CN104122250A - Method for rapid detection of lactose in milk - Google Patents
Method for rapid detection of lactose in milk Download PDFInfo
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- CN104122250A CN104122250A CN201410317843.4A CN201410317843A CN104122250A CN 104122250 A CN104122250 A CN 104122250A CN 201410317843 A CN201410317843 A CN 201410317843A CN 104122250 A CN104122250 A CN 104122250A
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- Prior art keywords
- lactose
- milk
- characteristic peak
- raman
- raman spectrum
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Links
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 title claims abstract description 73
- 239000008101 lactose Substances 0.000 title claims abstract description 73
- 235000013336 milk Nutrition 0.000 title claims abstract description 36
- 239000008267 milk Substances 0.000 title claims abstract description 36
- 210000004080 milk Anatomy 0.000 title claims abstract description 36
- 238000000034 method Methods 0.000 title claims abstract description 29
- 238000001514 detection method Methods 0.000 title abstract description 4
- 238000001237 Raman spectrum Methods 0.000 claims abstract description 26
- 238000004458 analytical method Methods 0.000 claims abstract description 16
- 238000001069 Raman spectroscopy Methods 0.000 claims abstract description 12
- 239000013558 reference substance Substances 0.000 claims abstract description 9
- 239000013078 crystal Substances 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 15
- 239000007864 aqueous solution Substances 0.000 claims description 10
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 6
- 229910052710 silicon Inorganic materials 0.000 claims description 6
- 239000010703 silicon Substances 0.000 claims description 6
- 239000011159 matrix material Substances 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 238000012113 quantitative test Methods 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 150000001720 carbohydrates Chemical class 0.000 abstract description 6
- 238000010223 real-time analysis Methods 0.000 abstract 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 9
- 239000008103 glucose Substances 0.000 description 9
- 229930182830 galactose Natural products 0.000 description 8
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 5
- 235000014633 carbohydrates Nutrition 0.000 description 5
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 4
- 238000003556 assay Methods 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 201000010538 Lactose Intolerance Diseases 0.000 description 2
- 239000006071 cream Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000001845 vibrational spectrum Methods 0.000 description 2
- 206010000087 Abdominal pain upper Diseases 0.000 description 1
- 208000019901 Anxiety disease Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010067715 Gastrointestinal sounds abnormal Diseases 0.000 description 1
- 238000004497 NIR spectroscopy Methods 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000036506 anxiety Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 235000008939 whole milk Nutrition 0.000 description 1
Landscapes
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a method for rapid detection of lactose in milk based on Raman spectra, that is to say, the milk and a reference substance solution are subjected to isovolumetric mixed dissolving according to a ratio of 1:1, and after Raman spectrum analysis, the concentration of lactose in the milk can be conveniently and rapidly detected through contrasting with a standard curve. The determination method not only is rapid, but also is not disturbed by other saccharides; and in addition, with cooperation of a hand-held Raman analyzer, on-site and outdoor real-time analysis can be realized.
Description
Technical field
What the present invention relates to is the method in a kind of Food Engineering Development field, specifically a kind of method that adopts Raman spectrum fast detecting lactose.
Background technology
China produces the liquid milk that exceedes more than 2,000 ten thousand tons per year at present, exceedes every year in addition in addition the import milk of 200,000 tons.Milk is a kind of nutritious food, contains rich in protein, mineral matter, vitamin and carbohydrates.In milk, the principal ingredient of carbohydrates is lactose, and the content of lactose is up to 4-5%.Generally, lactose can be directly absorbed by the body, but for the crowd's (tool statistics ratio is up to 75%) who suffers from " lactose intolerance ", the milk of drinking normal contents lactose can cause the symptoms such as belly anxiety, borborygmus, stomachache, exhaust and diarrhoea.For the crowd who suffers from lactose intolerance, need to drink the even milk of free from lactose of low lactose.Therefore for the production of milk, convenient, Determination of lactose method fast and accurately, is to control lactose content important guarantee.
The methods such as the conventional titrimetry of the mensuration of lactose and stratographic analysis, time-consuming and can not realize on-line determination, therefore, be applicable to the rapid assay methods of lactose in the urgent need to exploitation.Also reported in recent years the fast method of some mensuration lactose, as, Enzyme Electrode and near infrared spectroscopy etc.But these methods are applied to the detection of lactose and more or less all have some problems, as: Enzyme Electrode is easily subject to the interference of other sugar (glucose etc.); Because hydrone has a very strong absorption to infrared, near infrared spectrum is also not too applicable to the Fast Measurement of lactose in liquid milk.
The present invention has developed one and has utilized Raman spectrum, the method for express-analysis lactose.Raman spectrum analysis because of economical, simply, feature fast, in the analysis of biological sample, application is more and more extensive in recent years.Because the Raman spectrum of hydrone is very little, thereby compare with infrared spectrum, Raman spectrum is the analysis of where applicable fluid sample more.This external cause Raman spectrum is vibrational spectrum, and the nuance of the structure of matter can both reflect from the difference of its vibrational spectrum, therefore different carbohydrates, as: the Raman spectrum of lactose, glucose, galactose is also easy to distinguish.At present, according to our understanding, also do not apply the research report of Raman spectrum express-analysis lactose.In addition, do not find publication or the granted patent identical with this patent protection request through retrieval data center of State Intellectual Property Office (www.sipo.gov.cn), World Intellectual Property Organization (www.wipo.int), EUROPEAN PATENT OFFICE (www.espacenet.com) and United States Patent (USP) trademark office (www.uspto.gov) yet.
Summary of the invention
The invention provides a kind of method that adopts Raman spectrum fast detecting lactose.Compare with existing technology, this method is not subject to the interference of other carbohydrates (glucose and galactose), and it is quick, accurate to measure.In addition utilize Handheld Raman spectrometer, adopt method of the present invention can realize easily real-time, quick, the Accurate Determining of scene, outdoor lactose content.
The present invention is achieved by the following technical solutions: after milk is mixed by 1:1 equal-volume with reference substance solution, get 15 microlitres and drop in Raman and strengthen in matrix and carry out Raman spectrum analysis, then obtain quickly and accurately the content of lactose in milk sample according to the peak height ratios reference standard curve of lactose and reference substance characteristic peak.
Described reference substance solution refers to: the crystal violet aqueous solution of 0.02mmol/L.
Described Raman strengthens matrix and refers to: the silicon chip of surfacing, it is for subsequent use that silicon chip cleans up post-drying with alcohol and water before using.
Described Raman spectrum analysis condition refers to: Raman spectroscopy scans scope is 800-1300cm
-1wave-number range; The Raman spectrum raw data that obtains, deduction is except the quantitative test for lactose content after fluorescence background.
Described lactose and the characteristic peak of reference substance refer to: lactose characteristic peak is 1085cm
-1, the characteristic peak of crystal violet is 1173cm
-1.
Described typical curve refers to: the lactose aqueous solution of the mixing various criterion concentration (0.028-0.1mol/L) containing 0.01mmol/L crystal violet is carried out to Raman spectrum analysis (800-1300cm
-1wave number).The relative peak height of the lactose characteristic peak (lactose characteristic peak (1085cm of the various criterion lactose solution obtaining
-1) peak height and crystal violet characteristic peak (1173cm
-1) ratio of peak height) and with the normal linearity relation curve of corresponding lactose concn.
Advantage of the present invention is: 1) without complicated sample preparation, mensuration process is simple, quick; 2) adopt crystal violet as interior mark, measurement result is reproducible, reliable; 3) adopt 1085cm
-1as the signature analysis peak of lactose, not only avoid the interference of other materials in milk (as albumen etc.), also get rid of the interference that the existence of other carbohydrates is analyzed lactose, realize the specificity analyses of lactose.
brief description of the drawings
Below in conjunction with accompanying drawing and example, the present invention is further described.
Fig. 1 is the Raman spectrogram of lactose (a) glucose (b) and galactose (c).
Fig. 2 is the Raman spectrogram that the normal milk (b) of lactose (a) does not contain lactose milk (c).
Fig. 3 is the Raman spectrogram of lactose (a) lactose and crystal violet (b) milk and crystal violet (c) crystal violet (d).
Fig. 4 is the Raman spectrum that 0.1mmol/L crystal violet is mixed variable concentrations lactose aqueous solution.
Fig. 5 is lactose concn and the linear relationship of the relative peak height of raman characteristic peak.
embodiment
Below in conjunction with specific embodiment, the present invention is described in further detail.Should be understood that the scope that following examples are to illustrate the invention and not to limit the present invention.
Embodiment 1,1085cm
-1and 1173cm
-1it is respectively the characteristic peak of lactose and crystal violet
Configure respectively lactose, glucose, the galactose aqueous solution of 0.1mol/L, and 50% milk (bright full-cream), 50% free from lactose milk (permanent natural) aqueous solution.Getting respectively above-mentioned solution 15 microlitres drops on silicon chip in 800-1300cm
-1in wave-number range, obtain Raman spectrum, result is if accompanying drawing 1 is with as shown in accompanying drawing 2.
From accompanying drawing 1, can find out, although lactose has many characteristic peaks, 1085cm
-1characteristic peak does not appear in glucose and galactose; Further from the result of accompanying drawing 2, also can find out the 1085cm of lactose
-1characteristic peak not by background interference, and does not also occur, can be directly used in the analysis of lactose completely in milk in free from lactose milk.
Configure respectively the lactose (a) of 0.1mol/L, the aqueous solution that 0.1mol/L lactose contains 0.1mmol/L crystal violet (b) and 0.1mmol/L (d) and 0.2mmol/L crystal violet, then the aqueous solution of 0.2mmol/L crystal violet is mixed in the ratio of 1:1 with milk (bright full-cream), obtain milk crystal violet solution to be measured (c).The Raman spectrum of above-mentioned solution a, b, c, d is shown in accompanying drawing 3.
From accompanying drawing 3, can find out 1173cm
-1be the characteristic peak of crystal violet, and be not subject to the interference of other materials in lactose and milk.In milk, add after crystal violet 1173cm
-1can be used as the reference peak of measuring lactose in milk completely.
Lactose concn in embodiment 2, solution has extraordinary linear relationship with the relative peak height of its characteristic peak
The aqueous solution of mixing variable concentrations lactose in configuration 0.1mmol/L crystal violet.Wherein the concentration of lactose be respectively 0.028,0.043,0.065,0.083,0.1mol/L.Getting respectively above-mentioned 5 kinds of solution 15 microlitres drops on silicon chip in 800-1300cm
-1in wave-number range, obtain Raman spectrum, after spectral data deduction fluorescence background, see accompanying drawing 4.By the relative peak height (H of lactose characteristic peak in each solution Raman spectrum
1085/ H
1173) see accompanying drawing 5 with the lactose concn mapping in solution.
From accompanying drawing 5, can find out, the relative peak height ([L]) of lactose characteristic peak has good correlativity with the lactose concn (I) in solution.Dependent equation is I=14.518 × [L]+0.0167, and related coefficient is 0.999.
In embodiment 3, actual milk system, measure the interference that accurately and is not subject to glucose and galactose
Table 1: the comparison of method of the present invention and other common method measurement results
Milk is selected bright whole milk, adds respectively glucose and the galactose of 0.1mol/L in milk, is configured to that glucose is rich in milk and galactose is rich in milk.Respectively to the wherein concentration of lactose of these three kinds of milk testings.Assay method adopts two kinds, and the one, Raman spectra detection process of the present invention, the 2nd, conventional film method.Two kinds of assay methods the results are shown in Table 1.
As can be seen from Table 1, for normal milk.Adopting this method is consistent with the result that additive method obtains.But in the time having other sugar to exist, adopt the method for this method not to be interfered.
Claims (6)
1. one kind is utilized the method for Raman spectrum fast detecting lactose, it is characterized in that, after milk is mixed by 1:1 equal-volume with reference substance solution, get 15 microlitres and drop in Raman and strengthen in matrix and carry out Raman spectrum analysis, then obtain quickly and accurately the content of lactose in milk sample according to the peak height ratios reference standard curve of lactose and reference substance characteristic peak.
2. method according to claim 1, is characterized in that, described reference substance solution refers to: the crystal violet aqueous solution of 0.02mmol/L.
3. method according to claim 1, is characterized in that, described Raman strengthens matrix and refers to: the silicon chip of surfacing, it is for subsequent use that silicon chip cleans up post-drying with alcohol and water before using.
4. method according to claim 1, is characterized in that, described Raman spectrum analysis condition refers to: Raman spectroscopy scans scope is 800-1300cm
-1wave-number range; The Raman spectrum raw data that obtains, deduction is except the quantitative test for lactose content after fluorescence background.
5. method according to claim 1, is characterized in that, described lactose and the characteristic peak of reference substance refer to: lactose characteristic peak is 1085cm
-1, the characteristic peak of crystal violet is 1173cm
-1.
6. method according to claim 1, it is characterized in that, described typical curve refers to: the lactose aqueous solution of the mixing various criterion concentration (0.028-0.1mol/L) containing 0.01mmol/L crystal violet is carried out to Raman spectrum analysis (800-1300cm
-1wave number); The relative peak height of the lactose characteristic peak (lactose characteristic peak (1085cm of the various criterion lactose solution obtaining
-1) peak height and crystal violet characteristic peak (1173cm
-1) ratio of peak height) and with the normal linearity relation curve of corresponding lactose concn.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104777148A (en) * | 2015-04-17 | 2015-07-15 | 华东理工大学 | Method for rapidly detecting total protein in milk |
| CN105572096A (en) * | 2015-12-16 | 2016-05-11 | 新希望双喜乳业（苏州）有限公司 | Rapid detection and identification method for adulteration of milk |
| CN106290297A (en) * | 2016-07-30 | 2017-01-04 | 江南大学 | A kind of method of lipid composition in Fast Evaluation difference butter oil ball |
| CN107179296A (en) * | 2017-04-26 | 2017-09-19 | 深圳市赛亿科技开发有限公司 | The milk cup of lactose content in a kind of detection dairy produce |
| CN108802003A (en) * | 2018-05-25 | 2018-11-13 | 浙江大学 | A method of quick, Nondestructive Identification silk product and content |
| CN109342393A (en) * | 2018-11-15 | 2019-02-15 | 中牧实业股份有限公司 | A method of utilizing glucose content in Raman spectrum detection cell culture medium |
| CN109425589A (en) * | 2017-09-05 | 2019-03-05 | 北京化工大学 | A kind of fast and accurately Cortex Eucommiae gel content measuring method |
| CN118671048A (en) * | 2024-08-21 | 2024-09-20 | 唐山市食品药品综合检验检测中心(唐山市农产品质量安全检验检测中心、唐山市检验检测研究院) | Food quality detection method, device and system based on optical analysis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106290297B (en) * | 2016-07-30 | 2019-01-11 | 江南大学 | A kind of method of lipid composition in Fast Evaluation difference butter oil ball |
| CN107179296A (en) * | 2017-04-26 | 2017-09-19 | 深圳市赛亿科技开发有限公司 | The milk cup of lactose content in a kind of detection dairy produce |
| CN109425589A (en) * | 2017-09-05 | 2019-03-05 | 北京化工大学 | A kind of fast and accurately Cortex Eucommiae gel content measuring method |
| CN109425589B (en) * | 2017-09-05 | 2020-06-02 | 北京化工大学 | Rapid and accurate eucommia gum content measuring method |
| CN108802003A (en) * | 2018-05-25 | 2018-11-13 | 浙江大学 | A method of quick, Nondestructive Identification silk product and content |
| CN108802003B (en) * | 2018-05-25 | 2020-10-30 | 浙江大学 | Method for rapidly and nondestructively identifying silk product and content |
| CN109342393A (en) * | 2018-11-15 | 2019-02-15 | 中牧实业股份有限公司 | A method of utilizing glucose content in Raman spectrum detection cell culture medium |
| CN118671048A (en) * | 2024-08-21 | 2024-09-20 | 唐山市食品药品综合检验检测中心(唐山市农产品质量安全检验检测中心、唐山市检验检测研究院) | Food quality detection method, device and system based on optical analysis |
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