CN103954575A - Method for identifying fresh pork and unfrozen pork - Google Patents
Method for identifying fresh pork and unfrozen pork Download PDFInfo
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- CN103954575A CN103954575A CN201410187773.5A CN201410187773A CN103954575A CN 103954575 A CN103954575 A CN 103954575A CN 201410187773 A CN201410187773 A CN 201410187773A CN 103954575 A CN103954575 A CN 103954575A
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Abstract
The invention discloses a method for identifying fresh pork and unfrozen pork. According to the method, a sample is divided into two parts, one part is analyzed directly, and the other part is analyzed after being frozen and unfrozen. According to the method, the same treatment is performed, the ratios of the light absorption values of the two parts at 340 nanometers are compared, and the whether the sample is fresh pork or unfrozen pork is judged according to the ratios. The method is simple, the time is short, results are reliable and the cost is low.
Description
Technical field
The present invention relates to pork authentication method field, relate in particular to a kind of discrimination method of fresh and freezing rear solution cold pork.
Background technology
Along with the problem of food safety and hygiene is more and more serious, people expect to have bought all wool and a yard wide commodity.Such as in European & American Market, green meat is approximately more expensive 20% than separating frozen meat, valuable 50% when the highest, and separates frozen meat mouthfeel and can't wait green meat.But some retailer, for individual interest, pretends to be fresh pork to sell with the solution cold pork after freezing, and consumer's interests are suffered damage.Therefore, how to differentiate that green meat and solution frozen meat become at present one of problem demanding prompt solution on the market, but existing similar method of discrimination process is loaded down with trivial details in the world at present, accuracy is low, and does not still have a kind of method to differentiate fresh pork reconciliation cold pork.
Summary of the invention
The object of the present invention is to provide a kind of authentication method qualification pork easy, quick, with low cost, that accuracy rate is high, required sample size is few whether fresh.
For achieving the above object, the invention provides the discrimination method of a kind of live fresh pork and freezing rear solution cold pork, it is characterized in that contrasting the ratio of pork sample light absorption value before and after freeze-thaw, comprise the steps.
Get two parts of testing samples, every part of 2-4g, puts into a copy of it-20 DEG C of freezing 12h left and right, the 20min post analysis that thaws under room temperature, another part of Direct Analysis;
Analytical approach: the pork of 2-4g is put into 10ml 100mM pH=6 phosphate buffered saline(PBS) and soak after 10 minutes, get in soak solution to the 600 μ L 100mM pH=6 phosphate buffered saline(PBS) of 23 μ L, add 50 μ L 34.4mM EDTA reactions after 3 minutes simultaneously, continue to add 1.5mM NADH and the 10 μ L 5.9mM biacetyl coacetylases of 27 μ L, mix rear horse back and survey its light absorption value at 340nm place, a numerical value of every 10s record is set, record 2min, taking the time as transverse axis, the numerical value of every 10s record is that the longitudinal axis calculates its pace of change;
Determination methods: the ratio of frozen meat and green meat light absorption value pace of change is separated in contrast after freezing, in the time that ratio is greater than 1.8, determines that raw sample is green meat; In the time that ratio is less than 1.4, determine that raw sample is to separate frozen meat; In the time that ratio falls between, need again judge, until result is greater than 1.8 or be less than 1.4.
Testing sample is required to be lean meat part.
Inventive principle:
HADH is a kind of cyclophorase, and this enzyme can be that under 6 left and right, room temperature condition, catalyzing N ADH changes NAD at environment pH
+, concrete reaction is as follows:
Therefore, HADH enzyme live height can pass through NADH(reactant) minimizing speed or NAD
+the calculating of advancing the speed of (product), reactant NADH has maximum light absorption value at 340nm place, therefore HADH enzyme is lived, height can be by the minimizing speed judgement of NADH, the i.e. rate of change (reducing speed) of 340nm place light absorption value.
Advantage of the present invention:
The present invention is the ratio alive of the HADH enzyme after freeze-thaw by comparative sample and sample, and other method needs the HADH enzyme of measuring a large amount of solution frozen meat and green meat to live just, and need utilize statistics rule judgement sample source.Therefore, the method has been saved judgement time and medicine, and result is clear, understandable.
Authentication method of the present invention is by EDTA(34.4mM) directly mix with 600 μ L according to 50 μ L with the phosphate-buffered salt of 0.1mol/L.Instead of the general phosphate-buffered salt that first adds, after add EDTA, therefore, the present invention has saved the time.
The meat that authentication method of the present invention is differentiated need is divided into two parts that size approaches, and every part of about 2-4g, without the special concrete weight of considering pork, without the concrete position of considering pork.
The present invention determines that from starting to identify result only need be less than 120min, and conventional method needs even tens of hours a few hours.
Authentication method of the present invention only needs 27 μ L NADH and 10 μ L biacetyl coacetylases (Acetoacetyl-CoA).General approach need add the above NADH of 30 μ L and 15 μ L left and right biacetyl coacetylases, and the method has reduced appraisal cost.
Authentication method of the present invention only needs 2 groups of samples (≤10 samples) just can distinguish the source of pork, and accuracy is high.Other HADH enzyme activity determination need do a large amount of data statisticss and do statistical analysis the source that could differentiate meat.
These advantages mainly come from 1, ratioing technigue judgement.Determination methods is to measure sample and the HADH enzyme of sample through the freezing front and back that thaw again ratio alive, greatly reduces workload and work difficulty, the difference of having avoided different parts to cause; 2, the reasonable value of rational soak time and soak solution volume.The method can accurately judge the source of pork sample, and uses instrument very simple.
In addition, pork sampling can only be got thin, can not get fat meat and bone.Because: 1, theoretically, fat meat and bone contain less mitochondria, so cannot be with HADH(cyclophorase) the enzyme height of living differentiates the source of meat; 2, find from a large amount of sample analyses, fat meat and bone cannot judge the true source of meat.
Embodiment
Describe embodiments of the invention below in detail, the example of described embodiment is intended to for explaining the present invention, and can not be interpreted as limitation of the present invention.Unreceipted concrete technology or condition person in embodiment, according to the described technology of the document in this area or condition or carry out according to product description.The unreceipted person of production firm of agents useful for same or instrument, being can be by the conventional products of commercial acquisition.
embodiment 1: the qualification of fresh pork and freezing rear pork
Solution preparation:
Phosphate buffered saline(PBS): the sodium dihydrogen phosphate (NaH of (1) 1 mol/L
2pO
4) storage liquid: dissolve 13.8g in enough water, making final volume is 0.1L; (2) 1mol/L sodium hydrogen phosphate (Na
2hPO
4) storage liquid: dissolve 14.2g in enough water, making final volume is 0.1L.The sodium hydrogen phosphate of the 1mol/L of 13.8ml is mixed with the sodium dihydrogen phosphate of the 1mol/L of 87.7ml, adding water (being about 900mL) to the overall solution volume of a certain amount of volume is 1L, then regulates the pH value to 6 of the rear phosphate buffered saline(PBS) of dilution with 1 mol/L phosphate-buffered salt mother liquor.
EDTA: the EDTA that is 0.5M by 1,040 μ L concentration is dissolved in the water of 14,000 μ L, joins to obtain the EDTA solution of 34.4mM.
NADH: the NADH powder that takes 0.106g adds water and is settled to 100ml and joins to obtain the concentration NAHD solution that is 1.5mM.
Biacetyl coacetylase (Acetoacetyl-CoA): the biacetyl coacetylase (Acetoacetyl-CoA) of 8mg is dissolved in 1,600 μ L water, joins to obtain 5.9mM biacetyl coacetylase solution.
Get respectively the lean meat part of two groups of samples, one group of solution frozen meat, one group of green meat.Green meat is divided into two groups (A1 group and A2 group), solution frozen meat is divided into two groups (B1 group and B2 group), 50 parts of every groups, every part of about 3g.A1 group is directly measured, and A2 group is placed in-20 DEG C of freezing 12h, after the 20min that thaws, measures, and B1 group is directly measured, and B2 group is placed in-20 DEG C of freezing 12h, after the 20min that thaws, measures under room temperature.
Measuring method: pork is put into 10ml(100mM pH=6) phosphate buffered saline(PBS) soaks 10 minutes, soak solution to the 600 μ L phosphate buffered saline(PBS) (100mM pH=6) of then getting 23 μ L and 50 μ L EDTA(34.4mM) in the mixed liquor of solution, react after 3 minutes, continue to add the NADH(1.5mM of 27 μ L) and 10 μ L biacetyl coacetylases (5.9mM Acetoacetyl-CoA), mix the pace of change of rear horse back at its light absorption value of 340nm place survey.Find by great amount of samples:
1. separating frozen meat (A2 group) and green meat (A1 group) the HADH enzyme ratio of living has 49 groups to be greater than 1.8, and only having 1 group, to be less than 1.8(be 1.77).A1 group has utmost point significant difference (P<0.001) with A2 group numerical value.The results are shown in Table 1
Table 1 is separated frozen meat and green meat HADH enzyme ratio table alive
Frozen meat and green meat HADH enzyme ratio alive are separated in note: A2/A1 representative, choose altogether 4 characteristic portions and 1 other position, and 10, each position is parallel, and wherein the 8th parallel ratio of fore shank is less than 1.8.
2. very approaching with normal solution frozen meat (B1 group) HADH enzyme value alive ratio through twice freezing solution frozen meat (B2 group) HADH enzyme value alive, its ratio is less than 1.4, has 2 groups of sample HADH enzymes to live not this scope (being respectively 1.41 and 1.53).B1 group and B2 group numerical value there was no significant difference (P>0.05).The results are shown in Table 2.
Twice freeze thawing meat of table 2 and solution frozen meat HADH enzyme ratio table alive
Note: B2/B1 represents twice freeze thawing meat and separates the frozen meat HADH enzyme ratio of living, and choose altogether 4 characteristic portions and 1 other position, 10, each position is parallel, and wherein the 10th of lean pork taken under the spinal column of a hog is parallel is greater than 1.4 with the parallel ratio in the 10th, other position.
Two times result has in conjunction with statistical analysis analysis: separate live 48 groups of ratios of frozen meat (A2 group) and green meat (A1 group) HADH enzyme and be less than 1.4 probability lower than 0.1%, through twice freezing solution frozen meat (B2 group) with normal separate frozen meat (B1 group) HADH enzyme live value ratio higher than 1.8 probability lower than 0.01%.Comprehensive explanation the method accurately, reliably.
Although illustrated and described embodiments of the invention above, be understandable that, above-described embodiment is exemplary, can not be interpreted as limitation of the present invention, those of ordinary skill in the art can change above-described embodiment within the scope of the invention in the situation that not departing from principle of the present invention and aim, amendment, replacement and modification.
Claims (2)
1. a discrimination method for live fresh pork and freezing rear solution cold pork, is characterized in that contrasting the ratio of pork sample through freeze-thaw front and back light absorption value, comprises the steps;
Get two parts of testing samples, every part of 2-4g, puts into a copy of it-20 DEG C of freezing 12h, the room temperature 20min post analysis that thaws, another part of Direct Analysis;
Analytical approach: the pork sample of 2-4g is put into 10ml 100mM pH=6 phosphate buffered saline(PBS) and soak after 10 minutes, get in soak solution to the 600 μ L 100mM pH=6 phosphate buffered saline(PBS) of 23 μ L, add 50 μ L 34.4mM EDTA reactions after 3 minutes simultaneously, continue to add 1.5mM NADH and the 10 μ L5.9mM biacetyl coacetylases of 27 μ L, mix rear horse back and survey its light absorption value at 340nm place, a numerical value of every 10s record is set, record 2min, taking the time as transverse axis, the numerical value of every 10s record is that the longitudinal axis calculates its pace of change;
Determination methods: the ratio of frozen meat and raw sample light absorption value pace of change is separated in contrast after freezing, in the time that ratio is greater than 1.8, determines that raw sample is green meat; In the time that ratio is less than 1.4, determine that raw sample is to separate frozen meat; In the time that ratio falls between, need again judge, until result is greater than 1.8 or be less than 1.4.
2. the discrimination method of fresh and freezing rear solution cold pork described in claim 1, is characterized in that contrasting the ratio of pork sample through freeze-thaw front and back light absorption value.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103163084A (en) * | 2013-03-14 | 2013-06-19 | 广西工学院 | Characterization method for detecting freshness of shrimp product |
| CN103424384A (en) * | 2012-05-23 | 2013-12-04 | 北京和信非凡生物技术有限公司 | Enzyme activity detection method for mitochondrial respiratory chain compound I and reagents |
-
2014
- 2014-05-06 CN CN201410187773.5A patent/CN103954575A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103424384A (en) * | 2012-05-23 | 2013-12-04 | 北京和信非凡生物技术有限公司 | Enzyme activity detection method for mitochondrial respiratory chain compound I and reagents |
| CN103163084A (en) * | 2013-03-14 | 2013-06-19 | 广西工学院 | Characterization method for detecting freshness of shrimp product |
Non-Patent Citations (3)
| Title |
|---|
| EDUARDO M. RAMOS ET AL.,: "Effect of stunning methods on the differentiation of frozen-thawed bullfrog meat based on the assay of β-hydroxyacyl-CoA-dehydrogenase", 《FOOD CHEMISTRY》, vol. 87, 31 December 2004 (2004-12-31), pages 607 - 611 * |
| MANUELA FERNÁNDEZ ET AL.,: "Use of β-hydroxyacyl-CoA-dehydrogenase (HADH) activity to differentiate frozen from unfrozen fish and shellfish", 《EUROPEAN FOOD RESEARCH AND TECHNOLOGY》, vol. 209, no. 3, 31 August 1999 (1999-08-31), pages 205 - 208 * |
| N.Z. BALLIN ET AL.,: "Analytical methods for authentication of fresh vs. thawed meat – A review", 《MEAT SCIENCE》, vol. 80, no. 2, 31 December 2008 (2008-12-31), pages 151 - 158, XP023980057, DOI: doi:10.1016/j.meatsci.2007.12.024 * |
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Application publication date: 20140730 |