CN107561111B - The NMR spectrum method of cassava syrup is mixed in a kind of quick identification honey - Google Patents
The NMR spectrum method of cassava syrup is mixed in a kind of quick identification honey Download PDFInfo
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Abstract
Honey sample is configured to the solution of 0.03-0.5g/L by a kind of NMR spectrum method for quickly identifying incorporation cassava syrup in honey of the invention with heavy water after filtering；The phosphate buffer solution of the sodium azide containing 3- trimethylsilyl -1- propyl sulfonic acid sodium inner mark solution and 0.1-3.0 mM that volume fraction is 0.05-0.3% is added, mixing, adjustment pH is 2.5-4.0, measures to obtain nuclear magnetic resoance spectrum through nuclear magnetic resonance spectrometer；The chemical shift of honey sample marker is analyzed, is contained1H chemical shift δ is that two of 2.65ppm ± 0.02-0.05ppm and 2.09ppm ± 0.02-0.05ppm are 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, quickly identify and whether be mixed with cassava syrup in honey.
Description
Technical field
The present invention relates to field of detection of food safety, and in particular to a kind of quickly to identify the core that cassava syrup is mixed in honey
Magnetic resonant wave spectral method.
Background technique
According to regulation in standard GB/T 14963-2011 " national food safety standard honey ", honey is honeybee acquisition
Nectar/the secretion or honeydew of plant, in conjunction with itself secretion after, natural sweet substance made of sufficiently being made.Natural bee
The main component of honey is glucose (25.2%~35.3%), fructose (33.3%~43.0%), amino acid, minerals, dimension life
Element and multiple active enzymes.Honey deep favor by consumer because of its certain nutritive value and health-care effect.In recent years, bee
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 bee
Fructose and glucose ratio in honey in these syrup fructose and glucose ratio it is quite similar, can make to mix these syrup
The indices of honey afterwards comply fully with national standard, cause very big difficulty to detection.
Adulterated honey detects common method and 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 eliminates major part in manufacture craft
The carbon isotope value of glucide and most of oligosaccharides, carbon isotope value and honey is in same range, the above others side
Method is also difficult to detect the honey mixed with such syrup.Therefore, adulterated to solve the problems, such as to occur in honey production, establish sensitivity
Height, detection cycle are short, adapt to the authentication technique that cassava syrup is adulterated in the honey of new 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, occur energy level splitting, when absorption energy and atomic nucleus transition energy level
When equal, atomic energy order transition will occur to generate a kind of power spectrum, to acquire the nuclear-magnetism for carrying all information of honey
Resonate wave spectrum, final to analyze quick detection relevant to honey adulteration.Due to nuclear magnetic resonance technique tool rapidity, without it is destructive,
The advantages that repeatable, therefore carry out detection of the nuclear magnetic resonance technique 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 by law enforcement agency
False aspect has large effect.
Summary of the invention
The present invention provides a kind of NMR spectrum method for quickly identifying incorporation cassava syrup in honey, accuracys rate
Height, analysis time are short, can make up defect existing for current honey adulteration discrimination method, greatly improve and mix in adulterated honey
Enter the efficiency and accuracy of the detection of cassava syrup.
To solve the above-mentioned problems, the technical scheme adopted by the invention is that such, a kind of quickly identify is mixed in honey
The NMR spectrum method for entering cassava syrup, includes the following steps,
1) honey sample that will be stirred evenly, filter-cloth filtering are removed in honey after solid impurity, are configured to heavy water
The solution of 0.03-0.5g/L；The honey sample is chaste honey, rape honey and acacia honey；
2) the above-mentioned solution of certain volume is taken, the phosphate buffer solution that concentration is 0.5-2.0mol/L is added, the two
Volume ratio is 800-1500:100-300, wherein the 3- front three for being 0.05-0.3% containing volume fraction in phosphate buffer solution
The sodium azide of base silane base -1- propyl sulfonic acid sodium inner mark solution 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 placed in nuclear magnetic resonance spectrometer, detection obtains nuclear magnetic resoance spectrum；The analysis of NMR spectrum
Condition are as follows: popped one's head in using BBI；Measuring temperature: 20-35 DEG C of room temperature；Ambient humidity: 20-35%；Observed frequency 400MHz；Using
Presaturation method suppresses water peak；Pulse protocol are as follows: noesygppr1d or 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) chemical shift of honey sample marker is analyzed, is contained1H chemical shift 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 not have.
Preferably for the honey sample of crystallization, before stirring evenly, in closed situation, it is placed in no more than 40-60 DEG C
It warms, vibrates in water-bath, all melt to sample, after mixing evenly, then be rapidly cooled to room temperature.
Preferably, honey sample filtering uses aperture for the nylon filtering cloth of 0.10mm-0.14mm.
Preferably, concentration used by the pH value of step 2) adjustment final solution is 1.0mol/L hydrochloric acid or Strong oxdiative sodium.
This method is 10%w/w for the cassava syrup limit of identification mixed in honey.
The utility model has the advantages that compared with prior art, method accuracy rate of the invention is high, analysis time is short, can make up at present
Defect existing for honey adulteration discrimination method.Greatly improve the efficiency that the detection of cassava syrup is mixed in adulterated honey and accurate
Property.
Detailed description of the invention
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 for mixing the Gansu acacia honey of 10% cassava syrup.
Specific embodiment
By following specific embodiments to further explanation of the present invention, and advantage of the present invention is proved.
Experimental facilities includes: 400 type Nuclear Magnetic Resonance of Advance (Brooker,Switzerland company), 14.1T superconducting magnet,
5mm double-core z- gradient probe and 2.3 controlling test of Topspin and data processing software；5mm nuclear magnetic resonance sample pipe；High speed from
Scheming (Sigma, German company)；Turbine mixer (XW-80A type, Instrument Factory, Shanghai Medical Science Univ.)；(the match of LP403 assay balance
More benefits this, German company)；
Test solvent includes: that deuterated water (deuterium band width is 99.8% also known as heavy water) is purchased from Cambrideg Isotope
Laboratories company；3- trimethylsilyl -1- propyl sulfonic acid sodium (TSPSA) is purchased from Aldich-Sigma company；Phosphoric acid
Hydrogen dipotassium and sodium dihydrogen phosphate (excellent pure grade) are purchased from Aldich-Sigma company.
1. the confirmation of cassava syrup marker
1, the pre-treatment of 1 sample
Pure cassava syrup, pure honey are chosen respectively and mixes the honey of 10% cassava syrup as laboratory sample, wherein bee
Honey is Gansu acacia honey, for nodeless mesh in sample, is stirred for before filtering uniformly, and to there is crystallization, close
It in the case of closing, is placed in the water-bath no more than 60 DEG C and warms, vibrate, stirred evenly after sample all melts, be rapidly cooled to room temperature；
It by the sample after thawing, is filtered through the aperture 0.10mm-0.14mm nylon filtering cloth, removes solid impurity in honey.Take 0.1g sample
In centrifuge tube, 1mL heavy water is added, dissolution is complete；
Above-mentioned 800 μ l of solution is taken, the phosphate buffer solution (pH=2.0) that 100 μ l concentration are 0.5mol/L is added, wherein
In buffer solution containing volume fraction be 0.05% 3- trimethylsilyl -1- propyl sulfonic acid sodium inner mark solution and 0.1mM it is folded
Sodium nitride, vortex oscillation to uniform mixing, then uses the hydrochloric acid and Strong oxdiative sodium of 1.0mol/L, the pH value for adjusting final solution is
2.5, take wherein 600 μ l in nuclear magnetic tube.
1, the foundation of 2 nuclear-magnetism fingerprints databases
Adjust magnetic resonance spectroscopy instrument and nuclear magnetic tube be placed in nuclear magnetic resonance chemical analyser, rotation nuclear magnetic tube in case1HNMR
Detection, obtains the nuclear magnetic signal of sample；It is popped one's head in using BBI；Measuring 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 are as follows: 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.
1,3 spectrograms pre-process
Carrying out Fourier to resulting NMR signal with the data processing software that nuclear magnetic resonance spectrometer carries becomes to change,
Transformation points are 131072, and index linewidth factor is 1Hz, obtain the nuclear magnetic spectrum of sample.
With topspin software, (each parameter setting is as follows in Topspin software: 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 are tired
Add 16 times) the original fingerprint map 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 as 0.00 carry out spectral peak alignment；Baseline correction selection
Polynomial fit fit approach；Mutually correction selection global and metabonomics algorithm elder generation Automatic Optimal, it is rear for spy
Determine region and carry out manual correction, makes integrated value positive value as much as possible；Resulting Gansu acacia honey, cassava syrup and incorporation
The nuclear magnetic resonance map of the Gansu acacia honey of 10% cassava syrup, as shown in Figure 1, Figure 2 and Figure 3.
1, the comparative analysis of 4 nmr spectrums
By comparing research Gansu acacia honey, cassava syrup and the Gansu acacia honey sample for mixing 10% cassava syrup
Nmr spectrum, find to contain two in the honey sample of 10% cassava syrup of detected cassava syrup sample and incorporation
A higher spectral peak,1H chemical shift δ is respectively that two of 2.65ppm and 2.09ppm are unimodal, and pure sweet kind of a sample is changed herein
It is noiseless at displacement study.
In order to confirm the accuracy of looked for cassava syrup marker, the honey for having collected domestic different sources and nectar source is total
150 kinds of meter (including Chinese scholartree honey, rape honey, eucalyptus honey, chaste honey, clover honey, honey of lychee flowers, honey of lungan flowers, Mel Jujubae, fructus lycii honey, certain herbaceous plants with big flowers
Nectar, Radix Astragali honey, motherwort honey, loquat honey, Radix Codonopsis honey, fennel seeds honey and miscellaneous 16 kinds of nectar sources of nectar honey, 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 the result shows that, the cassava syrup marker looked for is all noiseless in all pure honey, day
Cassava syrup characteristic body is not present in right honey；Also by the cassava for adding various concentration in the honey of different sources and nectar source
Syrup starches marker to confirm final cassava syrup, is ultimately determined to1H chemical shift δ is1H chemical shift δ be 2.65ppm and
Two of 2.09ppm are unimodal, are the characteristic differences substances of cassava syrup significant difference Yu pure honey, this two spectral peak can be used
As the characteristic indication object of cassava syrup, while the detection of the cassava syrup honey sample by adding various concentration in honey,
It confirmed that the cassava syrup limit of identification mixed in honey is 10%.
2. actual sample detects
Unknown honey sample is detected with the method for mixing cassava syrup in the quick identification honey of foundation.It will be to
The 42 parts of honey samples identified are operated by the method for above-mentioned steps 1.2~1.4, obtain the nuclear magnetic resoance spectrum of all samples
Figure judges that unknown sample, the marker 1H chemical shift in cassava syrup is according to the cassava syrup brand object found
2.65ppm and 2.09ppm, the chemical shift △ δ of the sample detected under identical experiment condition and pure cassava syrup marker
Within 0.02-0.05ppm range, then it is assumed that be mixed with cassava syrup in the sample, detected according to this to unknown sample
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: it is not detected in all true honey1The cassava sugar that H chemical shift is 2.65ppm and 2.09ppm
The characteristic indication object of slurry, and detected in all cassava syrup spectrograms1H chemical shift is two of 2.65ppm and 2.09ppm
Spectral peak has also detected the marker of cassava syrup 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 in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (5)
1. a kind of quickly identify the NMR spectrum method for mixing cassava syrup in honey, which is characterized in that including walking as follows
Suddenly, the 1) honey sample that will be stirred evenly, filter-cloth filtering remove in honey after solid impurity, are configured to 0.03- with heavy water
The solution of 0.5g/L；The honey sample is chaste honey, rape honey and acacia honey；
2) the above-mentioned solution of certain volume is taken, the phosphate buffer solution that concentration is 0.5-2.0mol/L, the volume of the two is added
Than for 800-1500:100-300, wherein the 3- trimethyl silicane for being 0.05-0.3% containing volume fraction in phosphate buffer solution
The sodium azide of alkyl -1- propyl sulfonic acid sodium inner mark solution 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 placed in nuclear magnetic resonance spectrometer, detection obtains nuclear magnetic resoance spectrum；The analysis condition of NMR spectrum
Are as follows: it is popped one's head in using BBI；Measuring 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 are as follows: noesygppr1d or 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) analyze the chemical shift of honey sample marker, containing 1H chemical shift δ be 2.65ppm ± 0.02-0.05ppm with
Two of 2.09ppm ± 0.02-0.05ppm are unimodal, then can determine whether to be mixed with cassava syrup in the sample, otherwise not have.
2. a kind of NMR spectrum method for quickly identifying incorporation cassava syrup in honey according to claim 1,
It is characterized in that, for the honey sample of crystallization, before stirring evenly, in closed situation, is placed in 40-60 DEG C of water-bath and warms,
Oscillation all melts to sample, after mixing evenly, then is rapidly cooled to room temperature.
3. a kind of NMR spectrum method for quickly identifying incorporation cassava syrup in honey according to claim 1,
It is characterized in that, honey sample filtering uses aperture for the nylon filtering cloth of 0.10mm-0.14mm.
4. a kind of NMR spectrum method for quickly identifying incorporation cassava syrup in honey according to claim 1,
It is characterized in that, it is 1.0mol/L hydrochloric acid or Strong oxdiative sodium that step 2), which adjusts concentration used by the pH value of final solution,.
5. a kind of NMR spectrum method for quickly identifying incorporation cassava syrup in honey according to claim 1,
It is characterized in that, this method is 10%w/w for the cassava syrup limit of identification mixed in honey.
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| US11774383B2 (en) | 2020-07-01 | 2023-10-03 | Indian Oil Corporation Limited | Quick NMR method for identification and estimation of components in hand-rub formulations |
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| CN109444200B (en) * | 2018-12-21 | 2022-02-11 | 钛和中谱检测技术(江苏)有限公司 | Method for identifying northeast basswood honey by adopting nuclear magnetic resonance hydrogen spectrum fingerprint |
| 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 |
| CN110702832B (en) * | 2019-12-13 | 2020-06-23 | 中国农业科学院蜜蜂研究所 | Application of high-content 4-hydroxyquinoline as characteristic marker of jujube honey |
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