CN107505349A - A kind of nuclear magnetic resonance for differentiating true and false honey is uncoupled hydrogen spectral method - Google Patents
A kind of nuclear magnetic resonance for differentiating true and false honey is uncoupled hydrogen spectral method Download PDFInfo
- Publication number
- CN107505349A CN107505349A CN201710886371.8A CN201710886371A CN107505349A CN 107505349 A CN107505349 A CN 107505349A CN 201710886371 A CN201710886371 A CN 201710886371A CN 107505349 A CN107505349 A CN 107505349A
- Authority
- CN
- China
- Prior art keywords
- honey
- nuclear magnetic
- magnetic resonance
- uncoupled
- data
- 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.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
- G01N24/08—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
Landscapes
- Physics & Mathematics (AREA)
- High Energy & Nuclear Physics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
A kind of nuclear magnetic resonance for differentiating true and false honey is uncoupled hydrogen spectral method, is related to nuclear magnetic resonance spectroscopy.Known true and false honey sample is added into buffer solution, the sample of preparation pipettes supernatant liquor after centrifugation and is transferred in nuclear magnetic tube；H NMR spectroscopy pulse train of uncoupling compiled in advance is imported on nuclear magnetic resonance chemical analyser；Pulse train parameters and sampling parameter are set, carry out data acquisition；After the completion of sampling, the data processing of correlation is carried out, obtains the spectrogram of uncoupling of all samples, the subsection integral data for spectrogram of uncoupling are obtained by the pretreatment of corresponding experimental data；Subsection integral data are imported into statistical analysis software, judge to analyze the compound for causing group difference with group difference size, judge the adulterated mode of adulterated honey between two groups of groups by principal component analysis and orthogonal Partial Least Squares Method.Pulse train used is a kind of pure chemistry displacement spectral method of same core wide-band decoupling, can obtain and eliminate the high-resolution that coupling is split point1HNMR is composed.
Description
Technical field
The present invention relates to nuclear magnetic resonance spectroscopy detection method, more particularly, to the one-dimensional homonuclear decoupling of utilization1H H NMR spectroscopies
A kind of nuclear magnetic resonance for differentiating true and false honey is uncoupled hydrogen spectral method.
Background technology
Honey is honeybee adopts to obtain nectar from the spending of flowering plant, brewageed after being combined with itself secretion rich in big
Measure the wholefood of the sugar such as fructose and glucose, a small amount of sucrose, maltose being easily absorbed by the human body and mcg vitamin, tool
There is very high nutrition and health care effect, enjoy consumers.Huge market brings huge economic interests, therefore just has
Many illegal retailers pretend to be the honey to obtain interests by adulterated into honey, and adulterated means are rich and changeful, have and mix thereto
Enter cheap sucrose, plant syrup or glucose syrup to pretend to be honey etc., higher honey (the acacia nectar of also oriented price
Deng) in the relatively low honey (rape nectar etc.) of incorporation price come the means such as adulterate.These behaviors consumption in serious infringement
Sound development while person's interests also to honey industry generates great negative effect[1]。
For the adulterated mode to emerge in an endless stream, detection means be also evolving.Observed from traditional sense organ pair
The detection of physical and chemical index, then differentiated to by modern analytical technique.Method more commonly used at present is carbon isotope ratio
Rate method[2-3], chromatography (gas-chromatography[4-5], chromatography of ions[6-7]With liquid phase separation-isotope mass spectrometry[8]Deng) and its combination skill
Art, spectrum analysis[9], display scanning calorimetry[10]Deng.These methods respectively have length, and some is limited to experiment condition, and some are only
Can be for certain a kind of adulterated mode.
In recent years, mutually tied with statistical and analytical tool using nuclear magnetic resonance (nuclear magnetic resonance, NMR)
The method of conjunction has gradually entered into the visual field of people[11-13]。
Nuclear magnetic resonance technique is one of conventional means for parsing molecular structure, is obtained in the field such as chemistry, biology, medical science
Very extensive application is arrived.Believe by scalar coupling caused by the interaction between bond electron pair caused by nuclear spin
Breath (J couplings) can obtain intramolecular structural information[14].However, hydrogen spectrum chemical shift range it is very narrow (about 10~
15ppm), spectral peak caused by being acted on due to scalar coupling splits point often so that spectral peak overlaps, and causes1H H NMR spectroscopy resolution ratio
Low result[15-17].How important parameter of the resolution ratio as nuclear magnetic resoance spectrum plot quality, improve spectra resolution rate all the time
All it is the study hotspot in nuclear magnetic resonance technique.In recent years, by eliminating scalar coupling --- the spectral peak split point will be coupled and become
The unimodal pure chemistry displacement spectral technology to reduce peak overlap, significantly improve spectral resolution has obtained significant progress.From 1976
Two dimension (two dimensional, 2D) J is composed projection acquisition One-Dimensional Pure chemical shift spectrum and opened by year Ernst et al. first passage
Begin[18], constantly there are many initiative pure chemistry displacement spectral methods to be suggested.For example, by Bax et al. propose perseverance when develop
(constant time evolution) method[19], by separation from different C (12C/13C) chemical bond is connected1H completes to uncouple
BIRD (bilinear rotational decoupling) method[20], by Zangger and Sterk propositions based on layer choosing
ZS (Zangger-Sterk) method uncoupled[21]And the utilization proposed by Morris two frequency scans in the opposite direction small
The pulsed PSYCHE of angle chirp (pure shift yielded by chirp excitation) method[22]Deng.Its
In, it is by during indirect dimension develops, applying 180 ° of pulsus durus punchings and one and gradient that very representational ZS, which goes dual method,
Simultaneously apply 180 ° of soft pulses of selectivity, make in each layer be chosen core overturn 360 ° (unaffected, non-selected core turns over
Turn 180 °), therefore dimension only has chemical shift evolution indirectly, scalar coupling is met again.The freedom of uncoupling obtained due to unitary sampling
Sensing deamplification FID (free induction decay) signal is very short, and signal resolution is inadequate, therefore, is adopted by two dimension
Sample, i.e., to t1Evolution obtain it is a series of comprising the FID of spectrum information of uncoupling.Recombinated by the FID to be uncoupled to these by splicing
A complete FID signal is obtained, pure chemistry displacement spectra is obtained after Fourier transformation, this method sequence is realized more simple
Single, effect of uncoupling is good, can significantly lift the resolution ratio of spectrogram[5][10]。
Honey discrimination method based on nuclear magnetic resonance, conventional method such as electronic nose method and chromatographic process are compared, having can
The advantages that repeatability, Noninvasive, at the same it is high sensitivity, reproducible[1].With nuclear magnetic resonance associated with analysis method be mainly
Principal component analysis (principalcomponent analysis, PCA) and orthogonal offset minimum binary in multivariate statistical analysis
Method discriminant analysis (orthogonalpartial least squares discriminant analysis, OPLS-DA).PCA
Method is to carry out the Linear Mapping method of dimension-reduction treatment to higher-dimension variable in the case where keeping data message to lose minimum principle.
OPLS-DA methods are then to establish a PLS model by response variable Y of sample class.OSC filtered judge to classification it is uncorrelated
Variable information, only retain the variable relevant with classification judgement so that classification discriminant analysis can concentrate on it is related to classification
On variable, the judgement of mode identification method is improved[23][24].In proton nmr spectra, the integral area at proton signal peak
It is directly proportional with the number of hydrogen atoms for producing the signal, therefore spectrogram integration data is analyzed using multivariate statistical analysis method
Can obtain it is different detection samples between different compounds relative amount difference medical diagnosis on disease, food authenticity etc. obtain
Extensive use[13,23-24].At present, ZS goes dual method to be only used for simple chemical example to prove its spectrum effect of uncoupling, and does not exist
It is applied in the complicated Natural Product Samples of actual composition.
The content of the invention
It is an object of the invention to provide can obtain what is uncoupled1H H NMR spectroscopies, spectra resolution rate is improved, with reference to multivariate statistics
The nuclear magnetic resonance that a kind of its true and false honey of discriminating applied in the true and false discriminating of honey can be achieved in analysis is uncoupled hydrogen spectral method.
The present invention comprises the following steps：
1) prepared by honey sample：
In step 1), prepared by the honey sample, can prepare to turn into the honey of required detection and use nuclear magnetic resoance spectrum
The solution example of instrument detection；Selected honey can be by Jiangsu Entry-Exit Inspection and Quarantine Bureau of Shanxi Province animals and plants and food inspection center institute
There is provided, Hua Yuan is acacia and rape, and the place of production there are the ground such as Jiangsu, Sichuan and Henan；False honey be inspection center have confirmed by
What in the market was collected into mixes the adulterated honey of different proportion rice syrup, conversion syrup etc.；It is to weigh honey with quadrat method
100mg, add the phosphate buffer that 500 μ L contain 0.05%TSP (2,2,3,3- tetra- deuterated trimethyl silane acrylic acid)
(0.60mol/L,K2HPO4/NaH2PO4, pH=4.92,99.9%D2O), mixed using oscillator；Stand at room temperature
After 5min, through centrifuging 10min, rotating speed 10000rpm, its supernatant liquor 500 μ L are taken to be added in 5mm nuclear magnetic tubes and seal.
2) the NMR detections of honey solution example；
In step 2), the NMR detections of the honey solution example, sequence compiled in advance can be imported and carry out experiment number
According to collection, obtain1H NMR uncouple spectrogram and conventional spectrogram；The honey solution example nuclear magnetic tube prepared is placed in
In 500MHz AgilentNMRSystem nuclear magnetic resonance spectrometers, opened with conventional single-pulse sequence acquisition one1H H NMR spectroscopies, are composed
The line width of line, foundation is provided for spectrum width parameter setting, while line width values reflect field homogeneity implementations；To nuclear magnetic resonance spectrometer
Operate and compiled in advance ZS sequences and tnnoesy sequences are imported in software, Reasonable Parameters are set, carry out data sampling, its
In, the free induction decay signal obtained by ZS sequences needs to carry out free induction by data processor compiled in advance
Deamplification (FID) splicing restructuring obtains new FID, by the FID obtained by this spliced FID and tnnoesy sequence points
Fourier transformation is not carried out, obtains uncouple spectrogram and the conventional spectrogram of detected honey sample.
3) experimental data pre-processes；
In step 3), the experimental data pretreatment, the spectrogram obtained by testing can be carried out to the adjustment of phase baseline, gone
Follow-up modeling analysis is supplied except solvent peak, subsection integral and normalized obtain integration data；
Resulting ZS from step 2) is uncoupled to compose to use with conventional tnnoesy spectrums and uses MestReNova (Version respectively
9.0) software carries out the data prediction behaviour such as manual phase correction, multiple spot Baseline wander, calibration, spectral peak alignment and subsection integral
Make；In an experiment, the spectrogram obtained can become irregular in water peak part (δ=4.75~4.90), therefore by this partial spectrum
Figure is removed in order to avoid influenceing subsequent analysis；Remainder region (δ=0.50~9.0) are integrated at equal intervals, wherein between integration
△ δ=0.01 is divided into, the integration data of acquisition is normalized, the integration data unloading after normalization is the lattice of ASC II
Formula models for subsequent statistical.
4) experimental result Treatment Analysis.
In step 4), the experimental result Treatment Analysis, integration data can be imported in SIMCA-P+ softwares and be led
Constituent analysis and orthogonal Partial Least Squares Method, true and false honey difference thing is found out, judges adulterated mode；Modeling analysis mainly exists
Carried out on SIMCA-P+ softwares, using principal component analysis (principalcomponentanalysis, PCA) and orthogonal minimum partially
Square-Discriminant Analysis (orthogonapartial least squares discriminant analysis, OPLS-DA)；
Gained ASC II integration datas in step 3) are imported into SIMCA-P+ softwares, carry out data using Median Normal Ctr methods locates in advance
Reason, carries out PCA, obtains the clustering information of each group, two groups of the aggregations that can be relatively good of true honey and adulterated honey, while group afterwards
Between also have obvious difference, i.e., group difference is small and group difference is big, in order to further get be originally inputted in variable it is related
The maximum composition of degree, using initial data as X variables, group carries out OPLS-DA as response variable Y, removes initial data X
The information unrelated with dependent variable Y in variable, preferable discriminant classification effect can be obtained；By OPLS data MALTLAB softwares
The load diagram (loading plot) of analysis model is drawn after being handled, is thrown by load diagram and the OPLS variable for analyzing to obtain
Shadow importance (variable importance for the projection, VIP) this parameter can be obtained poor group
The different maximum integrating range of differentia influence i.e. between true and false honey, corresponding to being corresponding to corresponding chemical shift value on spectrogram
Spectral peak, carry out Spectra peak recognition and determine difference metabolin；Acquired results are secondly α-Portugal to cause difference metabolin to be mainly fructose
Grape sugar, β-glucose, it is judged as mixing syrup, is coincide with the adulterated situation of actual sample, tests credible reliable.
The invention has the advantages that：
(1) ZS spectral methods of uncoupling are combined by the present invention with multivariate statistical analysis method, and applied to the natural of complexity
In the composition analysis of food honey.By carrying out PCA and an orthogonal most young waiter in a wineshop or an inn partially to the pure chemistry displacement spectrogram of acquisition
Squares analysis, the discriminating of true and false honey is realized, meanwhile, it have found the difference material of true honey and adulterated honey, it is determined that mix
False mode.
(2) Zangger-Sterk of the present invention goes dual method to remove broadband same core1H-1H is coupled, and is only retained
Chemical shift information, obtains the higher hydrogen spectrogram of uncoupling of quality applied to complicated honey sample, improves spectra resolution rate.
ZS pulse trains are simple, easily operated, and conventional spectrometer conditions can meet requirement of experiment.
(3) compared with existing honey discrimination method, NMR methods proposed by the present invention have repeatability, Noninvasive
And spectra resolution rate it is high the advantages that.Using this method can to the kind of honey, source, it is true and false detect, be food inspection
And the analysis of the complex sample such as biological tissue's metabolin provides a kind of simple and effective method.
Brief description of the drawings
Fig. 1 is that ZS used in the present invention uncouples pulse train.
Fig. 2 be ZS used in the present invention uncouple pulse layer choosing splice schematic diagram.
Fig. 3 is the true honey collected respectively using uncouple spectral method ZS sequences and tnnoesy sequences and adulterated honey
Uncouple hydrogen spectrum and conventional hydrogen spectrum.
Fig. 4 be adulterated honey sample (being shown as in figure square) and true honey sample (star is shown as in figure) carry out it is main into
Shot chart obtained by analysis (PCA).
Fig. 5 is that adulterated honey sample (being shown as in figure square) and true honey sample (triangle is shown as in figure) are carried out just
Hand over the shot chart obtained by Partial Least Squares Method (OPLS-DA).
Fig. 6 is the load diagram obtained by OPLS analysis results by MATLAB processing.
Embodiment
ZS spectral methods of uncoupling are combined by the present invention with multivariate statistical analysis method, pass through the purifying displacement study to acquisition
Spectrogram carries out PCA and orthogonal Partial Least Squares Method, realizes the discriminating of true and false honey, have found true honey
With the difference material of adulterated honey, it is determined that adulterated mode.
Fig. 1 provides ZS used in the present invention and uncoupled pulse train, and Fig. 2 provides ZS used in the present invention and uncoupled pulse layer
Choosing splicing schematic diagram.
Each step in specific implementation process of the present invention is as follows：
(1) preparation of honey source and solution example
The honey studied is provided by Jiangsu Entry-Exit Inspection and Quarantine Bureau of Shanxi Province animals and plants and food inspection center, true and false honey
Judgement be mainly that test is detected by the honey adulteration of standard technique Services Co., Ltd according to SGS, test applies mass spectrum
And chromatographic technique.The source of true honey is mainly acacia and rape, and mainly there are the ground such as Jiangsu, Sichuan and Henan in the place of production.False honey
The adulterated honey having been found commercially being collected into for inspection center, adulterated form are mainly different proportion incorporation rice sugar
Slurry, corn syrup and glucose syrup etc..The compound method of nmr experiments sample is：Honey 100mg is taken, it is added to 500 μ L and contains
(0.60mol/L, K in 0.05%TSP (the deuterated trimethyl silane acrylic acid of 2,2,3,3- tetra-) phosphate buffer2HPO4/
NaH2PO-4, pH=4.92,99.9%D2O), mixed using oscillator.After standing 5min at room temperature, through 10000rpm
10min is centrifuged, takes its supernatant liquor 500 μ L to be added in 5mm nuclear magnetic tubes and seal.
(2) nmr spectrum gathers
The present invention is using Zangger-Sterk (hereinafter referred to as ZS) sequence (delay-90 ° of-t1/2-180°-180°
(sel)-t1/2- acq), this method is to be based on layer choosing decoupling (slice- by the people of Klaus Zangger and Heinz Sterk two
Selective decoupling) method proposition.When experiment starts, the magnetization vector of all protons in the presence of 90 ° of pulses
XOY plane is turned to from Z axis, afterwards t1/2Time in proceed by evolution under J couplings, an and after-applied weak ladder
Spend field, in the presence of weak gradient fields, in sample cell the spin of varying level aspect marked by locus, now apply one
180 ° of pulsus durus punching overturns all proton magnetization vectors, while 180 ° of arteries and veins of selectivity with one with gradient fields application simultaneously
Collective effect is rushed, makes to be chosen core in each layer to overturn 360 °, equivalent to not overturning, and non-selected core has only overturn 180 °,
Therefore, for being chosen core, dimension only has chemical shift evolution indirectly, reaches the purpose for eliminating J couplings.It is and different in different layers
Core will be chosen, equally reach the purpose uncoupled.
Because the FID signal of uncoupling that unitary sampling obtains is very short, signal resolution is inadequate, therefore, is sampled by two dimension
To a series of FID for including spectrum information of uncoupling.Before the detection phase starts, because the effect scalar coupling of 180 ° of assembled pulses is drilled
Change and meet again by complete, each FID first data point must be continuously permutated and combined as a new FID, real
Now this is very simple, and can cut each FID data point and be spliced turns into a new FID.FID is led to again afterwards
Crossing Fourier transformation turns into required spectrogram of uncoupling.
Used in the present invention is 500MHz AgilentNMR System nuclear magnetic resonance spectrometers, in order to show that ZS uncouples spectrum
Effect, this experiment acquires conventional hydrogen spectrum and compareed, and sequence used is tnnoesy (delay-90 ° of-t1-90°-tm-90°-
Acq) sequence.Specific parameter setting is respectively that ZS removes even sequence：Relaxation delay 2s, it is 6000Hz directly to tie up spectrum width SW, indirectly
It is 60Hz to tie up spectrum width SW1, and it is 25 to tie up sampling number ni indirectly, and accumulative frequency nt is 8, and sampling number np is 32000, and sky sweeps number
Ss is 16, and sampling time at is 2.5s；Tnnoesy sequences：Relaxation delay 2s, spectrum width sw are 10000Hz, and accumulative frequency nt is
128, sampling number 64000, it is 8 that sky, which sweeps number ss, and sampling time at is 2.2s, in t1(4 μ s) and tmEnter during (100ms)
Row water suppression.
(3) experimental data pre-processes
By the ZS of acquisition uncouple spectrum with MestReNova softwares (Version 9.0) spectrogram is carried out manual phase correction,
The data predictions such as multiple spot Baseline wander operate, and are calibrated with the TSP peaks obtained in tnnoesy spectrograms as δ=0.In an experiment,
Because the water peak part water peak part (δ=4.75~4.90) in spectrogram can become irregular, thus this part gone divided by
Exempt to influence subsequent analysis.Remainder is integrated at equal intervals with region (δ=0.50~9.0), wherein integration interval is △ δ
=0.01.The integration data obtained by each spectrogram is normalized afterwards, arrangement obtains all by normalization
The integration data of processing, its unloading is entered into excel forms and supplies follow-up modeling analysis.
(4) experimental result and analysis
A. one-dimensional hydrogen spectrum compares
As shown in figure 3, uncouple spectrogram and the conventional spectrogram of respectively true and false honey.It is from conventional spectrogram as can be seen that exhausted
Most of spectral peaks are all concentrated between δ 3.0~4.2, and spectral peak is very crowded in this section of spectrogram, very seriously overlapping, are corresponded to
Contained substantial amounts of glucide, is mainly made up of glucose and fructose in honey.Compare uncouple spectrogram and conventional spectrogram,
, it is apparent that doublet of doublets (dd) of the β glucose at δ 3.25 and δ 4.64 double peak (d) after uncoupling
Respectively obtain one unimodal (s), double peak (d) of the α glucose at δ 5.23 also obtained unimodal (s) after uncoupling, for spectrogram
In crowded overlapping part δ 3.0~4.2, also be apparent from being easily recognized after uncoupling, spectrogram is simplified, and is significantly increased
The resolution ratio of spectrogram.The relatively low signal to noise ratio of ZS is limited to, the weaker spectral peak of intensity can not obtain, and the higher spectral peak of intensity is all
It can belong in ZS spectrums and obtain.Contained component is obtained by Spectra peak recognition and its corresponding to conventional spectrogram and spectrogram of uncoupling
In spectral peak chemical displacement value it is as shown in table 1., can be with figure 3 it will also be seen that the comparison diagram of true honey and adulterated honey
It was found that because honey sample constituent itself is complicated, after still being uncoupled before either uncoupling, all can not directly be seen from naked eyes
Measure difference.
Table 1
Peak type：s:Singlet, it is unimodal；d:Doublet, it is bimodal；t:Triplet, triplet；dd:doublet of
Doublets, doublet of doublets；m:Multriplet, multiplet.
B. the ZS obtained before is composed into normalization integral data and imports progress statistics of variable analysis in SIMCA-P+ softwares,
Need to be standardized data before analysis, common standardized method there are Median Normal (mean center
Scaling, Ctr), self-adaption specification (unit variance scaling, UV) and Pa Laituo it is upscaled
(paretoscaling, Par), need to use different standardized methods for different data models, judgment models quality
Parameter has R2X、R2Y and Q2, wherein, R2X and R2Y represents interpretability of the model for X, Y matrix, Q respectively2Then represent model
Predictive ability, R2And Q2Show that model is more reliable and more stable closer to 1.This example selects to enter using Ctr standardized methods after
Row analysis.Accompanying drawing 4 is principal component analysis (PCA) shot chart, the R of model2And Q2Respectively 0.884 and 0.793, illustrate that model can
Lean on, the 1st group is adulterated honey in figure, is presented in figure with black square point, second group is true honey, is presented with star point.Two
Aggregation in group group, distinguish between group, represent that group difference is smaller, and group difference is larger, and there are two samples in true one group of honey
Point is not gathered in group, verified sample source, it is found that the two samples belong to the source honey that do not suit with other samples in group
Prepared, judge to spend the difference in source to result in peeling off for the two samples.
In order to further obtain, distinguishing the constituent with conspicuousness contribution group, continuing OPLS- to model
DA, it is accompanying drawing 5 to obtain model score figure, while the load diagram of model, i.e. accompanying drawing 6 are made using MATLAB softwares.Will in analysis
Obtained NMR data is as X variables, and grouping information is as Y variables.As can be seen that in original argument's X information is eliminated with
Behind the incoherent parts of dependent variable Y, classifying quality is more preferable.It is poor that load diagram can be used for distinguishing true and false honey metabolism in model
Foreign matter.It is typically the material away from X-axis in load diagram that different groups of sample classifications are contributed with larger metabolite, with reference to
This parameter of VIP, finally selects the chemical displacement value corresponding to forward 10 numerical value of VIP values from big to small, i.e., and 4.00,
3.72,4.11,3.71,3.59,3.90,5.24,4.03,4.65.By these chemical displacement values and Spectra peak recognition result phase in table 1
Compare, it can be found that the material for causing true and false honey difference is mainly fructose, it is secondly phlorose, β-glucose.And adopted
Adulterated honey is that the material mainly containing fructose and glucose such as true honey different proportion incorporation rice syrup, plant syrup changes
The relative amount of fructose and glucose is become.Because honey inherently contains fructose and glucose, this kind of adulterated discriminating is more tired
Difficulty, but ZS spectrums of uncoupling have distinguished true and false honey with reference to the detection method of multivariate statistical analysis, have found difference thing source, demonstrate,prove
The validity for the method for understanding.
Claims (10)
- The hydrogen spectral method 1. a kind of nuclear magnetic resonance for differentiating true and false honey is uncoupled, it is characterised in that comprise the following steps：1) prepared by honey sample：2) the NMR detections of honey solution example；3) experimental data pre-processes；4) experimental result Treatment Analysis.
- The hydrogen spectral method 2. a kind of nuclear magnetic resonance for differentiating true and false honey as claimed in claim 1 is uncoupled, it is characterised in that in step 1) in, prepared by the honey sample, and the honey of required detection, which is prepared, turns into the solution example detected with nuclear magnetic resonance spectrometer.
- The hydrogen spectral method 3. a kind of nuclear magnetic resonance for differentiating true and false honey as claimed in claim 1 is uncoupled, it is characterised in that in step 1) in, selected honey is provided by Jiangsu Entry-Exit Inspection and Quarantine Bureau of Shanxi Province animals and plants and food inspection center, and Hua Yuan is ocean There are Jiangsu, Sichuan and Henan in Chinese scholartree and rape, the place of production.
- The hydrogen spectral method 4. a kind of nuclear magnetic resonance for differentiating true and false honey as claimed in claim 1 is uncoupled, it is characterised in that in step 1) in, false honey is the incorporation different proportion rice syrup being collected into by the market, the conversion syrup that inspection center has confirmed Adulterated honey；With quadrat method to weigh honey 100mg, 500 μ L (2,2,3,3- tetra- deuterated trimethyls containing 0.05%TSP are added Silane acrylic acid) phosphate buffer, mixed using oscillator；After standing 5min at room temperature, through centrifuging 10min, turn Fast 10000rpm, its supernatant liquor 500 μ L are taken to be added in 5mm nuclear magnetic tubes and seal, the composition of the phosphate buffer is 0.60mol/L,K2HPO4/NaH2PO4, pH=4.92,99.9%D2O。
- The hydrogen spectral method 5. a kind of nuclear magnetic resonance for differentiating true and false honey as claimed in claim 1 is uncoupled, it is characterised in that in step 2) in, the NMR detections of the honey solution example, import sequence compiled in advance and carry out experimental data collection, obtain1H NMR uncouples spectrogram and conventional spectrogram.
- The hydrogen spectral method 6. a kind of nuclear magnetic resonance for differentiating true and false honey as claimed in claim 1 is uncoupled, it is characterised in that in step 2) in, the honey solution example nuclear magnetic tube prepared is placed in 500MHz AgilentNMRSystem nuclear magnetic resonance spectrometers In, opened with conventional single-pulse sequence acquisition one1H H NMR spectroscopies, the line width of spectral line is obtained, provide foundation for spectrum width parameter setting, together When line width values reflect field homogeneity implementations；Operated to nuclear magnetic resonance spectrometer imported in software in advance compiled ZS sequences with And tnnoesy sequences, Reasonable Parameters are set, carry out data sampling, wherein, the free induction decay signal obtained by ZS sequences needs Free induction decay signal splicing restructuring is carried out by data processor compiled in advance and obtains new FID, by this FID obtained by spliced FID and tnnoesy sequences carries out Fourier transformation respectively, obtains going for detected honey sample Even spectrogram and conventional spectrogram.
- The hydrogen spectral method 7. a kind of nuclear magnetic resonance for differentiating true and false honey as claimed in claim 1 is uncoupled, it is characterised in that in step 3) in, the experimental data pretreatment, the adjustment of spectrogram progress phase baseline, removal solvent peak, segmentation obtained by testing are accumulated Divide and normalized obtains integration data and supplies follow-up modeling analysis.
- The hydrogen spectral method 8. a kind of nuclear magnetic resonance for differentiating true and false honey as claimed in claim 1 is uncoupled, it is characterised in that in step 3) in, the ZS obtained by from step 2) is uncoupled into spectrum and conventional tnnoesy spectrums with carrying out hand with MestReNova softwares respectively The data prediction operations such as dynamic phase correction, multiple spot Baseline wander, calibration, spectral peak alignment and subsection integral, to the integration of acquisition Data are normalized, and the integration data unloading after normalization is the forms of ASC II so that subsequent statistical models.
- The hydrogen spectral method 9. a kind of nuclear magnetic resonance for differentiating true and false honey as claimed in claim 1 is uncoupled, it is characterised in that in step 4) in, the experimental result Treatment Analysis, will integration data import SIMCA-P+ softwares in carry out principal component analysis and it is orthogonal partially Least-square analysis, true and false honey difference thing is found out, judges adulterated mode；Modeling analysis is carried out on SIMCA-P+ softwares, Using principal component analysis and orthogonal PLS discriminant analysis.
- The hydrogen spectral method 10. a kind of nuclear magnetic resonance for differentiating true and false honey as claimed in claim 1 is uncoupled, it is characterised in that in step 4) in, gained ASC II integration datas in step 3) is imported into SIMCA-P+ softwares, enter line number using Median Normal Ctr methods Data preprocess, PCA being carried out afterwards, obtaining the clustering information of each group, the two groups of aggregations of true honey and adulterated honey, i.e. group difference is small And group difference is big, the composition for being originally inputted that degree of correlation is maximum in variable is got, X variables, group are used as using initial data OPLS-DA is carried out as response variable Y, information unrelated with dependent variable Y in initial data X variables is removed, OPLS data is used MALTLAB softwares draw the load diagram of analysis model after being handled, the variable drop for analyzing to obtain by load diagram and OPLS This parameter of importance is obtained to the maximum integrating range of differentia influence between the i.e. true and false honey of group difference, corresponding on spectrogram Spectral peak as corresponding to corresponding chemical shift value, carry out Spectra peak recognition and determine difference metabolin；Acquired results are to cause difference Metabolin is mainly fructose, is secondly phlorose, β-glucose, is judged as mixing syrup, situation kiss adulterated with actual sample Close.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710886371.8A CN107505349B (en) | 2017-09-27 | 2017-09-27 | A kind of nuclear magnetic resonance identifying true and false honey is uncoupled hydrogen spectral method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710886371.8A CN107505349B (en) | 2017-09-27 | 2017-09-27 | A kind of nuclear magnetic resonance identifying true and false honey is uncoupled hydrogen spectral method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107505349A true CN107505349A (en) | 2017-12-22 |
| CN107505349B CN107505349B (en) | 2019-02-26 |
Family
ID=60698988
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710886371.8A Active CN107505349B (en) | 2017-09-27 | 2017-09-27 | A kind of nuclear magnetic resonance identifying true and false honey is uncoupled hydrogen spectral method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107505349B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108614000A (en) * | 2018-04-13 | 2018-10-02 | 江苏出入境检验检疫局动植物与食品检测中心 | Manuka honey discrimination method based on SIMCA models |
| CN110044944A (en) * | 2019-04-22 | 2019-07-23 | 中国科学院山西煤炭化学研究所 | A kind of magnetic resonance detection method for F- T synthesis waste water component analysis |
| CN110361680A (en) * | 2019-06-21 | 2019-10-22 | 厦门大学 | A kind of super-resolution nuclear magnetic resonance two dimension J decomposition spectral method |
| CN110470806A (en) * | 2019-08-26 | 2019-11-19 | 安徽泓顺源生物科技有限公司 | A method of distinguishing that honey is true and false according to honey component |
| CN112132028A (en) * | 2020-09-23 | 2020-12-25 | 厦门大学 | Method for obtaining high-quality nuclear magnetic resonance pure chemical shift spectrum |
| CN112816515A (en) * | 2020-12-29 | 2021-05-18 | 中国食品发酵工业研究院有限公司 | Method for identifying adulterated syrup in honey by utilizing nuclear magnetic resonance hydrogen spectrum |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004236876A (en) * | 2003-02-06 | 2004-08-26 | Natl Inst Of Radiological Sciences | Human body equivalent dielectric gel, its manufacturing method and use of gel |
| JP2015028493A (en) * | 2014-10-02 | 2015-02-12 | 独立行政法人物質・材料研究機構 | Chiral shift reagent for nmr and method for determining optical purity using the same |
| CN104713895A (en) * | 2015-03-13 | 2015-06-17 | 中国科学院武汉物理与数学研究所 | Method for distinguishing between pure and syrup-adulterated honey based on combination of hydrogen nuclear magnetic resonance and partial least square method |
| CN105510372A (en) * | 2016-01-27 | 2016-04-20 | 江苏出入境检验检疫局动植物与食品检测中心 | Method for building model for DPLS-BS-UVE rapid honey identification |
| CN105548233A (en) * | 2015-10-30 | 2016-05-04 | 中国科学院武汉物理与数学研究所 | Method for discriminating acacia honey and rape honey on basis of H-nuclear magnetic resonance |
-
2017
- 2017-09-27 CN CN201710886371.8A patent/CN107505349B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004236876A (en) * | 2003-02-06 | 2004-08-26 | Natl Inst Of Radiological Sciences | Human body equivalent dielectric gel, its manufacturing method and use of gel |
| JP2015028493A (en) * | 2014-10-02 | 2015-02-12 | 独立行政法人物質・材料研究機構 | Chiral shift reagent for nmr and method for determining optical purity using the same |
| CN104713895A (en) * | 2015-03-13 | 2015-06-17 | 中国科学院武汉物理与数学研究所 | Method for distinguishing between pure and syrup-adulterated honey based on combination of hydrogen nuclear magnetic resonance and partial least square method |
| CN105548233A (en) * | 2015-10-30 | 2016-05-04 | 中国科学院武汉物理与数学研究所 | Method for discriminating acacia honey and rape honey on basis of H-nuclear magnetic resonance |
| CN105510372A (en) * | 2016-01-27 | 2016-04-20 | 江苏出入境检验检疫局动植物与食品检测中心 | Method for building model for DPLS-BS-UVE rapid honey identification |
Non-Patent Citations (2)
| Title |
|---|
| R. CONSONNI ET AL.: "NMR Characterization of Saccharides in Italian Honeys of Different Floral Sources", 《J. AGRIC. FOOD CHEM.》 * |
| 刘芸 等: "核磁共振技术结合化学计量学方法用于蜂蜜的掺假鉴别", 《分析测试学报》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108614000A (en) * | 2018-04-13 | 2018-10-02 | 江苏出入境检验检疫局动植物与食品检测中心 | Manuka honey discrimination method based on SIMCA models |
| CN110044944A (en) * | 2019-04-22 | 2019-07-23 | 中国科学院山西煤炭化学研究所 | A kind of magnetic resonance detection method for F- T synthesis waste water component analysis |
| CN110361680A (en) * | 2019-06-21 | 2019-10-22 | 厦门大学 | A kind of super-resolution nuclear magnetic resonance two dimension J decomposition spectral method |
| CN110470806A (en) * | 2019-08-26 | 2019-11-19 | 安徽泓顺源生物科技有限公司 | A method of distinguishing that honey is true and false according to honey component |
| CN112132028A (en) * | 2020-09-23 | 2020-12-25 | 厦门大学 | Method for obtaining high-quality nuclear magnetic resonance pure chemical shift spectrum |
| CN112132028B (en) * | 2020-09-23 | 2023-12-26 | 厦门大学 | Method for obtaining high-quality nuclear magnetic resonance pure chemical shift spectrum |
| CN112816515A (en) * | 2020-12-29 | 2021-05-18 | 中国食品发酵工业研究院有限公司 | Method for identifying adulterated syrup in honey by utilizing nuclear magnetic resonance hydrogen spectrum |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107505349B (en) | 2019-02-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107505349B (en) | A kind of nuclear magnetic resonance identifying true and false honey is uncoupled hydrogen spectral method | |
| Hatzakis | Nuclear magnetic resonance (NMR) spectroscopy in food science: A comprehensive review | |
| CN105510372B (en) | Set up the true and false model method of DPLS BS UVE quick discriminating honey | |
| Dais et al. | Quality assessment and authentication of virgin olive oil by NMR spectroscopy: A critical review | |
| CN106950241B (en) | A kind of method of other adulterated oil type and contents in prediction tea oil | |
| Santone et al. | Saliva metabolomics by NMR for the evaluation of sport performance | |
| CN108508055B (en) | Metabonomics-based diabetes resistance potential marker metabolic pathway of Guangxi Yaoshan sweet tea and research method | |
| Nagana Gowda et al. | Overview of NMR spectroscopy-based metabolomics: opportunities and challenges | |
| Bo et al. | High-resolution pure shift NMR spectroscopy offers better metabolite discrimination in food quality analysis | |
| CN106199474B (en) | A kind of low-field nuclear magnetic resonance two-dimensional spectrum inversion algorithm | |
| CN106855552A (en) | A kind of method for differentiating honey types using non-target metabonomic technology | |
| CN105548233A (en) | Method for discriminating acacia honey and rape honey on basis of H-nuclear magnetic resonance | |
| Choze et al. | Distinction between a transgenic and a conventional common bean genotype by 1H HR-MAS NMR | |
| CN107300565A (en) | The NMR spectrum method of rice syrup is mixed in a kind of quick discriminating honey | |
| Li et al. | Metabolic discrimination of different Rhodiola species using 1H-NMR and GEP combinational chemometrics | |
| CN107543838A (en) | A kind of adulterated magnetic resonance detection method for planting butter cream in dilute cream | |
| Viskić et al. | NMR in the service of wine differentiation | |
| Sobolev et al. | NMR-based approaches in the study of foods | |
| CN107340348A (en) | A kind of method for building up of Bletilla striata medicinal materials HPLC finger-prints | |
| CN106018452A (en) | Peanut variety nondestructive testing method based on nuclear magnetic resonance technology | |
| CN106645254A (en) | Method for identifying grape variety and year of wine | |
| Alcantara et al. | Chemometric analysis applied in 1H HR-MAS NMR and FT-IR data for chemotaxonomic distinction of intact lichen samples | |
| Rebiai et al. | Quality assessment of medicinal plants via chemometric exploration of quantitative NMR data: a review | |
| Lemus Ringele et al. | NMR Metabolite Profiling in the Quality and Authentication Assessment of Greek Honey—Exploitation of STOCSY for Markers Identification | |
| Lin et al. | Water dynamics of Mung bean (Vigna radiata) sprouts treated with 6-benzylaminopurine: discrimination by low-field nuclear magnetic resonance and spectrometry |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |