CN107389722A - The method that ESR characterizes beef fat quality comparison during multigelation - Google Patents
The method that ESR characterizes beef fat quality comparison during multigelation Download PDFInfo
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- CN107389722A CN107389722A CN201710641582.5A CN201710641582A CN107389722A CN 107389722 A CN107389722 A CN 107389722A CN 201710641582 A CN201710641582 A CN 201710641582A CN 107389722 A CN107389722 A CN 107389722A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
- G01N24/10—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using electron paramagnetic resonance
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
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Abstract
The invention discloses the method that a kind of electron spin resonance (ESR) characterizes beef fat quality comparison during multigelation, it includes：(1) electron spin resonance running parameter is set, and described running parameter includes central magnetic field intensity, field sweep width, microwave power, modulation amplitude, receiver gain, time constant, conversion time, x frequency bands；(2) multigelation processing is carried out to beef, then the extraction of fat is carried out to the beef after defrosting, the fat of extraction is measured using electron spin resonance afterwards, the object of measure includes free radical intensity, free radical quantity or induction time；(3) the electron spin resonance spectroscopy figure of acquisition is analyzed and processed, so as to realize the sign for beef fat quality comparison during multigelation.The method of the present invention has the advantages that detection sensitivity is high, and analysis solvent consumption is few, and sample treatment is simple, and quickly and efficiently beef fat quality during multigelation can be detected.
Description
Technical field
The present invention relates to the method that beef quality during a kind of detection multigelation changes, and in particular to one kind is with electronics
The method that spin resonance (ESR) characterizes lipid deterioration in beef quality during multigelation.
Background technology
Chilled storage is still most widely used meat and meat products store method in current long-distance transport, and it can preferably be protected
Hold product quality and product is had compared with long shelf life.Because the current cold chain transportation system in China is poor, cause the beef in China
It is easier to undergo freeze-thaw (freeze thawing) process repeatedly.Many researchs confirm that multigelation accelerates the quality of meat, flavor,
The deterioration of color, microbial activity and nutritive value.Most of researchers think, the rotten master of meat caused by multigelation
Want the formation that reason is ice crystal in freezing process.The ice crystal of formation causes the rupture of cell membrane, so that being catalyzed lipid in cell
The dominant catalyst release of oxidation, accelerates lipid oxidation.And caused free radical and many chemistry are anti-during fatty deterioration
Should be closely related, it is the mark of Food Oxidation process, the chain reaction that free radical triggers is the essence of fatty autoxidation, freely
Base can also influence flavour of food products.
Mainly there are peroxide value, TBARS production of thiobarbituric acid reactive to the index evaluated on Fat quality at present
(Thiobarbituric Acid Reactive Substances, TBARS) and acid value etc..And main detection method is main
There are high performance liquid chromatography, chemoluminescence method and fluorescence detection etc., but reagent, the consumptive material needed for these methods are more, sample
Pre-treatment is cumbersome, analysis time length.
The content of the invention
It is a primary object of the present invention to provide a kind of electron spin resonance (ESR) characterize multigelation during beef fat
The method of fat quality comparison, it has high sensitivity, easy to operate, analyzes the features such as rapid, so as to overcome in the prior art
Deficiency.
For achieving the above object, present invention employs following technical scheme：
The embodiments of the invention provide a kind of ESR characterize multigelation during beef fat quality comparison method, its
Including：
(1) set electron spin resonance running parameter, described running parameter include central magnetic field intensity, field sweep width,
Microwave power, modulation amplitude, receiver gain, time constant, conversion time, x frequency bands；
(2) multigelation processing is carried out to beef, then the extraction of fat is carried out to the beef after defrosting, afterwards using electronics
Spin resonance is measured to the fat of extraction, and the object of measure includes free radical intensity, free radical quantity or induction time；
(3) the electron spin resonance spectroscopy figure of acquisition is analyzed and processed, so as to realize for ox during multigelation
The sign of meat fat quality comparison.
In some preferred embodiments, step (1) the central magnetic field intensity is arranged to 3350.00G, field sweep width
It is arranged to 100.00G, microwave power is arranged to 20mW, and modulation amplitude is arranged to 1.000G, and receiver gain is set
For 30dB, time constant is arranged to 0.01ms, and conversion time is arranged to 40.96ms, and x frequency bands are arranged to
9.42GHz.In some embodiments, in the step (2), to beef carry out multigelation processing during, be by
It is that a freeze thawing follows that beef is thawed to beef central temperature as 2~4 DEG C using after -18~-20 DEG C of freezing more than 24h at 2~4 DEG C
Ring.
Exist in some embodiments, the step (2) includes：Beef after defrosting is carried out to blend processing, then will extraction
Agent is mixed with hamburger and shaken overnight, is removed extractant in 40~50 DEG C of rotary evaporations afterwards, is realized the extraction of fat；It is described
Extractant include petroleum ether, its boiling point is 30~60 DEG C.
In some preferred embodiments, the step (2) includes：Radical scavenger is fatty according to 1 with extracting:
10 volume ratios mix in ESR is managed, and timing entering ESR test chambers since ESR pipes, and every 2~5min is between time
Every record once, electron spin resonance detection collection of illustrative plates is obtained.
Preferably, the step (2) includes：Every 2~5min is that a time interval records once, and each fat sample will
The electron spin resonance of acquisition is asked to detect collection of illustrative plates as 15~25, that is, the time interval number for requiring each fat sample measure is
15~25.In some embodiments, the information of electron spin resonance detection collection of illustrative plates include free radical peak height value, from
By base resultant spin number, spin concentration and induction time.
Further, foregoing free radical resultant spin number, the computational methods of spin concentration are to take the spin in Xenon softwares
Quantitative function carries out Integral Processing to spectral line, and spin sum and spin concentration is calculated by computer.And induction time
Free radical is calculated as the time corresponding to the flex point in slow growth interval and rapid growth section.
In some embodiments, the step (3) includes：Determine and selected certainly in the electron spin resonance detection collection of illustrative plates
By the peak intensity situation of change of base peak, the sign for beef fat quality comparison during multigelation is realized.
Further, the selected free base peak is the central peak of multiplet.More specifically, if the peak number of multiplet is
Odd number, take central peak.If the peak number of multiplet is even number, the higher peak in center both sides is taken.
In some embodiments, the step (3) includes：Defined with the size of free base peak peak height value in seclected time
For free radical intensity, induction time or free radical quantity, so as to realize for beef fat quality comparison during multigelation
Sign.
In some preferred embodiments, the temperature in ESR detection process in ESR cavitys is 95~100 DEG C.
Preferably, the step (2) includes：The toluene solution of radical scavenger is fatty in ESR pipes with extracting
Mix, insert ESR test chambers afterwards and detected.
Preferably, the concentration of the toluene solution of the radical scavenger is 0.1wt%~0.7wt%.
Further, the radical scavenger includes the N- tert-butyl groups -- phenyinitrone (PBN) or 5,5- dimethyl -1- pyrroles
Cough up quinoline nitrogen oxides (DMPO) etc., and not limited to this.
Than prior art, the present invention is using beef fat product during electron spin resonance (ESR) sign multigelation
Qualitative change, there is the advantages that detection sensitivity is high, and analysis solvent consumption is few, and sample treatment is simple, can be quickly and efficiently to anti-
Beef fat quality is detected in multiple frozen-thaw process.
Brief description of the drawings
Fig. 1 a- Fig. 1 c respectively illustrate peroxide value fatty in beef after different multigelations in the embodiment of the present invention 1,
TBARS and acid value situation of change.
Fig. 2 shows ESR Free Radical Signal spectrograms fatty in beef after seven Frozen-thawed cycleds in the embodiment of the present invention 1
Change, wherein at interval of 5 minutes records once until 50 minutes.
Fig. 3 shows in the embodiment of the present invention 1 free radical of the fat when heating 50min in the beef of different Frozen-thawed cycleds
Signal intensity.Fig. 4 shows the measure collection of illustrative plates of free radical intensity in the embodiment of the present invention 1.
Fig. 5 shows the measure collection of illustrative plates of induction time in the embodiment of the present invention 1.
Embodiment
As it was previously stated, in view of the defects of prior art, inventor is able to propose this through studying for a long period of time and largely putting into practice
The technical scheme of invention, it is mainly that a kind of electron spin resonance (Electron Spin Resonance, abbreviation ESR) characterizes
The method of beef fat quality comparison during multigelation, it has the features such as method is sensitive, and sample treatment is simple.
In some more specific embodiment, described method comprises the following steps：
(1) determination of testing conditions：
The parameter setting of electron spin resonance (ESR)；
(2) sample detection
A, multigelation processing is carried out to beef, fat-extraction is carried out to the beef after defrosting；
B, the fat of extraction is subjected to the measure of free radical intensity, quantity and induction time etc.；
(3) electron spin resonance (ESR) spectrum analysis
Electron spin resonance (ESR) spectrogram of acquisition is analyzed and processed accordingly.
Further speaking, in abovementioned steps (1), the parameter of electron spin resonance (ESR) can be configured to：Center magnetic
Field (Center Field) is 3350.00G；Field sweep width (Sweep Width) is 100.00G；Microwave power (Microwave
Power) it is 20m W；Modulation amplitude (Modulation Amplitude) is 1.000G；Receiver gain (receiver
gain)30dB；Time constant (time constant) 0.01ms；Conversion time (conversion time) 40.96ms；X frequencies
Band frequency (x-band frequency) 9.42GHz.
Further speaking, abovementioned steps (2) can include：It is (cold with -18~-20 DEG C that multigelation processing is carried out to beef
Freeze after more than 24h and to thaw that to be 2~4 DEG C to beef central temperature be a Frozen-thawed cycled at 2~4 DEG C, specifically for example with -20 DEG C
More than defrosting 20h is a Frozen-thawed cycled at 4 DEG C after freezing more than 24h).Beef after defrosting is carried out to blend processing, utilized
Petroleum ether (30~60 DEG C of boiling point) mixes with hamburger, is placed in shaking table concussion overnight, utilizes vacuum rotary evaporator recovery stone afterwards
Oily ether, obtain fat.
Further, abovementioned steps (2) can also include：According to the toluene solution of radical scavenger, (concentration is about
0.1wt%~0.7wt%) and fatty volume ratio 1:10 mix, and start timing into ESR cavitys with ESR pipes, between per certain time
(recorded once for example, every 2~5min is a time interval, each fat sample requires the electronics obtained certainly every recording once
Rotation resonance detection collection of illustrative plates is 15~25, that is, the time interval number for requiring each fat sample measure is 15~25), obtain
All spectrograms, information therein include peak height value, free radical resultant spin number, spin concentration and induction time etc..
Further speaking, abovementioned steps (3) can include：
A, selection and fixation of the free radical intensity on peak are determined.
B, size, induction time length or the free radical of peak height value in characterizing method selection a period of time of free radical intensity
Quantity.
Technical scheme is described in further details with reference to embodiment and accompanying drawing, but the present invention and not only
It is limited only to following embodiments.
In the following example, stripping and slicing (4cm × 4cm × 4cm) encapsulation freezing processing is carried out to beef first.With -20 DEG C
More than defrosting 20h is a Frozen-thawed cycled at 4 DEG C again after freezing more than 24h.
In the following example, using following testing conditions：
The parameter setting of electron spin resonance (ESR)：Central magnetic field (Center Field) is 3350.00G；Field sweep width
(Sweep Width) is 100.00G；Microwave power (Microwave Power) is 20m W；Modulation amplitude (Modulation
Amplitude) it is 1.000G；Receiver gain (receiver gain) 30dB；Time constant (time constant)
0.01ms；Conversion time (conversion time) 40.96ms；X frequency bands (x-band frequency) 9.42GHz.
The present embodiment of embodiment 1 implements 7 Frozen-thawed cycleds (abbreviation freeze thawing group) to raw beef, to complete ox of thawing
Meat carries out blending processing, is mixed using petroleum ether (30~60 DEG C of boiling point) with hamburger, is placed in shaking table concussion overnight, rear using true
Empty Rotary Evaporators reclaim petroleum ether at 40~50 DEG C, obtain fat.The fat of 200 μ L extractions and 20 μ L PBN are added respectively
Into ESR pipes, shake until being sufficiently mixed.The measure of free radical intensity, quantity and induction time is carried out, and obtains beef fat
ESR spectrograms.
Beef during the present embodiment also demonstrates multigelation using peroxide value, TBARS and acid value as evaluation index
The mass change of fat.
Determination of POV determines according to the method in GB5009.227-2016：Beef sample 2g~3g (is accurate to
0.001g) it is placed in 250mL iodine flasks, adds 30mL chloroforms-glacial acetic acid liquid (volume ratio 40：60) mixed liquor, gently shake
Shake, be completely dissolved fat.It is accurate to add 1.00mL saturated solution of potassium iodide, bottle cap is stoppered, and the 0.5min that gently shakes, in the dark
Place 3min.Take out and add 100mL water, (peroxide value estimate is in 0.15g/ with sodium thiosulfate standard solution immediately after shaking up
100g and it is following when, with 0.002mol/L standard liquids, when peroxide value estimate is more than 0.15g/100g, use 0.01mol/L
Standard liquid) titration separate out iodine, when being titrated to faint yellow, add the starch indicators of 1mL 1%, continue titrate and shake strongly to
It is terminal that solution blueness, which disappears,.Enter line blank test simultaneously.Blank test consumes 0.01mo1/L hypo solution volumes
V0It must not exceed 0.1mL.
X1(mmol/kg)=C × (V-V0)×0.1269×100/m
X1- peroxide value, unit are gram every hectogram (g/100g)
The sodium thiosulfate standard solution volume of V-sample consumption, unit is milliliter (mL)
The sodium thiosulfate standard solution volume of V0-blank test consumption, unit is milliliter (mL)
The concentration of c-sodium thiosulfate standard solution, unit are mole every liter (mol/L)
0.1269-suitable with 1.00mL sodium thiosulfate standard titration solution [c (Na2S2O3)=1.000mol/L]
The quality of iodine
M-sample mass, unit are gram (g)
100-conversion coefficient.
TBARS measure according to：Take 5g meat samples mixed with 10mL 7.5%TCA-EDTA (0.1%) after high-speed homogenization, filtering,
Supernatant 5ml is taken, adds 5ml TBA (2.88g/L) solution.Taken out in boiling water bath after water-bath 40min, put room temperature cooling.So
Above-mentioned reaction solution is transferred to clean colorimetric cylinder afterwards, the shaking of 5ml chloroforms is added, takes supernatant after stratification, carry out ultraviolet inspection
Survey, obtain wavelength as the light absorption value at 532nm.Standard curve is drawn with 1,1,3,3- tetraethoxypropanes.
Acid value determination determines according to the method in GB5009.229-2016：Clean 250mL conical flask is taken, is pressed
According to 10g or so samples, its quality m units are gram.Add ether-isopropanol (volume ratio 1:1) mixed liquor 50mL~100mL and 3
The phenolphthalein indicator of the drop of drop~4, shake well dissolved samples.Titrated again with equipped with 0.1mol/L standard titration solutions, work as sample
Solution begins to show blush, and when in 15s without obvious fade, for the terminal of titration.Stop titration at once, record this titration and disappeared
The milliliter number of the standard titration solution of consumption, this numerical value are V.
The another conical flask for taking a clean 250mL is accurate to add same volume, identical type when being determined with said sample
Organic solvent mixed liquor and indicator, shaking mix.Titrated again with equipped with 0.1mol/L standard titration solutions, work as sample solution
Begin to show blush, and when in 15s without obvious fade, for the terminal of titration.Stop titration at once, record what this titration was consumed
The milliliter number of standard titration solution, this numerical value are V0。
XAV=(V-V0)*c*56.11/m
XAV- acid value, unit are every gram of milligram (mg/g)
The volume for the standard titration solution that V-Specimen Determination is consumed, unit are milliliter (mL)
V0The volume for the standard titration solution that-blank determination accordingly is consumed, unit is milliliter (mL)
The molar concentration of c-standard titration solution, unit are mole every liter (mol/L)
The molal weight of 56.1-potassium hydroxide, unit are gram every mole (g/mol)
The sample weighting amount of m-oil sample, unit are gram (g).
Acid value≤1mg/g, result of calculation retain 2 decimals；1mg/g<Acid value≤100mg/g, result of calculation retain 1 small
Number；Acid value>100mg/g, result of calculation retain to integer-bit.
In addition, Frozen-thawed cycled all corresponds to control group (beef for being constantly in freezing state) as a comparison every time, other
Experiment condition is identical with freeze thawing group.
Fig. 1 a- Fig. 1 c and table 1 are referred to, test result is shown, with the increase of number of freezing and thawing, fatty peroxide value,
Increase change is all presented in TBARS and acid value.Illustrate the increase with number of freezing and thawing, deterioration occurs for the fatty quality of beef.Together
When, Fig. 2, Fig. 3 and table 1 are referred to, with the increase of number of freezing and thawing, increase is also presented in the intensity of free radical.Fat quality is commented
Valency index carries out correlation analysis with free radical intensity to be obtained, and the free resonance (ESR) of electronics can accurately characterize multigelation
To beef fat quality comparison.
Correlation analysis between the Fat quality evaluation index of table 1 and ESR
Embodiment 2：It is identical with embodiment 1, but radical scavenger PBN is replaced with into DMPO.
It should be appreciated that above preferred embodiment is merely to illustrate present disclosure, in addition, the present invention also has other
Embodiment, as long as those skilled in the art because of technical inspiration involved in the present invention, and use equivalent substitution or equivalent deformation
The technical scheme that mode is formed is all fallen within protection scope of the present invention.
Claims (10)
1. a kind of method that ESR characterizes beef fat quality comparison during multigelation, it is characterised in that including：
(1) electron spin resonance running parameter is set, and described running parameter includes central magnetic field intensity, field sweep width, microwave
Power, modulation amplitude, receiver gain, time constant, conversion time, x frequency bands；
(2) multigelation processing is carried out to beef, then the extraction of fat is carried out to the beef after defrosting, afterwards using electron spin
Resonate and the fat of extraction is measured, the object of measure includes free radical intensity, free radical quantity or induction time；
(3) the electron spin resonance spectroscopy figure of acquisition is analyzed and processed, so as to realize for beef fat during multigelation
The sign of fat quality comparison.
2. according to the method for claim 1, it is characterised in that：Step (1) the central magnetic field intensity is arranged to
3350.00G, field sweep width are arranged to 100.00G, and microwave power is arranged to 20mW, and modulation amplitude is arranged to 1.000G,
Receiver gain is arranged to 30dB, and time constant is arranged to 0.01ms, and conversion time is arranged to 40.96ms, x frequency bands frequency
Rate is arranged to 9.42GHz.
3. according to the method for claim 1, it is characterised in that：In the step (2), carried out to beef at multigelation
During reason, be by beef using thawed after -18~-20 DEG C of freezing more than 24h at 2~4 DEG C to beef central temperature as 2~
4 DEG C are a Frozen-thawed cycled.
4. the method according to claim 1 or 3, it is characterised in that the step (2) includes：Beef after defrosting is entered
Row blends processing, then extractant is mixed with hamburger and shaken overnight, removes extractant in 40~50 DEG C of rotary evaporations afterwards,
Realize the extraction of fat；Described extractant includes the petroleum ether that boiling point is 30~60 DEG C.
5. according to the method for claim 1, it is characterised in that the step (2) includes：By radical scavenger and extraction
Fat according to 1:20~1:10 volume ratios ESR manage in mix, and since ESR pipes enter ESR test chambers timing, every 2
~5min is that a time interval records once, obtains electron spin resonance detection collection of illustrative plates；Preferably, in ESR detection process
Temperature in ESR cavitys is 95~100 DEG C.
6. according to the method for claim 5, it is characterised in that the step (2) includes：Every 2~5min is a time
Once, the electron spin resonance detection collection of illustrative plates that each fat sample requires to obtain is 15~25 to interval record, that is, requires each
The time interval number of fat sample measure is 15~25.
7. the method according to claim 5 or 6, it is characterised in that：The packet of the electron spin resonance detection collection of illustrative plates
Include peak height value, free radical resultant spin number, spin concentration and the induction time of free radical.
8. according to the method for claim 1, it is characterised in that the step (3) includes：Determine the electron spin resonance
The peak intensity situation of change that free base peak is selected in collection of illustrative plates is detected, is realized for beef fat quality comparison during multigelation
Sign.
9. according to the method for claim 1, it is characterised in that the step (3) includes：With free base peak in seclected time
The size of peak height value is defined as free radical intensity, induction time or free radical quantity, so as to realize for during multigelation
The sign of beef fat quality comparison.
10. according to the method for claim 5, it is characterised in that：The radical scavenger includes the N- tert-butyl groups -- phenyl
Nitrone or 5,5- dimethyl -1- pyrrolin nitrogen oxides；Preferably, the step (2) includes：By the toluene of radical scavenger
Solution and the fat of extraction mix in ESR is managed, and insert ESR test chambers afterwards and are detected；Preferably, the free radical is caught
The concentration for obtaining the toluene solution of agent is 0.1wt%~0.7wt%.
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| CN108535306A (en) * | 2018-03-05 | 2018-09-14 | 大连工业大学 | A method of detection seeweed polyphenol is to biscuit lipid oxidation inhibition |
| CN109655514A (en) * | 2018-12-29 | 2019-04-19 | 合肥工业大学 | A kind of method of quick detection beef freeze thawing degree |
| CN110220965A (en) * | 2019-07-17 | 2019-09-10 | 山东省分析测试中心 | A kind of method of quick identification multigelation meat |
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