CN107328767A - The quick determination method of peroxidase in a kind of guaranteed milk - Google Patents
The quick determination method of peroxidase in a kind of guaranteed milk Download PDFInfo
- Publication number
- CN107328767A CN107328767A CN201710542751.XA CN201710542751A CN107328767A CN 107328767 A CN107328767 A CN 107328767A CN 201710542751 A CN201710542751 A CN 201710542751A CN 107328767 A CN107328767 A CN 107328767A
- Authority
- CN
- China
- Prior art keywords
- milk
- cow
- peroxidase
- sample
- determination method
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
Landscapes
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of quick determination method of peroxidase in cow's milk, comprise the following steps：Pretreatment：Take cow's milk to be divided into many parts, carry out the sample that different heat treatments constitute multiple different disposals；Cooling：After the completion of sample treatment, less than 40 DEG C are cooled to；Colour developing record：Hydrogen peroxide and p-phenylenediamine reaction are added, record is changed over time, the different colours that each sample is presented, the condition of the processing of recording step 1 and the color of final milk sample, and correspond；Test and compare：Learnt from else's experience the cow's milk of pasteurize, and added hydrogen peroxide and p-phenylenediamine, the ratio for adding hydrogen peroxide and p-phenylenediamine is identical with step 3；Shake up, react 1 10 minutes；The color data and pretreatment condition obtained with step 3 is contrasted, and obtains the heating degree of cow's milk in pasteurization processes, and peroxidase residual quantity.
Description
Technical field
The present invention relates to a kind of peroxide enzyme assay method, lactoperoxidase in more particularly to a kind of pasteurized milk
Enzyme method for quickly detecting contents, belongs to dairy products detection and analysis field.
Present invention also contemplates that the quick analysis verification method of the pasteurize disposition for cow's milk, can be quick
Verify the treatment temperature and treatment effect of the pasteurize of cow's milk, it is ensured that the best results of pasteurize, nutrient component damages
Reduce as far as possible.
Background technology
Lactoperoxidase (Lactoperoxidase, LP) is naturally present in a kind of hemoprotein in milk,
Content is especially enriched in colostrum.LP has bacteriostatic activity, under conditions of no refrigeration, can be very good to extend guaranteeing the quality for fresh milk
Phase.But pasteurize, the Technology for Heating Processing of ultra high temperature sterilization can cause lactoperoxidase to inactivate, and cause normal temperature dairy products to exist
Sterilization is later to lose natural protection on the contrary.Therefore, the lactoperoxidase in monitoring dairy products can control dairy products
Quality, extend dairy products the securely held time limit.
Majority state judges that the indicant of the pasteurize upper limit is LP, and LP is a kind of interior protoenzyme more stable to heat.Ox
Peroxidase system not only can have inhibitory action to some bacteriums within a certain period of time in breast, while sramana can also be killed
The Gram-negative bacterias such as Salmonella, pseudomonas.Whether judge lactoperoxidase currently without corresponding quick determination method
The ratio withed a hook at the end and retained, also judges the heat intensity of pasteurization milk without method rapid comparison.
Although also having some determination methods of research report on the peroxidase in food, these detection sides
Method can substantially be divided into：ABTS detection methods, TMB detection methods and guaiacol detection method.Above-mentioned three kinds of detection methods belong to specification
The determination method of change, is respectively provided with preferable detection for peroxidase and acts on, with higher accuracy, but it is detected
Process is required for using large-scale plant equipment, and the process of detection and analysis is also extremely numerous and diverse, the need for being unfavorable for quick analysis.
Also have tried to design test paper detecting method carry out peroxidase detection and analysis, but test paper application composition
More complicated, existence and stability is poor, and Detection results are influenceed by test paper quality fluctuation.The Test paper for providing high-quality is usual
Mean higher cost, for largely carrying out the enterprise of fresh milk production of articles, it tends to be difficult to by such Test paper
It is applied among per a batch of Product checking.
In fact, also being closed in presence and cow's milk pasteurize heat treatment process of the nobody for lactoperoxidase
Connection research, although knowing in pasteurization processes can cause lactoperoxidase to inactivate, and enough attention are not proposed but.
The content of the invention
It is an object of the invention to overcome the quick detection analysis method of the lactoperoxidase in the presence of prior art
Missing, and there is provided peroxidase in a kind of cow's milk for the not enough defect of the application for the detection and analysis of lactoperoxidase
Quick determination method.
In order to realize foregoing invention purpose, the invention provides following technical scheme：
A kind of quick determination method of peroxidase in cow's milk, lactoperoxidase enzyme assay method comprises the following steps：
(1) pre-process：Take cow's milk to be divided into many parts, carry out the sample that different heat treatments constitute multiple different disposals.It is preferred that
Many parts of milk samples are not heated or are heated to respectively 60-85 DEG C by ground, constitute the sample of multiple different disposals.Preferably, heat
Mode is heating water bath.
(2) cool down：After the completion of sample treatment, less than 40 DEG C are cooled to.
(3) colour developing record：Hydrogen peroxide and p-phenylenediamine reaction are added, each sample is recorded and changes over time, presentation is not
Same color, the condition of the processing of recording step 1 and the color of final milk sample, and correspond.Preferably, it will can also test
As a result compared with standard color comparison card, be recorded as that data form can be inquired about.
(4) test and compare：Learnt from else's experience the cow's milk of pasteurize, and added hydrogen peroxide and p-phenylenediamine, add hydrogen peroxide and
The ratio of p-phenylenediamine is identical with step 3；Shake up, react 1-10 minutes.
The color data and pretreatment condition obtained with step 3 is contrasted, and obtains cow's milk in pasteurization processes
Heating degree, and peroxidase residual quantity.
Further, take milk/lactogenesis sample to add hydrogen peroxide and p-phenylenediamine, add the ratio of hydrogen peroxide and p-phenylenediamine
It is identical with step 3；Shake up, react 1-10 minutes；The color data and pretreatment condition obtained with step 3 is contrasted, and is obtained
To lactogenesis sample whether by Overheating Treatment and heat treatment degree, and peroxidase residual quantity.Obtained result is to be based on face
The qualitative conclusions whether sample that colour contrast is drawn is heat-treated, and heating strength (step 3 test sample known to some
Pretreatment) within strength range interval, it can instruct to optimize the intensity that milk pasteurize is heat-treated by this conclusion.
The quick determination method of the peroxidase of the present invention, has cast aside general purification of the prior art processing, large-scale
Equipment detection and analysis technique, it is directly anti-with p-phenylenediamine using peroxidase decomposition hydrogen peroxide active in raw milk
The characteristics of cyanic colours should be immediately turned into is tested and analyzed.
The content of the peroxidase obtained in pasteurized milk is calculated by color contrast, obtained result is probably
There is certain difference or difference in the result of one interval range, and traditional chromatography, instrumental method.Although result is probably one
Individual interval range (based on Colorimetric results before certain known two data colour developing result), but can be substantially reflect thing
The degree of the heat-treated of material, the residual quantity of peroxidase therein.
Certain temperature is heated to according to raw milk, peroxidase is destroyed to be inactivated, no color change, but during with placing
Between it is elongated, a small amount of peroxidase reaction of residual gradually becomes the key properties such as shallow cinerous again.Same raw material milk is taken to distinguish
Do not heat, be heated to different temperature, according to the color change and Coloring Time after cooling, record important feature and draw standard
Colorimetric scheme, then actually detected colour developing situation is compared with colorimetric card, whether qualitative reflection lactogenesis is by Overheating Treatment or heated
Degree for the treatment of, the heating degree of pasteurised milk and peroxidase residual quantity.Although the accuracy of testing result is not as good as large scale equipment
Result is tested and analyzed, but relative to large-scale detection device detection process is cumbersome, redundancy detection cycle, is realized rapidly and efficiently
Detection, therefore quick determination method can be referred to as, with more actual application value.
Further, step 1, take cow's milk to be divided into after many parts, be at least divided into more than 4-15 parts, wherein at least, which contains portion, is
Do not heat.Preferably, be divided into 7,8,9,10,11,12,13,14,15,16 or 17 parts, set at multiple different heating
The temperature spot of reason, carries out the pretreatment of extensive differentiation.
Further, step 1, take cow's milk to be divided into after many parts, respectively according to not heating, heating water bath to 60 DEG C, 70 DEG C, 75
DEG C, 79 DEG C, 80 DEG C, 81 DEG C, 82 DEG C, 85 DEG C set and samples and pre-processed.Temperature of the sample spot of setting in pasteurize
More treatment temperature parameter point is nearby set, the need for can preferably realizing for accurate control.The newborn mistake of emphasis monitoring
The denaturation temperature scope of oxide enzyme, realizes the accurate control for testing result.More temperature is set in 80-85 DEG C of interval
Point, cow's milk discoloration difference is big in this interval range, and the residual quantity of peroxidase is because temperature fluctuation is sensitive, more temperature
Point be more beneficial for subsequently for data differentiation, compare.
Further, step 2, processing is quickly cooled to less than 37-45 DEG C after completing, and is preferably cooled to less than 40 DEG C, prevents
Only single dwell course draws in temperature range of the cow's milk temperature reduction Process of Peroxide enzyme after heat treatment temperature to cooling
More uncontrollable rotten destructions are played, accuracy, uniformity that peroxidase is set up in detection are improved, it is to avoid the temperature of heat treatment
Du Genggao sample is destroyed by the thermal metamorphism of longer time.In the temperature range less than 40 DEG C, general peroxidase has
Good biological stability, is unlikely to deteriorate destruction.Being cooled to less than 40 DEG C of time should be no more than 5-300 seconds, preferably not surpass
10-200 seconds are spent, preferably cooldown rate is that sample is cooled to below target temperature within 10-60 seconds.For example cooldown rate can
To be to be cooled to 10, in 12,15,18,20,23,25,28,30,35 or 40 seconds below target temperature.
Preferably, preferably cooled down by the way of water cooling, the characteristics of water cooling has simple and convenient easy implementation, and
Cooling velocity is fast, effect is good.It is preferred that it is for instance possible to use the mode that flowing water washes away is cooled down.Flowing water washes away can be with originally
Water is washed away, and flowing water washes away cooling rate soon, simple and easy to apply.
Further, step 2, the rate of temperature fall of cooling procedure and the temperature of the pasteurize processing during production cow's milk become
Change speed to be consistent, or close to the control parameter condition in cow's milk production process.The speed of cooling is controlled, is allowed to and produces
It is consistent, it is preferably corresponding with production situation of change, improve the accuracy needed for quick detection in production process, it is ensured that
Produce the quality of cow's milk.Complete to be cooled to less than 40 DEG C accordingly according to the temperature fall time of pasteurize.
Further, every 1-6 months, step 1-3, the condition of trailer record processing and the color of final milk sample are updated
Corresponding relation.Different time or the cow's milk endoperoxide enzyme of different batches are denatured mistake present in pasteurization processes
Work there may be certain difference, and reproduction verification is carried out at regular intervals, the accuracy of detection method can be improved, carry simultaneously
Height monitoring analysis personnel improve the lookup and corrigendum for detection error for the sensitiveness of data.
Meanwhile, also provide a kind of according to the quick detection of lactoperoxidase analysis progress research checking pasteurize journey
The quick analysis verification method of the dairy products pasteurize situation (temperature, time) of degree/effect.
The main feature of pasteurize is that sterilization temperature is relatively low, can be very good keep cow's milk in nutrition into
Divide not to be destroyed and be lost in, improve the nutritive value of dairy products.But any heating can all cause the stream of some nutritional ingredients
Lose, so, it is generally desirable to the temperature control of pasteurize in alap scope, reduce the loss of nutritional ingredient.But,
The temperature of pasteurize again can not be too low, to ensure that the microorganism in cow's milk is killed in pasteurize completely.
In order to study pasteurize and above-mentioned cow's milk in peroxidase reserved collaboration, realize dairy products energy
Enough to retain peroxidase as far as possible, there is provided more for intrinsic antibacterial/bactericidal property during the intrinsic storage of raising cow's milk
The shelf-life of good fresh milk, reduce the requirement of preservation condition or reduce the use of antipathogenic composition.Needing one kind badly can reversely divide
The detection method of pasteurize effect is analysed, the optimum condition parameter of pasteurize is studied.Or, accurate reversely analysis production process
The method of middle pasteurizing temperature.
A kind of pasteurizing temperature detection method, is built using the step 1-3 of the quick determination method of above-mentioned peroxidase
The colour developing comparison chart of cow's milk for erecting the temperature of reaction and finally handling, by analyzing the later cow's milk of pasteurize
Colour developing situation, draws the temperature of pasteurize, verifies the true temperature of sterilization, improve production effect and product it is consistent
Property.
A kind of pasteurizing temperature detection method, comprises the following steps：
(1) pre-process：Take cow's milk to be divided into many parts, carry out the sample that different heat treatments constitute multiple different disposals.It is preferred that
Many points of milk samples are separately heated to 60-85 DEG C by ground, constitute the sample of multiple different disposals.Preferably, mode of heating is water
Bath heating.Preferably, the processing time in heat treatment process is identical with the time of pasteurize.Or heat treatment time and Pasteur
The time phase difference of sterilization is within 1-10 seconds, and preferably error is controlled in 2-8 seconds, such as 4, the error within 5,6,7 seconds.
(2) cool down：After the completion of sample treatment, less than 40 DEG C are cooled to.Preferably, when cooling, the speed of cooling and
Cooldown rate in pasteurization processes is consistent, or error is in below 1%-40%, preferably cooldown rate difference 1%-
Less than 30%.Preferably, cooled down using water-cooling pattern.
(3) colour developing record：Hydrogen peroxide and p-phenylenediamine reaction are added, record is changed over time, and each sample is presented not
Same color, the condition of the processing of recording step 1 and the color of final milk sample, and correspond.
(4) test and compare：Learnt from else's experience the cow's milk of pasteurize, and added hydrogen peroxide and p-phenylenediamine, add hydrogen peroxide and
The ratio of p-phenylenediamine is identical with step 3；Shake up, react 1-10 minutes；
The color data and pretreatment condition obtained with step 3 is contrasted, and extrapolates the temperature of pasteurize.
Detection method of the invention for pasteurizing temperature, according to the modifying characteristics of peroxidase, first design sample
The milk sample of accurate known heat treatment temperature is realized in pretreatment, and sample then is carried out into hydrogen peroxide, to benzene two
Amine chromogenic reaction, record obtains the response parameter of characteristic.Response parameter and actual Pasteur finally according to these characteristics
Sterilize cow's milk and carry out visual comparison, extrapolate the intensity of pasteurize in production process.Can be with by the projectional technique of the present invention
The accurate intensity (temperature, time) for knowing pasteurize, and then the parameter of pasteurize is adjusted, realize and processed for dairy products
There is provided more preferable high-quality milk product for the further optimization of technique.
Compared with prior art, beneficial effects of the present invention：
1st, the quick determination method of peroxidase of the invention eliminates existing large scale equipment determination method
Drawback, the reserved of the peroxidase in cow's milk is quickly verified using colorimetric method, with good specific aim, mistake in cow's milk
The detection efficiency of oxide enzyme is significantly improved lifting.
2nd, the quick determination method of peroxidase of the invention can be used for the pasteurize intensity for studying guaranteed milk, more
Good optimization pasteurize parameter so that the nutriment in cow's milk is preferably remained, is reduced in high-temperature process
Nutrient loss.
3rd, whether detection method of the invention can be determined that lactogenesis sample by Overheating Treatment and heat treatment degree, and peroxide
Compound enzyme residual quantity, new reliable detection method is provided for detection milk quality.
Brief description of the drawings：
Fig. 1 is the quick detection result of peroxidase in embodiment 1.
Fig. 2 is the quick detection result of peroxidase in embodiment 2.
Fig. 3 is the quick detection result of peroxidase in embodiment 3.
Fig. 4 is the quick detection result of peroxidase in embodiment 4.
Fig. 5 is the quick detection result of peroxidase in embodiment 5.
Fig. 6 is the quick detection result of peroxidase in embodiment 6.
Embodiment
With reference to test example and embodiment, the present invention is described in further detail.But this should not be understood
Following embodiment is only limitted to for the scope of above-mentioned theme of the invention, it is all that this is belonged to based on the technology that present invention is realized
The scope of invention.
【Embodiment 1】
The quick detection of peroxidase in cow's milk
Opening relationships formula：Take 90ml cow's milk to be divided into 9 equal portions, be placed in 9 test tubes, do not heat or heating water bath to 60 DEG C,
70 DEG C, 75 DEG C, 79 DEG C, 80 DEG C, 81 DEG C, 82 DEG C, 85 DEG C, kept for 15 seconds.Then, water tap down-flow water, which washes away, is cooled to 40
Below DEG C.0.5ml hydrogen peroxide and the reaction of 0.5ml p-phenylenediamine are separately added into, record is changed over time, and each sample is presented not
Same color, record color status are corresponding with heat treatment temperature before with developing time.
Table 1：Peroxidase testing result
| Heating-up temperature (DEG C) | Color change | Peroxidase result | Coloring Time |
| Do not heat | Become navy blue | It is positive | Change colour immediately |
| 60 | Become navy blue | It is positive | Change colour immediately |
| 70 | Become navy blue | It is positive | Change colour immediately |
| 75 | Become light blue | It is positive | Change colour immediately |
| 79 | Become light blue | It is positive | Change colour immediately |
| 80 | Nondiscolouring | It is negative | 3-4min is put to start to become light grey |
| 81 | Nondiscolouring | It is negative | 5min is put to start to become light grey |
| 82 | Nondiscolouring | It is negative | 10min is put to start to become light grey |
| 85 | Nondiscolouring | It is negative | 20min is put to start to become light grey |
| Pasteurised milk * * | Nondiscolouring | It is negative | 20min is put to start to become light grey |
*80-85℃：It is unified after reagent adding to place 10min or 20min observation color distortions.
* bars kill milk：Through 85 ± 2 DEG C, 15s bars are killed.
As a result it is as shown in Figure 1.
Test and compare：Learnt from else's experience the cow's milk or raw milk 10mL of pasteurize, and added 0.5ml hydrogen peroxide and 0.5ml pairs
Phenylenediamine, shakes up, and reacts 1-20 minutes；Color status and the time data progress pair of observation period colour developing situation and above-mentioned record
Than obtaining the heating degree of cow's milk in pasteurization processes, and peroxidase residual quantity.The milk for the treatment of of different temperature,
Color after reagent adding reaction, color and colorimetric card can be contrasted when actually detected the quick intensity that judge pasteurised milk heat-treated and
The level of deactivation of the peroxidase beneficial to pasteurised milk.
【Embodiment 2】
The quick detection of peroxidase in cow's milk
Opening relationships formula：Take 80ml cow's milk to be divided into 8 equal portions, be placed in 8 test tubes, do not heat or heating water bath to 60 DEG C,
65 DEG C, 72 DEG C, 78 DEG C, 80 DEG C, 82 DEG C, 85 DEG C, kept for 15 seconds, while detecting commercially available sterile milk.Then, water tap flows down
Water, which washes away, is cooled to less than 40 DEG C.0.2ml hydrogen peroxide and the reaction of 0.2ml p-phenylenediamine are added, record is changed over time, each sample
The different colours that product are presented, record color status are corresponding with heat treatment temperature before with developing time.
Table 2：Peroxidase testing result
| Heating-up temperature (DEG C) | Color change | Peroxidase result | Coloring Time |
| Do not heat | Become navy blue | It is positive | Change colour immediately |
| 60 | Become navy blue | It is positive | Change colour immediately |
| 65 | Become navy blue | It is positive | Change colour immediately |
| 72 | Become light blue | It is positive | Change colour immediately |
| 78 | Become light blue | It is positive | Change colour immediately |
| 80 | Nondiscolouring | It is negative | 3-4min is put to start to become light grey |
| 82 | Nondiscolouring | It is negative | 10min is put to start to become light grey |
| 85 | Nondiscolouring | It is negative | 20min is put to start to become light grey |
| UHT milk * * | Nondiscolouring | It is negative | 20min is put to start to become light grey |
* UHT milk：Commercially available sterile milk (UHT milk).
Test and compare：Learnt from else's experience the cow's milk or raw milk 10mL of high temperature sterilization, and added 0.2ml hydrogen peroxide and 0.2ml pairs
Phenylenediamine, shakes up, and reacts 1-20 minutes；Color status and the time data progress pair of observation period colour developing situation and above-mentioned record
Than obtaining the heating degree of cow's milk in pasteurization processes, and peroxidase residual quantity.
As a result it is as shown in Figure 2.
【Embodiment 3】
The quick detection of peroxidase in cow's milk
Opening relationships formula：Take 50ml cow's milk to be divided into 10 equal portions, be placed in 10 test tubes, do not heat or heating water bath is to 60
DEG C, 70 DEG C, 75 DEG C, 77 DEG C, 79 DEG C, 81 DEG C, 83 DEG C, 85 DEG C and 90 DEG C, keep 10 seconds.Then, water tap down-flow water is rushed
Brush is cooled to less than 40 DEG C.0.2ml hydrogen peroxide and the reaction of 0.2ml p-phenylenediamine are separately added into, record is changed over time, each sample
The different colours that product are presented, record color status are corresponding with heat treatment temperature before with developing time.
Table 3：Peroxidase testing result
Test and compare：Learnt from else's experience the cow's milk or raw milk 10mL of pasteurize, and added 0.2ml hydrogen peroxide and 0.2ml pairs
Phenylenediamine, shakes up, and reacts 1-20 minutes；Color status and the time data progress pair of observation period colour developing situation and above-mentioned record
Than obtaining the heating degree of cow's milk in pasteurization processes, and peroxidase residual quantity.
As a result it is as shown in Figure 3.
【Embodiment 4】
The quick detection of peroxidase in cow's milk
Opening relationships formula：Take 50ml cow's milk to be divided into 10 equal portions, be placed in 10 test tubes, do not heat or heating water bath is to 70
DEG C, 75 DEG C, 77 DEG C, 79 DEG C, 81 DEG C, 83 DEG C, 85 DEG C, 87 DEG C, 90 DEG C, keep 5 seconds.Then, water tap down-flow water washes away
It is cooled to less than 40 DEG C.0.5ml hydrogen peroxide and the reaction of 0.5ml p-phenylenediamine are separately added into, record is changed over time, each sample
The different colours of presentation, record color status are corresponding with heat treatment temperature before with developing time.
Table 4：Peroxidase testing result
| Heating-up temperature (DEG C) | Color change | Peroxidase result | Coloring Time |
| Do not heat | Become navy blue | It is positive | Change colour immediately |
| 70 | Become navy blue | It is positive | Change colour immediately |
| 75 | Become light blue | It is positive | Change colour immediately |
| 77 | Become light blue | It is positive | Change colour immediately |
| 79 | Become light blue | It is positive | Change colour immediately |
| 81 | Nondiscolouring | It is negative | 3-4min is put to start to become light grey |
| 83 | Nondiscolouring | It is negative | 5min is put to start to become light grey |
| 85 | Nondiscolouring | It is negative | 10min is put to start to become light grey |
| 87 | Nondiscolouring | It is negative | 20min is put to start to become light grey |
| 90 | Nondiscolouring | It is negative | 20min is put to start to become light grey |
Test and compare：Learnt from else's experience the cow's milk of pasteurize, and added hydrogen peroxide and p-phenylenediamine, add hydrogen peroxide and to benzene
The ratio of diamines is consistent with adding usage ratio during opening relationships formula；Shake up, react 1-20 minutes；Observation period colour developing situation
Contrasted with the color status and time data of above-mentioned record, obtain the heating degree of cow's milk in pasteurization processes, and
Peroxidase residual quantity.
As a result it is as shown in Figure 4.
【Embodiment 5】
The quick detection of peroxidase in cow's milk
Opening relationships formula：Take 60ml cow's milk to be divided into 6 equal portions, be placed in 6 test tubes, do not heat or heating water bath to 77 DEG C,
79 DEG C, 81 DEG C, 82 DEG C, 85 DEG C, kept for 10 seconds, be cooled to less than 40 DEG C, while detecting UHT sterile milks.It is separately added into 0.5ml
Hydrogen peroxide and the reaction of 0.5ml p-phenylenediamine, record are changed over time, and the different colours that each sample is presented record color status
It is corresponding with heat treatment temperature before with developing time.
Table 5：Peroxidase testing result
| Heating-up temperature (DEG C) | Color change | Peroxidase result | Coloring Time |
| Do not heat | Become navy blue | It is positive | Change colour immediately |
| 77 | Become light blue | It is positive | Change colour immediately |
| 79 | Become light blue | It is positive | Change colour immediately |
| 81 | Nondiscolouring | It is negative | 3-4min is put to start to become light grey |
| 82 | Nondiscolouring | It is negative | 5min is put to start to become light grey |
| 85 | Nondiscolouring | It is negative | 10min is put to start to become light grey |
| UHT sterile milks | Nondiscolouring | It is negative | 20min is put to start to become light grey |
Test and compare：Learnt from else's experience the cow's milk or raw milk 10mL of high temperature sterilization, and added 0.5ml hydrogen peroxide and 0.5ml pairs
Phenylenediamine, shakes up, and reacts 1-20 minutes；Color status and the time data progress pair of observation period colour developing situation and above-mentioned record
Than obtaining the heating degree of cow's milk in pasteurization processes, and peroxidase residual quantity.
As a result it is as shown in Figure 5.
【Embodiment 6】
The quick detection of peroxidase in cow's milk
Opening relationships formula：Take 90ml cow's milk to be divided into 9 equal portions, be placed in 9 test tubes, do not heat or heating water bath to 70 DEG C,
75 DEG C, 77 DEG C, 78 DEG C, 80 DEG C, 82 DEG C, 83 DEG C, 85 DEG C, kept for 10 seconds, be cooled to less than 40 DEG C, while detecting UHT sterilizings
Breast.0.5ml hydrogen peroxide and the reaction of 0.5ml p-phenylenediamine are separately added into, record is changed over time, the different face that each sample is presented
Color, record color status are corresponding with heat treatment temperature before with developing time.
Table 6：Peroxidase testing result
| Heating-up temperature (DEG C) | Color change | Peroxidase result | Coloring Time |
| Do not heat | Become navy blue | It is positive | Change colour immediately |
| 70 | Become navy blue | It is positive | Change colour immediately |
| 75 | Become navy blue | It is positive | Change colour immediately |
| 77 | Become navy blue | It is positive | Change colour immediately |
| 78 | Become light blue | It is positive | Change colour immediately |
| 80 | Nondiscolouring | It is negative | 3-4min is put to start to become light grey |
| 82 | Nondiscolouring | It is negative | 5min is put to start to become light grey |
| 83 | Nondiscolouring | It is negative | 10min is put to start to become light grey |
| 85 | Nondiscolouring | It is negative | 20min is put to start to become light grey |
| UHT sterile milks | Nondiscolouring | It is negative | 20min is put to start to become light grey |
Test and compare：Learnt from else's experience the cow's milk or raw milk 10mL of high temperature sterilization, and added 0.5ml hydrogen peroxide and 0.5ml pairs
Phenylenediamine, shakes up, and reacts 1-20 minutes；Color status and the time data progress pair of observation period colour developing situation and above-mentioned record
Than obtaining the heating degree of cow's milk in pasteurization processes, and peroxidase residual quantity.
As a result it is as shown in Figure 6.
Above-mentioned multiple embodiments test result indicates that can be by taking cow's milk/milk advance killed similar to Pasteur
Analog heat-treating is carried out in bacterium temperature range, chromogenic reaction is carried out with hydrogen peroxide and p-phenylenediamine, it is established that reliable sample shows
Color change contrast relationship table.Then, same chromogenic reaction is carried out to milk/milk sample of feature, contrasts colour developing situation, point
The residual quantity of the peroxidase in milk/milk sample is analysed, there is important directive significance to lifting milk/Milk Quality.
The data collected using above-described embodiment, same color development treatment is carried out to existing pasteurized milk, is compared
Color change situation, analyzes the heating strength of pasteurized milk, optimization pasteurize intensity and fluctuating change situation, carries
Stable sterilization effect and the uniformity of product in height production.
Claims (9)
1. the quick determination method of peroxidase, comprises the following steps in a kind of cow's milk：
（1）Pretreatment：Take cow's milk to be divided into many parts, carry out the sample that different heat treatments constitute multiple different disposals；
（2）Cooling：After the completion of sample treatment, less than 40 DEG C are cooled to；
（3）Colour developing record：Hydrogen peroxide and p-phenylenediamine reaction are added, each sample is recorded and changes over time, the different face of presentation
Color, the condition of the processing of recording step 1 and the color of final milk sample, and correspond；
（4）Test and compare：Learnt from else's experience the cow's milk of pasteurize, and added hydrogen peroxide and p-phenylenediamine, add hydrogen peroxide and to benzene
The ratio of diamines is identical with step 3；Shake up, react 1-10 minutes；
The color data and pretreatment condition obtained with step 3 is contrasted, and obtains the heated of cow's milk in pasteurization processes
Degree, and peroxidase residual quantity.
2. the quick determination method of peroxidase in cow's milk as claimed in claim 1, it is characterised in that by detecting different heat
The different colours that processing sample is obtained, set up standard control color, lactogenesis or pasteurization milk can be detected in the same manner,
By color contrast, fast qualitative judges the heating degree of cow's milk in pasteurization processes, obtains whether lactogenesis sample is overheated
Processing and heat treatment degree, and peroxidase residual quantity.
3. the quick determination method of peroxidase in cow's milk as claimed in claim 1, it is characterised in that step 1, takes cow's milk point
Into after many parts, wherein at least contains portion and not heated.
4. the quick determination method of peroxidase in cow's milk as claimed in claim 1, it is characterised in that step 1, takes cow's milk point
Into after many parts, respectively according to not heating, heating water bath is set to 60 DEG C, 70 DEG C, 75 DEG C, 79 DEG C, 80 DEG C, 81 DEG C, 82 DEG C, 85 DEG C
Put sample and pre-processed.
5. the quick determination method of peroxidase in cow's milk as claimed in claim 1, it is characterised in that step 2, processing is completed
It is quickly cooled to less than 37-45 DEG C later.
6. the quick determination method of peroxidase in cow's milk as claimed in claim 1, it is characterised in that step 2, cool time
It should be no more than 5-300 seconds.
7. the quick determination method of peroxidase in cow's milk as claimed in claim 1, it is characterised in that step 2, using water cooling
Mode cooled down.
8. the quick determination method of peroxidase in cow's milk as claimed in claim 1, it is characterised in that step 2, cooling procedure
Rate of temperature fall and production cow's milk during pasteurize processing rate temperature change be consistent, or close to cow's milk life
Control parameter condition during production.
9. the quick determination method of peroxidase in cow's milk as claimed in claim 1, it is characterised in that every 1-6 months, more
The corresponding relation of new step 1-3, the condition of trailer record processing and the color of final milk sample.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710380703 | 2017-05-25 | ||
| CN2017103807035 | 2017-05-25 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107328767A true CN107328767A (en) | 2017-11-07 |
Family
ID=60196188
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710542751.XA Pending CN107328767A (en) | 2017-05-25 | 2017-07-05 | The quick determination method of peroxidase in a kind of guaranteed milk |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107328767A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110514654A (en) * | 2018-05-21 | 2019-11-29 | 中国农业科学院北京畜牧兽医研究所 | The rapid detection method of lactoperoxidase in a kind of cow's milk |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1595119A (en) * | 2004-07-06 | 2005-03-16 | 陕西科技大学 | Method for rapid detection of ammonium salts in fresh milk |
| CN101256168A (en) * | 2008-04-22 | 2008-09-03 | 内蒙古蒙牛乳业（集团）股份有限公司 | Method for testing peroxidase content in cow's milk |
| CN101290323A (en) * | 2008-05-20 | 2008-10-22 | 内蒙古蒙牛乳业（集团）股份有限公司 | Cow milk and milk powder nitrite qualitative checking method and reagent kit |
| CN101329280A (en) * | 2008-07-31 | 2008-12-24 | 内蒙古蒙牛乳业（集团）股份有限公司 | Fast testing method of hydrogen peroxide residue in milk-like liquid and testing reagent kit thereof |
| CN101576484A (en) * | 2009-06-08 | 2009-11-11 | 浙江理工大学 | Reagent and method for detecting concentration of hydrogen peroxide |
| CN101793838A (en) * | 2010-04-01 | 2010-08-04 | 陕西科技大学 | Rapid detection method for hydrogen peroxide in fresh milk |
| CN104406977A (en) * | 2014-12-19 | 2015-03-11 | 东北农业大学 | Method for detecting lactoperoxidase in raw milk using test paper |
| CN106008704A (en) * | 2016-08-05 | 2016-10-12 | 内蒙古伊利实业集团股份有限公司 | Method for producing lactoferrin and lactoperoxidase |
-
2017
- 2017-07-05 CN CN201710542751.XA patent/CN107328767A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1595119A (en) * | 2004-07-06 | 2005-03-16 | 陕西科技大学 | Method for rapid detection of ammonium salts in fresh milk |
| CN101256168A (en) * | 2008-04-22 | 2008-09-03 | 内蒙古蒙牛乳业（集团）股份有限公司 | Method for testing peroxidase content in cow's milk |
| CN101290323A (en) * | 2008-05-20 | 2008-10-22 | 内蒙古蒙牛乳业（集团）股份有限公司 | Cow milk and milk powder nitrite qualitative checking method and reagent kit |
| CN101329280A (en) * | 2008-07-31 | 2008-12-24 | 内蒙古蒙牛乳业（集团）股份有限公司 | Fast testing method of hydrogen peroxide residue in milk-like liquid and testing reagent kit thereof |
| CN101576484A (en) * | 2009-06-08 | 2009-11-11 | 浙江理工大学 | Reagent and method for detecting concentration of hydrogen peroxide |
| CN101793838A (en) * | 2010-04-01 | 2010-08-04 | 陕西科技大学 | Rapid detection method for hydrogen peroxide in fresh milk |
| CN104406977A (en) * | 2014-12-19 | 2015-03-11 | 东北农业大学 | Method for detecting lactoperoxidase in raw milk using test paper |
| CN106008704A (en) * | 2016-08-05 | 2016-10-12 | 内蒙古伊利实业集团股份有限公司 | Method for producing lactoferrin and lactoperoxidase |
Non-Patent Citations (3)
| Title |
|---|
| L.W.AURAND.ETAL: "A METHOD FOR THE ESTIMATION OF PEROXIDASE ACTIVITY IN MILK", 《JOURNAL OF DAIRY SCIENCE》 * |
| NICHOLETTE E MARKS.ETAL: "Use of hydrogen peroxide detection strips to determine the extent of pasteurization in whole milk", 《ORIGINAL RESEARCH》 * |
| 李清等: "速率法测定辣根过氧化物酶", 《大连大学学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110514654A (en) * | 2018-05-21 | 2019-11-29 | 中国农业科学院北京畜牧兽医研究所 | The rapid detection method of lactoperoxidase in a kind of cow's milk |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| MUIR | The shelf‐life of dairy products: 1. Factors influencing raw milk and fresh products | |
| Dash et al. | A comprehensive review on heat treatments and related impact on the quality and microbial safety of milk and milk-based products | |
| European Food Safety Authority (EFSA) et al. | The use of alkaline phosphatase and possible alternative testing to verify pasteurisation of raw milk, colostrum, dairy and colostrum‐based products | |
| Tripaldi et al. | Chemical and microbiological characteristics of homogenised ricotta cheese produced from buffalo whey | |
| Hovjecki et al. | Rheological and textural properties of goat's milk set‐type yoghurt as affected by heat treatment, transglutaminase addition and storage | |
| Sosnowski et al. | Alkaline phosphatase activity and microbiological quality of heat-treated goat milk and cheeses | |
| CN107328767A (en) | The quick determination method of peroxidase in a kind of guaranteed milk | |
| Hassan et al. | Microbiological and physicochemical analysis of different UHT milks available in market | |
| Rashtchi et al. | Comparison of the microbial, physicochemical, and sensorial properties of raw and pasteurized Lighvan cheeses during ripening time | |
| US4175141A (en) | Inactivation of heat resistant bacterial proteases in ultra-high temperature treated milk | |
| Stănciuc et al. | γ-Glutamyl transferase inactivation in milk and cream: A comparative kinetic study | |
| Farag et al. | Implementation of HACCP in the production of Egyptian cheeses: A review | |
| US2886450A (en) | Process for preparing sterile evaporated milk | |
| US20160058024A1 (en) | Improved milk and milk-related products | |
| US20140335256A1 (en) | Heat-treatment of milk | |
| CN109060709A (en) | A method of cow's milk heat treatment degree is detected based on infrared spectrum technology | |
| Khayrullin et al. | Study on the effects of different sterilization methods and storage conditions on milk quality | |
| Poushi et al. | State-of-the-art in milk processing for improvement of the quality of pasteurized milk and UHT milk | |
| Aydemir et al. | The effect of heat treatment and starter culture on colour intensity and sensory properties of Kulek cheese | |
| Medveďová et al. | Microbial and senzory quality of raw milk cheeses from the milk vending machines | |
| Skura | Standards, control, and future outlook | |
| Partovi et al. | Technological properties of Lactobacillus plantarum strains isolated from Siahmazgi cheese | |
| Merin et al. | Standards for camel milk | |
| CN112868760B (en) | Method for reducing UHT sterilized milk fat floating degree and complaint rate | |
| CN112244086B (en) | Method for producing high-activity shelf-life-prolonged milk by ultraviolet sterilization and microfiltration |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20171107 |