uses it: official food control and public health laboratories, contract testing labs, agricultural residue monitoring programs, cereal and produce quality control teams, and research groups studying plant growth regulator residues.
• Practical use cases:
  • Routine monitoring of chlormequat and mepiquat residues in fruit, vegetable and cereal matrices.
  • Regulatory compliance testing against maximum residue limits (MRLs).
  • Method transfer, validation and inter‑laboratory studies for residue laboratories.
  • Confirmatory analysis when screening methods indicate potential positives.

Related standards

• EN 15055 - Non fatty foods: determination of chlormequat and mepiquat by LC‑MS/MS (alternative, higher‑selectivity method).
• ISO 5725 series - referenced for precision study design and interpretation.

This standard provides a validated, reproducible LC‑MS workflow for sensitive determination of chlormequat and mepiquat in non‑fatty food matrices and supports regulatory monitoring and laboratory quality assurance.

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Non fatty foods - Determination of chlormequat and mepiquat - LC-MS method&06Aliments non gras - Détermination du chlorméquat et du mépiquat - Méthode CL-SMFettarme Lebensmittel - Bestimmung von Chlormequat und Mepiquat - LC-MS-VerfahrenTa slovenski standard je istoveten z:EN 15054:2006SIST EN 15054:2006en67.050ICS:SLOVENSKI
STANDARDSIST EN 15054:200601-september-2006

EUROPEAN STANDARDNORME EUROPÉENNEEUROPÄISCHE NORMEN 15054May 2006ICS 67.060; 67.080.01 English VersionNon fatty foods - Determination of chlormequat and mepiquat -LC-MS methodAliments non gras - Détermination du chlorméquat et dumépiquat - Méthode CL-SMFettarme Lebensmittel - Bestimmung von Chlormequat undMepiquat - LC-MS-VerfahrenThis European Standard was approved by CEN on 20 April 2006.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the Central Secretariat or to any CEN member.This European Standard exists in three official versions (English, French, German). A version in any other language made by translationunder the responsibility of a CEN member into its own language and notified to the Central Secretariat has the same status as the officialversions.CEN members are the national standards bodies of Austria, Belgium, 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 STANDARDIZATIONCOMITÉ EUROPÉEN DE NORMALISATIONEUROPÄISCHES KOMITEE FÜR NORMUNGManagement Centre: rue de Stassart, 36
B-1050 Brussels© 2006 CENAll rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 15054:2006: E

Examples of experimental conditions.9 Annex B (informative)
Precision data.16 Bibliography.18

3.4 Glacial acetic acid, mass fraction w at least 96 g/100 g 3.5 Acetonitrile, HPLC quality 3.6 Methanol, HPLC quality 3.7 Water, suitable for HPLC 3.8 Chlormequat chloride stock solution, mass concentration (C5H13NCl2) = 4,30 µg/ml in methanol This stock solution contains 3,33 µg/ml chlormequat cation. 3.9 Mepiquat chloride stock solution, (C7H16NCl) = 4,37 µg/ml in methanol This stock solution contains 3,33 µg/ml mepiquat cation.
1) Celite 545 is a product supplied by Celite Corp. (World Minerals Inc., Santa Barbara, CA, USA). This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of the product named. Equivalent products may be used if they can be shown to lead to the same results.

Prepare six calibration solutions (no. 1 to no. 6) in 10 ml volumetric flasks as follows. Into the flasks, pipette 10 µl, 50 µl, 100 µl, 300 µl, 1 000 µl or 3 000 µl each of chlormequat chloride stock solution (3.8) and of mepiquat chloride stock solution (3.9). To all flasks add 60 µl of the internal standard solution 1 (3.10) and 60 µl of the internal standard solution 2 (3.11). Dilute the solutions no. 1 to no. 5 to the mark with a methanol/water mixture 1 + 1 (V/V) and solution no. 6 with water. All six calibration solutions contain 100 ng/ml of d4-chlormequat cation and 100 ng/ml of d3-mepiquat cation. Solutions no. 1 to no. 6 contain 3,3 ng/ml, 16,7 ng/ml, 33,3 ng/ml, 100 ng/ml, 333 ng/ml and 1 000 ng/ml each of chlormequat cation and of mepiquat cation respectively. On the basis of a 20 g test portion, this corresponds to 0,01 mg/kg, 0,05 mg/kg, 0,10 mg/kg, 0,30 mg/kg, 1,00 mg/kg or 3,00 mg/kg each of chlormequat cation and mepiquat cation. 4 Apparatus Usual laboratory apparatus and, in particular, the following: 4.1 Homogenizer or high speed blender, fitted with jar
4.2 Centrifuge, capable of producing a centrifugal acceleration of at least 3 000 g at the bases of the centrifuge tubes 4.3 Syringe, capacity of at least 2 ml 4.4 Syringe filter, pore size 0,45 µm (polyamide or polytetrafluoroethylene) 4.5 Glass vials, 1,8 ml volume; applicable for automatic injection into an autosampler 4.6 LC-MS system, equipped with electrospray interface 5 Procedure 5.1 Preparation of the samples Where possible, carry out the analysis of samples immediately on their arrival in the laboratory. Do not analyse a laboratory sample which is wholly or extensively spoiled. For analysis take only the portion of the laboratory sample to which the maximum residue level applies. No further plant-parts may be removed. A record of the plant-parts which have been removed shall be kept. The sample thus prepared is the test sample.
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