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Standard substance containing free-state and conjugated-state clenbuterol in swine urine and preparation method of standard substance

A standard substance, free state technology, used in material separation, measurement devices, instruments, etc., can solve the problems of inaccuracy, affect the accuracy, reliability and comparability of test results, and exceed standards, and achieve good application prospects and good social benefits. And economic benefits, good stability effect

Active Publication Date: 2021-06-04
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if the samples obtained by artificial exogenous addition of "temporary preparation" are not subjected to biological enzyme pretreatment, the conventional kits or test strips are used to detect the results on site, which is often inaccurate, resulting in false negative results
Therefore, the so-called positive samples of pig urine simulated to contain clenbuterol obtained through "temporary preparation" will seriously affect the detection results in the food safety monitoring work, especially in the field environment where the experimental conditions cannot meet strict conditions. Accuracy, reliability and comparability
Urine samples that are considered not to exceed the standard by the on-site test results are tested by liquid chromatography-tandem mass spectrometry after pretreatment by enzymatic hydrolysis in the laboratory, and the result may exceed the standard

Method used

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  • Standard substance containing free-state and conjugated-state clenbuterol in swine urine and preparation method of standard substance
  • Standard substance containing free-state and conjugated-state clenbuterol in swine urine and preparation method of standard substance
  • Standard substance containing free-state and conjugated-state clenbuterol in swine urine and preparation method of standard substance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Standard substance candidate preparation

[0031] In a pig farm with a scale of about 1000 pigs, 12 representative ordinary fattening pigs (4-5 months old, 2-4 weeks before slaughter, body weight about 120kg) were randomly selected as representative samples for parallel experiments. After formulating feed containing a certain concentration of clenbuterol, feed pigs regularly. At the same time, pig urine was continuously collected in clean sample bottles to determine the content of clenbuterol. Through the continuous and uninterrupted detection of pig urine samples obtained from 12 samples, it was found that the following general rules were found. After 3 hours of feeding, clenbuterol could be detected in pig urine; during 3-72 hours, The concentration of clenbuterol in pig urine was in a continuous rising period; the content was stable after 72h. After stopping feeding, it was in a rapid decline period within 3-48 hours; after 48 hours, the content of clenbu...

Embodiment 2

[0041] Embodiment 2 homogeneity test

[0042] Randomly select 11 packaging units according to the whole packaging process before, during and after, and the randomly selected samples are numbered from 1 to 11, and each randomly selected unit takes 3 sub-samples in parallel, and the record numbers are 1-1, 1-2 , 1-3, 2-1, 2-2, 2-3, ..., 11-1, 11-2, 11-3. The homogeneity test method adopts liquid chromatography-isotope dilution mass spectrometry to measure the results (see the standard substance determination part for specific method parameters), and the results are statistically analyzed by analysis of variance, and judged by comparing the F test value with the F critical value. See Table 1 for the results of the homogeneity test and statistical analysis of the clenbuterol components in pig urine.

[0043] Table 1 Pig urine matrix standard substance homogeneity test results containing clenbuterol (ng / mL)

[0044]

[0045]

[0046] The above experimental data show that th...

Embodiment 3

[0047] Embodiment 3 Stability

[0048] Long-term stability monitoring studies were carried out at 0, 1, 3, 6, 9, and 12 months, respectively. Three packaging units are randomly selected each time, and each unit is measured in parallel three times. The measurement method is the same as that used in the homogeneity test, both of which are liquid chromatography-isotope dilution chromatography mass spectrometry. The average value of the measurement results of the three packaging units is taken as the long-term stability monitoring result. The trend analysis method is used to analyze the results, and the monitoring time and the results are fitted with a straight line, and the results are statistically analyzed. The long-term stability monitoring results of the pig urine freeze-dried powder containing clenbuterol are shown in Table 2. A straight line was fitted with the detection time and results, and the trend analysis method was used to statistically analyze the stability test res...

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Abstract

The invention discloses a standard substance containing free-state and conjugated-state clenbuterol in swine urine and a preparation method of the standard substance. The method includes: feeding animals to obtain a swine urine raw material containing free-state and conjugated-state clenbuterol concentration ranges, fixing the value, and freeze-drying to obtain the free-state and conjugated-state clenbuterol standard substance in the swine urine freeze-dried powder of which the uniformity stability meets the requirements; adding water for redissolving into liquid, pre-treating the liquid by using a mixed biological enzyme enzymolysis method, and determining the content of clenbuterol in the matrix standard substance by using a liquid chromatography-tandem mass spectrometry / isotope internal standard method for combined valuing. The standard substance provided by the invention reflects the real condition of the clenbuterol standard substance in swine urine metabolized in an animal body, the fixed value is more accurate, and the uniformity and the stability are good; therefore, the standard substance can be widely applied to many fields of clenbuterol residue detection in animal urine, quality verification of rapid screening products, instrument detection method evaluation, laboratory internal quality control, laboratory capability verification and comparison and the like.

Description

technical field [0001] The invention relates to the technical field of preparation of veterinary drug residue detection standard substances, in particular to a condensed clenbuterol standard substance in pig urine freeze-dried powder metabolized by animals and a preparation method thereof. Background technique [0002] clenbuterol beta 2 - One of the receptor agonists, it is also one of the earliest and most typical representatives of toxic and harmful compounds commonly known as "leanbuterol". It is characterized by low cost, quick effect, and many It was exposed for the first time that it was illegally used in animal breeding, which brought serious risks to human health and public health safety. Clenbuterol has always been included in the key monitoring parameters of animal food. At present, there are many methods for the determination of clenbuterol residues in edible animal tissues such as pork and pig liver and pig urine. The instrument method is usually liquid chromat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/02G01N30/72
CPCG01N30/06G01N30/02G01N30/72
Inventor 单吉浩李建勋马康薛晓锋邢富国隋福顺卢培培赵玉乐徐迪
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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