CN114181176B - Shishan kunmiki A and preparation method thereof, and identification method of bauhinia championii honey - Google Patents
Shishan kunmiki A and preparation method thereof, and identification method of bauhinia championii honey Download PDFInfo
- Publication number
- CN114181176B CN114181176B CN202111131616.9A CN202111131616A CN114181176B CN 114181176 B CN114181176 B CN 114181176B CN 202111131616 A CN202111131616 A CN 202111131616A CN 114181176 B CN114181176 B CN 114181176B
- Authority
- CN
- China
- Prior art keywords
- honey
- mobile phase
- increased
- shishan
- percent
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D301/00—Preparation of oxiranes
- C07D301/32—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D303/00—Compounds containing three-membered rings having one oxygen atom as the only ring hetero atom
- C07D303/02—Compounds containing oxirane rings
- C07D303/12—Compounds containing oxirane rings with hydrocarbon radicals, substituted by singly or doubly bound oxygen atoms
- C07D303/32—Compounds containing oxirane rings with hydrocarbon radicals, substituted by singly or doubly bound oxygen atoms by aldehydo- or ketonic radicals
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention relates to the technical field of honey detection, and particularly discloses a shishan kunmamesu A, a preparation method thereof and an identification method of bauhinia championii honey. The kukukoamine A has a structure shown in a formula (1):
Description
Technical Field
The invention relates to the technical field of honey detection, in particular to Shishan kunmamesu A, a preparation method thereof and an identification method of bauhinia championii honey.
Background
Bauhinia championii is a flowering plant of bauhinia of the subfamily caesalpiniaceae of the family leguminosae, and is mainly distributed in indonesia, india, vietnam and southern china. The flowering period is 9-10 months, and the flower honey is collected by bees and brewed to obtain rare bitter honey. The bauhinia championii honey is neutral and bitter and has medicinal value, and is derived from a medicinal plant bauhinia championii and is recently attracted by people as rare bitter honey. The bauhinia championii honey has high market value and adulteration phenomenon, seriously influences the benefits of beekeepers and consumers and restricts the development of characteristic economy of bauhinia championii honey industry.
Therefore, it is necessary and urgent to establish a set of method for identifying the true and false of bauhinia championii honey.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide a method for identifying bauhinia championii honey.
In order to achieve the purpose, the technical scheme of the invention is as follows:
in a first aspect, the present invention provides a kukoamine a having a structure represented by formula (1):
according to the invention, through the research on the bauhinia championii honey, a new compound contained in the bauhinia championii honey is discovered, and the compound can be used for judging the authenticity of the bauhinia championii honey by combining Shishan kunmamesu B, trans-abscisic acid, photochromic acid and cis-trans-abscisic acid.
In a second aspect, the invention provides a method for preparing the kukoamine A, which is specifically obtained by separating phenolic components in honey through high performance liquid chromatography, wherein the honey comprises bauhinia championii honey.
In the invention, when the high performance liquid chromatography separation is carried out, the mobile phase A is an aqueous solution of acetic acid with the concentration of 0.18-0.22 percent, and the mobile phase B is a methanol solution of the acetic acid with the concentration of 0.18-0.22 percent;
the gradient elution procedure was: the mobile phase B is increased from 9 percent to 14 percent in 0-11 min; the time is 11-14 min, and the mobile phase B is increased from 14% to 15%; 14-17 min, keeping the mobile phase B at 15%; the time is 17-24 min, and the mobile phase B is increased from 15% to 16%; the mobile phase B is increased from 16% to 17% in 24-28 min; the mobile phase B is increased from 17 percent to 22 percent within 28 to 30 min; 30-41 min, keeping the mobile phase B at 22%; the time is 41-46 min, and the mobile phase B is increased from 22% to 23%; 46-55 min, the mobile phase B rises from 23% to 25%; the time is 55-60 min, and the mobile phase B is increased from 25% to 28%; the mobile phase B is increased from 28 percent to 30 percent in 60-70 min; the mobile phase B is increased from 30 percent to 40 percent in 70-80 min; the time is 80-90 min, and the mobile phase B is increased from 40% to 45%; the time is 90-100 min, and the mobile phase B is increased from 45% to 52%; the mobile phase B is increased from 52% to 57% in 100-110 min; 110-120 min, the mobile phase B rises from 57% to 65%; the mobile phase B is increased from 65% to 70% in 120-130 min; 130-135 min, and the mobile phase B is increased from 70% to 80%.
The mobile phase A varies with the volume of the mobile phase B, and the sum of the two is 100%.
In the invention, the chromatographic column is a Phenomenex Gemini C18 chromatographic column, the flow rate is 0.6-0.8mL/min, the sample injection amount is 20 mu L, the column temperature is 34-36 ℃, and the detection wavelength is 270nm.
Preferably, the mobile phase A is a 0.2% acetic acid aqueous solution, and the mobile phase B is a 0.2% acetic acid methanol solution; the flow rate was 0.7mL/min and the column temperature was 35 ℃.
In the invention, the high performance liquid chromatography also comprises a step of extracting phenolic components in honey by using a solid phase extraction column before the high performance liquid chromatography is carried out; specifically, the method comprises the steps of extracting by using a Strata-X-A solid phase extraction column, using methanol as an activating agent, water as a balancing agent and an eluting agent, using a methanol solution of formic acid with the concentration of 9-11% (preferably 10%) as an eluent, blow-drying the obtained eluent, redissolving by using methanol, and filtering to obtain the eluent;
and/or the aperture of the filter membrane is 0.22 μm during filtration;
preferably, the eluent is a 10% strength methanoic acid solution in methanol.
The extraction method can effectively remove interfering components in the bauhinia championii honey and realize effective enrichment of target components.
In the invention, before extracting the phenolic components in the honey by using the solid phase extraction column, the method also comprises the step of pretreating the honey, and specifically comprises the following steps: mixing Mel and water, adjusting pH to 6.5-7, centrifuging at 8800-9200rpm for 18-22min, and collecting supernatant;
preferably, the pH is adjusted to 6.8 and the mixture is centrifuged at 9000rpm for 20min.
The amount of water added for pretreatment in the present invention is such that the honey is sufficiently dissolved and is suitable for subsequent operations.
In the invention, after phenolic components in honey are separated by high performance liquid chromatography, target peak eluent is collected, and the retention time of the target peak is 26.93min.
In a third aspect, the invention provides application of Shishan kunmamesu A in identifying bauhinia championii honey.
The method comprises the steps of detecting phenolic components in honey to be detected through a high performance liquid chromatography, and judging the honey to be detected to be the bauhinia championii honey when the phenolic components simultaneously comprise kukosmin A, kukosmin B, trans-abscisic acid, photochromic element and cis-abscisic acid;
the Shishan bitter honey element A is as described above, and the Shishan bitter honey element B has a structure shown in a formula (2):
the chemical name of kukukoamine B is 2- (1, 2-dihydroxypropyl) -4, 8-trimethy-1-oxapipro [2.5] oct-4-en-6-one, which is commercially available.
In the identification method, the method for detecting the honey to be detected by the high performance liquid chromatography is as described above.
The high performance liquid chromatography can effectively separate the Shishan kunmamesu A, the Shishan kunmamesu B, the trans-abscisic acid, the photochromic pigment and the cis-abscisic acid from the honey to be detected, thereby realizing the accurate judgment of authenticity.
Preferably, when the content of the Shishan bitter honey A in the honey to be detected is 5.40-9.81mg/kg, the content of the Shishan bitter honey B in the honey to be detected is 17.10-32.87mg/kg, the content of trans-abscisic acid is 6.37-11.01mg/kg, the content of photochromic pigment is 3.11-28.34mg/kg, and the content of cis-abscisic acid and trans-abscisic acid is 6.65-14.11mg/kg, the honey to be detected is determined to be the bauhinia championii honey.
In a fourth aspect, the invention provides a finger print of bauhinia championii honey, which is obtained by processing bauhinia championii honey through high performance liquid chromatography, wherein the high performance liquid chromatography is as described above. The method specifically comprises the steps of pretreatment of the honey, extraction of phenolic components in the honey by a solid-phase extraction column and high performance liquid chromatography detection.
The invention has the beneficial effects that:
the invention establishes a solid-phase extraction and high performance liquid chromatography separation detection method for plant compounds in bauhinia championii honey, and identifies 5 representative plant compounds contained in the bauhinia championii honey by an HPCL-QTOF method, wherein the 5 representative plant compounds are Shishankukosmei A, shishankusmei B, trans-abscisic acid, photochromism, cis-abscisic acid and trans-abscisic acid respectively. Introducing the obtained bauhinia championii honey plant compound HPLC chromatogram into a traditional Chinese medicine chromatogram fingerprint similarity evaluation system, and preliminarily simulating to generate a bauhinia championii honey standard fingerprint with representative significance.
According to the invention, through comparison of HPLC (high performance liquid chromatography) spectra of four large single-flower honey, namely locust honey, linden honey, vitex honey and rape honey and analysis and comparison of collected plant compounds of other honey of Xiaozhong, such as safflower honey, chinese gall honey, buckwheat honey, motherwort honey, medlar honey and the like, the compounds Shishankukonmi A, shishankunmi B, trans-abscisic acid, photochromic elements and cis-trans abscisic acid are not simultaneously detected in other single-flower honey except bauhinia championii honey. The five components can be used as a characteristic plant compound marker of the bauhinia championii honey, are applied to evaluating the authenticity and quality of the bauhinia championii honey, and have important practical significance for protecting the legal rights and interests of honey consumers and maintaining the healthy development of the honey consumption industry.
Drawings
FIG. 1 is a primary mass spectrum of Shishan kunmamesu A, wherein the upper diagram is a primary mass spectrum of Shishan kunmamesu A in positive ion mode; the lower figure is a primary mass spectrum of Shishan kunmamesu A under the negative ion mode.
FIG. 2 is a secondary mass spectrum of Shishan kunmamectin A, wherein the upper half part of the diagram is a primary mass spectrum of Shishan kunmamectin A in positive ion mode; the lower half part is a secondary mass spectrum of Shishan bitter honey A under a positive ion mode.
FIG. 3 shows the fragmentation pathway of Shishan bitter honey A by mass spectrometry.
FIG. 4 is a chromatogram of the phenolic components of bauhinia championii honey in example 1 of the present invention.
FIG. 5 is an HPLC overlay spectrum of the raw honey of bauhinia championii from ten different production areas in example 2 of the present invention. In the figure, R represents a standard fingerprint spectrum generated based on the Chinese alpine rush raw material honey in ten different producing areas, and S1-S10 represent the Chinese alpine rush raw material honey in the ten different producing areas.
Detailed Description
Preferred embodiments of the present invention will be described in detail with reference to the following examples. It is to be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention. Various modifications and alterations of this invention will become apparent to those skilled in the art without departing from the spirit and scope of this invention.
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1
The experimental example provides a shishan kunmiki A and a preparation method thereof, and establishment of finger print of characteristic components of bauhinia championii honey.
The operation method comprises the following steps:
(1) preparing a bauhinia championii honey solution: accurately weighing 20.0g of bauhinia championii honey into a beaker, adding 80mL of deionized water, stirring with a glass rod until the honey is fully dissolved, adding 5% ammonia water solution to adjust the pH value to about 6.8, centrifuging at 9000rpm for 20min, and taking the supernatant for later use.
(2) Extracting and enriching Chinese alpine rush honey plant compounds: extracting phenols in Mel with Strata-X-A solid phase extraction column, activating with 3mL methanol, balancing the extraction column with 3mL deionized water, adding the above supernatant, eluting with deionized water, eluting with 10% formic acid methanol solution, and collecting eluate. The eluent was blown dry with nitrogen, redissolved in 2.0mL of methanol, and filtered through a 0.22 μm filter for use.
(3) Separating and detecting the bauhinia championii honey plant compound: separating and detecting the phenolic components in the honey by using high performance liquid chromatography, wherein the chromatographic column is a Phenomenex Gemini C18 chromatographic column, the mobile phase A is a 0.2% acetic acid aqueous solution, the mobile phase B is a 0.2% acetic acid methanol solution, and the gradient elution procedure is as follows: the mobile phase B is increased from 9 percent to 14 percent in 0-11 min; the mobile phase B is increased from 14 percent to 15 percent in 11-14 min; 14-17 min, keeping the mobile phase B at 15%; the time is 17-24 min, and the mobile phase B is increased from 15% to 16%; the mobile phase B is increased from 16% to 17% in 24-28 min; the time is 28-30 min, and the mobile phase B is increased from 17% to 22%; 30-41 min, keeping the mobile phase B at 22%; the time is 41-46 min, and the mobile phase B is increased from 22% to 23%; 46-55 min, the mobile phase B rises from 23% to 25%; the time is 55-60 min, and the mobile phase B is increased from 25% to 28%; the mobile phase B is increased from 28 percent to 30 percent in 60-70 min; the time is 70-80 min, and the mobile phase B is increased from 30% to 40%; the time is 80-90 min, and the mobile phase B is increased from 40% to 45%; the mobile phase B is increased from 45% to 52% in 90-100 min; the mobile phase B is increased from 52% to 57% in 100-110 min; 110-120 min, the mobile phase B rises from 57% to 65%; the mobile phase B is increased from 65% to 70% in 120-130 min; 130-135 min, the flow rate of the mobile phase B is 0.7mL/min, the sample injection amount is 20 mu L, the column temperature is 35 ℃, and the detection wavelength is 270nm. Detection, component separation, and chromatogram recording were performed according to the above conditions, see fig. 4. The eluates at each peak time were collected and identified.
The method for detecting the tandem diode array detector (HPLC-PDA) and the method for detecting the tandem quadrupole/time-of-flight mass spectrum (HPLC-QTOF) by the liquid chromatography are particularly adopted to identify the classes of the plant compounds obtained by effective separation and enrichment.
The specific identification method is detailed as follows:
1. carrying out structural identification on the plant compound components in the bauhinia championii honey detected under the liquid chromatography conditions:
liquid phase mass spectrometry conditions: the ion source was an ESI source (electrospray), the ion source spray voltage was 4kV, the exit voltage was 130V, the heating temperature was 350 ℃, the nitrogen (N2) flow rate was 11L/min, the collision gas was helium, the atomizing gas flow rate was 80kPa, the atomizer pressure was 40psi, and the mass scan range m/z =100-900Da.
2. The structure identification and analysis process and the result of the plant compound in the bauhinia championii honey are as follows:
(1) The structure analysis process of the compound in the eluent with the retention time of 26.93min is as follows, and a specific primary mass spectrogram is shown in figure 1, and a secondary mass spectrogram is shown in figure 2:
the compound has 241.1444M/z [ M + H ] in positive ion mode]+ and 263.1266M/z [ M + Na []+ has a molecular ion peak of 239.0567M/z [ M-H ] in negative ion mode]Molecular ion peak, molecular weight of 240 as judged from the first mass spectral information, the compound having a maximum uv absorption peak at a wavelength of 246 nm. The molecular formula is determined to be C by combining the secondary mass spectrum result and the maximum ultraviolet absorption wavelength of the compound 13 H 20 O 4 According to nuclear magnetic identification, the structural formula of the compound is as follows:
Wherein the secondary mass spectrometry process is as follows: firstly, the calculated unsaturation degree omega =4 according to the ion composition of the low-mass end, no aromatic ring is contained in the analysis structure, and according to the neutral loss of continuous dehydration of the excimer ions, m/z 241.1426 generates m/z 223.1326 and further generates m/z 205.1219, and the analysis structure at least contains two hydroxyl groups. By the excimer ion m/z 241.1426 through the decarbonization 2 H 4 O was found to have an m/z of 197.1170 and was analyzed to contain-CH (OH) CH in the molecular structure 3 "this segment, in positive ion mode, can undergo a hydrogen rearrangement reaction to achieve the removal of C 2 H 4 And O. M/z 109.1015 is obtained by CO removal from fragment ion m/z 137.0960, and m/z 57.0340 is obtained by CO removal from fragment ion m/z 85.0287, and the analysis molecular structure contains carbonyl. By fragment ion m/z 179.1064 through decoc 4 H 8 M/z 123.0437 was obtained, the structure was analyzed as either cyclic olefin, with the removal of part or as 2-methylpropene. The ethylenic bond or conjugation to the carbonyl group is presumed to be based on the maximum ultraviolet absorption wavelength provided. The elemental composition of m/z 153.0908 can be used to judge that a hydroxyl group is on the ring, and the mass spectrum fragmentation path of the compound is analyzed by combining the RDA fragmentation path and the information obtained by the previous analysis as shown in FIG. 3.
(2) The structure of the compound in the eluent with the retention time of 28.34min is resolved as follows:
the mass spectrum information of the compound is consistent with that of the shishan kunmametin A, the molecular weight of the compound is judged to be 240, and the compound has a maximum ultraviolet absorption peak at the wavelength of 246 nm. And (3) combining mass spectrum data, the retention time and the maximum ultraviolet absorption wavelength of a standard substance map, and finally determining that the structural formula of the compound is as follows according to the verification of the standard substance liquid chromatogram:
(3) The structure of the compound in the eluent with the retention time of 58.78min is resolved as follows:
the compound has a molecular ion peak of 265.1451M/z [ M + H ] + in a positive ion mode, a molecular ion peak of 263.1164M/z [ M-H ] -, which can be observed in a negative ion mode, the molecular weight of the compound is judged to be 264 according to the known mass spectrum information in the positive and negative ion modes, and the compound has a maximum ultraviolet absorption peak at the wavelength of 262 nm. And finally determining the compound as trans-abscisic acid and trans-abscisic acid by combining mass spectrum data, the retention time of a trans-abscisic acid standard substance map and the maximum ultraviolet absorption wavelength and verifying according to the liquid chromatogram of the trans-abscisic acid standard substance and the trans-abscisic acid standard substance.
(4) The structure of the compound in the eluent with the retention time of 61.43min is resolved as follows:
the compound has a molecular ion peak of 243.0883M/z [ M + H ] + in a positive ion mode, and a molecular ion peak of 241.0641M/z [ M-H ] -in a negative ion mode, the molecular weight of the compound is judged to be 242 according to first-order mass spectrum information, and the compound has a maximum ultraviolet absorption peak at the wavelength of 260 nm. And finally determining the compound as the photochromic element by combining mass spectrum data, retention time of a photochromic element standard substance map and the maximum ultraviolet absorption wavelength and verifying according to the photochromic element standard substance liquid chromatogram.
(5) The structure analysis process of the compound in the eluent with the retention time of 65.66min is as follows:
the compound has molecular ion peaks of 287.1269M/z [ M + Na ] +, 247.1339M/z [ M-H2O ] +, 529.2823M/z [2M + H ] +, 551.2635M/z [2M + Na ] +, and a molecular ion peak observed in a negative ion mode has 263.1203M/z [ M-H ] -, so that the molecular weight of the compound is judged to be 264, and the compound has a maximum ultraviolet absorption peak at a wavelength of 262 nm. And finally determining the compound to be cis-abscisic acid and trans-abscisic acid according to the retention time and the maximum ultraviolet absorption wavelength of mass spectrum data, the spectrum of the cis-abscisic acid standard substance and the spectrum of the trans-abscisic acid standard substance and the liquid chromatography verification of the cis-abscisic acid standard substance and the trans-abscisic acid standard substance.
Example 2
This example measured the content of Ten Chinese alpine beehives honey by HPLC of example 1, and quantitatively analyzed the content of the measured Chinese alpine honey by external standard method.
The specific honey source information is shown in the following table 1:
TABLE 1 bauhinia championii raw material honey
The obtained chromatogram superposition spectrum of Chinese alpine rush honey in ten different producing areas is shown in figure 5.
In ten honey samples, the average content of the Shishan kunmamesu A is 7.21mg/kg; the average content of the Shishan bitter honey element B is 24.39mg/kg; conversely, the average content of trans-abscisic acid is 8.15mg/kg; the average content of the light pigment is 8.68mg/kg; the average content of cis-abscisic acid and trans-abscisic acid is 9.87mg/kg. The detailed results are shown in Table 2.
TABLE 2 determination of the content of the respective bauhinia championii honey compounds (mg/kg)
Example 3
In this example, the high performance liquid chromatography of example 1 is adopted, and the acacia honey, tilia amurensis honey, vitex honey, rape honey, safflower honey, chinese gall honey, buckwheat honey, motherwort honey and medlar honey are subjected to component analysis by a high performance liquid chromatography (the source information of the specific raw honey is shown in table 3), and the HPLC spectrum analysis of the honey and the bauhinia honey in example 1 is compared. The results show that the compounds Shishan bitter honey A, shishan bitter honey B, trans-abscisic acid, photochromic element and cis-trans abscisic acid are not simultaneously detected in other single-flower honey except the bauhinia championii honey. The specific test results are shown in Table 4.
TABLE 3 source information table of other raw material honey
Table 4 table of information on whether other honey contains five kinds of marker compounds
Example 4
In this example, four commercially available (different brands from different sales platforms) bauhinia championii honey was subjected to detection analysis using hplc using the hplc method of example 1. Through HPLC chromatogram comparison analysis, the quality of the bauhinia championii honey of three brands is better, the compounds of the bauhinia championii honey A, the bauhinia championii honey B, the trans-abscisic acid, the photochromic acid and the cis-abscisic acid and the trans-abscisic acid are detected, the contents are respectively in the ranges recorded in Table 2, the specific average contents are respectively 6.76mg/kg, 24.39mg/kg, 8.62mg/kg, 6.07mg/kg and 9.88mg/kg, and the sum of the bauhinia championii honey A and the bauhinia championii honey B is respectively 23.00mg/kg, 29.71mg/kg and 40.74mg/kg. And if the five compounds are not detected simultaneously, judging that the product is not the bauhinia championii honey.
Example 5
Sensory identification is carried out on the bauhinia championii honey, the bauhinia championii honey has certain bitter taste, and the bitter taste of the bauhinia championii honey is predicted by combining VirtualTaste (charite. De), so that the bauhinia championii honey is from the bauhinia championii honey; in addition, the color of the bauhinia championii honey is bright yellow and is close to the color of the photochromic substance, so that the color of the bauhinia championii honey is known to be derived from the photochromic substance.
Example 6
This example provides more qualitative determinations of phenolic components in bauhinia championii raw honey. In this example, the high performance liquid chromatography of example 1 was used to analyze the components of 52 bauhinia championii raw material honey originated from Guangxi by using a high performance liquid chromatograph, and the analysis was compared with the HPLC chromatogram analysis of bauhinia championii honey in example 1. The results show that the compound Shishan bitter honey A, shishan bitter honey B, trans-abscisic acid, photochromic acid and cis-abscisic acid and trans-abscisic acid can be simultaneously detected in other 52 cases of the bauhinia championii raw material honey.
In conclusion, through comparison of high performance liquid chromatography spectrums of four common large single nectars, namely bauhinia honey, acacia honey, schizonepeta honey and linden honey and analysis and comparison of collected plant compound components of other honey of the minority, such as leonurus honey, chinese gall honey, safflower honey, buckwheat honey, medlar honey and the like, the compound Shishan bitter nectin A, shishan bitter nectin B, trans-abscisic acid, photochromic elements and cis-abscisic acid and trans-abscisic acid are not simultaneously detected in other single nectars except bauhinia honey. Therefore, the Shishan kunmiki A, the Shishan kunmiki B, the trans-abscisic acid, the photochromic substance and the cis-abscisic acid and the trans-abscisic acid can be used as the characteristic plant compound markers of the bauhinia championii honey and are applied to the construction of an authenticity and quality evaluation system of the bauhinia championii honey.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (7)
1. Shishan kunmamesu A is characterized by having a structure shown in a formula (1):
2. a method for preparing Shishan kukokukosu A as claimed in claim 1, which is obtained by separating phenolic components of honey including bauhinia championii honey by high performance liquid chromatography;
when the high performance liquid chromatography is carried out, the mobile phase A is an aqueous solution of acetic acid with the concentration of 0.18-0.22%, and the mobile phase B is a methanol solution of acetic acid with the concentration of 0.18-0.22%;
the gradient elution procedure was: the mobile phase B is increased from 9 percent to 14 percent in 0-11 min; the mobile phase B is increased from 14 percent to 15 percent in 11-14 min; 14-17 min, keeping the mobile phase B at 15%; the time is 17-24 min, and the mobile phase B is increased from 15% to 16%; the time is 24-28 min, and the mobile phase B is increased from 16% to 17%; the mobile phase B is increased from 17 percent to 22 percent within 28 to 30 min; 30-41 min, keeping the mobile phase B at 22%; the time is 41-46 min, and the mobile phase B is increased from 22% to 23%; 46-55 min, the mobile phase B rises from 23% to 25%; the time is 55-60 min, and the mobile phase B is increased from 25% to 28%; the mobile phase B is increased from 28 percent to 30 percent in 60-70 min; the mobile phase B is increased from 30 percent to 40 percent in 70-80 min; the time is 80-90 min, and the mobile phase B is increased from 40% to 45%; the mobile phase B is increased from 45% to 52% in 90-100 min; the mobile phase B is increased from 52% to 57% in 100-110 min; 110-120 min, the mobile phase B rises from 57% to 65%; the mobile phase B is increased from 65% to 70% in 120-130 min; 130-135 min, and the mobile phase B is increased from 70% to 80%;
the chromatographic column is a Phenomenex Gemini C18 chromatographic column, the flow rate is 0.6-0.8mL/min, the sample injection amount is 20 mu L, the column temperature is 34-36 ℃, and the detection wavelength is 270nm;
the high performance liquid chromatography further comprises a step of extracting the phenolic components in the honey with a solid phase extraction column before the high performance liquid chromatography is carried out; specifically, extracting by using a Strata-X-A solid phase extraction column, taking methanol as an activating agent, water as a balancing agent and an eluting agent, taking a methanol solution of formic acid with the concentration of 9-11% as an eluent, blow-drying the obtained eluent, redissolving the eluent by using methanol, and filtering to obtain the eluent; the aperture of the filter membrane is 0.22 μm during filtration;
before extracting the phenolic components in the honey by using a solid phase extraction column, the method also comprises the step of honey pretreatment, and specifically comprises the following steps: mixing Mel and water, adjusting pH to 6.5-7, centrifuging at 8800-9200rpm for 18-22min, and collecting supernatant;
separating phenol components in honey by high performance liquid chromatography, and collecting target peak eluate, wherein the retention time of the target peak is 26.93min.
3. The method according to claim 2, wherein the mobile phase A is an aqueous solution of acetic acid with a concentration of 0.2%, and the mobile phase B is a methanol solution of acetic acid with a concentration of 0.2%; the flow rate was 0.7mL/min and the column temperature was 35 ℃.
4. The method according to claim 2, wherein the eluent is a 10% strength methanoic acid in methanol.
5. The method of claim 2, wherein the step of pre-treating the honey comprises: mixing Mel and water, adjusting pH to 6.8, centrifuging at 9000rpm for 20min, and collecting supernatant.
6. A method for identifying bauhinia championii honey is characterized in that phenolic components in honey to be detected are detected through a high performance liquid chromatography, and when the phenolic components simultaneously comprise shishan bitter honey A, shishan bitter honey B, trans-abscisic acid, photochromic element and cis-abscisic acid, the honey to be detected is judged to be bauhinia championii honey;
the kukomashin a as described in claim 1, and said kukomashin B has a structure represented by formula (2):
7. the identification method according to claim 6, wherein when the content of the Shishan bitter honey A in the honey to be detected is 5.40-9.81mg/kg, the content of the Shishan bitter honey B is 17.10-32.87mg/kg, the content of trans-abscisic acid is 6.37-11.01mg/kg, the content of photochromic pigment is 3.11-28.34mg/kg, and the content of cis-abscisic acid is 6.65-14.11mg/kg, the honey to be detected is determined to be bauhinia championii honey.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111131616.9A CN114181176B (en) | 2021-09-26 | 2021-09-26 | Shishan kunmiki A and preparation method thereof, and identification method of bauhinia championii honey |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111131616.9A CN114181176B (en) | 2021-09-26 | 2021-09-26 | Shishan kunmiki A and preparation method thereof, and identification method of bauhinia championii honey |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114181176A CN114181176A (en) | 2022-03-15 |
| CN114181176B true CN114181176B (en) | 2023-04-18 |
Family
ID=80601363
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111131616.9A Active CN114181176B (en) | 2021-09-26 | 2021-09-26 | Shishan kunmiki A and preparation method thereof, and identification method of bauhinia championii honey |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114181176B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115611840B (en) * | 2022-09-20 | 2024-03-26 | 广西梧州甜蜜家蜂业有限公司 | Cold winter honey element and preparation method and application thereof |
| CN117233292B (en) * | 2023-11-13 | 2024-03-08 | 中国农业科学院蜜蜂研究所 | Identification method of nine-dragon rattan honey |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105044230A (en) * | 2015-06-30 | 2015-11-11 | 西北大学 | Method for identifying linden honey, vicia villosa Roth honey and rape honey |
-
2021
- 2021-09-26 CN CN202111131616.9A patent/CN114181176B/en active Active
Non-Patent Citations (1)
| Title |
|---|
| 丁平等.鸡血藤化学成分的指纹图谱研究.《华西药学杂志》.2010,第25卷(第4期),461-463. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114181176A (en) | 2022-03-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114181176B (en) | Shishan kunmiki A and preparation method thereof, and identification method of bauhinia championii honey | |
| Sawaya et al. | Electrospray ionization mass spectrometry fingerprinting of propolis | |
| Crozier et al. | Analysis of picogram quantities of indole-3-acetic acid by high performance liquid chromatography-fluorescence procedures | |
| CN111220763B (en) | Application of high-content DSM as characteristic marker of linden honey | |
| CN111398499B (en) | Application of 3-amino-2-naphthoic acid in identifying apis cerana honey and apis mellifera honey | |
| KR102202225B1 (en) | Biomarker for the Discriminating Geographical Origins of Sesame and Method for Discriminating Geographical Origin Using the Same | |
| CN111693597A (en) | Identification method of ganglioside with high coverage and application thereof | |
| García‐Estévez et al. | Effect of the type of oak barrels employed during ageing on the ellagitannin profile of wines | |
| CN108152389A (en) | A kind of method for differentiating honey of lychee flowers | |
| CN111398498B (en) | Application of indole-3-methyl acetate in identifying apis cerana honey and apis mellifera honey | |
| CN111337605B (en) | Method for evaluating authenticity of lotus bee pollen | |
| CN111426776B (en) | Application of HQR as characteristic marker of schefflera octophylla honey | |
| CN116908352A (en) | Detection and identification method for loquat honey | |
| CN107462656A (en) | A kind of method of galanthamine content in quick detection amrallid | |
| CN115950979B (en) | Method for tracing origin of complex matrix tobacco extract | |
| CN109828052B (en) | Method for rapidly identifying species, authenticity or existence of notopterygium root | |
| CN111398497B (en) | Application of kynurenine in identifying Chinese bee honey and apis mellifera honey | |
| CN115611840B (en) | Cold winter honey element and preparation method and application thereof | |
| CN107561186B (en) | Method for simultaneously detecting contents of three alkaloids in lycoris plants | |
| CN113820435B (en) | Application of stachydrine and levo-stachydrine as characteristic markers of Leucosceptrum Nostoc honey | |
| Zhao et al. | Characteristic component profiling and identification of different Uncaria species based on high‐performance liquid chromatography–photodiode array detection tandem ion trap and time of flight mass spectrometry coupled with rDNA ITS sequence | |
| CN112129856B (en) | Method for identifying ellagic acid from gallnut | |
| CN117402197A (en) | Vitex negundo extract and preparation method thereof and identification method of Vitex negundo honey | |
| CN117263790A (en) | Longan essence, preparation method thereof and identification method of longan honey | |
| CN116930378A (en) | Detection and identification method for wild osmanthus honey |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |