CN101349684B - Method for forecasting UHT milk shelf life - Google Patents
Method for forecasting UHT milk shelf life Download PDFInfo
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- CN101349684B CN101349684B CN2008101466272A CN200810146627A CN101349684B CN 101349684 B CN101349684 B CN 101349684B CN 2008101466272 A CN2008101466272 A CN 2008101466272A CN 200810146627 A CN200810146627 A CN 200810146627A CN 101349684 B CN101349684 B CN 101349684B
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Abstract
The invention relates to a method for predicting shelf period of UHT milk goods, which comprises the following steps: setting the content of free amino nitrogen in UHT milk under normal temperate to be a shelf period reference value, and the predicting coefficient under the shelf period reference value is K, detecting the content of free amino nitrogen in UHT milk which is actually stored, when the content value of free amino nitrogen which is detected reaches the shelf period reference value, using the actual storage time of the UHT milk to divide the predicting coefficient K, and getting the shelf period of the UHT milk which is stored under normal temperature. The invention establishes a method for using the content of free amino nitrogen to detect the shelf period of UHT milk, when the new products of UHT milk are developed, the method can be used to assure the shelf period in shorter times, reduces the development period and the investment of human and material resources, and leads the developmental achievement to be conversed into economic benefits as soon as possible.
Description
Technical field
The present invention relates to a kind of method of predicating UHT milk shelf life, especially a kind of method of utilizing the content that detects UHT Ruzhong FAN to come predicating UHT milk shelf life.
Background technology
UHT (Ultra High Temperature) breast is a kind of product that occurs in the sixties in 20th century, and it is meant the dairy product that the technological process that adopts UHTS and sterile filling processes.
Compare with Pasteur's breast, the UHT breast has solved the problem of liquid newborn transportation, storage, fresh-keeping difficulty, has kept nutriments such as protein, calcium effectively.Its characteristic is: product can reach commercial sterilization, only needs to preserve at normal temperatures, and distribution does not need cold chain, shelf life long (generally can store 1～6 month), and the nutrition in Ruzhong (except indivedual water soluble vitamins) is identical with pasteurization milk basically.UHT compares with traditional pasteurize, can significantly reduce the loss of cow's milk color, local flavor and nutritive value, thereby becomes present the best and processing mode with fastest developing speed.
The time that food shelf index futures product can be stored, the time span of client's quality requirements can accepted and satisfied to product quality in period of storage.Shelf life accelerated tests (ASLT, Accelerated shelf-lifetesting) mainly is to the product of long shelf life long inefficient problem of cycle and a kind of experimental technique of growing up under study for action.Be product to be placed quicken its reaction under the fierce condition, make the real shelf life that (is shorter than the time of its real shelf life) in the short period of time and dopes product.The condition that accelerated test is used mainly contains: the temperature cycles of high temperature, high humidity, illumination, more strictness and high concussion etc., use high humidity, high temperature and illumination in food service industry more, and wherein the use of temperature accelerated test is comparatively extensive.
In recent years, businessman is devoted to study the product of long shelf life in order to obtain to sell interests, for example receives shelf life extension (the Extended shelf-life ESL) breast of domestic and international extensive concern in recent years, i.e. the ESL breast.The product of research long shelf life has become a kind of development trend.
At present; All do not detect the particularly method of UHT milk shelf life of cow's milk in GB and the industry standard, for the judgement of UHT new product shelf life, mainly rely on factors such as the comprehensive sterilization mode of research and development person, packaged form now; Provide empirical value; Afterwards with product in the normal temperature condition held, routine observation is confirmed the shelf life of product with this.The result that this method is judged has very strong subjectivity, and judges chronicly, needs to verify over 6-8 month the accuracy of judgement usually.The time that research needs in the process of improving the UHT breast and developing new product is long, and drain on manpower and material resources is bigger.
Fibrinolysin (plasmin in the cow's milk; PL) be mainly derived from blood, in the Ruzhong casein played hydrolytic action, it can be hydrolyzed to β-CN, the α s-CN in Ruzhong the peptide chain of γ-CN and some weak points; Its optimal pH is 7.4～7.5, and optimum temperature is 37 ℃.This enzyme in the Ruzhong major part with the proenzyme of non-activity (plasminogen, PG) form exists, breast store or processing procedure in, PG can change activated PL under the effect of activator.The Ruzhong also exists the suppressant of this enzyme and proenzyme, and they are limiting the activity of PL to a certain extent.
Research shows that PL and PG have higher thermotolerance, and pasteurize and UHT sterilization all can not make their complete deactivations.UHT breast through direct pasteurization has 19% PL activity residual after sterilization, obvious at the proteolysis in the UHT of direct pasteurization breast lay up period Ruzhong simultaneously.(138 ℃, UHT Ruzhong 2.4s) still finds to have the PL of small amount of residual active handling through indirect heating.
Because the high-fire resistance of PL makes it in the storage process of dairy products, can continue the performance hydrolytic action, and the quality in the dairy products storage process is impacted.UHT breast lay up period PL hydrolysis Ruzhong albumen discharges the FAN that produces bitter taste, can cause harmful effect to the local flavor of breast.
Summary of the invention
The fundamental purpose of the present invention's research is to set up a kind of method with FAN content prediction UHT milk shelf life, through this method, but judges the shelf life that UHT is newborn within a short period of time.
The invention provides a kind of method of predicating UHT milk shelf life, comprise the steps:
1) UHT Ruzhong FAN content value is as the shelf life reference value under normal temperature of setting, and the predictive coefficient under the above-mentioned shelf life reference value is K;
2) the free amine group nitrogen content in the actual UHT Ruzhong of storing of detection; When the FAN content value that obtains when detection reaches above-mentioned shelf life reference value; With this moment UHT breast actual period of storage divided by above-mentioned predictive coefficient K, can obtain the UHT breast shelf life of being stored at normal temperatures.
Said predictive coefficient is measured by following steps:
1.1) the regular content that detects the UHT Ruzhong FAN of storing under actual UHT breast of storing and the normal temperature simultaneously;
1.2) the UHT breast of noting actual storage respectively and normal temperature time T 1, the T2 when the UHT Ruzhong FAN content value of storage reaches above-mentioned shelf life reference value down;
1.3) predictive coefficient K=T1/T2 under the said shelf life reference value.
Abovementioned steps 2) and/or step 1.1) in, detected the free amine group nitrogen content in the UHT Ruzhong once stored in every 3-5 days.
Predictive coefficient K under the said shelf life reference value is 0.165-0.185, is preferably 0.17-0.18.
Said shelf life reference value is 1.95-2.00mmol/L.
The storage temperature of the said actual UHT breast of storing is 40-50 ℃, is preferably 45 ℃, and said normal temperature is 25 ℃.
Among the present invention, the method that detects the free amine group nitrogen content in UHT Ruzhong is:
A.) prepare the FAN standard solution of a plurality of variable concentrations, and detect the absorbance of each standard solution respectively,, draw the content of FAN and detect the typical curve between the absorbance according to measured data;
B.) absorbance of the UHT breast of the said storage of detection with absorbance and the comparison of above-mentioned typical curve that detection obtains, promptly obtains the content of the UHT Ruzhong FAN of above-mentioned storage.
During the preparation standard curve, adopt the L-phenylalanine standard substance of FAN as a reference in the step a), the preparation L-phenylalanine WS; Concentration is respectively: 0mmol/L, 0.5mmol/L, 1.0mmol/L, 1.5mmol/L, 2.0mmol/L, as the standard solution of drawing standard curve, respectively get the standard solution 150 μ L of above-mentioned variable concentrations; Add 3mL OPA (OPA) reagent of deriving respectively; Mixing at room temperature reacts 3min, and 340nm measures each absorbance; According to the data of gained, draw the content of FAN and the typical curve between the absorbance.OPA must be at present with join at present.
Above-mentioned steps b) sample treatment of UHT breast is: get the UHT breast of 5.00mL, add the trichloroacetic acid 10.00mL of 12% (0.75N), add 1.00mL distilled water, vibration shakes up, and centrifugal (2000g/min) 15min gets supernatant liquor.Above-mentioned steps b) assay method of UHT Ruzhong FAN is in: get the supernatant liquor 150 μ Ls of UHT breast to be measured after centrifugal and directly add in the cuvette; Add 3mL OPA reagent, shake mixing repeatedly, at room temperature react 3min; 340nm measures its absorbance; Absorbance of measuring and above-mentioned typical curve are contrasted,, calculate the content of UHT Ruzhong FAN.
At lay up period, plasmase can continuous decomposition of protein at cow's milk, produces FAN, so the free amine group nitrogen content can increase along with the increase of period of storage, and among the present invention, the terminal point concentration of said FAN is 1.95-2.00mmol/L.Under 25 ℃ of normal temperature and 45 ℃ of two temperature conditions, store the UHT breast respectively; When its free amine group nitrogen content reaches 1.95-2.00mmol/L, the record period of storage, with the storage time under the normal temperature condition divided by period of storage under 45 ℃ of conditions; Obtain coefficient 0.165-0.185, preferred 0.17-0.18.When the content of FAN reached terminal point concentration, the record period of storage divided by 0.165-0.185, preferably promptly obtained the shelf life of the said UHT breast under 25 ℃ with said period of storage divided by 0.17-0.18.
The present invention when exploitation UHT breast new product, predicts its shelf life with FAN content detection UHT milk shelf life within a short period of time in this way, reduces the input of R&D cycle and manpower and materials, makes the research and development achievement be converted into economic benefit as early as possible.Method of the present invention can also be used to detecting the shelf life of existing procucts.Method detection speed of the present invention is fast, needs to accomplish usually in 10-20 days, and general needs 2-6 individual month with existing method, and error is less than 10 days, and related coefficient is higher, R
2>=0.75, computing method are simple.
Description of drawings
Fig. 1 is the typical curve of free ammonia nitrogen in one embodiment of the present of invention.
Embodiment
Embodiment of the present invention is intended to explain invention; Rather than restriction protection scope of the present invention; Any those of ordinary skill in this area all should be able to know clearly and understand; Disclosed proportional range can complete embodiment of the present invention inerrably to specifications; Any difference slightly on the numerical value that causes because of the difference in cow's milk breast source should so the content that the present invention is intended to protect utilization to detect UHT Ruzhong FAN comes the method for predicating UHT milk shelf life, rather than will especially strictly not be limited to ratio on the some concrete numerical value as the different slightly reason of testing result of the present invention.
Embodiment 1:
From producing, get the UHT breast sample of two different batches, numbering A and B, respectively at 45 ℃ and normal temperature (25 ℃) preservation, per five days difference sample thiefs detect the content of FAN, and it is following to detect step:
1, drawing standard curve.Adopt the L-phenylalanine standard substance of FAN as a reference.Prepare the WS of L-phenylalanine respectively, concentration is respectively: 0mmol/L, 0.5mmol/L, 1.0mmol/L, 1.5mmol/L, 2.0mmol/L, and as the standard solution of drawing standard curve.The standard solution 150 μ L that respectively get above-mentioned variable concentrations directly add in the cuvette; Add 3mL OPA (OPA) reagent of deriving; Shake mixing repeatedly, at room temperature react 3min, 340nm measures its absorbance (adopting the Japanese SHIMADZU UV-1700 of company type spectrophotometer).Wherein, the collocation method of OPA is: accurately take by weighing the 40mg OPA and be dissolved in the 1mL methyl alcohol, add the 25mL100mmol/L sodium tetraborate, and 2.5mL10%SDS (sodium dodecylsulphonate), 100 μ L beta-mercaptoethanols, adding distil water is settled to 50mL.OPA must be at present with join at present.The gained typical curve is as shown in Figure 1.
2, the pre-treatment mode of UHT breast to be measured is: the UHT breast of accurately getting 5.00mL; The trichloroacetic acid 10.00mL that adds 0.75mol/L adds 1.00mL distilled water, and vibration shakes up; Centrifugal (LXJ-IIB type low speed high capacity multitube hydro-extractor) be 15min (2000g/min), and it is subsequent use to get supernatant liquor.
3, the assay method of UHT Ruzhong FAN is: the supernatant liquor 150 μ L that get testing sample directly add in the cuvette; Add 3mL OPA reagent; Shake mixing repeatedly; At room temperature react 3min, 340nm measures its absorbance (adopting the SHIMADZU UV-1700 of company type spectrophotometer).The comparison typical curve with absorbance substitution typical curve equation, calculates the content of UHT Ruzhong FAN.
4, measure the predictive coefficient K of UHT milk shelf life: set 1.95-2.00mmol/L as a shelf life reference value; Measured in per five days one time 45 ℃ store the newborn free amine group nitrogen content of UHT with normal temperature (25 ℃); When the UHT Ruzhong of 45 ℃ of storages free amine group nitrogen content reaches 1.96mmol/L; Write down time T 1:20 days that have stored; When the UHT Ruzhong of storage at normal temperature free amine group nitrogen content reached 1.95mmol/L, time T 2:115 days of storing of record were about 0.17 with 20 days divided by the predictive coefficient K=T1/T2 that can obtain under the above-mentioned shelf life reference value in 115 days.
The UHT of two storage temperatures breast was measured once free amine group nitrogen content separately in per five days; After product arrives the shelf life terminal point; Value and period of storage with the free amine group nitrogen content of all mensuration are done linear regression analysis, and two groups of UHT milk products have been chosen in this experiment, and group is respectively A, B.Linear regression analysis result between UHT breast lay up period free amine group nitrogen content and the period of storage is as shown in table 1:
Table 1
Firstenberg-Eden thinks, when regression coefficient more than 0.85, i.e. R
2>=0.7225 o'clock, it was good to can be considered correlativity, can be used for Food Inspection.Thereby the UHT lactoprotein percent hydrolysis that obtains 45 ℃ of storages is about 6 times of UHT lactoprotein percent hydrolysis of storage at normal temperature.
From producing, respectively at 45 ℃ and normal temperature (25 ℃) preservation, per three days difference sample thiefs detect the content of FAN with sample, and it is following to detect step:
1, preparation standard curve.Adopt the L-phenylalanine standard substance of FAN as a reference.Prepare the L-phenylalanine WS respectively, concentration is respectively: 0mmol/L, 0.5mmol/L, 1.0mmol/L, 1.5mmol/L, 2.0mmol/L, and as the standard solution of drawing standard curve.The standard solution 150 μ L that respectively get above-mentioned variable concentrations directly add in the cuvette; Add 3mL OPA (OPA) reagent of deriving; Shake mixing repeatedly, at room temperature react 3min, 340nm measures its absorbance (adopting the Japanese SHIMADZU UV-1700 of company type spectrophotometer).Wherein, the collocation method of OPA is: accurately take by weighing the 40mg OPA and be dissolved in the 1mL methyl alcohol, add the 25mL100mmol/L sodium tetraborate, and 2.5mL10%SDS (sodium dodecylsulphonate), 100 μ L beta-mercaptoethanols, adding distil water is settled to 50mL.OPA must be at present with join at present.
2, the pre-treatment mode of UHT breast to be measured is: the UHT breast of accurately getting 5.00mL; The trichloroacetic acid 10.00mL that adds 0.75mol/L adds 1.00mL distilled water, and vibration shakes up; Centrifugal (LXJ-IIB type low speed high capacity multitube hydro-extractor) be 15min (2000g/min), and it is subsequent use to get supernatant liquor.
3, the assay method of UHT Ruzhong FAN is: the supernatant liquor 150 μ L that get testing sample directly add in the cuvette; Add 3mL OPA reagent; Shake mixing repeatedly; At room temperature react 3min, 340nm measures its absorbance (adopting the SHIMADZU UV-1700 of company type spectrophotometer).The comparison typical curve with absorbance substitution typical curve equation, calculates the content of UHT Ruzhong FAN.
4, measure the predictive coefficient K of UHT milk shelf life: set 1.95-2.00mmol/L as a shelf life reference value; Measured in per three days one time 45 ℃ store the newborn free amine group nitrogen content of UHT with normal temperature (25 ℃); When the UHT Ruzhong of 45 ℃ of storages free amine group nitrogen content reaches 1.97mmol/L; Write down time T 1:9 days that store; When the UHT Ruzhong of storage at normal temperature free amine group nitrogen content reached 1.99mmol/L, time T 2:51 days of storing of record were about 0.18 with 9 days divided by the predictive coefficient K=T1/T2 that can obtain under the above-mentioned shelf life reference value in 51 days.
The UHT of two storage temperatures breast was measured once free amine group nitrogen content separately in per three days, after product arrives the shelf life terminal point, do linear regretional analysis with the value and the period of storage of the free amine group nitrogen content of all mensuration.Linear regression analysis result between UHT breast new product lay up period free amine group nitrogen content and the period of storage is as shown in table 2:
Table 2
Firstenberg-Eden thinks, when regression coefficient more than 0.85, i.e. R
2>=0.7225 o'clock, it was good to can be considered correlativity, can be used for Food Inspection.Thereby the UHT lactoprotein percent hydrolysis that obtains 45 ℃ of storages is about 6 times of UHT lactoprotein percent hydrolysis of storage at normal temperature.
Embodiment 3
Adopt the method identical, the free amine group nitrogen content in detection UHT Ruzhong with embodiment 1.
Measured in per five days one time 45 ℃ store the newborn free amine group nitrogen content of UHT with normal temperature (25 ℃); When the UHT Ruzhong of 45 ℃ of storages free amine group nitrogen content reaches 1.96mmol/L; Fallen in the shelf life reference value 1.95-2.00mmol/L scope that sets the 20 days time that record is stored.
The shelf life of prediction shelf life UHT breast: with 20 days predictive coefficient K=0.17-0.18 divided by setting; Can infer and obtain under the normal temperature condition; When the shelf life reference value was set at 1.95-2.00mmol/L, the shelf life of this UHT breast was approximately 111-118 days under the normal temperature.Through detection validation, the actual shelf life under this UHT breast normal temperature is 115 days.
Embodiment 4
Adopt the method identical, the free amine group nitrogen content in detection UHT Ruzhong with embodiment 2.
Measured in per three days one time 45 ℃ store the newborn free amine group nitrogen content of UHT with normal temperature (25 ℃); When the UHT Ruzhong of 45 ℃ of storages free amine group nitrogen content reaches 1.97mmol/L; Fallen in the shelf life reference value 1.95-2.00mmol/L scope that sets the 9 days time that record is stored.
The shelf life of prediction shelf life UHT breast: with 9 days predictive coefficient K=0.17-0.18, can infer to obtain under the normal temperature condition that when the shelf life reference value was set at 1.95-2.00mmol/L, the shelf life of this UHT breast was approximately 50-53 days under the normal temperature divided by setting.Through detection validation, the actual shelf life under this UHT breast normal temperature is 51 days.
Embodiment 5
Adopt the method identical, the free amine group nitrogen content in detection UHT Ruzhong with embodiment 1.
Measured in per five days one time 40 ℃ store the newborn free amine group nitrogen content of UHT with normal temperature (25 ℃); When the UHT Ruzhong of 40 ℃ of storages free amine group nitrogen content reaches 1.98mmol/L; Fallen in the shelf life reference value 1.95-2.00mmol/L scope that sets the 15 days time that record is stored.
The shelf life of prediction shelf life UHT breast: with 15 days predictive coefficient K=0.165-0.185 divided by setting; Can infer and obtain under the normal temperature condition; When the shelf life reference value was set at 1.95-2.00mmol/L, the shelf life of this UHT breast was approximately 81-91 days under the normal temperature.Through detection validation, the actual shelf life under this UHT breast normal temperature is 86 days.
Embodiment 6
Adopt the method identical, the free amine group nitrogen content in detection UHT Ruzhong with embodiment 2.
Measured in per three days one time 50 ℃ store the newborn free amine group nitrogen content of UHT with normal temperature (25 ℃); When the UHT Ruzhong of 50 ℃ of storages free amine group nitrogen content reaches 1.99mmol/L; Fallen in the shelf life reference value 1.95-2.00mmol/L scope that sets the 11 days time that record is stored.
The shelf life of prediction shelf life UHT breast: with 11 days predictive coefficient K=0.165-0.185 divided by setting; Can infer and obtain under the normal temperature condition; When the shelf life reference value was set at 1.95-2.00mmol/L, the shelf life of this UHT breast was approximately 59-67 days under the normal temperature.Through detection validation, the actual shelf life under this UHT breast normal temperature is 63 days.
With the method for FAN content detection UHT milk shelf life, but confirm UHT breast new product shelf life within a short period of time, reduce the input of R&D cycle and manpower and materials, make the research and development achievement be converted into economic benefit as early as possible, computing method are simple.
Claims (7)
1. the method for a predicating UHT milk shelf life comprises the steps:
1) sets under the normal temperature UHT Ruzhong FAN content value as the shelf life reference value; Said shelf life reference value is 1.95-2.00mmol/L; Predictive coefficient under the above-mentioned shelf life reference value is K; Predictive coefficient K under the said shelf life reference value is 0.165-0.185, and said predictive coefficient is measured by following steps:
1.1) the regular content that detects the UHT Ruzhong FAN of storing under actual UHT breast of storing and the normal temperature simultaneously;
1.2) the UHT breast of noting actual storage respectively and normal temperature time T 1, the T2 when the UHT Ruzhong FAN content value of storage reaches above-mentioned shelf life reference value down;
1.3) predictive coefficient K=T1/T2 under the said shelf life reference value;
2) the free amine group nitrogen content in the actual UHT Ruzhong of storing of detection; When the FAN content value that obtains when detection reaches above-mentioned shelf life reference value; With this moment UHT breast actual period of storage divided by above-mentioned predictive coefficient K, can obtain the UHT breast shelf life of being stored at normal temperatures.
2. the method for claim 1 is characterized in that, the storage temperature of the said actual UHT breast of storing is 40-50 ℃, and said normal temperature is 25 ℃.
3. the method for claim 1 is characterized in that, said step 2) and/or step 1.1) in, the free amine group nitrogen content in the UHT Ruzhong that detection in every 3-5 days is once stored.
4. like the described method of one of claim 1-3, it is characterized in that the method for the free amine group nitrogen content in the UHT Ruzhong that said detection is stored is:
A.) prepare the FAN standard solution of a plurality of variable concentrations, and detect the absorbance of each standard solution respectively,, draw the content of FAN and detect the typical curve between the absorbance according to measured data;
B.) absorbance of the UHT breast of the said storage of detection with absorbance and the comparison of above-mentioned typical curve that detection obtains, promptly obtains the content of the UHT Ruzhong FAN of above-mentioned storage.
5. method as claimed in claim 4 is characterized in that said step a) comprises the steps:
Adopt the L-phenylalanine standard substance of FAN as a reference, the preparation L-phenylalanine WS, concentration is respectively: 0mmol/L, 0.5mmol/L, 1.0mmol/L, 1.5mmol/L, 2.0mmol/L; Standard solution as the drawing standard curve; Respectively get the standard solution 150 μ L of above-mentioned variable concentrations, add 3mL OPA derivative reagent respectively, mixing; At room temperature react 3min; 340nm measures each absorbance, according to the data of gained, draws the content of FAN and the typical curve between the absorbance.
6. method as claimed in claim 4; It is characterized in that the sample treatment of UHT breast is in the said step b): get the UHT breast of 5.00mL, add the trichloroacetic acid 10.00mL of 0.75mol/L; Add 1.00mL distilled water; Vibration shakes up, and centrifugal 15min gets supernatant liquor under the centrifugal condition of 2000g/min.
7. method as claimed in claim 5 is characterized in that above-mentioned steps b) in the assay method of UHT Ruzhong FAN be: get the supernatant liquor 150 μ Ls of UHT breast to be measured after centrifugal and directly add in the cuvette; Add 3mL OPA derivative reagent; Mixing at room temperature reacts 3min, and 340nm measures its absorbance; With absorbance of measuring and the contrast of above-mentioned typical curve, calculate the content of UHT Ruzhong FAN.
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| CN103063660A (en) * | 2012-12-12 | 2013-04-24 | 内蒙古伊利实业集团股份有限公司 | Method for predicting shelf life of infant milk powder |
| CN106680270B (en) * | 2017-02-14 | 2020-04-10 | 中国科学院兰州化学物理研究所 | Colorimetric analysis method for detecting ornithine |
| CN109738374B (en) * | 2019-01-29 | 2022-07-01 | 云南中烟工业有限责任公司 | Method for predicting shelf life of gum-based chewing tobacco |
| CN110702673A (en) * | 2019-10-22 | 2020-01-17 | 中国科学院新疆理化技术研究所 | Ammonium-radical naked-eye colorimetric detection reagent |
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| US5824494A (en) * | 1994-05-01 | 1998-10-20 | Sirotech Ltd. | Method for enumerating bacterial populations |
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