CN109444200A - A method of northeast eucalyptus honey is identified using nuclear magnetic resonance spectroscopy finger-print - Google Patents
A method of northeast eucalyptus honey is identified using nuclear magnetic resonance spectroscopy finger-print Download PDFInfo
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- CN109444200A CN109444200A CN201811568084.3A CN201811568084A CN109444200A CN 109444200 A CN109444200 A CN 109444200A CN 201811568084 A CN201811568084 A CN 201811568084A CN 109444200 A CN109444200 A CN 109444200A
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- 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
Abstract
The present invention provides a kind of method for identifying northeast eucalyptus honey using nuclear magnetic resonance spectroscopy finger-print, belongs to honey detection technique field, comprising steps of weighing honey or syrup sample to be identified, is diluted with phosphate buffer, stood after vortex；The solution after dilution is titrated to pH=2.5-5.0 with hydrochloric acid or potassium hydroxide solution；Treated sample is measured, the heavy aqueous solution constant volume for containing 2,2,3,3- tetra- deuterated trimethyl silane acrylic acid and sodium azide is added, is packed into nuclear magnetic tube after being vortexed uniformly；Using the sample in Liquid NMR instrument measurement nuclear magnetic tube, nuclear magnetic resonance spectroscopy finger-print is obtained, three there are a chevron near 3.0 ± 0.02ppm displacement refer to peak, then prove that identified honey or syrup sample are northeast eucalyptus honey.Discrimination method economical and efficient provided by the invention, sample handling characteristics are flexible, can intuitively and accurately identify pure northeast eucalyptus honey.
Description
Technical field
The invention belongs to honey detection technique fields, and in particular to a kind of to identify east using nuclear magnetic resonance spectroscopy finger-print
The method of northern eucalyptus honey.
Background technique
Eucalyptus honey is the nectar that honeybee acquires from Flores tiliae, is clear, yellowish thick liquid under room temperature, under low temperature easily
The congealed fat shape that is creamy white crystallization, color such as grease, fragrant taste, taste and sweet mouthfeel is the characteristic that China Northeast Mountain Areas forest is abounded with
Wild honey kind.Northeast eucalyptus honey was once well-known as imperial tribute honey in the Qing Dynasty, because being rich in glucose and fructose, Duo Zhongwei
Raw element, inorganic salts, organic acid enzyme and the biotin for promoting growth in humans and vigor can more enrich blood moistening lung than general honey, promotion
Cytothesis increases appetite etc., also the therefore deep favor by consumer.However, under the driving of commercial interest, many illegal quotient
Dealer is adulterated to the progress of sterling northeast eucalyptus honey kind using various sugar, syrup and cheap rape honey etc., causes to sell in the market
Eucalyptus honey brand it is multifarious, price is irregular, seriously compromises the quality prestige of honey industry and the health of consumer
And interests.Therefore it is necessary to which inventing a kind of detection technique that is simple and efficient realizes Rapid identification to sterling honey kind, to guarantee
Consumer, which buys, trusts the authentic pure northeast eucalyptus honey of high-quality.
The research of country's honey context of detection focuses mostly on to mix in syrup and analyze at present, and the report of related honey types identification
Road is then relatively limited, a kind of method of quickly identification honey types and adulterated honey of patent (application No. is 201810462442.6)
A kind of method adulterated using fluorescence emission spectrum identification honey types and syrup is disclosed, method is easy quickly, but due to
It, which is analyzed, uses syrup and honey sample limited amount, can not unify the feature for measuring different cultivars honey or adulterated honey, because
And easily there is the phenomenon of result inaccuracy.Patent is a kind of to be used for chaste honey, rape honey and acacia honey based on nuclear magnetic resonance technique
Identify (application No. is 201710742918.7) and disclose a kind of method for identifying specific honey using nuclear magnetic resonance technique, accurately
Efficiently, but honey sample is related to complicated filtration treatment, and preparation of samples process uses a large amount of expensive heavy water (0.03-0.5g/
L honey heavy aqueous solution), the cost is relatively high.Nuclear magnetic resonance spectroscopy (1H NMR) fingerprint pattern technology is as a kind of advanced inspection
Survey means, have it is lossless, quick, accuracy is high, the advantage of favorable reproducibility, have been used for safety or the sugar of a variety of honey products
Adulterated research is starched, and the country identifies pure northeast eucalyptus honey using this technology and is not yet seen in report.
Summary of the invention
Identified in view of the deficiencies of the prior art, it is an object of the present invention to provide a kind of using nuclear magnetic resonance spectroscopy finger-print
The method of northeast eucalyptus honey, economical and efficient, sample handling characteristics are flexible, can intuitively and accurately identify pure northeast lime tree
Honey.
The present invention provides the following technical solutions:
A method of northeast eucalyptus honey is identified using nuclear magnetic resonance spectroscopy finger-print, comprising the following steps:
S1, honey or syrup sample to be identified are weighed, is diluted with phosphate buffer, after sufficient vortex at standing
Reason；
S2, the solution after diluting in S1 is titrated to pH=2.5-5.0 with the hydrochloric acid of 1mol/L or potassium hydroxide solution；
S3, it measures through S2 treated sample 500-900 μ L, is added and contains 0.1-0.2%2,2,3,3- tetra- deuterated trimethyls
The heavy aqueous solution of silane acrylic acid and 0.01-0.02% sodium azide, constant volume to 1mL measure 500-600 μ L dress after being vortexed uniformly
Enter nuclear magnetic tube；
S4, the sample in nuclear magnetic tube, determination condition are measured using Liquid NMR instrument are as follows:
Water peak is suppressed using presaturation method；
Pulse protocol are as follows: noesygppr1d；
O1:4.7-4.8；
NS:BBI probe 16 or 32, BBO probe 32 or 64；
DS=2 or 4；
D1=4s；
S5, nuclear magnetic resonance spectroscopy finger-print is obtained, there are a chevrons near 3.0 ± 0.02ppm displacement in map
Three refer to peaks, then prove that identified honey or syrup sample is northeast eucalyptus honey, if can regard as without this feature peak shape
Non- northeast eucalyptus honey, syrup or adulterated northeast eucalyptus honey, syrup.
Preferably, in the S1, when honey or syrup sample to be identified are crystal habit, 60 DEG C of perseverance need to be lower than
It is weighed after melting cooling in tepidarium.
Preferably, the potassium dihydrogen phosphate that phosphate buffer is the 0.5mol/L that ultrapure water is prepared in the S1.
Preferably, the 1mol/L hydrochloric acid in the S2 or potassium hydroxide solution are prepared by ultrapure water.
Preferably, the measuring temperature of the Liquid NMR instrument in the S3 is 20-30 DEG C.
The beneficial effects of the present invention are:
(1) method provided by the invention, deuterated reagent consumption is few, at low cost.
(2) in method provided by the invention, the processing mode of honey to be identified or syrup sample is flexible, is simple and efficient.
(3) by the related assays parameter of setting Liquid NMR instrument, map obtained is observed and analyzes, it can be intuitive
Accurately distinguish northeast eucalyptus honey and non-northeast eucalyptus honey.
Detailed description of the invention
Attached drawing is used to provide further understanding of the present invention, and constitutes part of specification, with reality of the invention
It applies example to be used to explain the present invention together, not be construed as limiting the invention.In the accompanying drawings:
Fig. 1 is nuclear magnetic spectrogram of 4 northeast eucalyptus honey samples in 2.68-3.12ppm in embodiment 1；
Fig. 2 is the nuclear magnetic spectrogram of rape honey, chaste honey and acacia honey sample in 2.68-3.12ppm in embodiment 1；
Fig. 3 is the nuclear magnetic spectrogram of sunflower honey, cotton honey and Mel Jujubae sample in 2.68-3.12ppm in embodiment 1；
Fig. 4 is the nuclear magnetic spectrogram of fennel honey, Chinese prickly ash honey and buckwheat honey sample in 2.68-3.12ppm in embodiment 1；
Fig. 5 is nuclear magnetic spectrogram of 4 northeast eucalyptus honey samples in 2.68-3.12ppm in embodiment 2；
Fig. 6 is the nuclear magnetic spectrogram of lemon honey, citrus honey and honey of lungan flowers sample in 2.68-3.12ppm in embodiment 2；
Fig. 7 is the nuclear-magnetism spectrum of Radix Codonopsis honey in embodiment 2, rubber honey and the sweet sample of Acacia mangiumwilld tree in 2.68-3.12ppm
Figure；
Fig. 8 is nuclear magnetic spectrogram of 8 syrup samples in 2.68-3.12ppm in embodiment 2.
Specific embodiment
Embodiment 1
1, key instrument:
400MHz Liquid NMR spectrometer (Brooker,Switzerland company), BBI probe, equipped with suitable for microtitration
BTpH Unit；IT0079 electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd.)；5mm nuclear magnetic tube (the U.S.
Wilmad-labglass company).
2, reagent:
Sodium azide (NaN3, analyze pure) and it is purchased from Tianjin good fortune morning chemical reagent factory.Heavy water (D2O, 99.9% is deuterated) it is purchased from
Hai'an Nai Ji Chemical Co., Ltd..2,2,3,3- deuterated trimethyl silane propionic acid sodium salts (TSP, 98% is deuterated), potassium dihydrogen phosphate
(KH2PO4, analyze pure) and it is purchased from Shanghai Sinopharm Chemical Reagent Co., Ltd..37%HCl solution (excellent pure grade) is purchased from Shanghai state
Chemical reagent Co., Ltd, medicine group.KOH (analysis is pure) is purchased from Shanghai Sinopharm Chemical Reagent Co., Ltd..Wherein,
The KH of 0.5mol/L2PO4 buffer, the HCl and KOH solution of 1mol/L are formulated by ultrapure water respectively.0.15%TSP and
0.015%NaN3Mixed solution configured with heavy water.
3, sample collection
The acquisition of 3.1 pure northeast eucalyptus honeys: eucalyptus honey raw material in northeast used picks up from Jilin Province, China, Liaoning and black dragon respectively
River area, saves 1-2 in the environment of 20-25 DEG C；
The acquisition of 3.2 other single pure honey: rape honey, chaste honey, acacia honey, sunflower sweet (Xinjiang), cotton honey, jujube
Nectar, fennel honey, Chinese prickly ash honey and buckwheat honey；The places of origin of raw materials is arrived in person to acquire or provided by the well-known manufacturing enterprise of Chinese honey, is protected
It is true and reliable to demonstrate,prove source, and every kind of honey kind includes at least 20 samples, Storage period is 1-2.
4, sample detection
S1, the northeast 0.1g eucalyptus honey (eucalyptus honey 1, eucalyptus honey 2, eucalyptus honey 3 and eucalyptus honey 4) and other honey are weighed respectively
Sample, crystallized sample weigh after need to melting cooling under the water bath condition lower than 60 DEG C, and the KH of 0.5mol/L is added2PO4Buffer
1mL, sufficient vortex are stood；
S2, with 1mol/L hydrochloric acid or potassium hydroxide solution by the sample solution diluted in S1 be titrated to pH=2.8 ±
0.1；
Processed 800 μ L of sample is titrated in S3, measurement S2, is added and contains 0.15%2,2,3,3- tetra- deuterated trimethyl silanes
Acrylates (TSP) and 0.015% sodium azide (NaN3) 200 μ L of heavy aqueous solution, be vortexed uniformly after measure 550 μ L be packed into core
Magnetic tube；
S4, the sample in nuclear magnetic tube, determination condition are measured using Liquid NMR instrument are as follows:
Measuring temperature: 25 DEG C；
Water peak is suppressed using presaturation method；
Pulse protocol are as follows: noesygppr1d；
O1:4.7-4.8 (depending on water suppression condition)；
P1=10.4 μ s；
TD=64K；
NS=16 or 32；
DS=4；
SWH=8023Hz；
D1=4s；
D8=0.01s；
S5, analyzed using software Topspin3.5 all raw materials honey kind nuclear magnetic resonance spectroscopy (1H NMR) data, it will be interior
The chemical shift of mark TSP is set as 0, then carries out baseline correction and phasing；The nuclear magnetic resonance spectroscopy finger-print of acquisition
In, there are three fingers peak (see attached drawing 1) of a chevron, oil near 3.0 ± 0.02ppm displacement for only pure northeast eucalyptus honey
Dish honey, chaste honey, acacia honey, sunflower sweet (Xinjiang), cotton honey, Mel Jujubae, fennel honey, Chinese prickly ash honey and buckwheat honey are without this feature
Peak shape (see attached drawing 2,3,4), therefore the presence at this feature peak can be used as the mark for identifying northeast eucalyptus honey.
Embodiment 2
1, key instrument:
400MHz Liquid NMR spectrometer (German Brooker company), BBO probe；IT0079 electronic balance (Mei Tele-
Hold in the palm benefit instrument (Shanghai) Co., Ltd.)；ET-18 potentiometric titrimeter (Mettler-Toledo Instrument (Shanghai) Co., Ltd.)；
5mm nuclear magnetic tube (Wilmad-labglass company, the U.S.).
2, reagent:
Sodium azide (NaN3, analyze pure) and it is purchased from Tianjin good fortune morning chemical reagent factory.Heavy water (D2O, 99.9% is deuterated) it is purchased from
Hai'an Nai Ji Chemical Co., Ltd..2,2,3,3- deuterated trimethyl silane propionic acid sodium salts (TSP, 98% is deuterated), potassium dihydrogen phosphate
(KH2PO4, analyze pure) and it is purchased from Shanghai Sinopharm Chemical Reagent Co., Ltd..37%HCl solution (excellent pure grade) is purchased from Shanghai state
Chemical reagent Co., Ltd, medicine group.KOH (analysis is pure) is purchased from Shanghai Sinopharm Chemical Reagent Co., Ltd..Wherein,
The KH of 0.5mol/L2PO4Buffer, the HCl and KOH solution of 1mol/L are formulated by ultrapure water respectively.0.15%TSP and
0.015%NaN3Mixed solution configured with heavy water.
3, sample collection
The acquisition of 3.1 pure northeast eucalyptus honeys: eucalyptus honey raw material in northeast used picks up from Jilin Province, China, Liaoning and black dragon respectively
River area, saves 1-2 in the environment of 20-25 DEG C；
The acquisition of 3.2 other single pure honey: honey of lungan flowers, citrus honey, lemon honey, Acacia mangiumwilld tree honey, rubber honey, party
Ginseng honey；The places of origin of raw materials is arrived in person to acquire or provided by the well-known manufacturing enterprise of Chinese honey, guarantees that source is true and reliable, and every kind of honey
Kind includes at least 20 samples, and Storage period is 1-2；
The collection of 3.3 all kinds of syrup: including beet class, starch and other 8, all types of syrup, respectively from domestic each
Syrup manufacturer.
4, sample detection
S1, weigh respectively the northeast 8g eucalyptus honey (eucalyptus honey 5, eucalyptus honey 6, eucalyptus honey 7 and eucalyptus honey 8) and other honey and
All kinds of syrup samples, crystallized sample weigh after need to melting cooling under the water bath condition lower than 60 DEG C, are added 0.5mol/L's
KH2PO4Buffer 25mL, sufficient vortex are stood；
S2, with 1mol/L hydrochloric acid or potassium hydroxide solution by the sample solution diluted in S1 be titrated to pH=2.8 ±
0.1；
Processed 700 μ L of sample is titrated in S3, measurement S2, is added and contains 0.15%2,2,3,3- tetra- deuterated trimethyl silanes
Acrylates (TSP) and 0.015% sodium azide (NaN3) 300 μ L of heavy aqueous solution, be vortexed uniformly after measure 600 μ L be packed into core
Magnetic tube；
S4, the sample in nuclear magnetic tube, determination condition are measured using Liquid NMR instrument are as follows:
Measuring temperature: 20 DEG C；
Water peak is suppressed using presaturation method；
Pulse protocol are as follows: noesygppr1d；
O1:4.7-4.8 (depending on water suppression condition)；
P1=10.5 μ s；
TD=64K；
NS=32 or 64；
DS=2；
SWH=8012Hz；
D1=4s；
D8=0.01s；
S5, analyzed using software Topspin3.5 all raw materials honey kind nuclear magnetic resonance spectroscopy (1H NMR) data, it will be interior
The chemical shift of mark TSP is set as 0, then carries out baseline correction and phasing；The nuclear magnetic resonance spectroscopy finger-print of acquisition
In, there are three fingers peak (see attached drawing 5) of a chevron, dragons near 3.0 ± 0.02ppm displacement for only pure northeast eucalyptus honey
Eye honey, citrus honey, lemon honey, Acacia mangiumwilld tree honey, rubber honey, Radix Codonopsis honey and 8 syrup samples are without this feature peak shape (see attached
Fig. 6,7,8), therefore the presence at this feature peak can be used as identify northeast eucalyptus honey mark.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, although referring to aforementioned reality
Applying example, invention is explained in detail, for those skilled in the art, still can be to aforementioned each implementation
Technical solution documented by example is modified or equivalent replacement of some of the technical features.It is 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 method for identifying northeast eucalyptus honey using nuclear magnetic resonance spectroscopy finger-print, which is characterized in that including following step
It is rapid:
S1, honey or syrup sample to be identified are weighed, is diluted with phosphate buffer, the stewing process after sufficient vortex；
S2, the solution after diluting in S1 is titrated to pH=2.5-5.0 with the hydrochloric acid of 1mol/L or potassium hydroxide solution；
S3, it measures through S2 treated sample 500-900 μ L, is added and contains 0.1-0.2%2,2,3,3- tetra- deuterated trimethyl silanes
The heavy aqueous solution of acrylic acid and 0.01-0.02% sodium azide, constant volume to 1mL measure 500-600 μ L after being vortexed uniformly and are packed into core
Magnetic tube；
S4, the sample in nuclear magnetic tube, determination condition are measured using Liquid NMR instrument are as follows:
Water peak is suppressed using presaturation method；
Pulse protocol are as follows: noesygppr1d；
O1:4.7-4.8；
NS:BBI probe 16 or 32, BBO probe 32 or 64；
DS=2 or 4；
D1=4s；
S5, nuclear magnetic resonance spectroscopy finger-print is obtained, there are the three of a chevron near 3.0 ± 0.02ppm displacement in map
Refer to peak, then proves that identified honey or syrup sample are northeast eucalyptus honey, if non-east can be regarded as without this feature peak shape
Northern eucalyptus honey, syrup or adulterated northeast eucalyptus honey, syrup.
2. a kind of method for identifying northeast eucalyptus honey using nuclear magnetic resonance spectroscopy finger-print according to claim 1,
It is characterized in that, in the S1, when honey or syrup sample to be identified are crystal habit, 60 DEG C of water bath with thermostatic control need to be lower than
It is weighed after middle thawing is cooling.
3. a kind of method for identifying northeast eucalyptus honey using nuclear magnetic resonance spectroscopy finger-print according to claim 1,
It is characterized in that, phosphate buffer is the potassium dihydrogen phosphate for the 0.5mol/L that ultrapure water is prepared in the S1.
4. a kind of method for identifying northeast eucalyptus honey using nuclear magnetic resonance spectroscopy finger-print according to claim 1,
It is characterized in that, the 1mol/L hydrochloric acid or potassium hydroxide solution in the S2 are prepared by ultrapure water.
5. a kind of method for identifying northeast eucalyptus honey using nuclear magnetic resonance spectroscopy finger-print according to claim 1,
It is characterized in that, the measuring temperature of the Liquid NMR instrument in the S3 is 20-30 DEG C.
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| CN114088827A (en) * | 2021-10-29 | 2022-02-25 | 中国农业科学院蜜蜂研究所 | Application of shikimic acid and quinic acid as characteristic markers of linden honey |
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Address after: 210000 building B1-3, building 16, Shuwu, No. 73, tanmi Road, Jiangbei new district, Nanjing City, Jiangsu Province Applicant after: Titanium and spectrum detection technology (Jiangsu) Co.,Ltd. Address before: 5 and 6 / F, block C, Dingye biological building, No.10 Xinghuo Road, Jiangbei new district, Nanjing, Jiangsu Province 210000 Applicant before: JIANGSU SINOGRAPHY TESTING Co.,Ltd. |
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