CN107561111A - The NMR spectrum method of cassava syrup is mixed in a kind of quick discriminating honey - Google Patents
The NMR spectrum method of cassava syrup is mixed in a kind of quick discriminating honey Download PDFInfo
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Abstract
The NMR spectrum method of cassava syrup is mixed in a kind of quick discriminating honey of the present invention, by honey sample, is configured to 0.03 0.5g/L solution after filtering with heavy water；The phosphate buffer solution containing the propyl sulfonic acid sodium inner mark solution of 3 trimethylsilyl 1 that volume fraction is 0.05 0.3% and 0.1 3.0 mM sodium azide is added, mixing, adjustment pH is 2.5 4.0, determines to obtain nuclear magnetic resoance spectrum through nuclear magnetic resonance spectrometer；Honey sample label chemical shift is analyzed, is contained1H chemical shifts δ is two of the 0.05ppm of ± 0.02 0.05ppm and 2.09ppm of 2.65ppm ± 0.02 unimodal, it then can determine whether to be mixed with cassava syrup in the sample, otherwise do not have, this method can analyze a large amount of samples in the short time, whether quick discriminating goes out is mixed with cassava syrup in honey.
Description
Technical field
The present invention relates to field of detection of food safety, and in particular to the core of cassava syrup is mixed in a kind of quick discriminating honey
Magnetic resonant wave spectral method.
Background technology
According to standard GB/T 14963-2011《National food safety standard honey》Middle regulation, honey are honeybee collections
Nectar/the secretion or honeydew of plant, after being combined with itself secretion, the natural sweet substance through fully brewageing.Natural bee
The main component of honey is glucose (25.2%~35.3%), fructose (33.3%~43.0%), amino acid, mineral matter, dimension life
Element and various active enzyme.Honey is deep because of its certain nutritive value and health-care effect to be favored by consumer.In the last few years, honeybee
Sweet adulteration is further serious, and some glucides are mainly mixed into honey, reduces the quality of honey, compromises consumption
The interests of person.
The syrup mainly mixed in recent years is four kinds：Sugar beet molasses, rice syrup, cassava syrup, wheat syrup.Due to honeybee
Fructose and glucose ratio and the fructose in these syrup and glucose ratio in honey is quite similar, can make these syrup of incorporation
The indices of honey afterwards comply fully with national standard, and very big difficulty is caused to detection.
The conventional method of adulterated honey detection has in recent years：C4 (corn syrup detection), SMR (rice syrup detection), SMB
(sugar beet molasses detection), C3/C4 (carbon isotope ratio method), TLC (high fructose starch syrup thin-layer method) and external enzyme process etc. are several
Kind detection method.But the conversion syrup of the C3 plants such as the cassava syrup occurred now, major part is eliminated in its manufacture craft
The carbon isotope value of glucide and most of oligosaccharides, its carbon isotope value and honey is in same scope, above others side
Method also is difficult to honey of the detection mixed with such a syrup.Therefore, to solve the adulterated problem occurred in honey production, sensitivity is established
Height, detection cycle are short, adapt to the authentication technique that cassava syrup is adulterated in the newly honey of adulterated means just seems particularly significant and tight
Compel.
NMR spectrum technology is exactly the radio-frequency pulse art with certain frequency, is radiated at the inside of honey, utilizes honey
In contained all substances atomic nucleus under magnetic fields, energy level splitting occurs, when energy and the atomic nucleus transition energy level of absorption
When equal, atomic energy order transition will occur so as to produce a kind of power spectrum, so as to gather the nuclear-magnetism for carrying all information of honey
Resonate wave spectrum, the final analysis quick detection related to honey adulteration.Due to nuclear magnetic resonance technique tool rapidity, without it is destructive,
The advantages that repeatable, therefore carry out the detection that nuclear magnetic resonance technique is used for cassava syrup in honey, for honey manufacturing enterprise
Raw material collect inspection and control of product quality in terms of, and market product quality testing and strike are mixed for law enforcement agency
False aspect has large effect.
The content of the invention
The invention provides a kind of NMR spectrum method that cassava syrup is mixed in quick discriminating honey, accuracy rate
Height, analysis time are short, can make up the defects of current honey adulteration discrimination method is present, drastically increase and mixed in adulterated honey
Enter the efficiency and accuracy of cassava syrup detection.
In order to solve the above problems, the technical solution adopted in the present invention is such, is mixed in a kind of quick discriminating honey
Enter the NMR spectrum method of cassava syrup, comprise the following steps,
1) honey sample that will be stirred, filter-cloth filtering, remove in honey after solid impurity, be configured to heavy water
0.03-0.5g/L solution；Described honey sample is chaste honey, rape honey and acacia honey；
2) the above-mentioned solution of certain volume is taken, adds the phosphate buffer solution that concentration is 0.5-2.0mol/L, both
Volume ratio is 800-1500：100-300, wherein, containing the 3- front threes that volume fraction is 0.05-0.3% in phosphate buffer solution
The sodium azide of base silane base -1- propyl sulfonic acid sodium inner mark solutions and 0.1-3.0mM, vortex oscillation is to uniform mixing, and adjustment is most
The pH value of whole solution is 2.5-4.0, is placed in nuclear magnetic tube；
3) nuclear magnetic tube is positioned in nuclear magnetic resonance spectrometer, detection obtains nuclear magnetic resoance spectrum；The analysis of NMR spectrum
Condition is：Popped one's head in using BBI；Measurement temperature:20-35 DEG C of room temperature；Ambient humidity:20-35%；Observed frequency 400MHz；Using
Presaturation method suppresses water peak；Pulse protocol is：Noesygppr1d either zg30 or zgpr；Spectrum width:6410Hz；90 ° of pulses
Width:Pl=6.45 μ s；Pulse delay time:Dl=15s；Launching centre:O1=4.7；Accumulative frequency:16 or 64；
4) honey sample label chemical shift is analyzed, is contained1H chemical shifts δ be 2.65ppm ± 0.02-0.05ppm and
Two of 2.09ppm ± 0.02-0.05ppm are unimodal, then can determine whether to be mixed with cassava syrup in the sample, otherwise do not have.
Preferably for the honey sample of crystallization, before stirring, in the case of closed, it is placed in no more than 40-60 DEG C
Warm, vibrate in water-bath, treat that sample all melts, after stirring, then be rapidly cooled to room temperature.
Preferably, honey sample filtering uses nylon filtering cloth of the aperture for 0.10mm-0.14mm.
Preferably, concentration is 1.0mol/L hydrochloric acid or Strong oxdiative sodium used by the pH value of step 2) adjustment final solution.
This method is 10%w/w for the cassava syrup limit of identification mixed in honey.
Beneficial effect：Compared with prior art, method accuracy rate of the invention is high, analysis time is short, can make up at present
The defects of honey adulteration discrimination method is present.Drastically increase efficiency that the detection of cassava syrup is mixed in adulterated honey and accurate
Property.
Brief description of the drawings
Fig. 1 is the nuclear magnetic spectrogram of Gansu acacia；
Fig. 2 is the nuclear magnetic spectrogram of cassava syrup；
Fig. 3 is the nuclear magnetic spectrogram of the Gansu acacia honey of 10% cassava syrup of incorporation.
Embodiment
By detailed description below to further explanation of the present invention, and line justification is entered to advantage of the present invention.
Experimental facilities includes：The type NMRs of Advance 400 (Brooker,Switzerland company), 14.1T superconducting magnets,
5mm double-core z- gradient probes and the controlling tests of Topspin 2.3 and data processing software；5mm nuclear magnetic resonance sample pipes；At a high speed from
Scheming (Sigma, German company)；Turbine mixer (XW-80A types, Instrument Factory, Shanghai Medical Science Univ.)；(the match of LP403 assay balances
More profits this, German company)；
Test solvent includes：Deuterated water (deuterium band width is 99.8%, also known as heavy water) is purchased from Cambrideg Isotope
Laboratories companies；3- trimethylsilyl -1- propyl sulfonic acids sodium (TSPSA) is purchased from Aldich-Sigma companies；Phosphoric acid
Hydrogen dipotassium and sodium dihydrogen phosphate (top pure grade) are purchased from Aldich-Sigma companies.
1. the confirmation of cassava syrup mark
1st, the pre-treatment of 1 sample
The honey of 10% cassava syrup of pure cassava syrup, pure honey and incorporation is chosen respectively as laboratory sample, wherein, honeybee
Honey is Gansu acacia honey, for nodeless mesh in sample, is stirred for before filtering uniformly, and to there is crystallization, close
In the case of closing, it is placed in the water-bath no more than 60 DEG C and warms, vibrate, is stirred evenly after sample all melts, be rapidly cooled to room temperature；
By the sample after thawing, filtered through 0.10mm-0.14mm apertures nylon filtering cloth, remove solid impurity in honey.Take 0.1g samples
In centrifuge tube, 1mL heavy water is added, dissolving is complete；
The above-mentioned μ l of solution 800 are taken, add the phosphate buffer solution (pH=2.0) that 100 μ l concentration are 0.5mol/L, wherein
Containing the folded of the 3- trimethylsilyl -1- propyl sulfonic acid sodium inner mark solutions and 0.1mM that volume fraction is 0.05% in cushioning liquid
Sodium nitride, vortex oscillation is to uniform mixing, and then with 1.0mol/L hydrochloric acid and Strong oxdiative sodium, the pH value for adjusting final solution is
2.5, wherein 600 μ l are taken in nuclear magnetic tube.
1st, the foundation of 2 nuclear-magnetism fingerprints databases
Adjust magnetic resonance spectroscopy instrument and nuclear magnetic tube be positioned in nuclear magnetic resonance chemical analyser, rotation nuclear magnetic tube in case1HNMR
Detection, obtains the nuclear magnetic signal of sample；Popped one's head in using BBI；Measurement temperature (probe temperature):20-35 DEG C of room temperature；Ambient humidity:
20-35%；Observed frequency 400MHz；Water peak is suppressed using presaturation method；Pulse protocol is：Noesygppr1d or zg30
Or zgpr；Spectrum width:6410Hz；90 ° of pulse widths:Pl=6.45 μ s；Pulse delay time:Dl=15s；Launching centre:O1
=4.7；Accumulative frequency:16 or 64.
1st, 3 spectrograms pre-process
Carrying out Fourier to the NMR signal of gained with the data processing software that nuclear magnetic resonance spectrometer carries becomes to change,
Conversion points are 131072, and index linewidth factor is 1Hz, obtains the nuclear magnetic spectrum of sample.
With topspin softwares, (each parameter setting is as follows in Topspin softwares：PULPROG=Jresgpprqf, AQ_mod
=DQD, TD=32K, NS=8*N, DS=4, TD0=1, SW=6000Hz, D1=3s, DE=4 μ s, D8=0.1s, signal tire out
Add 16 times) the original fingerprint collection of illustrative plates of acquisition is pre-processed, including spectral peak alignment, baseline correction and phasing.With 3- tri-
Methyl-monosilane base -1- propyl sulfonic acid sodium does internal standard compound, and chemical shift is set to 0.00 carry out spectral peak alignment；Baseline correction selects
Polynomial fit fit approach；Mutually correction selection global and metabonomics algorithms elder generation Automatic Optimal, it is rear for spy
Determine region carry out manual correction, make integrated value as much as possible be on the occasion of；Gansu acacia honey, cassava syrup and the incorporation of gained
The nuclear magnetic resonance map of the Gansu acacia honey of 10% cassava syrup, as shown in Figure 1, Figure 2 and Figure 3.
1st, the comparative analysis of 4 nmr spectrums
Pass through comparative studies Gansu acacia honey, cassava syrup and the Gansu acacia honey sample for mixing 10% cassava syrup
Nmr spectrum, find detected cassava syrup sample and mix 10% cassava syrup honey sample in contain two
Individual higher spectral peak, its1H chemical shifts δ is respectively two of 2.65ppm and 2.09ppm unimodal, and pure honey kind sample is changed herein
It is noiseless at displacement study.
In order to confirm the accuracy of looked for cassava syrup mark, the honey that have collected domestic different sources and nectar source is total to
150 kinds of meter (including Chinese scholartree honey, rape honey, Mel, chaste honey, clover honey, honey of lychee flowers, honey of lungan flowers, Mel Jujubae, matrimony vine honey, certain herbaceous plants with big flowers
Nectar, Radix Astragali honey, motherwort honey, loquat honey, Radix Codonopsis honey, the honey in the sweet and miscellaneous 16 kinds of nectar sources of nectar of fennel seeds, respectively from the Liao Dynasty
Rather, Hebei, the Inner Mongol, Shandong, Hunan, Anhui etc. amount to 26 provinces and cities), analysis find containing chemical shift 2.65ppm and
It is noiseless at 2.09ppm, illustrate that result shows, the cassava syrup mark looked for is all noiseless in all pure honey, day
Cassava syrup characteristic body is not present in right honey；Also by the cassava that various concentrations are added in the honey of different sources and nectar source
Syrup starches mark to confirm final cassava syrup, is ultimately determined to1H chemical shifts δ is1H chemical shifts δ be 2.65ppm and
Two of 2.09ppm are unimodal, are the characteristic differences materials of cassava syrup significant difference and pure honey, can use this two spectral peak
As the characteristic indication thing of cassava syrup, while the detection of the cassava syrup honey sample by adding various concentrations in honey,
The cassava syrup limit of identification for confirming to be mixed in honey is 10%.
2. actual sample detects
Unknown honey sample is detected with the method that cassava syrup is mixed in the quick discriminating honey of foundation.It will treat
The 42 parts of honey samples differentiated are operated by the method for above-mentioned steps 1.2~1.4, obtain the nuclear magnetic resoance spectrum of all samples
Figure, judge that unknown sample the label 1H chemical shifts in cassava syrup are according to the cassava syrup brand thing found
2.65ppm and 2.09ppm, the chemical shift △ δ of the sample detected under identical experiment condition and pure cassava syrup mark
Within 0.02-0.05ppm scopes, then it is assumed that cassava syrup is mixed with the sample, unknown sample is detected according to this
Analysis, concrete outcome are as shown in table 1.
Table 1
| Sample type | Legitimate reading | Detect result |
| True honey sample (16) | It is negative | It is negative |
| Cassava syrup sample (10) | It is positive | It is positive |
| Mix the honey sample (16) of cassava syrup | It is positive | It is positive |
Interpretation of result：Do not detected in all true honey1The cassava sugar that H chemical shifts are 2.65ppm and 2.09ppm
The characteristic indication thing of slurry, and detected in all cassava syrup spectrograms1H chemical shifts are two of 2.65ppm and 2.09ppm
Spectral peak, the mark of cassava syrup has also been detected in the adulterated honey spectrograms of other incorporation cassava syrup, has illustrated the standard of this method
True rate is 100%.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
God any modification, equivalent substitution and improvements made etc., should be included in the scope of the protection with principle.
Claims (5)
1. the NMR spectrum method of cassava syrup is mixed in a kind of quick discriminating honey, it is characterised in that including following step
Suddenly,
1) honey sample that will be stirred, filter-cloth filtering, remove in honey after solid impurity, 0.03- is configured to heavy water
0.5g/L solution；Described honey sample is chaste honey, rape honey and acacia honey；
2) the above-mentioned solution of certain volume is taken, adds the phosphate buffer solution that concentration is 0.5-2.0mol/L, both volumes
Than for 800-1500：100-300, wherein, containing the 3- trimethyl silicanes that volume fraction is 0.05-0.3% in phosphate buffer solution
The sodium azide of alkyl -1- propyl sulfonic acid sodium inner mark solutions and 0.1-3.0mM, vortex oscillation to uniform mixing, adjustment are final molten
The pH value of liquid is 2.5-4.0, is placed in nuclear magnetic tube；
3) nuclear magnetic tube is positioned in nuclear magnetic resonance spectrometer, detection obtains nuclear magnetic resoance spectrum；The analysis condition of NMR spectrum
For：Popped one's head in using BBI；Measurement temperature:20-35 DEG C of room temperature；Ambient humidity:20-35%；Observed frequency 400MHz；Using pre- full
Water peak is suppressed with method；Pulse protocol is：Noesygppr1d either zg30 or zgpr；Spectrum width:6410Hz；90 ° of pulses are wide
Degree:Pl=6.45 μ s；Pulse delay time:Dl=15s；Launching centre:O1=4.7；Accumulative frequency:16 or 64；
4) honey sample label chemical shift is analyzed, is contained1H chemical shifts δ be 2.65ppm ± 0.02-0.05ppm and
Two of 2.09ppm ± 0.02-0.05ppm are unimodal, then can determine whether to be mixed with cassava syrup in the sample, otherwise do not have.
2. the NMR spectrum method of cassava syrup is mixed in a kind of quick discriminating honey according to claim 1, its
It is characterised by, for the honey sample of crystallization, before stirring, in the case of closed, is placed in the water-bath no more than 40-60 DEG C
Warm, vibration, treat that sample all melts, after stirring, then be rapidly cooled to room temperature.
3. the NMR spectrum method of cassava syrup is mixed in a kind of quick discriminating honey according to claim 1, its
It is characterised by, honey sample filtering uses nylon filtering cloth of the aperture for 0.10mm-0.14mm.
4. the NMR spectrum method of cassava syrup is mixed in a kind of quick discriminating honey according to claim 1, its
It is characterised by, concentration is 1.0mol/L hydrochloric acid or Strong oxdiative sodium used by the pH value of step 2) adjustment final solution.
5. the NMR spectrum method of cassava syrup is mixed in a kind of quick discriminating honey according to claim 1, its
It is characterised by, this method is 10%w/w for the cassava syrup limit of identification mixed in honey.
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| RU2696772C1 (en) * | 2019-01-22 | 2019-08-06 | Федеральное государственное автономное образовательное учреждение высшего образования "Северо-Восточный федеральный университет имени М.К.Аммосова" | Method for determining the thermal effect on honey |
| CN110702832A (en) * | 2019-12-13 | 2020-01-17 | 中国农业科学院蜜蜂研究所 | Application of high-content 4-hydroxyquinoline as characteristic marker of jujube honey |
| EP3933392A1 (en) | 2020-07-01 | 2022-01-05 | Indian Oil Corporation Limited | Quick nmr method for identification and estimation of components in hand-rub formulations |
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