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    • 专利摘要:
    The present invention provides a method for identifying the authenticity of linden honey. The method includes the following steps: 1): mixing a honey to be identified with dissolution water, and adjusting the pH value to 6-7 to obtain a sample solution; 2): pass the sample solution through a solid phase extraction column by reverse phase anion exchange, and elution from the extraction column with a methanol solution containing 10% by weight of acid formic, and collection of an eluent; 3) detecting the eluent by high performance liquid chromatography, and comparing a chromatogram of the substance detected with a fingerprint of a phytochemical component of lime blossom honey; the fingerprint of the phytochemical component of lime blossom honey is a fingerprint including lindenine, trans, trans-abscisic acid and cis, trans-abscisic acid.
    公开号:BE1026676B1
    申请号:E20195732
    申请日:2019-10-22
    公开日:2020-11-30
    发明作者:Fumin Yu；Hongcheng Zhang；Hailong Zhu；Jiangtao Qiao
    申请人:Shanghai Senfengyuan Bee Ind Co Ltd；Institute Of Apicultural Res Chinese Academy Of Agricultural Sciences；
    IPC主号:
    专利说明:

    PROCESS FOR IDENTIFYING THE AUTHENTICITY OF TILLEUL HONEY
    TECHNICAL FIELD The present invention belongs to the technical field of food inspection, and particularly relates to a method for identifying the authenticity of lime blossom honey.
    BACKGROUND As one of the most widely produced monofloral honeys in China, lime blossom honey is deeply appreciated by consumers. However, false and adulterated linden honey is very common in the market, and counterfeiting of linden honey is taking place on a large scale and becoming specialized. A large amount of falsified lime blossom honey is introduced to the market, which seriously threatens the health of consumers and affects the development of Chinese beekeeping.
    Currently, the methods for identifying the authenticity of honey mainly include sensory identification, analysis of pollen and testing for physical and chemical indices. In recent years, regarding the adulteration of honey, advanced modern instruments and equipment have been used to develop a variety of techniques and methods for detection and analysis, including a method for measuring the ratio of stable carbon isotopes (GB / T 18932.1-2002), a spectral analysis (near infrared absorption spectrum, fluorescence, Raman, ultraviolet / visible) chromatography (GB / T 18932.1-2002), a method for the detection of amylase , and others. These methods present problems such as unstable results, complicated operation, and high cost.
    SUMMARY In view of this, the present invention provides a method for identifying the authenticity of linden honey, and the method of the present invention gives reliable and stable results and a high degree of determination and is easy to operate, simple, practical. and low cost.
    To achieve the above objective, the present invention provides a method for identifying the authenticity of lime blossom honey, including the following steps: step 1): mixing of a honey to be identified with water of dissolution, and adjusting the pH value to 6-7 to obtain a sample solution; step 2): passing the sample solution through a solid phase extraction column by reverse phase anion exchange, eluting the extraction column with a methanol solution containing 10% by weight formic acid, and collecting the eluent; and step 3): detection of the eluent by high performance liquid chromatography, comparison of a detected liquid chromatogram with an imprint of a phytochemical component of genuine lime blossom honey, where if the type of a substance detected corresponds to the imprint of the phytochemical component of authentic linden honey, honey is authentic linden honey; and the imprint of the phytochemical component of genuine lime blossom honey is a fingerprint including lindenin (4- (2-hydroxypropan-2-yl) cyclohexa-1,3-diene-1-carboxylic acid), trans , trans-abscisic and cis acid, trans-abscisic.
    Preferably, step 3) further includes the calculation, after separation by high performance liquid chromatography, of the contents of lindenine, trans, trans-abscisic acid and cis, trans-abscisic acid detected in the honey to be detected. identified by chromatogram obtained by high performance liquid chromatography using an external standard method.
    Preferably, the content of lindenin in genuine lime honey is greater than 45 µg / 100 g.
    Preferably, the content of trans, trans-abscisic acid in authentic lime honey is greater than 15 µg / 100 g.
    Preferably, the content of cis, trans-abscisic acid in authentic lime blossom honey is greater than 30 µg / 100 g.
    Preferably, the conditions for high performance liquid chromatography detection in step 3) include: a flow rate of 0.7 ml / min; an ultraviolet detector having a detection wavelength of 280 nm; a C18 separation column having a specification of 150 x 4.6 mm, 5 µm; and a sample loading volume of 20 µl; where a mobile phase A is an aqueous solution of acetic acid having a mass fraction of 0.2%; a mobile phase B is an acetic acid-methanol solution having a mass fraction of 0.2%; a gradient elution schedule is 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 %] of mobile phase B; [30-36) min, (22% -25%] of mobile phase B; [36-41) min, (25% -30%] of 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 imprint of the phytochemical substance of the lime blossom honey is obtained using the following steps: taking 50 to 100 samples of authentic lime blossom honey and carrying out a detection by high performance liquid chromatography to obtain a chromatogram, respectively. phytochemicals from each of the samples, and importing the phytochemicals chromatograms from all samples into traditional Chinese medicine fingerprint software to obtain the phytochemical fingerprint in genuine lime honey.
    Preferably, a standard curve for calculating lindenine content according to the external standard method in step 3) is y = 1453.8x - 368132, where x is a peak area of a detected substance, y is the substance content detected; and the unit of y is ng.
    Preferably, a standard curve for the calculation of the content of trans, trans-abscisic acid and of cis, trans-abscisic acid according to the external standard method in step 3) is y = 4513x + 135049, where x is a peak area of a detected substance, and y is the content of the detected substance.
    Preferably, the ratio of the mass of honey to be identified to the mass of water is 1: (3.5-4.5). The beneficial effects of the present invention are as follows: According to the method for identifying the authenticity of linden honey provided by the present invention, a sample to be identified is mixed with water of dissolution and then the solution is passed through. a reverse phase anion exchange solid phase extraction column to enrich a phytochemical in the sample to be identified, the phytochemical in an eluent is more easily detected, the detection results are reliable and stable, the the degree of determination is high, the operation is easy, and the cost is low. The present invention establishes the sensitive, precise, highly efficient and universal method for identifying the authenticity of linden honey, which completely solves the problem of identifying the authenticity of linden honey, ensures the quality and edibility of the products. based on honey, protects the interests of consumers, and ensures the healthy development of the honey industry.
    BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is an imprint of a phytochemical component of lime blossom honey. Peak 1 to peak 3 represent lindenin, trans, trans-abscisic acid, cis, trans-abscisic acid.
    DETAILED DESCRIPTION The present invention provides a process for identifying the authenticity of lime blossom honey, including the following steps: step 1): mixing a honey to be identified with water of dissolution, and adjusting the pH value. at 6-7 to obtain a sample solution; step 2): passing the sample solution through a solid phase extraction column by reverse phase anion exchange, eluting the extraction column with a methanol solution containing 10% by weight d formic acid, and collection of the eluent; step 3): detection of the eluent by high performance liquid chromatography, and comparison of a detected liquid chromatogram with an imprint of a phytochemical component of genuine lime blossom honey, where if the type of a substance detected corresponds to the imprint of the phytochemical component of authentic linden honey, honey is authentic linden honey; and the phytochemical component fingerprint of authentic lime blossom honey is a fingerprint including lindenine, trans, trans-abscisic acid and cis, trans-abscisic acid.
    In the present invention, the honey to be identified is mixed with dissolution water, and the pH value is adjusted to 6-7 to obtain a sample solution. In the present invention, the ratio of the mass of honey to be identified to the mass of water is preferably 1: (3.5-4.5), more preferably 1: (3.8- 4.2), and most preferably 1: 4. In the present invention, the honey to be identified is any form of honey including, but not limited to, raw honey and commercial honey; and the water is preferably ultra pure water. In the present invention, the method further includes a centrifugation step after the sample to be identified has been mixed with water, where the rotational speed of the centrifugation is preferably 8000 to 12000 g, moreover. preferred 10,000 g; and the centrifugation time is preferably 8 to 12 min, more preferably 10 min. In the present invention, the effect of centrifugation is to remove impurities, and in the present invention, preferably, after centrifugation, a supernatant is collected for further processing. In the present invention, it is preferable to adjust the pH of the solution with an alkaline material, and it is more preferable to adjust the pH of the solution with an ammoniacal water; and the mass concentration of the ammonia water is preferably 4 to 6%, more preferably 5%.
    In the present invention, after the sample solution has been obtained, the sample solution is passed through a solid phase extraction column by reverse phase anion exchange, and an eluent is collected. In the present invention, the specification of the reverse phase anion exchange solid phase extraction column is RP-AE SPE. Prior to use, preferably, the reverse phase anion exchange solid phase extraction column of the present invention is activated with methanol and then equilibrated with ultra pure water. The amount of the methanol used in the present invention is preferably 0.8 to 1.2 ml, more preferably 1.0 ml; and the amount of the ultra pure water is preferably 0.8 to 1.2 ml, more preferably 1.0 ml.
    In the present invention, after the sample solution has passed through the column, the extraction column is preferably cleaned with ultra pure water, and then the extraction column is eluted with a methanol solution. containing 10% by weight of formic acid, and the eluent is collected.
    In the present invention, the amount of the ultra pure water used for cleaning the extraction column is preferably 0.8 to 1.2 ml, more preferably 1.0 ml; and the amount of the methanol solution containing 10% by weight of formic acid is 1.5 to 2.5 ml, more preferably 2.0 ml.
    In the present invention, after obtaining the eluent, the method preferably further includes drying the eluent and then further dissolving the eluent.
    In the present invention, the drying is preferably blow drying with nitrogen; the nitrogen flow rate and the nitrogen blowing drying time are not particularly limited, as long as drying can be achieved.
    In the present invention, after drying, preferably, further dissolution is carried out using a chromatographic grade solution of acetic acid-methanol (2:98 in V / V), and the amount of the acetic acid solution - chromatographic grade methanol used is preferably 1.5 to 2.5 ml, more preferably 2 ml.
    In the present invention, after a re-dissolved eluent is obtained, the eluent is filtered before high performance liquid chromatography.
    The filtration is preferably carried out by a filtration membrane, and the filtration membrane is preferably a 0.22 µm filtration membrane; and after the filtration, a high performance liquid chromatography sample loading solution is obtained.
    In the present invention, after the high performance liquid chromatography sample loading solution has been obtained, the sample loading solution is detected by high performance liquid chromatography, and a detected substance is compared with a fingerprint. 'a phytochemical component of linden honey, and if the type of the substance detected matches the imprint of the phytochemical component in linden honey, the honey is genuine linden honey.
    In the present invention, a mobile phase A of high performance liquid chromatography is water containing acetic acid having a mass fraction of 0.2%; a mobile phase B is methanol containing acetic acid having a mass fraction of 0.2%; a flow rate of high performance liquid chromatography is 0.7 ml / min; an ultraviolet detection wavelength is 280 nm; a separation column is a C18 column, and the specification of the C18 column is 150 x 4.6 mm, 5 µm; a sample loading volume is 20 µl; the elution gradient is [0-11) min, [0% -8%) of 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 %] of mobile phase B; [30-36) min, (22% -25%] of mobile phase B; [36-41) min, (25% -30%] of 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.
    In the present invention, after detection by high performance liquid chromatography, a chromatogram detected by high performance liquid chromatography is compared with an imprint of a phytochemical component of lime blossom honey to determine the type of a detected substance. In the present invention, the imprint of the phytochemicals of lime blossom honey is obtained by using the following steps: taking 50 to 100 samples of authentic lime blossom honey and carrying out a detection by high performance liquid chromatography to obtain a chromatogram, respectively. phytochemicals from each of the samples, and importing phytochemicals chromatograms from all samples into a traditional Chinese medicine fingerprint software to obtain the phytochemical fingerprint in lime honey. The authentic lime honey samples in the present invention are preferably from more than 30 apiaries in Jilin, Liaoning, Heilongjiang, etc. ; in the present invention, the imprint of the phytochemical component of linden honey is preferably as shown in FIG. 1, where lindenin 1, trans, trans-abscisic acid 2 and cis, trans-abscisic acid 3 are provided.
    In the present invention, linden honey refers to a sweet substance that is obtained by mixing linden blossom honey (including tilia amurensis and tilia mandshurica) collected by a honey bee with secretions from the bee's salivary gland melliferous and fully producing honey and is stored in a comb. Lindenine is a plant compound that has characteristics in linden honey. Abscisic acid is an endogenous plant hormone having a sesquiterpene structure in a plant, and has effects of regulating plant growth, inhibiting seed germination and promoting aging. With further research, abscisic acid plays an important role in the stress responses of plants in difficult situations such as drought, high salinity and low temperatures. Abscisic acid is a plant stressor and is therefore called a plant “stress hormone”. Linden honey contains abscisic acid.
    In the present invention, after the type of the detected substance has been determined, the content is calculated using an external standard method; a standard curve of trans, trans-abscisic acid and cis, trans-abscisic acid is y = 4513x + 135049, where x is the peak area of the substance detected, and y is the content of the substance detected; in the present invention, the unit of y is ng. In the present invention, a standard curve for lindenine is y = 1453.8x - 368132, where x is the peak area of the detected substance, y detects the substance content; and in the present invention, the unit of y is ng.
    In the present invention, the content of lindenin in authentic lime blossom honey is preferably greater than 45 µg / 100 g; the content of trans, trans-abscisic acid in authentic lime blossom honey is preferably greater than 15 µg / 100 g; and the content of cis, trans-abscisic acid in authentic lime honey is preferably greater than 30 µg / 100 g.
    The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but the technical solutions cannot be understood as limiting the scope of the protection of the present invention.
    Embodiment 1 An intermediary performed an assessment of the quality of a collected raw linden honey. Procedure: Step 1: accurately weigh 20 g of a sample of lime blossom honey, place the sample in a dry bottle, add 80 ml of ultra-pure dissolving water.
    Step 2: Perform a centrifugation at 10,000 x g for 10 min, and take a supernatant to use.
    Step 3: Adjust the pH to 6-7 with 5% ammonia water.
    Step 4: Activate a reverse phase anion exchange solid phase extraction column (RP-AE SPE) with 1 ml of methanol and equilibrate the column with 1 ml of ultra pure water (pH: 6-7) , and the supernatant passes through the column.
    Step 5: clean the extraction column with 1 ml of ultra pure water, elute the extraction column with 2 ml of methanol (containing 10% formic acid), and collect the eluent; Step 6: Blow dry the eluent with nitrogen at room temperature, dissolve again and mix the solid homogeneously with 2 ml of chromatographic grade acetic acid-methanol solution (2:98 in V / V ), and pass through a 0.22 µm filtration membrane to obtain a sample loading solution for HPLC.
    Step 7: Detect the HPLC Sample Loading Solution using an HPLC method; wherein according to a high performance liquid chromatography (HPLC) detection method, a mobile phase λ is water (containing 0.2% acetic acid); a mobile phase B is methanol (containing 0.2% acetic acid); a total flow rate is 0.7 ml / min; an ultraviolet detection wavelength is 280 nm; a separation column is a C18 column, (150 x 4.6 mm, 5 µm); a sample loading volume is 20 µl; the elution gradient is 0-11 min, 0% -8% B; 11-14 min, 8-10% B; 14-17 min, 10% -14% B; 17-24 min, 14-20% B; 24-28 min, 20-21% B; 28-30 min, 21-22% B; 30-36 min, 22-25% B; 36-41 min, 25-30% B; 41-46min, 30-33% B; 46-55 min, 33% B; 55-60min, 33-34% B; 60-70min, 34-36% B; 70-80min, 36-40% B; 80-90min, 40-45% B; 90-100 min, 45-52% B.
    Step 8: Calculate the content using an external standard method: A standard curve for lindenine is y = 1453.8x - 368132; and a standard curve for trans, trans-abscisic acid and cis, trans-abscisic acid is y = 4513x + 135049.
    Carry out the qualitative and quantitative analysis of a sample loading solution according to the HPLC method: Carry out a quantitative analysis according to the standard curves to obtain that the lindenin content was 80 µg / 100 g; the content of trans, trans-abscisic acid was 30 µg / 100 g; and the content of cis, trans-abscisic acid was 70 µg / 100 g. A chromatogram does not contain any other abnormal substance compared to a standard fingerprint.
    Embodiment 2 A company carried out a quality assessment of raw honey purchased. Procedure: Step 1: accurately weigh 20 g of a sample of lime blossom honey, place the sample in a dry bottle, add 80 ml of ultra-pure dissolving water.
    Step 2: Perform centrifugation at 10,000 x g for 10 min, take supernatant to use. Step 3: Adjust the pH to 6-7 with 5% ammonia water.
    Step 4: Activate a reverse phase anion exchange solid phase extraction column (RP-AE SPE) with 1 ml of methanol and equilibrate the column with 1 ml of ultra pure water (pH: 6-7) , and the supernatant passes through the column.
    Step 5: clean the extraction column with 1 ml of ultra pure water, elute the extraction column with 2 ml of methanol (containing 10% formic acid), and collect the eluent; Step 6: Blow dry the eluent with nitrogen at room temperature, dissolve again and mix the solid homogeneously with 2 ml of chromatographic grade acetic acid-methanol solution (2:98 in V / V ), and pass through a 0.22 µm filtration membrane to obtain a sample loading solution for HPLC.
    Step 7: Detect the sample loading solution for HPLC using an HPLC method; where according to a high performance liquid chromatography (HPLC) detection method, a mobile phase A is water (containing 0.2% acetic acid); a mobile phase B is methanol (containing 0.2% acetic acid); a total flow rate is 0.7 ml / min; an ultraviolet detection wavelength is 280 nm; a separation column is a C18 column, (150 x 4.6 mm, 5 µm); a sample loading volume is 20 µl; the elution gradient is 0-11 min, 9% -14% B; 11-14 min, 14-15% B; 14-17 min, 15% B; 17-24 min, 15-16% B; 24-28 min, 16-17% B; 28-30 min, 17-22% B; 30-38 min, 22-25% B; 38-41 min, 25-30% B; 41-46min, 30-33% B; 46-55 min, 33% B; 55-60min, 33-34% B; 60-70min, 34-36% B; 70-80min, 36-40% B; 80-90min, 40-45% B; 90-100 min, 45-52% B.
    Step 8: Calculate the content using an external standard method: A standard curve for lindenine is y = 1453.8x - 368132; and a standard curve of trans, trans-abscisic acid and cis, trans-abscisic acid is y = 4513x + 135049. Perform qualitative and quantitative analysis of a sample loading solution according to the method of HPLC: Carry out a quantitative analysis according to the standard curves to obtain that the content of lindenin was 130 µg / 100g; the content of trans, trans-abscisic acid was 42 µg / 100 g; and the content of cis-trans-abscissic acid was 60 µg / 100 g.
    A chromatogram does not contain any other abnormal substance compared to a standard fingerprint.
    In summary, raw honey is lime blossom honey.
    Embodiment 3 A third-party testing agency performed a quality inspection on a basic honey detected on order. Procedure: Step 1: accurately weigh 20 g of a sample of lime blossom honey, place the sample in a dry bottle, add 80 ml of ultra-pure dissolving water.
    Step 2: Perform a centrifugation at 10,000 x g for 10 min, and take a supernatant to use.
    Step 3: Adjust the pH to 6-7 with 5% ammonia water.
    Step 4: Activate a reverse phase anion exchange solid phase extraction column (RP-AE SPE) with 1 ml of methanol and equilibrate the column with 1 ml of ultra pure water (pH: 6-7) , and the supernatant passes through the column.
    Step 5: clean the extraction column with 1 ml of ultra pure water, elute the extraction column with 2 ml of methanol (containing 10% formic acid), and collect the eluent; Step 6: Blow dry the eluent with nitrogen at room temperature, dissolve again and mix the solid homogeneously with 2 ml of chromatographic grade acetic acid-methanol solution (2:98 in V / V ), and pass through a 0.22 µm filtration membrane to obtain a sample loading solution for HPLC.
    Step 7: Detect the sample loading solution for HPLC using an HPLC method; where according to a high performance liquid chromatography (HPLC) detection method, a mobile phase A is water (containing 0.2% acetic acid); a mobile phase B is methanol (containing 0.2% acetic acid); a total flow rate is 0.7 ml / min; an ultraviolet detection wavelength is 280 nm; a separation column is a C18 column, (150 x 4.6 mm, 5 µm); a sample loading volume is 20 µl; the elution gradient is 0-11 min, 9% -14% B; 11-14 min, 14-15% B; 14-17 min, 15% B; 17-24 min, 15-16% B; 24-28 min, 16-17% B; 28-30 min, 17-22% B; 30-38 min, 22-25% B; 38-41 min, 25-30% B; 41-46min, 30-33% B; 46-55 min, 33% B; 55-60 min, 33-
    34% B; 60-70min, 34-36% B; 70-80min, 36-40% B; 80-90min, 40-45% B; 90-100 min, 45-52% B. Step 8: Calculate the content using an external standard method: A standard curve for lindenine is y = 1453.8x - 368132; and a standard curve for trans, trans-abscisic acid and cis, trans-abscisic acid is y = 4513x + 135049.
    Carry out the qualitative and quantitative analysis of a sample loading solution according to the HPLC method: Carry out a quantitative analysis according to the standard curves to obtain that the content of lindenin was 94 µg / 100 g; the content of trans, trans-abscisic acid was 50 µg / 100 g; and the content of cis, trans-abscisic acid was 82 µg / 100 g. A chromatogram does not contain any other abnormal substance compared to a standard fingerprint. In summary, raw honey is lime blossom honey.
    As can be seen from the above embodiments, the method for identifying the authenticity of linden honey provided by the present invention gives reliable and stable detection results and a high degree of determination and is easy to perform. operate and low cost.
    The foregoing descriptions are only preferred methods of carrying out the present invention. It should be noted that for those of ordinary skill in the art, several improvements and modifications can furthermore be made without departing from the principle of the present invention. These improvements and modifications should also be interpreted as falling within the scope of protection of the present invention.
    权利要求:
    Claims (10)
    [1]
    1. Method for identifying the authenticity of lime blossom honey, comprising the following steps: step 1): mixing a honey to be identified with dissolution water, and adjusting the pH value to 6-7 for obtain a sample solution; step 2): passing the sample solution through a solid phase extraction column by reverse phase anion exchange, eluting the extraction column with a methanol solution containing 10% by weight d acid formigue, and collection of the eluent; and step 3): detecting the eluent by high performance liquid chromatography, comparing a detected liquid chromatogram with an imprint of a phytochemical component of genuine lime blossom honey, in which if the type of a substance detected matches with the imprint of the phytochemical component of authentic lime blossom honey, honey is authentic lime blossom honey; and the imprint of the phytochemical component of linden honey | authentic is an imprint comprising lindenine (4- (2-hydroxypropan-2-yl) cyclohexa-1,3-diene-1-carboxylic acid), trans, trans-abscisic acid and cis acid, trans-abscisic.
    [2]
    2. The method of claim 1, wherein step 3) further comprises calculating, after separation by high performance liquid chromatography, the contents of lindenine, trans, trans-abscisic acid and cis, trans-acid. abscissal line detected in honey to be identified according to a chromatogram obtained by high performance liquid chromatography using an external standard method.
    [3]
    3. A method according to claim 2, wherein the content of lindenin in authentic lime blossom honey is greater than 45 µg / 100 g.
    [4]
    4. Method according to claim 2 or 3, in which the content of trans, trans-abscisic acid in authentic lime blossom honey is greater than 15 µg / 100 g.
    [5]
    5. The method of claim 2, wherein the content of cis, trans-abscisic acid in authentic lime blossom honey is greater than 30 µg / 100 g.
    [6]
    The method of claim 1, wherein the conditions for detection by high performance liquid chromatography in step 3) comprise: a flow rate of 0.7 ml / min; an ultraviolet detector having a detection wavelength of 280 nm; a C18 separation column having a specification of 150 x 46 mm, 5 µm; and a sample loading volume of 20 µl; wherein a mobile phase A is an aqueous solution of acetic acid having a mass fraction of 0.2%; a mobile phase B is an acetic acid-methanol solution having a mass fraction of 0.2%; a gradient elution schedule is 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 %] of mobile phase B; [30-36) min, (22% -25%] of mobile phase B; [36-41) min, (25% -30%] of 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]
    7. The method of claim 6, wherein the imprint of the phytochemical substance of lime blossom honey is obtained by using the following steps: taking 50 to 100 samples of authentic lime blossom honey and carrying out a detection by liquid chromatography, respectively. performance to obtain a phytochemical component chromatogram of each of the samples, and import the phytochemical component chromatograms of all samples into a traditional Chinese medicine fingerprint software to obtain the phytochemical component fingerprint in authentic lime honey .
    [8]
    8. The method of claim 2, wherein a standard curve for calculating the lindenin content according to the external standard method in step 3) is y = 1453.8x - 368132, wherein x is a peak area. of a substance detected, y is the content of the substance detected; and the unit of y is ng.
    [9]
    9. The method of claim 1, wherein a standard curve for calculating the content of trans, trans-abscisic acid and cis, trans-abscisic acid according to the external standard method in step 3) is y = 4513x. + 135049, where x is a peak area of a detected substance, and y is the content of the detected substance.
    [10]
    10. The method of claim 1, wherein the ratio of the mass of honey to be identified to the mass of water is 1: (3.5-4.5).
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    同族专利:
    公开号 | 公开日
    BE1026676A1|2020-04-30|
    CN109752469A|2019-05-14|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    法律状态:
    2021-01-15| FG| Patent granted|Effective date: 20201130 |
    优先权:
    申请号 | 申请日 | 专利标题
    CN201811228827.2A|CN109752469A|2018-10-22|2018-10-22|A method of identifying eucalyptus honey authenticity|
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