Method for detecting patulin in apple juice

A patulin and apple juice technology, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of poor reproducibility of capillary electrophoresis, poor selectivity of high performance liquid chromatography, and penicillin is not suitable for electric field separation, etc. Accuracy and sensitivity, simple and convenient operation, short time-consuming effect

Active Publication Date: 2020-06-16
NAT INST OF METROLOGY CHINA
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Problems solved by technology

Thin-layer chromatography is an early detection method for patulin, but its disadvantages are low sensitivity, low recovery, and serious matrix interference
The capillary electrophoresis method itself has poor reproducibility, and patulin is electrically neutral and is not suitable for electric field separation, so there are few practical applications
In gas chromatography-mass spectrometry, patulin must be derivatized to increase volatility, but the active protons of phenolic compounds remaining in apple juice will interfere with the reaction and reduce the derivatization efficiency
At the same time, the derivatization step also complicates the sample preparation process, and insufficient reaction will lead to additional method errors
Poor selectivity of HPLC, retention of similarly behaving substances in reversed-phase chromatography such as 5-HMF can cause strong interference

Method used

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  • Method for detecting patulin in apple juice
  • Method for detecting patulin in apple juice
  • Method for detecting patulin in apple juice

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Embodiment 1

[0049] This embodiment provides a method for detecting patulin in apple juice. The method includes the following steps:

[0050] The patulin isotope internal standard was added to the apple juice to be tested, vortexed and oscillated for 5-10 minutes, and then centrifuged at 6000 r / min for 5-10 minutes, and the supernatant after centrifugation was absorbed to obtain clarified apple juice containing the internal standard.

[0051] First, use methanol and water to activate the HyperSep Retain AX solid-phase extraction column, then add the clarified apple juice containing internal standard into the activated HyperSep Retain AX solid-phase extraction column, and then use 0.1% volume fraction of acetic acid aqueous solution and water to rinse washing, and then eluted with methanol to obtain an eluate, which was blown dry with nitrogen, and redissolved in a solution with a volume ratio of water to methanol of 95:5 to prepare the sample to be tested.

[0052] Take patulin standard s...

Embodiment 2

[0057] This embodiment provides a method for detecting patulin in apple juice. The method includes the following steps:

[0058] 1. Apple juice sample and standard sample preparation:

[0059] Take 2 mL of apple juice with a pipette, add 100 μL of isotope internal standard with a concentration of 1 μg / mL, vortex and mix for 5 min, and then centrifuge at 6000 r / min for 5-10 min to separate the solid components and obtain clarified apple juice containing internal standard.

[0060] Standard sample preparation: Accurately weigh 100 mg of patulin standard substance, and prepare a standard stock solution with a mass concentration of 1 mg / mL with acetonitrile. A series of standard calibration solutions with different concentrations were prepared from the standard stock solution. Add an isotopic internal standard with a concentration of 1 μg / mL to make the concentration of the isotopic internal standard 50 ng / mL.

[0061] 2. Pretreatment of apple juice samples:

[0062] First, th...

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Abstract

The invention belongs to the technical field of patulin detection, and relates to a method for detecting patulin in apple juice. The method comprises the following steps: uniformly mixing apple juiceto be detected with a patulin isotope internal standard, and carrying out solid-liquid separation to obtain clarified apple juice containing the internal standard; pretreating the clarified apple juice containing the internal standard by using a quaternary amine polymer solid-phase extraction column to obtain a sample to be detected; preparing patulin standard working solutions that have differentconcentrations and contain the patulin isotope internal standards; carrying out UPLC-ID-MS / MS detection on the sample to be detected and the patulin standard working solutions with different concentrations to obtain mass spectrogram peak areas corresponding to the sample to be detected and the patulin standard working solutions with different concentrations; making a standard curve; and calculating the content of patulin in the sample to be detected according to the standard curve and the peak area of the mass spectrogram corresponding to the sample to be detected so as to obtain the contentof patulin in the apple juice to be detected. The method enables the detection accuracy and sensitivity of patulin in apple juice to be improved.

Description

technical field [0001] The invention belongs to the technical field of patulin detection, and more specifically relates to a method for detecting patulin in apple juice. Background technique [0002] Patulin is a poisonous fungal metabolite. Due to its strong electrophilic properties, it can interact with nucleophilic sulfhydryl and amino compounds in proteins and other substances, causing damage to the human central nervous system, digestive system and urinary system. Different degrees of toxic effects, with potential teratogenic, carcinogenic, mutagenic, fertility-affecting and immunosuppressive toxicity. The contamination of patulin in apple food, especially in apple juice, is a global problem and a serious threat to human health. The World Health Organization and the Food and Agriculture Organization of the United Nations set the maximum daily intake of patulin at 0.4 μg / kg body weight / day, and my country’s GB 2761-2017 national standard stipulates that the upper limit ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/86
CPCG01N30/02G01N30/06G01N30/8679G01N2030/045G01N2030/062
Inventor 李先江张庆合李红梅
Owner NAT INST OF METROLOGY CHINA
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