CN112557544A - Method for detecting lycium carotenoid metabolites - Google Patents
Method for detecting lycium carotenoid metabolites Download PDFInfo
- Publication number
- CN112557544A CN112557544A CN202011454821.4A CN202011454821A CN112557544A CN 112557544 A CN112557544 A CN 112557544A CN 202011454821 A CN202011454821 A CN 202011454821A CN 112557544 A CN112557544 A CN 112557544A
- Authority
- CN
- China
- Prior art keywords
- methanol
- solution
- mobile phase
- detecting
- water
- 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.)
- Granted
Links
Images
Classifications
-
- 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
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
-
- 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
Abstract
The invention discloses a detection method of a lycium carotenoid metabolite, and relates to the technical field of detection methods. Taking a proper amount of medlar, pretreating, preparing a test solution, and detecting by using a liquid chromatography; the liquid chromatography conditions are as follows: a chromatographic column: YM-C30 column (4.6X150mm, 5 μm); the mobile phase A is methanol/water solution, the mobile phase B is methyl tert-butyl ether/methanol solution, and gradient elution is adopted, wherein the volume ratio of methanol to water in the methanol water solution is 15: 85; the volume ratio of methyl tert-butyl ether to methanol in the methyl tert-butyl ether/methanol solution was 25: 75. The method has the advantages of rapidness, high efficiency, stability, accuracy and the like, and can be used for rapidly and effectively detecting the carotenoid metabolites of the medlar.
Description
Technical Field
The invention relates to the technical field of detection methods, in particular to a detection method of a lycium carotenoid metabolite.
Background
Carotenoids are fat-soluble pigments present in plants and microorganisms, and most carotenoids are yellow, orange or red in color. Numerous medical studies have shown that carotenoids have the effects of efficiently quenching singlet oxygen and scavenging free radicals, thereby reducing the damage of free radicals to cell genetic material and cell membranes. Epidemiological studies show that carotenoids play a more important role in protecting human health, such as preventing cardiovascular diseases and cancer. However, the human body cannot synthesize carotenoids by itself, and fruits and vegetables are important sources for obtaining carotenoids in humans.
The medlar is a functional characteristic plant resource with homology of medicine and food, is rich in various bioactive substances such as carotenoid, flavonoid substances, medlar polysaccharide and the like, and is widely applied to the health food industry. At present, the method for analyzing the carotenoids in the lycium barbarum is mainly High Performance Liquid Chromatography (HPLC), and the method is sensitive, good in repeatability and suitable for batch detection, wherein the selection of a mobile phase is a key factor influencing the separation effect of the carotenoids.
However, the detection methods of carotenoids disclosed in the prior art are not satisfactory for simultaneous detection of multiple carotenoids. In order to detect carotenoids in lycium barbarum more comprehensively and effectively, it is necessary to establish a rapid, simple, accurate and reliable analysis method for qualitative and quantitative analysis of index components of multiple carotenoids in lycium barbarum.
Disclosure of Invention
The invention provides a method for detecting a carotenoid metabolite of medlar, aiming at solving the problems in the prior art.
The technical scheme for solving the technical problems is as follows:
a method for detecting carotenoid metabolites of fructus Lycii comprises pretreating appropriate amount of fructus Lycii, preparing into test solution, and detecting by liquid chromatography; the liquid chromatography conditions are as follows: a chromatographic column: YM-C30 column (4.6X150mm, 5 μm); the mobile phase A is methanol/water solution, the mobile phase B is methyl tert-butyl ether/methanol solution, and gradient elution is adopted, wherein the volume ratio of methanol to water in the methanol water solution is 15: 85; the volume ratio of methyl tert-butyl ether to methanol in the methyl tert-butyl ether/methanol solution was 25: 75.
The invention has the beneficial effects that: the HPLC method is used for detecting carotenoid metabolites in the medlar, and has the advantages of convenient detection, high detection sensitivity and high sample recovery rate.
On the basis of the technical scheme, the invention can be further improved as follows.
In the method for detecting the carotenoid metabolites of the medlar, provided by the invention, the gradient elution procedure comprises the following steps:
| time (min) | A(％) | B(％) |
| 0 | 5 | 95 |
| 5 | 5 | 95 |
| 15 | 17 | 83 |
| 30 | 17 | 83 |
| 45 | 60 | 60 |
| 46 | 5 | 95 |
| 50 | 5 | 95 |
In the detection method of the lycium carotenoid metabolites, the detection wavelength is 450nm under the liquid chromatography condition.
In the detection method of the lycium carotenoid metabolites, the column temperature is 30 ℃ under the liquid chromatography condition.
In the detection method of the lycium carotenoid metabolites, the flow rate is 1ml/min under the liquid chromatography condition.
In the detection method of the lycium carotenoid metabolites, provided by the invention, the preparation method of the test solution comprises the following steps:
s1, grinding and crushing the medlar, then mixing with a proper amount of mobile phase B, and filtering until the extracting solution is colorless;
s2: mixing the extractive solutions, concentrating under reduced pressure, and drying to obtain crude extract;
s3: dissolving the crude extract of S2 with methanol, adding equal volume of 30% methanol-KOH, and saponifying for more than 4 hr to obtain saponified solution;
s4: concentrating the saponified solution under reduced pressure to a minimum volume, and adding a proper amount of mobile phase B for extraction until the extract is colorless;
and S5, combining the extraction liquid in the step S4, washing the alkali liquid during saponification by water, taking the upper extraction liquid, and concentrating the upper extraction liquid under reduced pressure to the minimum volume to obtain the concentrated carotenoid test solution.
The invention also provides a detection method of the lycium carotenoid metabolites, which comprises the following steps:
(1) and preparing a test solution:
s1, grinding and crushing the medlar, then mixing with a proper amount of mobile phase B, and filtering until the extracting solution is colorless;
s2: mixing the extractive solutions, concentrating under reduced pressure, and drying to obtain crude extract;
s3: dissolving the crude extract of S2 with methanol, adding equal volume of 30% methanol-KOH, and saponifying for more than 4 hr to obtain saponified solution;
s4: concentrating the saponified solution under reduced pressure to a minimum volume, and adding a proper amount of mobile phase B for extraction until the extract is colorless;
s5, combining the extracts in the step S4, washing the alkali liquor during saponification by water, taking the upper extract, concentrating the upper extract under reduced pressure to the minimum volume to obtain the concentrated carotenoid test solution;
(2) preparation of control solutions: taking a proper amount of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene reference substances, precisely weighing, and adding methanol to prepare a mixed solution containing 10 micrograms of neoxanthin, 10 micrograms of cordierite, 10 micrograms of epoxy zeaxanthin, 10 micrograms of lutein and 60 micrograms of beta-carotene per 1ml, so as to obtain the product;
(3) and (3) detection: injecting 10 mul of each of the reference substance and the test solution into a liquid chromatographic column for detection, wherein the chromatographic conditions are as follows: a chromatographic column: YM-C30 column (4.6X150mm, 5 μm); the mobile phase A is a methanol/water solution with the volume ratio of methanol to water being 15:85, the mobile phase B is a methyl tert-butyl ether/methanol solution with the volume ratio of methyl tert-butyl ether to methanol being 25:75, the detection wavelength is 450nm, the column temperature is 30 ℃, the flow rate is 1.0ml/min, and the gradient elution conditions are as follows:
the beneficial effect of adopting the further scheme is that:
1. the method can detect and separate 6 components of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene in the medlar, can comprehensively and truly reflect the inherent quality characteristics of the medlar, and saves time cost and reagent cost;
2. the detection time is reasonable, the peak-off time of the target substance is short, and the detection efficiency of the sample is greatly improved;
3. the separation degree of each target component is good, the baseline fluctuation is small, the background interference is small, and the components in the sample to be tested can be accurately detected;
4. through the invention investigation of repeatability, recovery rate, linearity and the like of the detection method, the result shows that the detection method has high stability, good repeatability, good linearity and high recovery rate.
Drawings
FIG. 1 high performance liquid chromatogram of example 1 sample
FIG. 2 high performance liquid chromatogram of the sample of example 2;
FIG. 3 is a high performance liquid chromatogram of the test sample of example 3;
FIG. 4 is a high performance liquid chromatogram of the test sample of example 4;
FIG. 5 high performance liquid chromatogram of the test sample of example 5;
in the figure: 1. neoxanthin, 2, cordierite, 3, epoxy zeaxanthin, 4, zeaxanthin, 5, lutein, 6, beta-carotene.
Detailed Description
The principles and features of this invention are described below in conjunction with specific embodiments, which are set forth merely to illustrate the invention and are not intended to limit the scope of the invention.
The experimental apparatus, the medicines and the reagents related to the invention are as follows:
LC-2010CHT high performance liquid chromatograph (Shimadzu Japan)
Milli-Q pure water system (American MA)
The methanol was chromatographically pure, the remaining reagents were analytically pure.
Preparation of the solution:
preparation of a reference solution: taking a proper amount of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene reference substances, precisely weighing, and adding methanol to prepare a mixed solution containing 10 micrograms of neoxanthin, 10 micrograms of cordierite, 10 micrograms of epoxy zeaxanthin, 10 micrograms of lutein and 60 micrograms of beta-carotene per 1ml, so as to obtain the product;
preparation of a test solution:
s1, grinding and crushing the medlar, then mixing with a proper amount of mobile phase B, and filtering until the extracting solution is colorless;
s2: mixing the extractive solutions, concentrating under reduced pressure, and drying to obtain crude extract;
s3: dissolving the crude extract of S2 with methanol, adding equal volume of 30% methanol-KOH, and saponifying for more than 4 hr to obtain saponified solution;
s4: concentrating the saponified solution under reduced pressure to a minimum volume, and adding a proper amount of mobile phase B for extraction until the extract is colorless;
s5, combining the extracts in the step S4, washing the alkali liquor during saponification by water, taking the upper extract, concentrating the upper extract under reduced pressure to the minimum volume to obtain the concentrated carotenoid test solution;
example one
And (3) detection: 10 mul of each of the reference solution and the sample solution is injected into a liquid chromatograph for detection. Wherein, the chromatographic conditions are as follows: the chromatographic column is YM-C30 chromatographic column (4.6X150mm, 5 μm); the mobile phase A is a methanol/water solution with a methanol-water volume ratio of 15:85, the mobile phase B is a methyl tert-butyl ether/methanol solution with a methyl tert-butyl ether-methanol volume ratio of 25:75, and the ratio of the mobile phase A to the mobile phase B is 16: 84; the detection wavelength is 450 nm; the number of theoretical plates should not be less than 3000 calculated according to the lutein peak.
The results of the HPLC analysis according to the above method are shown in FIG. 1. From the results in FIG. 1, it is clear that beta-carotene could not be detected, and that neoxanthin and cordierite peaks overlapped and the separation requirement could not be met.
Example two
And (3) detection: 10 mul of each of the reference solution and the sample solution is injected into a liquid chromatograph for detection. Wherein, the chromatographic conditions are as follows: the chromatographic column is YM-C30 chromatographic column (4.6X150mm, 5 μm); the mobile phase A is a methanol/water solution with the volume ratio of methanol to water being 15:85, and the mobile phase B is a methyl tert-butyl ether/methanol solution with the volume ratio of methyl tert-butyl ether to methanol being 25: 75; the detection wavelength is 450 nm; the number of theoretical plates is not less than 10000 calculated according to the lutein peak, and the following gradient elution is adopted.
| Time (min) | A(％) | B(％) |
| 0 | 10 | 90 |
| 5 | 10 | 90 |
| 20 | 20 | 80 |
| 30 | 40 | 60 |
| 45 | 75 | 25 |
| 55 | 10 | 90 |
| 60 | 10 | 90 |
The results of the HPLC analysis according to the above method are shown in FIG. 2. From the results of fig. 2, it can be seen that zeaxanthin and lutein cannot be completely separated, the separation requirement cannot be met, and the baseline shift of the detection gradient of β -carotene is large.
EXAMPLE III
And (3) detection: 10 mul of each of the reference solution and the sample solution is injected into a liquid chromatograph for detection. Wherein, the chromatographic conditions are as follows: the chromatographic column is YM-C30 chromatographic column (4.6X150mm, 5 μm); the mobile phase A is a methanol/water solution with the volume ratio of methanol to water being 15:85, and the mobile phase B is a methyl tert-butyl ether/methanol solution with the volume ratio of methyl tert-butyl ether to methanol being 15: 85; the detection wavelength is 450 nm; the number of theoretical plates is not less than 10000 calculated according to the lutein peak, and the following gradient elution is adopted.
| Time (min) | A(％) | B(％) |
| 0 | 15 | 85 |
| 5 | 15 | 85 |
| 20 | 30 | 70 |
| 30 | 40 | 60 |
| 45 | 75 | 25 |
| 55 | 10 | 90 |
| 60 | 10 | 90 |
The results of the HPLC analysis according to the above method are shown in FIG. 3. From the results in FIG. 3, it is clear that neoxanthin and cordierite peaks overlap, separation requirements are not satisfied, and β -carotene cannot be detected.
Example four
And (3) detection: 10 mul of each of the reference solution and the sample solution is injected into a liquid chromatograph for detection. Wherein, the chromatographic conditions are as follows: the chromatographic column is YM-C30 chromatographic column (4.6X150mm, 5 μm); the mobile phase A is a methanol/water solution with the volume ratio of methanol to water being 15:85, and the mobile phase B is a methyl tert-butyl ether/methanol solution with the volume ratio of methyl tert-butyl ether to methanol being 15: 85; the detection wavelength is 450 nm; the number of theoretical plates is not less than 10000 calculated according to the lutein peak, and the following gradient elution is adopted.
| Time (min) | A(％) | B(％) |
| 0 | 15 | 85 |
| 5 | 15 | 85 |
| 20 | 30 | 70 |
| 30 | 40 | 60 |
| 45 | 75 | 25 |
| 46 | 10 | 90 |
| 50 | 10 | 90 |
The results of the HPLC analysis according to the above method are shown in FIG. 4. From the results in FIG. 4, it is clear that neoxanthin and cordierite peaks overlap, separation requirements are not satisfied, and β -carotene cannot be detected.
EXAMPLE five
And (3) detection: 10 mul of each of the reference solution and the sample solution is injected into a liquid chromatograph for detection. Wherein, the chromatographic conditions are as follows: the chromatographic column is YM-C30 chromatographic column (4.6X150mm, 5 μm); the mobile phase A is a methanol/water solution with the volume ratio of methanol to water being 15:85, and the mobile phase B is a methyl tert-butyl ether/methanol solution with the volume ratio of methyl tert-butyl ether to methanol being 25: 75; the detection wavelength is 450 nm; the number of theoretical plates is not less than 10000 calculated according to the lutein peak, and the following gradient elution is adopted.
The results of the HPLC analysis according to the above method are shown in FIG. 5. According to the results in fig. 5, the separation degrees of the 6 components are all greater than 1.5, the detection effect is achieved, the gradient detection baseline of each component is the most stable, the interference is the least, and the detection effect is the best.
And (3) experimental verification:
the linear relationship is:
taking a mixed reference solution of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene (wherein the neoxanthin concentration is 0.22mg/ml, the cordierite concentration is 0.28mg/ml, the epoxy zeaxanthin concentration is 0.75mg/ml, the zeaxanthin concentration is 0.64mg/ml, the lutein concentration is 0.58mg/ml and the beta-carotene concentration is 0.68mg/ml), precisely measuring 0.5ml, 1ml, 2.5ml, 5ml and 10ml to 10ml volumetric flasks by using a pipette, adding methanol to dilute to a scale, and shaking up. Under the above chromatographic conditions, 10. mu.l of each sample was pipetted into a liquid chromatograph and the peak area was measured. Taking the sampling amount (mug) of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene as an abscissa and the peak area (A) of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene as an ordinate, and performing linear regression to obtain the standard curves of neoxanthin, cordierite, epoxy zeaxanthin, zeaxanthin and beta-carotene respectively. The results show that the components have good linear relation in the corresponding concentration range, and the linear range, the regression equation and the related coefficient are referred to the following table.
Linear relationship of carotenoid components in fructus Lycii
| Detecting the component | Linear relationship (ug) | Regression equation | Coefficient of correlation (r) |
| Novel flavins | 0.11-2.21 | Y＝965.7X-1.0876 | 0.9999 |
| Cordierite | 0.14-2.82 | Y＝726.2X-3.9512 | 0.9999 |
| Epoxy zeaxanthin | 0.375-7.34 | Y＝1652.4X+1.527 | 0.9998 |
| Zeaxanthin | 0.32-6.41 | Y＝1112.9X+2.445 | 0.9997 |
| Lutein (lutein) | 0.29-5.84 | Y＝1023.9X+3.109 | 0.9998 |
| Beta-carotene | 0.34-6.47 | Y＝1058.1X+1.477 | 1.0 |
Repeatability:
taking the same batch of medlar samples, precisely weighing, preparing 6 parts of test solution in parallel according to the method under the preparation item of the test solution, carrying out sample injection measurement according to the chromatographic conditions, recording a chromatogram, and calculating the RSD of the contents of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene to be 1.2%, 1.3%, 0.8%, 1.1%, 0.9% and 0.9% respectively, wherein the RSD is less than 2.0%, which shows that the method has good repeatability.
Stability:
preparing a test sample solution according to a test sample preparation method; respectively standing for 0, 2, 4, 8, 12 and 24h, injecting 10 μ l of sample, injecting into a liquid chromatograph, and respectively measuring peak areas of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene in the sample solution. RSD is calculated. The RSD for calculating the contents of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene are respectively 1.6%, 1.4%, 1.8%, 1.9%, 1.7% and 1.3%, and are all less than 2.0%, which indicates that the method has good stability.
And (3) sample recovery rate:
taking 6 parts of medlar samples, wherein the content of each part of medlar is about 0.1g, precisely weighing, placing the medlar samples in a conical flask with a stopper, respectively adding appropriate amounts of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene reference substances, preparing sample adding and recycling sample solution according to the preparation method of the sample solution, carrying out sample injection and measurement according to the chromatographic conditions, recording a chromatogram, and calculating the sample adding and recycling rates of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene, wherein the results are shown in the following table.
Recovery rate determination (n ═ 6)
| Detecting the component | Average recovery (%) | RSD(％) |
| Novel flavins | 99.8 | 1.9 |
| Cordierite | 101.8 | 1.8 |
| Epoxy zeaxanthin | 98.9 | 1.6 |
| Zeaxanthin | 102.4 | 2.1 |
| Lutein (lutein) | 99.7 | 1.8 |
| Beta-carotene | 97.9 | 1.7 |
As shown in the table above, the sample recovery rate of each component in the sample is between 95.0% and 105.0%, the RSD% is less than 3.0%, and the test result shows that the sample recovery rate is good.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (7)
1. A method for detecting a carotenoid metabolite of Chinese wolfberry is characterized by comprising the following steps: taking a proper amount of medlar, pretreating, preparing a test solution, and detecting by using a liquid chromatography; the liquid chromatography conditions are as follows: a chromatographic column: YM-C30 column (4.6X150mm, 5 μm); the mobile phase A is methanol/water solution, the mobile phase B is methyl tert-butyl ether/methanol solution, and gradient elution is adopted, wherein the volume ratio of methanol to water in the methanol water solution is 15: 85; the volume ratio of methyl tert-butyl ether to methanol in the methyl tert-butyl ether/methanol solution was 25: 75.
2. The method for detecting the carotenoids of Lycium barbarum as claimed in claim 1, wherein: the gradient elution procedure was:
3. The method for detecting the carotenoids of Lycium barbarum as claimed in claim 1, wherein: in the liquid chromatography condition, the detection wavelength is 450 nm.
4. The method for detecting the carotenoids of Lycium barbarum as claimed in claim 1, wherein: under the liquid chromatography condition, the column temperature is 30 ℃.
5. The method for detecting the carotenoids of Lycium barbarum as claimed in claim 1, wherein: in the liquid chromatography conditions, the flow rate was 1 ml/min.
6. The method for detecting the carotenoids of Lycium barbarum of claim 1, wherein the sample solution is prepared by the following steps:
s1, grinding and crushing the medlar, then mixing with a proper amount of mobile phase B, and filtering until the extracting solution is colorless;
s2: mixing the extractive solutions, concentrating under reduced pressure, and drying to obtain crude extract;
s3: dissolving the crude extract of S2 with methanol, adding equal volume of 30% methanol-KOH, and saponifying for more than 4 hr to obtain saponified solution;
s4: concentrating the saponified solution under reduced pressure to a minimum volume, and adding a proper amount of mobile phase B for extraction until the extract is colorless;
and S5, combining the extraction liquid in the step S4, washing the alkali liquid during saponification by water, taking the upper extraction liquid, and concentrating the upper extraction liquid under reduced pressure to the minimum volume to obtain the concentrated carotenoid test solution.
7. The method for detecting the carotenoids of Lycium barbarum as claimed in any one of claims 1-6, wherein the method comprises the steps of:
(1) and preparing a test solution:
s1, grinding and crushing the medlar, then mixing with a proper amount of mobile phase B, and filtering until the extracting solution is colorless;
s2: mixing the extractive solutions, concentrating under reduced pressure, and drying to obtain crude extract;
s3: dissolving the crude extract of S2 with methanol, adding equal volume of 30% methanol-KOH, and saponifying for more than 4 hr to obtain saponified solution;
s4: concentrating the saponified solution under reduced pressure to a minimum volume, and adding a proper amount of mobile phase B for extraction until the extract is colorless;
s5, combining the extracts in the step S4, washing the alkali liquor during saponification by water, taking the upper extract, concentrating the upper extract under reduced pressure to the minimum volume to obtain the concentrated carotenoid test solution;
(2) preparation of control solutions: taking a proper amount of neoxanthin, cordierite, epoxy zeaxanthin, lutein and beta-carotene reference substances, precisely weighing, and adding methanol to prepare a mixed solution containing 10 micrograms of neoxanthin, 10 micrograms of cordierite, 10 micrograms of epoxy zeaxanthin, 10 micrograms of lutein and 60 micrograms of beta-carotene per 1ml, so as to obtain the product;
(3) and (3) detection: injecting 10 mul of each of the reference substance and the test solution into a liquid chromatographic column for detection, wherein the chromatographic conditions are as follows: a chromatographic column: YM-C30 column (4.6X150mm, 5 μm); the mobile phase A is a methanol/water solution with the volume ratio of methanol to water being 15:85, the mobile phase B is a methyl tert-butyl ether/methanol solution with the volume ratio of methyl tert-butyl ether to methanol being 25:75, the detection wavelength is 450nm, the column temperature is 30 ℃, the flow rate is 1.0ml/min, and the gradient elution conditions are as follows:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011454821.4A CN112557544B (en) | 2020-12-09 | 2020-12-09 | Detection method of carotenoid metabolites of wolfberry |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011454821.4A CN112557544B (en) | 2020-12-09 | 2020-12-09 | Detection method of carotenoid metabolites of wolfberry |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112557544A true CN112557544A (en) | 2021-03-26 |
| CN112557544B CN112557544B (en) | 2023-06-30 |
Family
ID=75061843
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011454821.4A Active CN112557544B (en) | 2020-12-09 | 2020-12-09 | Detection method of carotenoid metabolites of wolfberry |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112557544B (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030129264A1 (en) * | 2001-06-29 | 2003-07-10 | Randal Hauptmann | Tagetes erecta marigolds with altered carotenoid compositions and ratios |
| US20040022881A1 (en) * | 2001-06-29 | 2004-02-05 | Randal Hauptmann | Mixed zeaxanthin ester concentrate and uses thereof |
| CN104749293A (en) * | 2015-04-21 | 2015-07-01 | 江苏省农业科学院 | Method for efficiently extracting carotenoids in yellow peach fruits and determining carotenoids in yellow peach fruits by liquid phase |
| CN105717209A (en) * | 2016-01-29 | 2016-06-29 | 甘肃农业大学 | Method for extracting carotenoid components in pepper leaves and quantitatively detecting content of carotenoid components |
| CN107561171A (en) * | 2017-07-06 | 2018-01-09 | 北京林业大学 | The high efficiency extraction and assay method of carotenoid in Chinese rose petal |
| CN108120788A (en) * | 2016-11-29 | 2018-06-05 | 康迎春 | HPLC analysis methods on five Carotenoids of detection |
| CN110412153A (en) * | 2019-06-26 | 2019-11-05 | 西南大学 | A method of carotenoid metabolin is marked based on LC-MS/MS technology screening honeysuckle yellow petal |
-
2020
- 2020-12-09 CN CN202011454821.4A patent/CN112557544B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030129264A1 (en) * | 2001-06-29 | 2003-07-10 | Randal Hauptmann | Tagetes erecta marigolds with altered carotenoid compositions and ratios |
| US20040022881A1 (en) * | 2001-06-29 | 2004-02-05 | Randal Hauptmann | Mixed zeaxanthin ester concentrate and uses thereof |
| CN104749293A (en) * | 2015-04-21 | 2015-07-01 | 江苏省农业科学院 | Method for efficiently extracting carotenoids in yellow peach fruits and determining carotenoids in yellow peach fruits by liquid phase |
| CN105717209A (en) * | 2016-01-29 | 2016-06-29 | 甘肃农业大学 | Method for extracting carotenoid components in pepper leaves and quantitatively detecting content of carotenoid components |
| CN108120788A (en) * | 2016-11-29 | 2018-06-05 | 康迎春 | HPLC analysis methods on five Carotenoids of detection |
| CN107561171A (en) * | 2017-07-06 | 2018-01-09 | 北京林业大学 | The high efficiency extraction and assay method of carotenoid in Chinese rose petal |
| CN110412153A (en) * | 2019-06-26 | 2019-11-05 | 西南大学 | A method of carotenoid metabolin is marked based on LC-MS/MS technology screening honeysuckle yellow petal |
Non-Patent Citations (9)
| Title |
|---|
| IRAKLI MARIA等: "Optimization and development of a high-performance liquid chromatography method for the simultaneous determination of vitamin E and carotenoids in tomato fruits", JOURNAL OF SEPARATION SCIENCE, vol. 39, no. 17, pages 3348 - 3356 * |
| 康迎春;尹跃;赵建华;刘兰英;: "HPLC法测定枸杞鲜果中主要类胡萝卜素组成", 食品工业, no. 12, pages 270 - 273 * |
| 胡丽松 等: "菠萝蜜类胡萝卜素检测方法研究及呈色物质分析", 《热带作物学报》 * |
| 胡丽松 等: "菠萝蜜类胡萝卜素检测方法研究及呈色物质分析", 《热带作物学报》, 31 December 2017 (2017-12-31), pages 950 - 956 * |
| 郑晓冬 等: "高效液相色谱法检测枸杞中类胡萝卜素研究进展", 《中国果菜》 * |
| 郑晓冬 等: "高效液相色谱法检测枸杞中类胡萝卜素研究进展", 《中国果菜》, 31 December 2013 (2013-12-31), pages 27 - 30 * |
| 陈丽;坤吉瑞;王静;童华荣;: "HPLC-PDA法分析不同茶树品种类胡萝卜素香气前体", 食品科学, no. 04, pages 193 - 198 * |
| 陈鹏飞 等: "大白菜的类胡萝卜素种类鉴定", 《中国农学通报》 * |
| 陈鹏飞 等: "大白菜的类胡萝卜素种类鉴定", 《中国农学通报》, 31 December 2011 (2011-12-31), pages 1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112557544B (en) | 2023-06-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11525815B2 (en) | Method for product quality control and fingerprint detection of epimedium brevicornu complex | |
| CN106770851B (en) | A kind of mobile phase formula of liquid chromatography for measuring carotenoid and its application | |
| CN113267588B (en) | Method for simultaneously detecting 7 acrylic acid and acrylic ester monomers in plastic product | |
| CN111089916B (en) | Method for detecting content of paeoniflorin, liquiritin and ammonium glycyrrhizinate in radix bupleuri and radix paeoniae alba oral liquid | |
| CN105021723A (en) | Method for simultaneous determination of content of geniposide, andrographolide and dehydroandrographolide in Zhimai tablets for clearing heat | |
| CN109632997A (en) | The extraction and measuring method of regallity glycosides B in a kind of lily medicinal material | |
| CN104897796A (en) | Method for determining oleanolic acid content and ursolic acid content in sesame oil through high performance liquid chromatography method | |
| CN112557544B (en) | Detection method of carotenoid metabolites of wolfberry | |
| CN101776659A (en) | Method for detecting anthocyanin in red radish through high-performance liquid chromatography | |
| CN116879424A (en) | Method for measuring content of terprivet glycoside in shengxuebao preparation | |
| CN111999411B (en) | High performance liquid chromatography detection method for cyanidin and 3 kinds of anthocyanin in fruits and vegetables | |
| CN112415104B (en) | Characteristic spectrum, construction method and detection method of pyrrosia pedunculata medicinal material, decoction pieces, standard decoction and formula granules thereof | |
| CN110687224B (en) | Method for measuring triptolide A in tripterygium wilfordii medicinal material and tripterygium wilfordii multi-glycoside tablet prepared from tripterygium wilfordii medicinal material | |
| CN113406241A (en) | Quality detection method of Xintong granules | |
| CN106053696A (en) | Method for identifying plant source of herbal rabdosia lophanthide | |
| CN108037200B (en) | Quality detection method of kidney nourishing and tranquilizing pills | |
| CN115372510B (en) | Hypocrellin fingerprint and establishment method and application thereof | |
| CN104897797A (en) | Method for determining oleanolic acid content and ursolic acid content in perilla frutescens oil through high performance liquid chromatography method | |
| CN105301122A (en) | Method for determination of contents of oleanolic acid and ursolic acid in kiwi fruit oil through high performance liquid chromatography | |
| CN110632198A (en) | HPLC fingerprint of inflammation diminishing and cough relieving tablets and construction method and application thereof | |
| CN113391002B (en) | Establishing method and application of Xintong granule UPLC fingerprint spectrum | |
| CN114778730B (en) | Method for simultaneously detecting 6 organic acids in aster medicinal material or aster product | |
| CN112903835B (en) | Fingerprint spectrum construction method and detection method of momordica grosvenori and red tangerine peel solid beverage | |
| CN117110491B (en) | Method for detecting ginsenoside in traditional Chinese medicine preparation containing American ginseng | |
| CN116577431A (en) | Method for simultaneously detecting content of six active ingredients of alisma orientale |
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 |