GB 31657.8-2025 National Food Safety Standard - Determination of residual levels of amitriptyline and its metabolites in bee products - Liquid chromatography tandem mass spectrometry methodis caused byMinistry of Agriculture and Rural Affairs, National Health Commission, and State Administration for Market Regulation of the People's Republic of ChinaImportant technical standards jointly issued by the three major ministries. This standard has been established inReleased on June 3, 2025And scheduledFormally implemented from September 1, 2025.

1、 Scope of application and detection objects

（1） Scope of application

Applicable tohoneyandroyal jellyThe standards may not be applicable to the testing of other bee products such as bee pollen and propolis for the two main types of bee products. In practical applications, the applicability of the methods needs to be confirmed based on specific circumstances.

（2） Detection object

DimethylformamideAnd its metabolites, its metabolitesDivided by the International Agency for Research on Cancer2B potential carcinogensThere are risks to human health.

2、 Operation process

(1)Method principles and technology selection

1. Standard adoptionLiquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) As a core technology, this method combines liquid chromatographyEfficient separation capabilityCombined with tandem mass spectrometryHigh sensitivity and high selectivity detection capabilityEspecially suitable for the analysis of trace residues in complex matrices;

The principle of the method is to monitor multiple reactions through characteristic ion pairs: amitriptyline (m/z 284.1 → 182.1) and its metabolites (m/z 122.1 → 92.1). This dual screening mechanism significantly improves the reliability of detection and can effectively avoid false positive results.

(II)Sample pre-treatment process

1. Sample preparation:Honey needs to be dissolved in a water bath at ≤ 60 ° C and stirred evenly; Royal jelly can be directly sampled, and all operations must be carried out in the dark. Honey should be stored at room temperature, and royal jelly should be stored at -18 ° C;

2. Extraction and purificationThe sample was subjected to ultrasonic extraction using acetonitrile water solution, and the extraction solution was purified using a C18 solid-phase extraction column to remove interfering substances such as sugars and proteins from honey and royal jelly;

3. Concentration and reconstitution:After purification, the eluent is dried with nitrogen gas, dissolved in an aqueous solution containing 0.1% formic acid, and finally filtered through a 0.22 μ m filter membrane before it can be analyzed on the machine.

(3)Instrument analysis conditions

1. Liquid chromatography conditions:Using a C18 chromatographic column, gradient elution was performed with 0.1% formic acid aqueous solution and methanol as mobile phases, column temperature controlled at 35 ° C, and injection volume of 10 μ L;

2. Mass spectrometry conditions:The electric spray ion source (ESI+) is used to operate in the positive ion mode. The ion source temperature is 290 ° C, and the spray voltage is 3500V. The detection sensitivity is improved through the multi reaction monitoring mode.

(IV)Quality Control and Verification

1. Standard requirements passedblank experimentandMatrix matching standard curveTo eliminate the interference of bee product matrix on quantitative results, the method islinear range0.1-50 μ g/L,correlation coefficientMust reach 0.999 or above;

2. Recovery rateControl between 80% -120%,relative standard deviationNot exceeding 10% ensures the accuracy and reliability of the method,methodDetection limit as low as 0.1 μ g/kgCan meet the strict residue limit requirements both domestically and internationally.

3、 Summary

GB 31657.8-2025, as a mandatory national standardStandardization and refinementThe testing process provides strong technical support for the quality and safety of bee products. As a manufacturer of standard materials, Xibiao supports GB 31657.8-2025. We welcome all major customers to come and purchase!