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GB/T 19540-2025Determination of zearalenone in feeds (English PDF)

饲料中玉米赤霉烯酮的测定

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Issued by

SAMR; SAC

Level / Type

National · Recommended

Issue date

June 30, 2025

Implementation date

January 1, 2026

Scope

GB/T 19540-2025 is the English-translated version of 饲料中玉米赤霉烯酮的测定.

GB/T 19540-2025 is the Chinese national standard covering measuring the fusarium toxin zearalenone in feed — the liquid chromatography with tandem mass spectrometry that is the new reference method and the high performance liquid chromatography kept alongside it, the acetonitrile extraction and the immunoaffinity column clean-up, the matrix-matched calibration that corrects for the feed itself suppressing the signal, the external standard quantification, and the detection and quantification limits in micrograms per kilogram. It replaces GB/T 19540-2004, and the thin layer chromatography and ELISA methods of that edition are dropped. In force from 1 January 2026. Issued on 30 June 2025, it has been in force since 1 January 2026, replacing GB/T 19540-2004.

Document preview — GB/T 19540-2025

National Standard of the People's Republic of China

ICS: 65.120
Classification: B 46
Replacing: GB/T 19540-2004

Issued by: State Administration for Market Regulation; Standardization Administration of the PRC

Foreword
1 Scope
2 Normative references
3 Terms and Definitions
4 Liquid chromatography-tandem mass spectrometry
4.1 Principle
4.2 Reagents or materials

Foreword

This document is in accordance with the provisions of GB/T 1.1-2020 "Guidelines for standardization work Part 1: Structure and drafting rules for standardization documents" Drafting is required.

This document replaces GB/T 19540-2004 “Determination of zearalenone in feeds” and GB/T 28716-2012 “Determination of zearalenone in feeds”.

Determination of mithralenone by immunoaffinity column cleanup - high performance liquid chromatography.

Compared with 2012, in addition to structural adjustments and editorial changes, the main technical changes are as follows.

a) The scope of application has been changed (see Chapter 1 of GB/T 19540-2004 and Chapter 1 of GB/T 28716-2012);

b) Thin layer chromatography and enzyme-linked immunosorbent assay (ELISA) have been deleted (see Chapter 3 and Chapter 4 of GB/T 19540-2004);

c) Liquid chromatography-tandem mass spectrometry has been added (see Chapter 4).

Please note that some of the contents of this document may involve patents. The issuing organization of this document does not assume the responsibility for identifying patents.

This document is proposed and coordinated by the National Feed Industry Standardization Technical Committee (SAC/TC 76).

This document was drafted by: Shanghai Academy of Agricultural Sciences and Shanghai Animal Disease Prevention and Control Center.

The main drafters of this document are: Han Zheng, Cao Ying, Zhao Zhihui, Nie Dongxia, Guo Dakai, Wang Xinyi, Wu Jianping, Yang Haifeng, Guo Wenbo, Zhang Jing, Yan Feng, Huang Qingwen, Fan Kai, Shang Jun, Wu Yushan.

The previous versions of this document and the documents it replaces are as follows.

--Published as GB/T 19540-2004 in 2004 and GB/T 28716-2012 in 2012; --This is the first revision.

Determination of Zearalenone in Feed

1 Scope

This document describes a liquid chromatography-tandem mass spectrometry and high performance liquid chromatography method for the determination of zearalenone in feed.

This document applies to the determination of zearalenone in compound feed, concentrate supplements, concentrate feed and plant feed ingredients.

The detection limit of this document is 2 µg/kg and the quantification limit is 10 µg/kg.

2 Normative references

GB/T 6682

GB/T 20195

3 Terms and Definitions

There are no terms or definitions that require definition in this document.

4 Liquid chromatography-tandem mass spectrometry

4.1 Principle

The analytes in the sample were extracted with acetonitrile solution, the extract was diluted with phosphate buffer solution, purified by immunoaffinity column, and analyzed by liquid chromatography-mass spectrometry.

The samples were determined by tandem mass spectrometry, calibrated by matrix-matched standard curve, and quantified by external standard method.

4.2 Reagents or materials

Warning - Zearalenone is highly toxic. Avoid contact with skin and clothing during operation and wear medical latex gloves.

Handle the reagents carefully and use them in a fume hood.

Unless otherwise specified, only analytical grade reagents were used.

4.2.1 Water. GB/T 6682, Grade 1.

4.2.2 Methanol. chromatographic grade.

4.2.3 Acetonitrile. chromatographic grade.

4.2.4 Formic acid. chromatographically pure. 4.2.5 Sodium chloride.

4.2.6 Sodium hydroxide solution (0.2 mol/L). Weigh 8.0 g of sodium hydroxide, dissolve it in water, transfer it to a 1000 mL volumetric flask, make up to volume, and mix thoroughly.

4.2.7 80% acetonitrile solution. Measure 800 mL of acetonitrile (4.2.3) into a 1000 mL volumetric flask, dilute with water, make up to volume, and mix thoroughly.

4.2.8 0.1% formic acid solution. Pipette 1 mL of formic acid (4.2.4) into a 1000 mL volumetric flask, dilute with water, make up to volume, and mix thoroughly.

4.2.9 Phosphate buffer solution (PBS). weigh 8.0 g sodium chloride, 1.16 g disodium hydrogen phosphate, 0.2 g potassium dihydrogen phosphate, and 0.2 g potassium chloride.

Dissolve and dilute with water to 1000 mL, and adjust the pH to 7.4 with sodium hydroxide solution (4.2.6).

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This preview omits tables, figures, formulas and parts of the technical clauses. The complete document — 15 pages — is available in the English PDF.

Referenced standards

Normative references

Editions of GB/T 19540

Edition	Title	Revision	Status
GB/T 19540-2025	Determination of zearalenone in feeds	current edition	Current
GB/T 19540-2004	Determination of zearalenone in feeds	previous edition	In force

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