CN104122250B - A kind of method of quick detection lactose - Google Patents
A kind of method of quick detection lactose Download PDFInfo
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- CN104122250B CN104122250B CN201410317843.4A CN201410317843A CN104122250B CN 104122250 B CN104122250 B CN 104122250B CN 201410317843 A CN201410317843 A CN 201410317843A CN 104122250 B CN104122250 B CN 104122250B
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- lactose
- raman spectrum
- milk
- raman
- crystal violet
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- 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 65
- 239000008101 lactose Substances 0.000 title claims abstract description 65
- 238000000034 method Methods 0.000 title claims abstract description 29
- 238000001514 detection method Methods 0.000 title abstract description 5
- 239000008267 milk Substances 0.000 claims abstract description 34
- 235000013336 milk Nutrition 0.000 claims abstract description 32
- 210000004080 milk Anatomy 0.000 claims abstract description 32
- 238000001237 Raman spectrum Methods 0.000 claims abstract description 27
- 238000004458 analytical method Methods 0.000 claims abstract description 13
- 238000001069 Raman spectroscopy Methods 0.000 claims abstract description 12
- 239000013078 crystal Substances 0.000 claims description 19
- 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
- 230000002708 enhancing effect Effects 0.000 claims description 4
- 239000011159 matrix material Substances 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 2
- 238000004445 quantitative analysis Methods 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- ZXJXZNDDNMQXFV-UHFFFAOYSA-M crystal violet Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1[C+](C=1C=CC(=CC=1)N(C)C)C1=CC=C(N(C)C)C=C1 ZXJXZNDDNMQXFV-UHFFFAOYSA-M 0.000 claims 1
- 239000000203 mixture Substances 0.000 claims 1
- 239000012086 standard solution Substances 0.000 claims 1
- 239000013558 reference substance Substances 0.000 abstract description 5
- 150000001720 carbohydrates Chemical class 0.000 abstract description 4
- 235000014633 carbohydrates Nutrition 0.000 abstract description 4
- 238000010223 real-time analysis Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 12
- 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 8
- 239000008103 glucose Substances 0.000 description 8
- 238000005259 measurement Methods 0.000 description 6
- 239000000523 sample Substances 0.000 description 3
- 238000001228 spectrum Methods 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
- 238000003556 assay Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 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 2
- 239000006071 cream Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010067715 Gastrointestinal sounds abnormal Diseases 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 238000004497 NIR spectroscopy Methods 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- FRYDSOYOHWGSMD-UHFFFAOYSA-N [C].O Chemical class [C].O FRYDSOYOHWGSMD-UHFFFAOYSA-N 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000013475 authorization Methods 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 235000020190 lactose-free milk Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- VUZPPFZMUPKLLV-UHFFFAOYSA-N methane;hydrate Chemical compound C.O VUZPPFZMUPKLLV-UHFFFAOYSA-N 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 235000018102 proteins Nutrition 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
- 239000000126 substance Substances 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
- 238000001845 vibrational spectrum 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
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- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The method of the invention discloses a kind of quick detection lactose based on Raman spectrum, that is: as long as milk and reference substance solution are pressed the isometric miscible of 1:1, after carrying out Raman spectrum analysis, reference standard curve can easily and quickly detect the lactose concn in milk.Measuring method of the invention is not only quickly nor by the interference of other carbohydrates；This fit on holds Raman analyser and is able to achieve scene and outdoor real-time analysis.
Description
Technical field
It is specifically a kind of quickly to be examined using Raman spectrum the present invention relates to a kind of method of food engineering development field
The method for surveying lactose.
Background technique
China produces ten thousand tons more than 2000 of liquid milk per year at present, and in addition there are the import milk per year over 200000 tons.Ox
Milk is a kind of food full of nutrition, protein, minerals, vitamin and carbohydrate rich in.Carbon water in milk
The main component of compound is lactose, and the content of lactose is up to 4-5%.Under normal circumstances, lactose can be direct by human body
It absorbs, but for crowd's (tool statistics ratio is up to 75%) with " lactose intolerance ", drinks the ox of normal contents lactose
Milk can cause the symptoms such as abdominal tension, borborygmus, abdominal pain, exhaust and diarrhea.For suffering from the crowd of lactose intolerance, need to drink
The even lactose-free milk of Low lactose milk.Therefore production for milk, convenient, fast and accurately Determination of lactose method, is control
Lactose content important guarantee.
The methods of titrimetry and chromatography are commonly used in the measurement of lactose, time-consuming and can not achieve online survey
Fixed, therefore, there is an urgent need to develop the rapid assay methods for being suitable for lactose.Some measurement lactose are also reported in recent years
Fast method, e.g., Enzyme Electrode and near infrared spectroscopy etc..But these methods be applied to lactose detection or
Mostly or less all there are some problems, such as: Enzyme Electrode is easy the interference by other sugared (glucose)；Because hydrone has to infrared
Very strong absorption, near infrared spectrum are also poorly suitable for the quick measurement of lactose in liquid milk.
The present invention develops a kind of using Raman spectrum, quickly analyzes the method for lactose.Raman spectrum analysis because
Economic, simple, quick feature, in recent years using more and more extensive in the analysis of biological sample.Due to the Raman of hydrone
Spectrum very little, thus compared with infrared spectroscopy, the analysis of Raman spectrum more where applicable fluid sample.This external cause Raman spectrum is vibration
Dynamic spectrum, the nuance of the structure of matter can be reflected from the difference of its vibrational spectrum, therefore different carbon hydrates
Object, such as: lactose, glucose, galactolipin Raman spectrum be also easily discriminated.Currently, there are no applications according to our understanding
Raman spectrum quickly analyzes the research report of lactose.In addition, by retrieval data center, State Intellectual Property Office
(www.sipo.gov.cn), World Intellectual Property Organization (www.wipo.int), EUROPEAN PATENT OFFICE (www.espacenet.com) and
U.S.Patent & Trademark Office (www.uspto.gov) is also without finding that publication identical with this patent protection request or authorization are special
Benefit.
Summary of the invention
The present invention provides a kind of method that lactose is quickly detected using Raman spectrum.It is compared with the prior art,
This method is not by the interference of other carbohydrates (glucose and galactolipin), and measurement is quickly, accurately.Also with Handheld Raman spectrum
Instrument, real-time, the quick, Accurate Determining at the realization scene, outdoor lactose content that can be convenient using method of the invention.
The present invention is achieved by the following technical solutions: milk is uniformly mixed with reference substance solution by 1:1 in equal volume
Afterwards, 15 microlitres of drops are taken to carry out Raman spectrum analysis in Raman enhancing matrix, then according to the peak of lactose and reference substance characteristic peak
Ratios reference standard curve quickly and accurately obtains the content of lactose in milk sample.
The reference substance solution refers to: the crystal violet aqueous solution of 0.02mmol/L.
The described Raman enhancing matrix refers to: the silicon wafer of surfacing, silicon wafer using it is preceding cleaned up with alcohol and water after
It is dried for standby.
The Raman spectrum analysis condition refers to: Raman spectroscopy scans range is 800-1300cm-1Wave-number range；It is obtained
Raman spectrum initial data is obtained, is deducted except the quantitative analysis for being used for lactose content after fluorescence background.
The characteristic peak of the lactose and reference substance refers to: lactose characteristic peak is 1085cm-1, the characteristic peak of crystal violet is
1173cm-1。
The standard curve refers to: by the mixing various criterion concentration (0.028- of the crystal violet containing 0.01mmol/L
Lactose aqueous solution 0.1mol/L) carries out Raman spectrum analysis (800-1300cm-1Wave number).The various criterion lactose solution obtained
Lactose characteristic peak with respect to peak height (lactose characteristic peak (1085cm-1) peak height and crystal violet characteristic peak (1173cm-1) peak height ratio
Value) with the normal linearity relation curve of corresponding lactose concn.
The invention has the advantages that 1) without complicated sample treatment, continuous mode is simple, quick；2) made using crystal violet
For internal standard, measurement result is reproducible, reliable；3) 1085cm is used-1As the signature analysis peak of lactose, milk is not only avoided
In other substances (such as albumen) interference, also eliminate interference of the presence to lactose assay of other carbohydrates, realize lactose
Specificity analysis.
Detailed description of the invention
The present invention is further described with example with reference to the accompanying drawing.
Fig. 1 is the Raman spectrogram of lactose (a) glucose (b) He galactolipin (c).
Fig. 2 is the Raman spectrogram that the normal milk (b) of lactose (a) is free of lactose milk (c).
Fig. 3 is the Raman spectrogram of lactose (a) lactose Yu crystal violet (b) milk and crystal violet (c) crystal violet (d).
Fig. 4 is the Raman spectrum of 0.1mmol/L crystal violet mixing various concentration lactose aqueous solution.
Fig. 5 is the linear relationship of lactose concn peak height opposite with raman characteristic peak.
Specific embodiment
Below in conjunction with specific embodiment, invention is further described in detail.It should be understood that following embodiment is only used for
The bright present invention, not for limiting the scope of the invention.
Embodiment 1,1085cm-1And 1173cm-1It is the characteristic peak of lactose and crystal violet respectively
Lactose, glucose, gala sugar aqueous solution and 50% milk (bright full-cream) 50% of 0.1mol/L is respectively configured
Agalasisa sugar-milk (permanent natural) aqueous solution.Take 15 microlitres of above-mentioned solution drops on silicon wafer in 800-1300cm respectively-1Wave-number range
Interior acquisition Raman spectrum, as a result as shown in attached drawing 1 and attached drawing 2.
Although can be seen that lactose from attached drawing 1, there are many characteristic peak, 1085cm-1Characteristic peak is not present in grape
In sugar and galactolipin；Further it can also be seen that the 1085cm of lactose from the result of attached drawing 2-1Characteristic peak does not have in milk
Also do not occur by background interference, and in agalasisa sugar-milk, is used directly for the analysis of lactose completely.
Be respectively configured the lactose (a) of 0.1mol/L, 0.1mol/L lactose crystal violet containing 0.1mmol/L (b) and
The aqueous solution of 0.1mmol/L (d) and 0.2mmol/L crystal violet, then by the aqueous solution of 0.2mmol/L crystal violet and milk (light
It is bright full-cream) it is mixed in the ratio of 1:1, obtain milk crystal violet solution (c) to be measured.The Raman spectrum of above-mentioned solution a, b, c, d are shown in attached
Fig. 3
As can be seen that 1173cm from attached drawing 3-1It is the characteristic peak of crystal violet, and not by other objects in lactose and milk
The interference of matter.After adding crystal violet in milk, 1173cm-1It can be used as the reference peak that lactose is measured in milk completely.
The opposite peak height of lactose concn and its characteristic peak in embodiment 2, solution has extraordinary linear relationship
Configure the aqueous solution that various concentration lactose is mixed in 0.1mmol/L crystal violet.Wherein the concentration of lactose is respectively
0.028,0.043,0.065,0.083,0.1mol/L.Take above-mentioned 15 microlitres of 5 kinds of solution drops on silicon wafer in 800- respectively
1300cm-1Raman spectrum is obtained in wave-number range, spectral data is shown in attached drawing 4 after deducting fluorescence background.By each solution Raman
Opposite peak height (the H of lactose characteristic peak in spectrum1085/H1173) map with the lactose concn in solution and see attached drawing 5.
Have very from the lactose concn (I) in the opposite peak height ([L]) and solution that can be seen that lactose characteristic peak in attached drawing 5
Good correlation.Dependent equation is I=14.518 × [L]+0.0167, related coefficient 0.999.
It measures in embodiment 3, practical milk system accurately and not by the interference of glucose and galactolipin
Table 1: method of the invention is compared with other common method measurement results
Milk selects bright whole milk, adds the glucose and galactolipin of 0.1mol/L in milk respectively, is configured to
Glucose is rich in milk rich in milk and galactolipin.Respectively to the concentration of these three milk testings wherein lactose.Measuring method is adopted
With two kinds, first is that Raman spectra detection process of the invention, second is that common film method.Two kinds of measuring methods the results are shown in Table 1.
From table 1 it follows that for normal milk.Using this method and other methods obtain the result is that consistent
's.But in the presence of there are other sugar, it is not interfered using the method for this method.
Claims (4)
1. a kind of method for quickly detecting lactose using Raman spectrum, which is characterized in that by milk and 0.02mmol/L
Crystal violet aqueous solution by 1:1 in equal volume after mixing, take 15 microlitres of drops to carry out Raman spectrum point in Raman enhancing matrix
Analysis, then according to lactose 1085cm- 1Wave number and crystal violet 1173cm- 1The feature peak height ratio reference standard curve of wave number quickly,
Accurately obtain the content of lactose in milk sample.
2. the method according to claim 1, wherein the Raman enhancing matrix refers to: the silicon of surfacing
Piece, silicon wafer using it is preceding cleaned up with alcohol and water after be dried for standby.
3. the method according to claim 1, wherein the Raman spectrum analysis condition refers to: Raman spectrum
Scanning range is 800-1300cm- 1Wave-number range；Obtained Raman spectrum initial data deducts newborn except being used for after fluorescence background
The quantitative analysis of sugared content.
4. the method according to claim 1, wherein the standard curve refers to: configuration 0.01mmol/L knot
Mix 0.028 in crystalviolet respectively, 0.043,0.065,0.083,5 normal concentration aqueous solutions of 0.1mol/L lactose carry out Raman
Spectrum analysis；The lactose 1085cm in 5 standard solution Raman spectrums obtained- 1Wavenumbers characteristic peak height and crystal violet 1173cm- 1
The standard relationship curve of wavenumbers characteristic peak height ratios and corresponding lactose concn.
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| CN104777148B (en) * | 2015-04-17 | 2018-05-01 | 华东理工大学 | A kind of method of total protein in quick detection 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 |
| 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 |
| CN109425589B (en) * | 2017-09-05 | 2020-06-02 | 北京化工大学 | Rapid and accurate eucommia gum content measuring method |
| 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 |
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| CN1157919A (en) * | 1995-06-28 | 1997-08-27 | 株式会社京都第一科学 | Method of optically measuring component in solution |
| EP1126757B1 (en) * | 1998-11-05 | 2002-11-27 | ChemoMetec A/S | A system for regulating the handling of milk during the milking process and a method for regulating said milking process |
| CN101034066A (en) * | 2007-02-03 | 2007-09-12 | 内蒙古伊利实业集团股份有限公司 | Method for testing lactose hydrolysis ratio in low lactose milk |
| CN101178365A (en) * | 2007-09-18 | 2008-05-14 | 中国农业大学 | Lactose content measuring method in milk powder |
| FR2924223A1 (en) * | 2007-11-23 | 2009-05-29 | Chiesi Farmaceutici S P A Soci | PROCESS FOR DETERMINING THE POLYMORPH PURITY OF MANIDIPINE DICHLORHYDRATE. |
| CN104777148A (en) * | 2015-04-17 | 2015-07-15 | 华东理工大学 | Method for rapidly detecting total protein in milk |
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2014
- 2014-07-04 CN CN201410317843.4A patent/CN104122250B/en not_active Expired - Fee Related
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| CN1157919A (en) * | 1995-06-28 | 1997-08-27 | 株式会社京都第一科学 | Method of optically measuring component in solution |
| EP1126757B1 (en) * | 1998-11-05 | 2002-11-27 | ChemoMetec A/S | A system for regulating the handling of milk during the milking process and a method for regulating said milking process |
| CN101034066A (en) * | 2007-02-03 | 2007-09-12 | 内蒙古伊利实业集团股份有限公司 | Method for testing lactose hydrolysis ratio in low lactose milk |
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