CN110296964A - A method of based on the mitogenetic cow's milk of fluorescence microscopy detection zone and full-cream reconstituted milk - Google Patents
A method of based on the mitogenetic cow's milk of fluorescence microscopy detection zone and full-cream reconstituted milk Download PDFInfo
- Publication number
- CN110296964A CN110296964A CN201910585109.9A CN201910585109A CN110296964A CN 110296964 A CN110296964 A CN 110296964A CN 201910585109 A CN201910585109 A CN 201910585109A CN 110296964 A CN110296964 A CN 110296964A
- Authority
- CN
- China
- Prior art keywords
- milk
- full
- cream
- reconstituted milk
- cream reconstituted
- 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
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/02—Food
- G01N33/04—Dairy products
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Pathology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a kind of methods based on fluorescence microscope detection technique detection zone mitogenetic cow's milk and full-cream reconstituted milk, are related to food science literature technical field.The method of the present invention are as follows: the fat globule in raw milk and full-cream reconstituted milk is marked with Nile red dye liquor, using fluorescence microscope, specific excitation wavelength and corresponding launch wavelength are set, acquire the image information of raw milk and full-cream reconstituted milk under specific amplification factor, by the difference of fat globule partial size and distribution in observation raw milk and full-cream reconstituted milk, the technology of quickly detection reconstituted milk is established.The invention has the advantages that low in cost, easy to operate quickly measurement result accuracy is high, has broad prospect of application in raw milk Quality Detection.
Description
Technical field
The invention belongs to food science literature technical fields, more particularly to a kind of detected based on fluorescence microscopy to distinguish
The method of raw milk and full-cream reconstituted milk.
Background technique
Raw milk nutritive value is abundant.But it considers the shortage of milk supply, and is not easy to store up without ultrapasteurized raw milk
Hiding and transport, thus have illegal businessman with cheap reconstituted milk to substitute raw milk.Full-cream reconstituted milk through ultra-high temperature sterilization, wherein
Vitamins and other nutritious components be destroyed, calcium ion can also form complex compound in conjunction with the casein in cow's milk, be not easy by human body
Digestion and absorption.
Current research is mostly based on cow's milk some ingredients that content changes during heat treatment (such as newborn fruit
Sugar, chaff propylhomoser, heat denatured protein etc.) measurement, or using the detection techniques such as near infrared spectrum, LC-MS combine chemistry meter
The method that amount is learned carries out non-targeted detection.Such method operation is relative complex, inconvenient for use, and testing cost is high.
Fluorescence microscopy is a kind of with high-resolution optical microscopy, observes the distribution situation of fluorescent material
Technology.Since reconstituted milk experienced the treatment processes such as sterilization, homogeneous and spray drying, the knot of fat globule membrane in process
Structure and composition change, to influence the particle diameter distribution of fat.Therefore, raw milk is distinguished using fluorescence microscope detection technique
There is certain feasibility with full-cream reconstituted milk.
Therefore, those skilled in the art is dedicated to developing a kind of reconstituted milk rapid detection method being simple and efficient.
Summary of the invention
For overcome the deficiencies in the prior art, the technical problem to be solved by the present invention is to how quickly detect to give birth to difference
Cow's milk and reconstituted milk.
The present invention provides a kind of methods for distinguishing raw milk and full-cream reconstituted milk based on fluorescence microscopy, with Buddhist nun sieve
The neutral lipid ingredient in raw milk and full-cream reconstituted milk is marked in red dye liquor, using fluorescence microscope acquisition raw milk and
Image information of the full-cream reconstituted milk under 40 times of object lens, and shooting saves to compare respectively, by observation raw milk and entirely
The difference of fat globule partial size and distribution in rouge reconstituted milk, to distinguish raw milk and full-cream reconstituted milk.
Specifically comprise the following steps:
S10: prepare raw milk and full-cream reconstituted milk；
S20: measuring samples preparation；
S30: the fluorescent image of fluorescence microscope acquisition raw milk and full-cream reconstituted milk is used.
Further, in step S10, full-cream reconstituted milk is dissolved in 60 DEG C of deionized waters of 200mL by 30g whole-fat milk powder and matches
It makes.
Further, step S20 further include:
S201: the preparation of Nile red dye liquor；
S202: Nile red dye liquor is mixed with raw milk and full-cream reconstituted milk respectively；
S203: the raw milk and full-cream reconstituted milk for taking 3~6 μ L step S202 to handle are placed on glass slide.
Further, Nile red dye liquor is to be dissolved in acetone by Nile red dye with the concentration of 42 μ g/mL to be formulated.
It further, is 1:9 according to the mixed proportion of volume ratio, raw milk or full-cream reconstituted milk and Nile red dye liquor.
Further, after raw milk or full-cream reconstituted milk are mixed with Nile red dye liquor, it is protected from light 30min at room temperature.
Further, in step S203, the raw milk and full-cream reconstituted milk for taking 5 μ L step S202 to handle are placed in load glass
On piece.
Further, in step S30, the excitation wavelength of fluorescence microscope is set as 510~550nm, launch wavelength setting
For 590nm.
Further, in step S30, the time for exposure of fluorescent image fluorescent image is 59.919ms, and sensitivity is
ISO1600, pixel are 1360 × 1024.
Further, in fluorescent image, the fat globule that the fat globule average grain diameter of full-cream reconstituted milk is less than raw milk is flat
Equal partial size, the partial size of the fat globule of full-cream reconstituted milk are unevenly distributed partial size of the degree higher than the fat globule of full-cream reconstituted milk
It is unevenly distributed degree.
Technical effect
The present invention is provided a kind of based on fluorescence using the difference of fat globule partial size and distribution in raw milk and full-cream reconstituted milk
The method of microscopy detection zone mitogenetic cow's milk and reconstituted milk.Compared to the prior art, the beneficial effects of the present invention are:
(1) sample pretreatment process is simple, detects fast speed；(2) testing cost is cheap, has potential application and popularization value.
Detailed description of the invention
Fig. 1 is a kind of method based on the mitogenetic cow's milk of fluorescence microscopy detection zone and reconstituted milk proposed by the present invention
Flow diagram；
Fig. 2 is fluorescent image of the raw milk under × 40 object lens, and picture have passed through reversion processing to print；
Fig. 3 is fluorescent image of the full-cream reconstituted milk under × 40 object lens, and picture have passed through reversion processing to print.
Specific embodiment
The present invention is further described below in conjunction with Figure of description, keep its technology contents more clear and is easy to understand.
The present invention can be emerged from by many various forms of embodiments, and protection scope of the present invention is not limited only to Wen Zhongti
The embodiment arrived.
Fig. 1 is a kind of method based on fluorescence microscopy detection zone mitogenetic cow's milk and reconstituted milk proposed by the present invention
Flow diagram.
The following steps are included:
One, prepare raw milk and full-cream reconstituted milk；
Two, Nile red dye liquor is prepared；
Three, 900 μ L raw milks or full-cream reconstituted milk are uniformly mixed with 100 μ L Nile red dye liquors, are protected from light at room temperature
30min；
Four, it takes mixed solution obtained by 5 μ L to drop to glass slide and prepares sample to be tested；
Five, the fluorescent image of raw milk and full-cream reconstituted milk is acquired under × 40 object lens.
Below by way of specific embodiment, the present invention will be further described.
S10: prepare raw milk and full-cream reconstituted milk
The verification sample that true raw milk is established as analysis method is acquired from Shanghai pasture.After raw milk extrusion immediately
- 2 DEG C are cooled to, and transports laboratory testing on the day of with incubator and ice bag.1 kind is had purchased from certain shopping online store to be used for
Prepare the whole-fat milk powder of full-cream reconstituted milk, room temperature storage.Full-cream reconstituted milk by 30g whole-fat milk powder be dissolved in 60 DEG C of 200mL go from
Sub- water is formulated, and is cooled to 4 DEG C, spare.
S20: measuring samples preparation
S201: the preparation of Nile red dye liquor: Nile red dye is dissolved in acetone with the concentration of 42 μ g/mL and is configured to Nile red
Dye liquor；
S202: the neutral lipid ingredient in raw milk and full-cream reconstituted milk is marked with Nile red dye liquor: 900 μ L 4
DEG C raw milk or full-cream reconstituted milk are uniformly mixed with 100 μ L Nile red dye liquors, are protected from light 30min at room temperature；
S203: the raw milk for taking 5 μ L marked and full-cream reconstituted milk prepare fluorescence detection sample.
S30: use the fluorescent image of fluorescence microscope acquisition raw milk and full-cream reconstituted milk: setting excitation wavelength is 510
~550nm, launch wavelength 590nm acquire raw milk under × 40 object lens using Olympus IX71 inverted fluorescence microscope
With the fluorescent image of full-cream reconstituted milk, time for exposure of image is 59.919ms, sensitivity ISO1600, pixel is 1360 ×
1024。
Fig. 2 and Fig. 3 is the fluorescent image of raw milk and full-cream reconstituted milk under × 40 object lens respectively, and two pictures pass through
Reversion processing is to print, and as seen from the figure, compared to raw milk, the fat globule average grain diameter of full-cream reconstituted milk is less than raw ox
Cream, partial size are unevenly distributed degree height, and there are the biggish fat globules of a small amount of partial size.For on the whole, full-cream reconstituted milk and
There are significant differences for the microstructure of raw milk fat globule, and this species diversity can be distinguished by common observation.Therefore, glimmering
Light microscope technology has feasibility in terms of distinguishing raw milk and full-cream reconstituted milk.
By above example, we use fluorescence microscope detection technique, quickly and effectively to distinguish raw milk and reconstituted milk
It provides way of thinking of theories and experiment is supported, and detection method is easy to operate, without equipment with high costs, having can be promoted
Property.
Although specific example and concrete analysis are illustrated that is be not intended to limit the invention is interior by the present invention one by one
Hold.Any researcher for being familiar with food inspection technology is not departing from main spirits and context of the invention, each when that can make
Kind change and polishing, including the sample for using agar glycopexis to dye.Because the protection scope of the invention should reality to apply for a patent
Subject to the scope of the claims of border.
Claims (10)
1. a kind of method for distinguishing raw milk and full-cream reconstituted milk based on fluorescence microscopy, which is characterized in that use Nile red
The neutral lipid ingredient in the raw milk and the full-cream reconstituted milk is marked in dye liquor, acquires institute using fluorescence microscope
The image information of raw milk and the full-cream reconstituted milk under 40 times of object lens is stated, by observation raw milk and full-cream reconstituted milk
The difference of fat globule partial size and distribution, to distinguish raw milk and full-cream reconstituted milk.
Specifically comprise the following steps:
S10: prepare raw milk and full-cream reconstituted milk；
S20: measuring samples preparation；
S30: the fluorescent image of fluorescence microscope acquisition raw milk and full-cream reconstituted milk is used.
2. a kind of method that raw milk and full-cream reconstituted milk are distinguished based on fluorescence microscopy according to claim 1,
It is characterized in that, in step S10, the full-cream reconstituted milk by 30g whole-fat milk powder be dissolved in 200mL60 DEG C of deionized water prepare and
At.
3. a kind of method that raw milk and full-cream reconstituted milk are distinguished based on fluorescence microscopy according to claim 1,
It is characterized in that, the step S20 further include:
S201: the preparation of Nile red dye liquor；
S202: the Nile red dye liquor is mixed with the raw milk and the full-cream reconstituted milk respectively；
S203: the raw milk and full-cream reconstituted milk for taking 3~6 μ L step S202 to handle are respectively placed on glass slide.
4. a kind of method that raw milk and full-cream reconstituted milk are distinguished based on fluorescence microscopy according to claim 3,
It is characterized in that, the Nile red dye liquor is to be dissolved in acetone by Nile red dye with the concentration of 42 μ g/mL to be formulated.
5. a kind of method that raw milk and full-cream reconstituted milk are distinguished based on fluorescence microscopy according to claim 3,
It is characterized in that, according to volume ratio, the mixed proportion of the raw milk or the full-cream reconstituted milk and the Nile red dye liquor is
1:9。
6. a kind of method that raw milk and full-cream reconstituted milk are distinguished based on fluorescence microscopy according to claim 5,
It is characterized in that, being protected from light at room temperature after the raw milk or the full-cream reconstituted milk are mixed with the Nile red dye liquor
30min。
7. a kind of method that raw milk and full-cream reconstituted milk are distinguished based on fluorescence microscopy according to claim 3,
It is characterized in that, the raw milk and full-cream reconstituted milk for taking 5 μ L step S202 to handle are placed in glass slide in the step S203
On.
8. a kind of method that raw milk and full-cream reconstituted milk are distinguished based on fluorescence microscopy according to claim 1,
It is characterized in that, the excitation wavelength of the fluorescence microscope is set as 510~550nm in step S30, launch wavelength is set as
590nm。
9. a kind of method that raw milk and full-cream reconstituted milk are distinguished based on fluorescence microscopy according to claim 1,
It is characterized in that, the time for exposure of the fluorescent image is 59.919ms in the step S30, sensitivity is ISO 1600, as
Element is 1360 × 1024.
10. a kind of method that raw milk and full-cream reconstituted milk are distinguished based on fluorescence microscopy according to claim 1,
It is characterized in that, the fat globule average grain diameter of the full-cream reconstituted milk is less than the fat globule of the raw milk in fluorescent image
Average grain diameter, the fat for being unevenly distributed degree and being higher than the full-cream reconstituted milk of the partial size of the fat globule of the full-cream reconstituted milk
The partial size of ball is unevenly distributed degree.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910585109.9A CN110296964A (en) | 2019-07-01 | 2019-07-01 | A method of based on the mitogenetic cow's milk of fluorescence microscopy detection zone and full-cream reconstituted milk |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910585109.9A CN110296964A (en) | 2019-07-01 | 2019-07-01 | A method of based on the mitogenetic cow's milk of fluorescence microscopy detection zone and full-cream reconstituted milk |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110296964A true CN110296964A (en) | 2019-10-01 |
Family
ID=68029773
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910585109.9A Pending CN110296964A (en) | 2019-07-01 | 2019-07-01 | A method of based on the mitogenetic cow's milk of fluorescence microscopy detection zone and full-cream reconstituted milk |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110296964A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114166784A (en) * | 2021-11-16 | 2022-03-11 | 华中农业大学 | Mid-infrared spectrum detection method for content of furfuryl amino acid in dairy product |
| CN116106231A (en) * | 2023-04-13 | 2023-05-12 | 北京三元食品股份有限公司 | Method for evaluating similarity between sample and breast milk in multiple dimensions |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107367494A (en) * | 2017-07-13 | 2017-11-21 | 上海交通大学 | The method for distinguishing recombined milk and fresh milk is detected with endogenous fluorescence spectral technique |
| CN108051421A (en) * | 2018-01-18 | 2018-05-18 | 上海交通大学 | The method of recombined milk is mixed based on two-dimentional external source fluorescence spectrum combination principal component analysis detection fresh milk |
-
2019
- 2019-07-01 CN CN201910585109.9A patent/CN110296964A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107367494A (en) * | 2017-07-13 | 2017-11-21 | 上海交通大学 | The method for distinguishing recombined milk and fresh milk is detected with endogenous fluorescence spectral technique |
| CN108051421A (en) * | 2018-01-18 | 2018-05-18 | 上海交通大学 | The method of recombined milk is mixed based on two-dimentional external source fluorescence spectrum combination principal component analysis detection fresh milk |
Non-Patent Citations (3)
| Title |
|---|
| 刘欢 等: "同步荧光技术检测鲜牛乳中掺杂复原乳的可行性研究", 《光谱学与光谱分析》 * |
| 迟雪露 等: "乳脂肪含量对牛乳理化性质的影响", 《食品科学》 * |
| 韩东海 等: "生鲜乳中还原乳的近红外光谱法鉴别", 《光谱学与光谱分析》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114166784A (en) * | 2021-11-16 | 2022-03-11 | 华中农业大学 | Mid-infrared spectrum detection method for content of furfuryl amino acid in dairy product |
| CN114166784B (en) * | 2021-11-16 | 2024-02-13 | 华中农业大学 | Mid-infrared spectrum detection method for furfuryl amino acid content in dairy product |
| CN116106231A (en) * | 2023-04-13 | 2023-05-12 | 北京三元食品股份有限公司 | Method for evaluating similarity between sample and breast milk in multiple dimensions |
| CN116106231B (en) * | 2023-04-13 | 2023-07-07 | 北京三元食品股份有限公司 | Method for evaluating similarity between sample and breast milk in multiple dimensions |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104956226B (en) | For the method for selective enrichment target cell, composition, kit and system | |
| Bicudo de Almeida-Muradian et al. | Standard methods for Apis mellifera honey research | |
| Dong et al. | Effect of microwave vacuum drying on the drying characteristics, color, microstructure, and antioxidant activity of green coffee beans | |
| Suh | Critical review: Metabolomics in dairy science–Evaluation of milk and milk product quality | |
| CN110296964A (en) | A method of based on the mitogenetic cow's milk of fluorescence microscopy detection zone and full-cream reconstituted milk | |
| CN105137073A (en) | Bovine Brucella colloidal gold antibody detection test paper strip | |
| Brodziak et al. | Effect of breed and feeding system on content of selected whey proteins in cow's milk in spring-summer and autumn-winter seasons | |
| Zhao et al. | Microwave vacuum drying of lotus seeds: Effect of a single-stage tempering treatment on drying characteristics, moisture distribution, and product quality | |
| CN107475365A (en) | Fluorescent PCR differentiates composition, kit and the method for sheep Niu Chengfen in sheep breast | |
| CN107607502A (en) | It is a kind of using multicolor fluorescence carbon point simultaneously and the method for Visual retrieval Multiple Classes of Antibiotics and the fluoroscopic examination instruction card of Multiple Classes of Antibiotics | |
| Jie et al. | Amino acid composition of royal jelly harvested at different times after larval transfer | |
| Cais-Sokolińska et al. | Freezing point and other technological properties of milk of the Polish Coldblood horse breed | |
| Kahl et al. | First tests of standardized biocrystallization on milk and milk products | |
| CN116858974B (en) | Identification method and application of pasteurized milk sterilization temperature | |
| CN105158463A (en) | Colloidal gold test paper bar for antibody detection of sheep Brucella | |
| Tzora et al. | The “Crosstalk” between microbiota and metabolomic profile of kefalograviera cheese after the innovative feeding strategy of dairy sheep by omega-3 fatty acids | |
| US7371565B2 (en) | Apparatus and method for measuring intracellular reactions | |
| CN111044695A (en) | Rapid assessment method for liquid milk heat treatment process | |
| Janiszewska-Turak et al. | Sustainable production and characteristics of dried fermented vegetables | |
| Gotti et al. | Determination of free amino acids in milk, colostrum and plasma of swine via liquid chromatography with fluorescence and UV detection | |
| Galdino et al. | Nutritional and physicochemical quality of formulations based on colostrum and bovine whey | |
| Watanabe et al. | An electrical discrimination method for rot in fresh cut apples using Cole–Cole plots | |
| Sheng et al. | Quantitative biochemical phenotypic heterogeneity of macrophages after myelin debris phagocytosis at a single cell level by synchrotron radiation fourier transform infrared microspectroscopy | |
| Liu et al. | Effects of the magnetic resonance imaging contrast agent Gd-DTPA on plant growth and root imaging in rice | |
| Ptáček et al. | Monitoring of milk performance of Wallachian sheep grazed under traditional Carpathian management in Western Beskids location. |
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 |