CN109752469A - A method of identifying eucalyptus honey authenticity - Google Patents
A method of identifying eucalyptus honey authenticity Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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Abstract
The present invention provides a kind of methods for identifying the eucalyptus honey true and false, belong to technical field of food detection, the described method comprises the following steps: 1) will honey be reflected and water mixed dissolution, adjusting pH value is 6~7, obtains sample solution；2) sample solution is crossed into reverse phase-anion-exchange solid phase extraction column, elutes extraction column with the methanol solution of the formic acid containing 10wt%, collects eluent；3) with eluent described in high performance liquid chromatography detection, the finger-print of the material pattern that will test and eucalyptus honey polyphenol components is compared, if detect substance type it is consistent with the finger-print of eucalyptus honey polyphenol components if be true eucalyptus honey；The finger-print of the eucalyptus honey polyphenol components be include lime tree element, anti-, trans- abscisic acid and suitable, the finger-print of trans- abscisic acid.The present invention establishes a set of sensitive, accurate, efficient, pervasive eucalyptus honey authenticity identification method, completely solves eucalyptus honey authenticity identification problem.
Description
Technical field
The invention belongs to Food Inspection technical field more particularly to a kind of methods for identifying eucalyptus honey authenticity.
Background technique
Eucalyptus honey is well received by consumers as one of maximum single flower honey of China's yield.However, of poor quality in the market mix
False eucalyptus honey honey is very universal, and the fraud of eucalyptus honey honey has formed scale, specialization.A large amount of vacation eucalyptus honeys are mixed into city
, the health of consumer is seriously threatened, the development of China's bee industry is influenced.
The method for identifying honey authenticity at present mainly has；Identify including sense organ, pollen analysis and physical and chemical index are tested.
In recent years, using advanced Modern Scientific Instruments in Chinese, developed a variety of detection and analysis technology and methods for honey adulteration, it is main to wrap
Include stable carbon isotope ratio method (GB/T 18932.1-2002), chromatography (GB/T 18932.1-2002) spectrum analysis (closely
Infrared, fluorescence, Raman, uv-visible absorption spectra), amylase detection method etc..There are unstable results, operation for these methods
Complexity, it is at high cost the problems such as.
Summary of the invention
In view of this, the present invention provides a kind of method for identifying eucalyptus honey authenticity, method of the present invention, as a result
Reliable and stable, degree of judgement is high, facilitates operation, is simple and easy, is low in cost.
Based on above-mentioned purpose, the present invention provides a kind of methods for identifying eucalyptus honey authenticity, comprising the following steps: 1) will
Honey to be reflected and water mixed dissolution, adjusting pH value is 6~7, obtains sample solution；2) by the sample solution cross reverse phase-yin from
Son exchange solid-phase extraction column elutes extraction column with the methanol solution of the formic acid containing 10wt%, collects eluent；3) high-efficient liquid phase color is used
Spectrum detects the eluent, and the finger-print of the liquid chromatogram that will test and true eucalyptus honey polyphenol components compares, if inspection
The type for measuring substance is consistent with the finger-print of true eucalyptus honey polyphenol components, is true eucalyptus honey；
The finger-print of the true eucalyptus honey polyphenol components be include lime tree element, anti-, trans- abscisic acid and suitable, it is trans- de-
Fall the finger-print of acid.
Preferably, after the step 3) further includes high performance liquid chromatography separation, according to high-efficient liquid phase chromatogram external standard method
Calculate the lime tree element detected, anti-, trans- abscisic acid and suitable, trans- abscisic acid is wait the content in the honey that reflects.
Preferably, content > 45 μ g/100g of the lime tree element in true eucalyptus honey.
Preferably, described anti-, content > 15 μ g/100g of the trans- abscisic acid in true eucalyptus honey.
Preferably, described suitable, content > 30 μ g/100g of the trans- abscisic acid in true eucalyptus honey.
Preferably, it is 0.7ml/min that the condition of high performance liquid chromatography detection described in step 3), which includes: flow velocity,；It is ultraviolet
Detector, Detection wavelength 280nm；Splitter is C18 column, and the specification of the C18 column is 150 × 4.6mm, 5 μm；Loading volume
For 20 μ l；
Mobile phase A is the acetic acid aqueous solution of mass fraction 0.2%；Mobile phase B is that the acetic acid methanol of mass fraction 0.2% is molten
Liquid；
Gradient program are as follows: [0~11) min, [0%~8%) Mobile phase B；[11~14) min, [8%~10%) stream
Dynamic phase B；[14~17) min, [10%~14%) Mobile phase B；[17~24) min, (14%~20%] Mobile phase B；[24~
28) min, (20%~21%] Mobile phase B；[28~30) min, (21%~22%] Mobile phase B；[30~36) min, (22%
~25%] Mobile phase B；[36~41) min, (25%~30%] Mobile phase B；[41~46) min, (30%~33%] mobile phase
B；[46~55) min, 33% Mobile phase B；[55~60) min, (33%~34%] Mobile phase B；[60~70) min, (34%~
36%] Mobile phase B；[70~80) min, (36%~40%] Mobile phase B；[80~90) min, (40%~45%] Mobile phase B；
[90~100) min, (45~52%] Mobile phase B.
Preferably, the eucalyptus honey polyphenols finger-print is prepared by the following: taking 50~100 parts of true lime trees
Sweet sample carries out high performance liquid chromatography detection respectively, the polyphenol components chromatogram of every a sample is obtained, by all samples
Polyphenol components chromatogram imports traditional Chinese medicine fingerprint software, obtains the finger-print of polyphenol components in true eucalyptus honey.
Preferably, the standard curve that external standard method calculates lime tree element in step 3) is y=7134x-6569, and wherein x is detection
The peak area of substance, y detect the content of substance, and the unit of y is ng.
Preferably, external standard method calculates anti-, trans- abscisic acid and suitable in step 3), and the standard curve of trans- abscisic acid is y=
4513x+135049, wherein x is the peak area for detecting substance, and y detects the content of substance.
Preferably, the mass ratio of the honey to be reflected and water is 1:(3.5~4.5).
Beneficial effects of the present invention: the method provided by the invention for identifying eucalyptus honey authenticity mixes sample to be reflected with water
Reverse phase-anion-exchange solid phase extraction column is crossed after closing dissolution, the polyphenols treated in mirror sample is enriched with, in eluent
Polyphenols be more easily detected, testing result is reliable and stable, degree of judgement is high, facilitates operation, is low in cost.The present invention
A set of sensitive, accurate, efficient, pervasive eucalyptus honey authenticity identification method is established, the knowledge of eucalyptus honey authenticity is completely solved
Other problem ensure that the quality and edible safety of honey product, protects consumer's interests and guarantees that honey industry develops in a healthy way.
Detailed description of the invention
Fig. 1 is the finger-print of eucalyptus honey polyphenol components.
Specific embodiment
The present invention provides a kind of methods for identifying eucalyptus honey authenticity, comprising the following steps: 1) will honey and water be reflected
Mixed dissolution, adjusting pH value is 6~7, obtains sample solution；2) sample solution reverse phase-anion exchange solid phase is crossed to extract
Column is taken, extraction column is eluted with the methanol solution of the formic acid containing 10wt%, collects eluent；3) 3) with high performance liquid chromatography detection described in
The finger-print of eluent, the liquid chromatogram that will test and true eucalyptus honey polyphenol components compares, if detecting substance
Type is consistent with the finger-print of true eucalyptus honey polyphenol components, is true eucalyptus honey；The finger of the true eucalyptus honey polyphenol components
Line map be include lime tree element, anti-, trans- abscisic acid and suitable, the finger-print of trans- abscisic acid.
In the present invention, will honey be reflected and water mixed dissolution, adjusting pH value is 6~7, obtains sample solution.In this hair
In bright, the mass ratio of the honey to be reflected and water is preferably 1:(3.5~4.5), more preferably 1:(3.8~4.2), most preferably
For 1:4.In the present invention, the honey to be reflected is any type of honey, including but not limited to raw material honey and commodity honey；It is described
Water is preferably ultrapure water.The present invention it is described preferably further include centrifugation step after the sample that reflects is mixed with water, the centrifugation
Revolving speed is preferably 8000~12000g, more preferably 10000g；The time of the centrifugation is preferably 8~12min, more preferably
10min.The effect of heretofore described centrifugation is removal impurity, and the present invention preferably collects supernatant after the centrifugation, is carried out
Subsequent processing.The pH value for preferably adjusting solution using alkaloids in the present invention, more preferably adjusts solution using ammonium hydroxide
PH value；The mass concentration of the ammonium hydroxide is preferably 4~6%, and more preferably 5%.
The present invention crosses reverse phase-anion-exchange solid phase extraction column after obtaining sample solution, by the sample solution, collects
Obtain eluent.In the present invention, the reverse phase-anion-exchange solid phase extraction column specifications and models RP-AE SPE.Institute of the present invention
The reverse phase stated-anion-exchange solid phase extraction column using preceding preferably after methanol activates with ultrapure water balance.In the present invention
Described in the dosage of methanol be preferably 0.8~1.2ml, more preferably 1.0ml；The dosage of the ultrapure water is preferably 0.8~
1.2ml, more preferably 1.0ml.In the present invention, by after the sample solution upper prop, extraction column preferably is cleaned with ultrapure water,
Then extraction column is eluted with the methanol solution of the formic acid containing 10wt%, collects eluent.In the present invention, the cleaning extraction column is used
The dosage of ultrapure water be preferably 0.8~1.2ml, more preferably 1.0ml；The dosage of the methanol solution of the formic acid containing 10wt%
For 1.5~2.5ml, more preferably 2.0ml.The present invention preferably further includes that the eluent is dry after obtaining eluent
After redissolve.In the present invention, the drying is preferably used and is dried with nitrogen；The present invention is to the flow velocity of the nitrogen and is dried with nitrogen
Time be not particularly limited, can be realized dry.The present invention preferably uses chromatographic grade acetic acid first after the drying
Alcoholic solution (2:98V/V) is redissolved, and the dosage of the chromatographic grade acetic acid methanol solution is preferably 1.5~2.5ml, more preferably
2ml.Before carrying out high performance liquid chromatography detection, the eluent is filtered after obtaining the eluent redissolved by the present invention.It is described
Filtering is preferably membrane filtration, and the filter membrane is preferably 0.22 μm of filter membrane, and high performance liquid chromatography sample solution is obtained after filtering.
After obtaining high performance liquid chromatography sample solution, the sample solution liquid described in high performance liquid chromatography detection will be examined the present invention
The finger-print of the substance measured and eucalyptus honey polyphenol components compares, and detects the type and eucalyptus honey polyphenol components of substance
Finger-print be unanimously then true eucalyptus honey.
The mobile phase A of the high performance liquid chromatography is the water of 0.2% acetic acid containing mass fraction in the present invention；Mobile phase B
For the methanol of 0.2% acetic acid containing mass fraction；The flow velocity of the high performance liquid chromatography is 0.7ml/min；Ultraviolet detection wavelength
For 280nm；Splitter is C18 column, and the specification of the C18 column is 150 × 4.6mm, 5 μm；Loading volume is 20 μ l；Gradient
For [0~11) min, [0%~8%) Mobile phase B；[11~14) min, [8%~10%) Mobile phase B；[14~17) min,
[10%~14%) Mobile phase B；[17~24) min, (14%~20%] Mobile phase B；[24~28) min, (20%~21%]
Mobile phase B；[28~30) min, (21%~22%] Mobile phase B；[30~36) min, (22%~25%] Mobile phase B；[36~
41) min, (25%~30%] Mobile phase B；[41~46) min, (30%~33%] Mobile phase B；[46~55) min, 33% stream
Dynamic phase B；[55~60) min, (33%~34%] Mobile phase B；[60~70) min, (34%~36%] Mobile phase B；[70~
80) min, (36%~40%] Mobile phase B；[80~90) min, (40%~45%] Mobile phase B；[90~100) min, (45~
52%] Mobile phase B.
The chromatogram and lime tree that the present invention arrives the high performance liquid chromatography detection after the high performance liquid chromatography detection
The finger-print of sweet polyphenol components compares, and confirmly detects the type of substance.In the present invention, the eucalyptus honey Polyphenols object
Matter finger-print is prepared by the following: 50~100 parts of true eucalyptus honey samples taken, carry out high performance liquid chromatography detection respectively,
The polyphenol components chromatogram of every a sample is obtained, the polyphenol components chromatogram of all samples is imported into traditional Chinese medicine fingerprint
Software obtains the finger-print of polyphenol components in eucalyptus honey.Heretofore described true eucalyptus honey sample source be preferably Jilin,
More than 30 a bee farms on the ground such as Liaoning and Heilungkiang；In the present invention, the finger-print of the eucalyptus honey polyphenol components is preferably such as
Shown in Fig. 1, wherein 1 is lime tree element, 2 be anti-, trans- abscisic acid, and 3 be suitable, trans- abscisic acid.
In the present invention, eucalyptus honey refers to that honeybee acquires lime tree nectar (including tilia amurensis and chaff linden), mixes with honeybee salivary gland
Secretion, through the sugariness substance for sufficiently brewing and being stored at honeycomb.Lime tree element is that have characteristic plant chemical combination in eucalyptus honey
Object.Abscisic acid is the existing plant endogenous hormones with sequiterpene structure in a kind of plant, has control plant growth, suppression
The effects such as aging are sprouted and promoted to production of hybrid seeds, and with the continuous deepening of research, abscisic acid is inverse in arid, the with high salt, low temperature of plant etc.
It plays an important role in the Stress responses of border, it is the anticounter-inducer of plant, thus " the stress hormone " of referred to as plant.Lime tree
Contain abscisic acid in honey.
The present invention calculates content after confirmly detecting the type of substance, using external standard method；It is described anti-, trans- abscisic acid and
Suitable, the standard curve of trans- abscisic acid is y=4513x+135049, and wherein x is the peak area for detecting substance, and y detects substance
Content；In the present invention, the unit of y is nanogram (ng).The standard curve of heretofore described lime tree element is y=7134x-6569,
Wherein x is the peak area for detecting substance, and y detects the content of substance；In the present invention, the unit of y is nanogram (ng).
In the present invention, the preferred 45 μ g/100g of > of content of the lime tree element in true eucalyptus honey；It is described anti-, it is trans- de-
Fall content preferred > 15 μ g/100g of the acid in true eucalyptus honey；Described suitable, content of the trans- abscisic acid in true eucalyptus honey is excellent
The 30 μ g/100g of > of choosing.
Technical solution provided by the invention is described in detail below with reference to embodiment, but they cannot be understood
For limiting the scope of the present invention.
Embodiment 1
Certain agent carries out quality evaluation to the raw material eucalyptus honey honey of collection.
Operating method:
1,20g eucalyptus honey honey sample is accurately weighed, is placed in dry bottle, the dissolution of 80ml ultrapure water is added；
2,10000 × g is centrifuged 10min, takes supernatant spare；
3, pH to 6-7 is adjusted with 5% ammonium hydroxide
4, reverse phase-anion-exchange solid phase extraction column (RP-AE SPE) through 1mL methanol activation after with 1mL ultrapure water (pH:
It 6-7) balances, supernatant crosses column；
5,1ml ultrapure water cleans extraction column, and 2ml methanol (containing 10% formic acid) elution extraction column collects eluent；
6, eluent is blown to drying with nitrogen at room temperature, multiple with 2mL chromatographic grade acetic acid methanol solution (2:98V/V)
It is miscible even, 0.22 μm of filter membrane is crossed, HPLC sample solution is obtained.
7, the HPLC sample solution is detected using HPLC method；The detection method of high performance liquid chromatography (HPLC) are as follows:
Mobile phase A is water (containing 0.2% acetic acid)；Mobile phase B is methanol (containing 0.2% acetic acid)；Overall flow rate is 0.7ml/min；Ultraviolet inspection
Survey wavelength is 280nm；Splitter is C18 column, (150 × 4.6mm, 5 μm)；Loading volume is 20 μ l；Gradient is 0-
11min, 0%-8%B；11-14min, 8-10%B；14-17min, 10%-14%B；17-24min, 14-20%B；24-
28min, 20-21%B；28-30min, 21-22%B；30-36min, 22-25%B；36-41min, 25-30%B；41-
46min, 30-33%B；46-55min, 33%B；55-60min, 33-34%B；60-70min, 34-36%B；70-80min,
36-40%B；80-90min, 40-45%B；90-100min, 45-52%B.
8, content is calculated using external standard method:
The standard curve of lime tree element is y=7134x-6569；Instead, trans- abscisic acid and suitable, the standard song of trans- abscisic acid
Line is y=4513x+135049；
Qualitative and quantitative analysis is carried out to sample solution according to HPLC method:
According to mark Qu Jinhang quantitative analysis, 80 μ g/100g of lime tree element are obtained；Instead, trans- abscisic acid is 30 μ g/100g；It is suitable, instead
Formula abscisic acid is 70 μ g/100g.And chromatogram is compared with standard finger-print, without other exotic matters.To sum up, raw material honey
It is eucalyptus honey.
Embodiment 2
Certain enterprise carries out quality evaluation to the raw material honey of buying.
Operating method:
1,20g eucalyptus honey honey sample is accurately weighed, is placed in dry bottle, the dissolution of 80ml ultrapure water is added；
2,10000 × g is centrifuged 10min, takes supernatant spare；
3, pH to 6-7 is adjusted with 5% ammonium hydroxide；
4, reverse phase-anion-exchange solid phase extraction column (RP-AE SPE) through 1mL methanol activation after with 1mL ultrapure water (pH:
It 6-7) balances, supernatant crosses column；
5,1ml ultrapure water cleans extraction column, and 2ml methanol (containing 10% formic acid) elution extraction column collects eluent；
6, eluent is blown to drying with nitrogen at room temperature, multiple with 2mL chromatographic grade acetic acid methanol solution (2:98V/V)
It is miscible even, 0.22 μm of filter membrane is crossed, HPLC sample solution is obtained.
7, the HPLC sample solution is detected using HPLC method；The detection method of high performance liquid chromatography (HPLC) are as follows:
Mobile phase A is water (containing 0.2% acetic acid)；Mobile phase B is methanol (containing 0.2% acetic acid)；Overall flow rate is 0.7ml/min；Ultraviolet inspection
Survey wavelength is 280nm；Splitter is C18 column, (150 × 4.6mm, 5 μm)；Loading volume is 20 μ l；Gradient is 0-
11min, 9%-14%B；11-14min, 14-15%B；14-17min, 15%B；17-24min, 15-16%B；24-28min,
16-17%B；28-30min, 17-22%B；30-38min, 22-25%B；38-41min, 25-30%B；41-46min, 30-
33%B；46-55min, 33%B；55-60min, 33-34%B；60-70min, 34-36%B；70-80min, 36-40%B；
80-90min, 40-45%B；90-100min, 45-52%B.
8, content is calculated using external standard method:
The standard curve of lime tree element is y=7134x-6569；Instead, trans- abscisic acid and suitable, the standard song of trans- abscisic acid
Line is y=4513x+135049；
Qualitative and quantitative analysis is carried out to sample solution according to HPLC method:
According to mark Qu Jinhang quantitative analysis, 130 μ g/100g of lime tree element are obtained；Instead, trans- abscisic acid is 42 μ g/100g；
Suitable, trans- abscisic acid is 60 μ g/100g.And chromatogram is compared with standard finger-print, without other exotic matters.To sum up, should
Raw material honey is eucalyptus honey.
Embodiment 3
Certain third party testing agency carries out quality testing to the commodity honey of commission detection.
Operating method:
Operating method:
1,20g eucalyptus honey honey sample is accurately weighed, is placed in dry bottle, the dissolution of 80ml ultrapure water is added；
2,10000 × g is centrifuged 10min, takes supernatant spare；
3, pH to 6-7 is adjusted with 5% ammonium hydroxide；
4, reverse phase-anion-exchange solid phase extraction column (RP-AE SPE) through 1mL methanol activation after with 1mL ultrapure water (pH:
It 6-7) balances, supernatant crosses column；
5,1ml ultrapure water cleans extraction column, and 2ml methanol (containing 10% formic acid) elution extraction column collects eluent；
6, eluent is blown to drying with nitrogen at room temperature, multiple with 2mL chromatographic grade acetic acid methanol solution (2:98V/V)
It is miscible even, 0.22 μm of filter membrane is crossed, HPLC sample solution is obtained.
7, the HPLC sample solution is detected using HPLC method；The detection method of high performance liquid chromatography (HPLC) are as follows:
Mobile phase A is water (containing 0.2% acetic acid)；Mobile phase B is methanol (containing 0.2% acetic acid)；Overall flow rate is 0.7ml/min；Ultraviolet inspection
Survey wavelength is 280nm；Splitter is C18 column, (150 × 4.6mm, 5 μm)；Loading volume is 20 μ l；Gradient is 0-
11min, 9%-14%B；11-14min, 14-15%B；14-17min, 15%B；17-24min, 15-16%B；24-28min,
16-17%B；28-30min, 17-22%B；30-38min, 22-25%B；38-41min, 25-30%B；41-46min, 30-
33%B；46-55min, 33%B；55-60min, 33-34%B；60-70min, 34-36%B；70-80min, 36-40%B；
80-90min, 40-45%B；90-100min, 45-52%B.
8, content is calculated using external standard method:
The standard curve of lime tree element is y=7134x-6569；Instead, trans- abscisic acid and suitable, the standard song of trans- abscisic acid
Line is y=4513x+135049.
Qualitative and quantitative analysis is carried out to sample solution according to HPLC method:
According to mark Qu Jinhang quantitative analysis, obtaining lime tree element is 94 μ g/100g；Instead, trans- abscisic acid is 50 μ g/100g；
Suitable, trans- abscisic acid is 82 μ g/100g.And chromatogram is compared with standard finger-print, without other exotic matters.To sum up, should
Raw material honey is eucalyptus honey.
As can be seen from the above embodiments, the method provided by the invention for identifying eucalyptus honey authenticity, testing result is reliable and stable,
Degree of judgement is high, facilitates operation, is low in cost.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of method for identifying eucalyptus honey authenticity, which comprises the following steps:
1) will honey be reflected and water mixed dissolution, adjusting pH value is 6~7, obtains sample solution；
2) sample solution is crossed into reverse phase-anion-exchange solid phase extraction column, is eluted with the methanol solution of the formic acid containing 10wt%
Extraction column collects eluent；
3) eluent described in high performance liquid chromatography detection, the liquid chromatogram that will test and true eucalyptus honey polyphenol components
Finger-print compare, if detect substance type it is consistent with the finger-print of true eucalyptus honey polyphenol components if be true lime tree
Honey；
The finger-print of the true eucalyptus honey polyphenol components be include lime tree element, anti-, trans- abscisic acid and suitable, trans- abscisic acid
Finger-print.
2. the method according to claim 1, wherein after the step 3) further includes high performance liquid chromatography separation,
Calculate the lime tree element detected, anti-according to high-efficient liquid phase chromatogram external standard method, trans- abscisic acid and suitable, trans- abscisic acid to
Content in mirror honey.
3. according to the method described in claim 2, it is characterized in that, content > 45 μ g/ of the lime tree element in true eucalyptus honey
100g。
4. according to the method in claim 2 or 3, which is characterized in that described anti-, trans- abscisic acid containing in true eucalyptus honey
Measure 15 μ g/100g of >.
5. according to the method described in claim 2, it is characterized in that, described suitable, content of the trans- abscisic acid in true eucalyptus honey
30 μ g/100g of >.
6. the method according to claim 1, wherein the condition of high performance liquid chromatography detection described in step 3)
Include: flow velocity be 0.7ml/min；UV detector, Detection wavelength 280nm；Splitter is C18 column, the specification of the C18 column
For 150 × 4.6mm, 5 μm；Loading volume is 20 μ l；
Mobile phase A is the acetic acid aqueous solution of mass fraction 0.2%；Mobile phase B is the acetic acid methanol solution of mass fraction 0.2%；
Gradient program are as follows: [0~11) min, [0%~8%) Mobile phase B；[11~14) min, [8%~10%) mobile phase
B；[14~17) min, [10%~14%) Mobile phase B；[17~24) min, (14%~20%] Mobile phase B；[24~28)
Min, (20%~21%] Mobile phase B；[28~30) min, (21%~22%] Mobile phase B；[30~36) min, (22%~
25%] Mobile phase B；[36~41) min, (25%~30%] Mobile phase B；[41~46) min, (30%~33%] Mobile phase B；
[46~55) min, 33% Mobile phase B；[55~60) min, (33%~34%] Mobile phase B；[60~70) min, (34%~
36%] Mobile phase B；[70~80) min, (36%~40%] Mobile phase B；[80~90) min, (40%~45%] Mobile phase B；
[90~100) min, (45~52%] Mobile phase B.
7. according to the method described in claim 6, it is characterized in that, the eucalyptus honey polyphenols finger-print is by following
Method obtains: taking 50~100 parts of true eucalyptus honey samples, carries out high performance liquid chromatography detection respectively, obtains the more of every a sample
The polyphenol components chromatogram of all samples is imported traditional Chinese medicine fingerprint software, obtains true eucalyptus honey by phenols component chromatogram
The finger-print of middle polyphenol components.
8. according to the method described in claim 2, it is characterized in that, the standard curve of external standard method calculating lime tree element is in step 3)
Y=7134x-6569, wherein x is the peak area for detecting substance, and y detects the content of substance；The unit of y is ng.
9. the method according to claim 1, wherein external standard method calculates anti-, trans- abscisic acid and suitable in step 3),
The standard curve of trans- abscisic acid is y=4513x+135049, and wherein x is the peak area for detecting substance, and y detection substance contains
Amount.
10. the method according to claim 1, wherein the mass ratio of the honey to be reflected and water be 1:(3.5~
4.5)。
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811228827.2A CN109752469A (en) | 2018-10-22 | 2018-10-22 | A method of identifying eucalyptus honey authenticity |
| BE20195732A BE1026676B1 (en) | 2018-10-22 | 2019-10-22 | Process to identify the authenticity of lime blossom honey |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811228827.2A CN109752469A (en) | 2018-10-22 | 2018-10-22 | A method of identifying eucalyptus honey authenticity |
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|---|---|---|---|---|
| CN116046954A (en) * | 2023-02-22 | 2023-05-02 | 秦皇岛海关技术中心 | Method for measuring content of callic acid in honey |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112461959A (en) * | 2020-11-16 | 2021-03-09 | 河南汇泉堂中药饮片科技有限公司 | Method for identifying selfheal honey and application thereof |
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2018
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116046954A (en) * | 2023-02-22 | 2023-05-02 | 秦皇岛海关技术中心 | Method for measuring content of callic acid in honey |
| CN116046954B (en) * | 2023-02-22 | 2023-09-05 | 秦皇岛海关技术中心 | Method for measuring content of callic acid in honey |
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| BE1026676A1 (en) | 2020-04-30 |
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