CN114324650A - Method for identifying bauhinia championii honey based on (Z) -EDTO characteristic marker - Google Patents
Method for identifying bauhinia championii honey based on (Z) -EDTO characteristic marker Download PDFInfo
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Abstract
The invention relates to the field of honey detection, in particular to a method for identifying bauhinia championii honey based on a (Z) -EDTO characteristic marker, which analyzes and contrasts different honey species and determines the characteristic marker of the bauhinia championii honey; after pretreatment, analyzing by adopting a liquid chromatography-tandem mass spectrometry method, and detecting whether the sample honey contains a (Z) -EDTO characteristic marker, thereby identifying whether the sample honey is the bauhinia championii honey. Compared with the prior art, the method for identifying the bauhinia championii honey based on the (Z) -EDTO characteristic marker provided by the invention takes the (Z) -EDTO characteristic marker as the characteristic marker in the bauhinia championii honey, has important significance for accurately identifying the bauhinia championii honey, is simple to operate, has high sensitivity and high accuracy, and has a great application prospect.
Description
Technical Field
The invention relates to the field of honey detection, and in particular relates to a method for identifying bauhinia championii honey based on a (Z) -EDTO characteristic marker.
Background
The honey is a natural health food with highly complex components, is rich in a plurality of nutrient substances beneficial to human bodies, and is deeply favored by consumers. The honey is natural sweet substance which is obtained by collecting nectar, honeydew or insect-secreted nectar secreted by honey source plants by bees, mixing with salivary gland secretion of the bees, and then sufficiently brewing and maturing in a honeycomb through multiple conversion actions. The honey contains abundant nutrients, such as water, amino acids, organic acids, proteins, minerals, vitamins, pigments, aromatic substances, bee pollen, hormones, etc., besides the main saccharides. Meanwhile, the Chinese medicinal composition has certain effects on resisting bacteria, enhancing the immune function, relieving cough, moistening the lung, dispelling wind, removing dampness, strengthening the spleen, regulating qi, resisting tumors and the like. The components and contents of honey vary with the variety of honey source, geographical environment, climate conditions, and bee-keeping techniques. Due to the difference of honey source plant species, honey source producing areas and bee species, the nutritional ingredients and the contents of different types of honey are different, and the characteristic ingredients of the honey are also key indexes for identifying the types of the honey.
The bauhinia championii honey is a natural sweet substance prepared by fully brewing and collecting nectar and secretion of bauhinia championii (the scientific name is bauhinia championii or bauhinia championii) which is a medicinal plant by bees and the secretion of the bauhinia championii. Most of them are produced in Guangxi provinces of China. The bauhinia championii honey is dark amber, glittering and translucent, has a special herb fragrance, tastes sweet, slightly bitter, is comfortable and tasty to the throat, and has a long aftertaste. The honey is natural and pollution-free, can clear away heat and toxic materials, has good curative effects on pharyngolaryngitis, cough due to lung heat and constipation, is an outstanding food therapy product, and is praised as 'top quality in south China and top quality in honey'. At present, no method for identifying the honey of bauhinia championii by using (Z) -EDTO as a characteristic marker is available.
Disclosure of Invention
In order to solve the problems mentioned in the background art, the invention aims to provide the method for identifying the bauhinia championii honey based on the (Z) -EDTO characteristic marker, which is simple to operate, high in sensitivity and high in accuracy, and can meet the requirement as a basis for identifying the bauhinia championii honey.
A method for identifying bauhinia championii honey based on (Z) -EDTO characteristic markers is characterized by comprising the following steps: identifying the honey of the bauhinia championii according to whether the honey to be detected contains the (Z) -8-ethylidene-6,7-dihydroxy-1,4, 4-trimethy-2-oxas ((Z) -EDTO) characteristic marker of the formula I;
preferably, the method comprises the following steps:
s1, determining a characteristic marker: analyzing and comparing the bauhinia championii honey in different regions, and determining that the characteristic marker of the bauhinia championii honey is (Z) -EDTO;
s2, sample pretreatment: mixing the sample honey seeds with water, performing centrifugal separation, taking a liquid phase, transferring the liquid phase to an HLB column, leaching with a leacheate, collecting the leacheate, fully and uniformly vortexing, and filtering;
s3, computer analysis: analyzing the filtrate by adopting a liquid chromatography-tandem mass spectrometry method, and detecting whether the characteristic marker of the bauhinia championii honey is contained;
s4, identifying whether the sample honey seeds are the bauhinia championii honey according to whether the sample honey seeds contain (Z) -EDTO.
9. Preferably, S1 is specifically: analyzing the bauhinia championii honey in different regions by adopting liquid chromatography-tandem mass spectrometry full-scan, characterizing fragment ion information of characteristic markers in the bauhinia championii honey, and determining that the (Z) -EDTO characteristic markers are shown in a formula I:
preferably, S2 is specifically: mixing the components in a mass ratio of 1: and (2-4) fully and uniformly vortexing the honey seeds of the sample with water, adjusting the pH value to 7-8, centrifuging for 10-30min at 10000r/min of 5000-.
Preferably, the chromatographic conditions of S3 are:
a chromatographic column: phenomenon Kinetex C18，150*4.6mm，2.6μm
Mobile phase: a: 0.1% methanoic acid methanol B: 0.1% formic acid water
Flow rate: 0.7mL/min
Column temperature: 30 deg.C
Sample introduction amount: 25 μ L.
Preferably, the mass spectrometry conditions of S3 are:
an ion source: heatable electric spray ion source (HESI-II)
Spraying voltage: 3.0kv
Capillary temperature: 280 deg.C
Heating temperature: 350 deg.C
Sheath gas (N)2) Flow rate: 50psi
Auxiliary gas (N)2) Flow rate: 8psi
Purge gas (N)2) Flow rate: 3 psi.
Has the advantages that: compared with the prior art, the method for identifying the bauhinia championii honey based on the (Z) -EDTO characteristic marker provided by the invention takes the (Z) -EDTO characteristic marker as the characteristic marker in the bauhinia championii honey, has important significance for accurately identifying the bauhinia championii honey, is simple to operate, has high sensitivity and high accuracy, and has a great application prospect.
Drawings
FIG. 1 is a characteristic ion fingerprint spectrum of various kinds of honey;
FIG. 2 shows characteristic ions of bauhinia championii honey1H NMR chart;
FIG. 3 shows characteristic ions of bauhinia championii honey13C NMR chart;
FIG. 4 is a mass spectrum of bauhinia championii honey.
Detailed Description
The present invention is described in detail below with reference to specific examples, which are given for the purpose of further illustrating the invention and are not to be construed as limiting the scope of the invention, and the invention may be modified and adapted by those skilled in the art in light of the above disclosure. Except for special description, the parts are parts by weight, the percentages are mass percentages, and the concentration is mass percentage concentration.
Example 1
Determination of characteristic markers: adopting liquid chromatography-tandem mass spectrometry to perform full-scan analysis on honey seeds in different regions to obtain fingerprint (figure 1), finding a characteristic marker (Z) -EDTO in the bauhinia championii honey through comparison, and determining that the structural formula of the characteristic marker is formula I through mass spectrum fragment ion scanning and nuclear magnetism characterization (table 1) of the characteristic marker;
table 1(Z) -EDTO marker ion related information is shown in table 2 below:
TABLE 2
Example 2
Accurately weighing 20.0g of honey to be measured in a 50mL centrifuge tube, adding 40.0mL of aqueous solution for extraction, fully and uniformly vortexing, adjusting the pH value to 7-8, and centrifuging for 10min at 5000 r/min; taking 15mL of methanol to balance the HLB solid-phase extraction column, addingPutting the honey into a 15mL aqueous solution activation extraction column, transferring the centrifuged honey sample liquid onto an HLB solid-phase extraction column, naturally draining, adding 15mL water to wash the solid-phase extraction column, finally leaching with 2mL methanol, collecting leacheate, fully and uniformly vortexing, filtering with a filter membrane with the pore diameter of 0.1 mu m, and putting the honey sample liquid on a machine to analyze by a liquid chromatography-tandem mass spectrometry method, wherein the chromatographic conditions are as follows: a chromatographic column: phenomenon Kinetex C18100 x 4.6, 2.6 μm; mobile phase: a: 0.1% methanoic acid methanol B: 0.1% formic acid water; flow rate: 0.7 mL/min; column temperature: 30 ℃; sample introduction amount: 25 mu L of the solution; the mass spectrum conditions are as follows: an ion source: a heatable electrospray ion source (HESI-II); spraying voltage: 3.0kv
Capillary temperature: 280 ℃; heating temperature: 350 ℃; sheath gas (N)2) Flow rate: 50 psi; auxiliary gas (N)2) Flow rate: 8 psi; purge gas (N)2) Flow rate: 3 psi; and detecting whether the sample honey contains the (Z) -EDTO characteristic marker, thereby identifying whether the sample honey is the bauhinia championii honey.
Example 3
Accurately weighing 12.5g of honey to be detected in a 50mL centrifuge tube, adding 50.0mL of aqueous solution for extraction, fully and uniformly vortexing, adjusting the pH value to 7-8, and centrifuging for 30min at 10000 r/min; taking 15mL of methanol to balance an HLB solid-phase extraction column, adding 15mL of aqueous solution to activate the extraction column, transferring the centrifuged honey sample liquid onto the HLB solid-phase extraction column, naturally draining, adding 15mL of water to wash the solid-phase extraction column, finally leaching with 2mL of methanol, collecting leacheate, filtering with a filter membrane with the pore diameter of 0.5 mu m after fully vortexing, putting the filter membrane on a machine, and analyzing by adopting a liquid chromatography-tandem mass spectrometry method, wherein the chromatographic conditions are as follows: a chromatographic column: phenomenon Kinetex C18100 x 4.6, 2.6 μm; mobile phase: a: 0.1% methanoic acid methanol B: 0.1% formic acid water; flow rate: 0.7 mL/min; column temperature: 30 ℃; sample introduction amount: 25 mu L of the solution; the mass spectrum conditions are as follows: an ion source: a heatable electrospray ion source (HESI-II); spraying voltage: 3.0kv
Capillary temperature: 280 ℃; heating temperature: 350 ℃; sheath gas (N)2) Flow rate: 50 psi; auxiliary gas (N)2) Flow rate: 8 psi; purge gas (N)2) Flow rate: 3 psi; detecting whether the honey species of the sample contains the (Z) -EDTO characteristic marker, thereby identifyingWhether the honey seeds are the bauhinia championii honey or not is sampled.
Example 4
Accurately weighing 20g of honey to be detected in a 50mL centrifuge tube, adding 50.0mL of aqueous solution for extraction, fully and uniformly vortexing, adjusting the pH value to 7-8, and centrifuging for 15min at 8000 r/min; taking 15mL of methanol to balance an HLB solid-phase extraction column, adding 15mL of aqueous solution to activate the extraction column, transferring the centrifuged honey sample liquid onto the HLB solid-phase extraction column, naturally draining, adding 15mL of water to wash the solid-phase extraction column, finally leaching with 2mL of methanol, collecting leacheate, filtering with a filter membrane with the pore diameter of 0.22 mu m after fully vortexing, and putting the honey sample liquid on a machine to analyze by adopting a liquid chromatography-tandem mass spectrometry method, wherein the chromatographic conditions are as follows: a chromatographic column: phenomenon Kinetex C18100 x 4.6, 2.6 μm; mobile phase: a: 0.1% methanoic acid methanol B: 0.1% formic acid water; flow rate: 0.7 mL/min; column temperature: 30 ℃; sample introduction amount: 25 mu L of the solution; the mass spectrum conditions are as follows: an ion source: a heatable electrospray ion source (HESI-II); spraying voltage: 3.0kv
Capillary temperature: 280 ℃; heating temperature: 350 ℃; sheath gas (N)2) Flow rate: 50 psi; auxiliary gas (N)2) Flow rate: 8 psi; purge gas (N)2) Flow rate: 3 psi; and detecting whether the sample honey contains the (Z) -EDTO characteristic marker, thereby identifying whether the sample honey is the honey of the bauhinia championii (shown in figure 4).
Finally, the above embodiments are only for illustrating the technical solutions of the present invention and not for limiting, although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all of them should be covered in the claims of the present invention.
Claims (8)
1. A method for identifying bauhinia championii honey based on (Z) -EDTO characteristic markers is characterized by comprising the following steps: identifying the honey of the bauhinia championii according to whether the honey to be detected contains the (Z) -8-ethylidene-6,7-dihydroxy-1,4, 4-trimethy-2-oxas ((Z) -EDTO) characteristic marker of the formula I;
2. the method for identifying bauhinia championii honey based on (Z) -EDTO characteristic markers, according to claim 1, wherein: analyzing and comparing different honey species, and determining characteristic markers of the bauhinia championii honey; after pretreatment, analyzing by adopting a liquid chromatography-tandem mass spectrometry method, and detecting whether the sample honey contains a (Z) -EDTO characteristic marker to identify whether the sample honey is the bauhinia championii honey, thereby evaluating the purity and quality of the bauhinia championii honey.
3. The method for identifying bauhinia championii honey based on (Z) -EDTO characteristic markers as claimed in claim 2, wherein: whether the bauhinia championii honey is adulterated or not is identified through (Z) -EDTO marker detection, and the method can be used for identifying the adulterated honey.
4. The method for identifying the honey of bauhinia championii based on the (Z) -EDTO characteristic markers as claimed in claim 2, wherein the determination process of the characteristic markers of the honey of bauhinia championii comprises the following specific steps: analyzing the bauhinia championii honey in different regions by adopting liquid chromatography-tandem mass spectrometry full-scan, characterizing fragment ion information of characteristic markers in the bauhinia championii honey, and determining that the (Z) -EDTO characteristic markers are shown in a formula I:
5. the method for identifying bauhinia championii honey based on (Z) -EDTO characteristic markers as claimed in claim 2, wherein the sample honey pretreatment process is as follows: mixing the sample honey seeds with water, performing centrifugal separation, taking a liquid phase, transferring the liquid phase to an HLB column, leaching with a leacheate, collecting the leacheate, fully and uniformly vortexing, and filtering.
6. The method for identifying bauhinia championii honey based on the (Z) -EDTO characteristic marker as claimed in claim 4, wherein the pretreatment process specifically comprises: mixing the components in a mass ratio of 1: and (2-4) fully and uniformly vortexing the honey seeds of the sample with water, adjusting the pH value to 7-8, centrifuging for 10-30min at 10000r/min of 5000-.
7. The method for identifying bauhinia championii honey based on (Z) -EDTO characteristic markers as claimed in claim 2, characterized in that the chromatographic conditions of the liquid chromatography-tandem mass spectrometry method are as follows:
a chromatographic column: phenomenon Kinetex C18，150*4.6mm，2.6μm
Mobile phase: a: 0.1% methanoic acid methanol B: 0.1% formic acid water
Flow rate: 0.7mL/min
Column temperature: 30 deg.C
Sample introduction amount: 25 μ L.
8. The method for identifying bauhinia championii honey based on (Z) -EDTO characteristic markers, according to claim 1, is characterized in that the mass spectrometry conditions of the liquid chromatography-tandem mass spectrometry method are as follows:
an ion source: heatable electrospray ion source HESI-II
Spraying voltage: 3.0kv
Capillary temperature: 280 deg.C
Heating temperature: 350 deg.C
Sheath gas (N)2) Flow rate: 50psi
Auxiliary gas (N)2) Flow rate: 8psi
Purge gas (N)2) Flow rate: 3 psi.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117233292A (en) * | 2023-11-13 | 2023-12-15 | 中国农业科学院蜜蜂研究所 | Identification method of nine-dragon rattan honey |
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