CN111337528B - Nuclear magnetic resonance hydrogen spectrometry for identifying beef, pork or duck meat mixed in mutton - Google Patents
Nuclear magnetic resonance hydrogen spectrometry for identifying beef, pork or duck meat mixed in mutton Download PDFInfo
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- CN111337528B CN111337528B CN202010053345.9A CN202010053345A CN111337528B CN 111337528 B CN111337528 B CN 111337528B CN 202010053345 A CN202010053345 A CN 202010053345A CN 111337528 B CN111337528 B CN 111337528B
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- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
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- G—PHYSICS
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Abstract
The invention discloses a nuclear magnetic resonance hydrogen spectrum method for identifying beef, pork or duck meat doped in mutton, which comprises the steps of weighing a proper sample, extracting by adopting an organic solvent, adding proper amount of ultrapure water, vortex centrifuging, blowing upper layer organic phase nitrogen, dissolving a small amount of treated sample by using deuterated chloroform containing TMS, and analyzing by adopting a 400MHz nuclear magnetic resonance analyzer. And judging whether the sample mutton is true mutton according to the obtained nuclear magnetic resonance hydrogen spectrum fingerprint spectrum. The identification method provided by the invention is simple, high in detection efficiency, high in result accuracy and good in reproducibility, and has reference significance for the authenticity research of the mutton doped with other meats or other meats and meat products.
Description
Technical Field
The invention relates to a food identification technology, in particular to a nuclear magnetic resonance hydrogen spectrum fingerprint spectrum method for identifying beef, pork or duck meat mixed in mutton.
Background
Mutton is tender in quality, delicious in taste, rich in various nutritional ingredients such as protein, essential amino acids and the like, low in cholesterol content, and very popular with consumers, but low in yield and relatively expensive in price. Many bad merchants are surprised to get the violence, are surprised to be disagreeable, and are filled with pork, duck, horse meat and other cheaper meats, so that the rights and interests of consumers are damaged, and the food market order is disturbed. Therefore, there is a strong need to develop a method for efficiently identifying real and false mutton, which provides technical support for government regulatory authorities.
The existing detection technology for meat adulteration comprises sensory detection, immunological technology, molecular biological technology, spectrum and chromatographic technology and the like. The molecular biology technology based on DNA detection is relatively mature for the false identification of mutton products, and has the advantages of strong specificity, high sensitivity and the like, but the operation is relatively complex, and the time and the labor are consumed.
A great deal of research has been done at home and abroadNuclear Magnetic Resonance (NMR) spectroscopy has proven to be a leading research and analytical testing technique worldwide and has a variety of applications in food safety, traceability and authenticity. The patent discloses a method for identifying real and false beef and mutton (application number 201310314554.4), which is to obtain carbon spectrum data of chloroform extract by using a high-field nuclear magnetic technique and realize meat identification according to fingerprint peaks in specific intervals and area algorithms of specific peaks. The method has the advantages of too many carbon spectrum scanning times and higher experiment cost. In fact, most of the food analysis studies by NMR have focused on nuclear magnetic fingerprint hydrogen spectroscopy, i.e 1 HNMR technology. Unlike the conventional analysis method, 1 the HNMR technology does not need to pay attention to specific components, can select a proper deuterated reagent to directly dissolve and analyze according to the chemical properties of food samples, can also utilize a specific extraction mode to separate and concentrate water phase or organic components in the samples and then dissolve and analyze, and can give out all proton information in the samples or in the extracted components at one time. In addition, the analytical identification research of meat often utilizes low-field nuclear magnetism, has the advantages of portability, convenient sample acquisition and direct use in industrial environment, but the magnetic field is difficult to be uniform, so that the sensitivity and the resolution of the instrument are low.
Disclosure of Invention
In order to solve the problems, the invention provides a simple and quick method for qualitatively distinguishing beef, pork or duck meat from mutton.
In order to achieve the above object, the present invention is realized by the following technical scheme:
the invention relates to a nuclear magnetic resonance hydrogen spectrum method for identifying beef, pork or duck meat mixed in mutton, which comprises the following steps:
s1, weighing a plurality of groups of samples, wherein each group of samples comprises a mutton sample, a beef sample, a pork sample and a duck sample, the weight of each sample in each group is 5-8g, each sample in each group is respectively crushed and independently loaded into a 50mL centrifuge tube, and the mixture is extracted for 15-20h by using 10-15mL of organic solvent;
s2, adding an equal volume of ultrapure water into each sample of the S1, and centrifuging for 5min at 4000rpm after vortex;
s3, transferring the upper organic phase of the sample treated by the S2 to a glass tube and blowing nitrogen at 50 ℃ for 1-2h;
s4, measuring 100 mu L of the sample processed in the step S3, adding 600 mu L of deuterated chloroform, measuring 500-600 mu L of the sample after vortex uniformity, and filling the sample into a nuclear magnetic tube;
s5, measuring a sample in the nuclear magnetic tube by adopting a 400MHz liquid nuclear magnetic resonance spectrometer, wherein the hydrogen spectrum measuring conditions are as follows: the pulse procedure is: zg30; p1=7.74 mus; td=64K; ds=2; ns=16; d1 =1s; the test temperature was 25 ℃;
s6, obtaining nuclear magnetic resonance hydrogen spectrum fingerprint spectra of samples of mutton, beef, pork or duck meat, and analyzing all the obtained samples by using software Topspin3.5 1 HNMR data, wherein the chemical shift of an internal standard TMS is set to be 0, then baseline correction and phase correction are carried out, samples are identified according to a map, and if the map simultaneously meets the following conditions: 1. three sets of multimodal were present within the 5.45-6.49ppm shift; 2. there are no three sets of multimodal between 6.90-7.68ppm shift; 3. no convex mountain-shaped peaks exist at two positions of 2.925ppm and 2.610ppm within the displacement of 2.45-3.23 ppm; the mutton sample is proved to be real mutton, otherwise the sample is not mutton or beef, pork or duck meat is added.
The invention further improves that: the meat sample in S1 is fresh meat purchased in market or meat roll which is frozen and preserved and has any part of head, neck, foot, viscera and skin removed, and the number of sample groups is not less than 5.
The invention further improves that: the organic solvent in S1 is one of normal hexane, low-boiling petroleum ether or high-boiling petroleum ether, wherein the boiling point of the low-boiling petroleum ether is 30-60 ℃, and the boiling point of the high-boiling petroleum ether is 60-90 ℃.
The invention further improves that: the deuterated chloroform in S4 contained 0.03% TMS.
The beneficial effects of the invention are as follows: the method is based on liquid nuclear magnetism 1 The HNMR fingerprint spectrum technology has the advantages of simple experimental operation, high efficiency in detection, high accuracy of results, good reproducibility and the like, and can be applied to the identification of whether beef, pork and duck are added in mutton. The pretreatment mode is simple and flexible, and the methodThe method has the advantages of less deuterated reagent consumption, high detection efficiency, accurate and reliable result and low detection cost. The method has reference significance for researching the authenticity of adding other meat or other meat products into the mutton.
Drawings
FIG. 1 is a nuclear magnetic resonance spectrum of each of the meat samples of examples 1-3 at 5.45-6.49 ppm.
FIG. 2 is a nuclear magnetic resonance spectrum of each of the meat samples of examples 1-3 at 6.90-7.68 ppm.
FIG. 3 is a nuclear magnetic resonance spectrum of each of the meat samples of examples 1-3 at 2.45-3.23 ppm.
Detailed Description
The technical scheme of the present invention will be further described with reference to specific examples, and the technical scheme of the present invention will be further described with reference to the specific examples. It will be appreciated by those skilled in the art that the examples are provided for illustration only and are not intended to limit the scope of the present invention.
As shown in fig. 1-3, the present invention is a nuclear magnetic resonance hydrogen spectrometry for identifying beef, pork or duck meat in mutton, comprising the steps of:
s1, weighing a plurality of groups of samples, wherein each group of samples comprises a mutton sample, a beef sample, a pork sample and a duck sample, the weight of each sample in each group is 5-8g, each sample in each group is respectively crushed and independently loaded into a 50mL centrifuge tube, and the mixture is extracted for 15-20h by using 10-15mL of organic solvent;
s2, adding an equal volume of ultrapure water into each sample of the S1, and centrifuging for 5min at 4000rpm after vortex;
s3, transferring the upper organic phase of the sample treated by the S2 to a glass tube and blowing nitrogen at 50 ℃ for 1-2h;
s4, measuring 100 mu L of the sample processed in the step S3, adding 600 mu L of deuterated chloroform, measuring 500-600 mu L of the sample after vortex uniformity, and filling the sample into a nuclear magnetic tube;
s5, measuring a sample in the nuclear magnetic tube by adopting a 400MHz liquid nuclear magnetic resonance spectrometer, wherein the hydrogen spectrum measuring conditions are as follows: the pulse procedure is: zg30; p1=7.74 mus; td=64K; ds=2; ns=16; d1 =1s; the test temperature was 25 ℃;
s6, obtaining mutton, beef, pork or duck meatNuclear magnetic resonance hydrogen spectrum finger print of sample, analysis of all samples obtained using software topspin3.5 1 HNMR data, wherein the chemical shift of an internal standard TMS is set to be 0, then baseline correction and phase correction are carried out, samples are identified according to a map, and if the map simultaneously meets the following conditions: 1. three sets of multimodal were present within the 5.45-6.49ppm shift; 2. there are no three sets of multimodal between 6.90-7.68ppm shift; 3. no convex mountain-shaped peaks exist at two positions of 2.925ppm and 2.610ppm within the displacement of 2.45-3.23 ppm; the mutton sample is proved to be real mutton.
The meat samples in S1 are fresh meat purchased in the market or meat rolls which are frozen and stored and removed from any parts after head, neck, feet, viscera and skin, the number of sample groups is not less than 5, and the samples can be put into a refrigerator for refrigeration and are subjected to experiments within 2 days. The organic solvent in S1 is one of normal hexane, low-boiling petroleum ether or high-boiling petroleum ether, wherein the low-boiling petroleum ether is avoided under the condition of high temperature weather, the boiling point of the low-boiling petroleum ether is 30-60 ℃, and the boiling point of the high-boiling petroleum ether is 60-90 ℃. The use of low boiling petroleum ether is avoided at room temperature greater than 30 ℃.
Example 1
S1, weighing a plurality of groups of samples, wherein each group of samples comprises a mutton sample, a beef sample, a pork sample and a duck sample, each group of fresh meat is 5g, crushed and put into a 50mL centrifuge tube, and extracted for 15h by 10mL of normal hexane;
s2, continuously adding 10mL of ultrapure water, and centrifuging at 4000rpm for 5min after vortex;
s3, transferring the upper organic phase to a glass tube and blowing nitrogen at 50 ℃ for 1.5 hours; wherein the deuterated chloroform contains 0.03% TMS;
s4, measuring 100 mu L of the nitrogen-blown extract, adding 600 mu L of deuterated chloroform, and measuring 500 mu L of the nitrogen-blown extract after vortex uniformity and filling the obtained mixture into a nuclear magnetic tube;
s5, measuring a sample in the nuclear magnetic tube by adopting a 400MHz liquid nuclear magnetic resonance spectrometer, wherein the hydrogen spectrum measuring conditions are as follows: the pulse procedure is: zg30; p1=7.74 mus; td=64K; ds=2; ns=16; d1 =1s; the test temperature was 25 ℃;
s6, obtaining nuclear magnetic resonance hydrogen spectrum fingerprints of samples of mutton, beef, pork or duckMap, analysis of all samples obtained using software topsapin 3.5 1 HNMR data, wherein the chemical shift of an internal standard TMS is set to be 0, then baseline correction and phase correction are carried out, and characteristic peaks of sample mutton are determined, if the patterns simultaneously meet the following conditions: 1. three sets of multimodal were present within the 5.45-6.49ppm shift; 2. there are no three sets of multimodal between 6.90-7.68ppm shift; 3. no convex mountain-shaped peaks exist at two positions of 2.925ppm and 2.610ppm within the displacement of 2.45-3.23 ppm; the mutton sample is proved to be real mutton, otherwise the sample is not mutton or beef, pork or duck meat is added.
Example 2
S1, weighing a plurality of groups of samples, wherein each group of samples comprises a mutton sample, a beef sample, a pork sample and a duck sample, each group of frozen meat or meat roll after thawing is 6g, crushed and filled into a 50mL centrifuge tube, and extracted for 18h by 12mL of normal hexane;
s2, continuously adding 12mL of ultrapure water, and centrifuging at 4000rpm for 5min after vortex;
the remaining steps were the same as in example 1.
Example 3
S1, weighing a plurality of groups of samples, wherein each group of samples comprises a mutton sample, a beef sample, a pork sample and a duck sample, 8g of each group of unfrozen frozen meat or meat roll is crushed and put into a 50mL centrifuge tube, and 15mL of normal hexane is used for extraction for 20 hours;
s2, continuously adding 15mL of ultrapure water, and centrifuging at 4000rpm for 5min after vortex;
the remaining steps were the same as in example 1.
S3, transferring the upper organic phase to a glass tube and blowing nitrogen at 50 ℃ for 2 hours;
the remaining steps were the same as in example 1.
The main instruments or containers used in the method are a 400MHz liquid nuclear magnetic resonance spectrometer, a BBI probe, an IT0079 electronic balance, a 5mm nuclear magnetic tube, a centrifuge and an EVA32 type full-automatic nitrogen blowing concentrator.
Claims (3)
1. A nuclear magnetic resonance hydrogen spectrometry method for identifying beef, pork or duck meat doped in mutton, which is characterized in that: the method comprises the following steps:
s1, weighing a plurality of groups of samples, wherein each group of samples comprises a mutton sample, a beef sample, a pork sample and a duck sample, the weight of each sample in each group is 5-8g, each sample in each group is respectively crushed and independently loaded into a 50mL centrifuge tube, and the mixture is extracted for 15-20h by using 10-15mL of organic solvent;
s2, adding an equal volume of ultrapure water into each sample of the S1, and centrifuging for 5min at 4000rpm after vortex;
s3, transferring the upper organic phase of the sample treated by the S2 to a glass tube and blowing nitrogen at 50 ℃ for 1-2h;
s4, measuring 100 mu L of the sample processed in the step S3, adding 600 mu L of deuterated chloroform, measuring 500-600 mu L of the sample after vortex uniformity, and filling the sample into a nuclear magnetic tube;
s5, measuring a sample in the nuclear magnetic tube by adopting a 400MHz liquid nuclear magnetic resonance spectrometer, wherein the hydrogen spectrum measuring conditions are as follows: the pulse procedure is: zg30; p1=7.74 mus; td=64K; ds=2; ns=16; d1 =1s; the test temperature was 25 ℃;
s6, obtaining nuclear magnetic resonance hydrogen spectrum fingerprint spectra of samples of mutton, beef, pork or duck meat, and analyzing all the obtained samples by using software Topspin3.5 1 HNMR data, wherein the chemical shift of an internal standard TMS is set to be 0, then baseline correction and phase correction are carried out, samples are identified according to a map, and if the map simultaneously meets the following conditions: (1) there are three sets of multimodal within a displacement of 5.45 to 6.49 ppm; (2) No three sets of multimodal were present between 6.90-7.68ppm shift; (3) No convex mountain-shaped peaks at the positions 2.925ppm and 2.610ppm within the displacement of 2.45-3.23 ppm; the mutton sample is proved to be real mutton; otherwise the sample is not mutton or is mixed with beef, pork or duck;
the organic solvent in S1 is one of normal hexane, low-boiling petroleum ether or high-boiling petroleum ether, wherein the boiling point of the low-boiling petroleum ether is 30-60 ℃, and the boiling point of the high-boiling petroleum ether is 60-90 ℃.
2. The nmr hydrogen spectrometry for identifying beef, pork or duck meat in mutton as claimed in claim 1, wherein: the meat samples in S1 are fresh meat purchased in the market or meat rolls which are removed from any part after head, neck, feet, viscera and skin and are stored in a freezing way, and the number of sample groups is not less than 5.
3. The nmr hydrogen spectrometry for identifying beef, pork or duck meat in mutton as claimed in claim 1, wherein: the deuterated chloroform in S4 contained 0.03% TMS.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102269720A (en) * | 2011-04-28 | 2011-12-07 | 王士桥 | Method for identifying edible oil quality through combination of nuclear magnetic resonance and pattern recognition method |
| CN103389318A (en) * | 2013-07-24 | 2013-11-13 | 厦门大学 | Method for identifying true and false beef and mutton |
| CN105021643A (en) * | 2014-04-18 | 2015-11-04 | 中国石油化工股份有限公司 | Method for measuring water contents of organic matters by virtue of nuclear magnetic resonance instrument |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102269720A (en) * | 2011-04-28 | 2011-12-07 | 王士桥 | Method for identifying edible oil quality through combination of nuclear magnetic resonance and pattern recognition method |
| CN103389318A (en) * | 2013-07-24 | 2013-11-13 | 厦门大学 | Method for identifying true and false beef and mutton |
| CN105021643A (en) * | 2014-04-18 | 2015-11-04 | 中国石油化工股份有限公司 | Method for measuring water contents of organic matters by virtue of nuclear magnetic resonance instrument |
Non-Patent Citations (1)
| Title |
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| 李伟妮等.冷藏山羊肉品质变化的核磁共振研究.食品工业科技.2010,第第31卷卷(第第31卷期),125-127. * |
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