CN108008036A - A kind of method for measuring Flos Tagetis Erectae extract Lutein content - Google Patents
A kind of method for measuring Flos Tagetis Erectae extract Lutein content Download PDFInfo
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- CN108008036A CN108008036A CN201711184785.2A CN201711184785A CN108008036A CN 108008036 A CN108008036 A CN 108008036A CN 201711184785 A CN201711184785 A CN 201711184785A CN 108008036 A CN108008036 A CN 108008036A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
- G01N30/8634—Peak quality criteria
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
- G01N2030/146—Preparation by elimination of some components using membranes
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Abstract
The invention discloses a kind of method for measuring Flos Tagetis Erectae extract Lutein content, comprise the following steps：The preparation of standard curve；Mobile phase：For A phases by acetonitrile, ethanol, ethyl acetate mixing gained, weight ratio is 1,000 800 successively：500‑450：300‑100；For B phases by water, acetone, triethylamine mixing gained, weight ratio is 800 650 successively：450‑100：3‑0.5；Gradient elution：The peak area of each concentration of lutein standard solution is recorded, makes the equation of linear regression of lutein；Flos Tagetis Erectae extract is dissolved in organic solvent, membrane filtration, obtain lutein sample to be tested, lutein sample to be tested is measured, record the peak area of lutein sample to be tested, the peak area is substituted into the equation of linear regression of lutein, the content of lutein sample to be tested Lutein is calculated.Advantage：Minute is short, and flowing the scientific matching of phase constituent makes mobile phase good fluidity, improves the accuracy and repeatability of measure.
Description
Technical field
The present invention relates to lutein content field of measuring technique, especially Flos Tagetis Erectae extract Lutein content measurement skill
Art.
Background technology
Marigold is also known as smelly chrysanthemum, tagetes erecta, annual herb plant.Lutein in marigold is in human eye retina
A kind of neccessary composition, it is main to play filter blue light, suppress the free radical that blue light produces, degenerate to veteran form macular pigment,
Cataract is inhibited, and significant role is played in terms of protect eyes.Because marigold Lutein content is higher, from ten thousand longevity
The technique of chrysanthemum extraction lutein gradually grows up, and whether the lutein content of raw material is enriched, the lutein purity after preparation
Deng being all factor that lutein class product preparation process needs clearly to control, a kind of measure Flos Tagetis Erectae extract Lutein is invented
Content method to improving the reliability of lutein content measure, accuracy, repeatability are considerable.
The content of the invention
It is an object of the invention to provide a kind of marigold to extract lutein preparation process.
The present invention adopts the following technical scheme that in order to achieve the above object：
A kind of method for measuring Flos Tagetis Erectae extract Lutein content, comprises the following steps：
(1) preparation of standard curve；
Lutein standard items are prepared into series of standards liquid with organic solvent, carry out high performance liquid chromatography test：
C30Performance liquid chromatographic column；
Mobile phase：
For A phases by acetonitrile, ethanol, ethyl acetate mixing gained, weight ratio is 1000-800 successively：500-450：300-
100；
For B phases by water, acetone, triethylamine mixing gained, weight ratio is 800-650 successively：450-100：3-0.5；
Gradient elution：
0-1min：The volume fraction of mobile phase A is 100%, and the volume fraction of Mobile phase B is 0；
1-10min：The volume fraction of mobile phase A is 70%, and the volume fraction of Mobile phase B is 30%；
10-17min：The volume fraction of mobile phase A is 50%, and the volume fraction of Mobile phase B is 50%；
17-20min：The volume fraction of mobile phase A is 0%, and the volume fraction of Mobile phase B is 100%；
20-23min：The volume fraction of mobile phase A is 100%, and the volume fraction of Mobile phase B is 0；
(2) peak area of each concentration of lutein standard solution is recorded, it is yellow with leaf using the peak area of lutein as ordinate
The concentration of element is abscissa, makes the equation of linear regression of lutein；
(3) Flos Tagetis Erectae extract Lutein assay：
Flos Tagetis Erectae extract is dissolved in organic solvent, membrane filtration, obtains lutein sample to be tested, according to step (1)
Chromatographic condition lutein sample to be tested is measured, record lutein sample to be tested peak area, the peak area substitute into
In the equation of linear regression of lutein, the content of lutein sample to be tested Lutein is calculated.
Further, the organic solvent of the step (1) and step (3) is that the ratio between acetone and methanol, both volumes are
4-2：1-3.
Further, the membrane filtration of the step (3) is to use fenestra as 0.30-0.40 microns of film.
Further, Flos Tagetis Erectae extract is dissolved in organic solvent in the step (3), by ultrasonic vibration, then
Carry out film process.
Further, the ultrasonic power is 100-200W, and supersonic frequency is 15-35KHz, concussion duration 15-20min.
Further, the performance liquid chromatographic column specification used in the high performance liquid chromatography test is 4.6mm*100mm,
Filler diameter is 2.0um.
Further, the column temperature of the performance liquid chromatographic column is 32 DEG C, and sampling volume is 6ul, and flow rate of mobile phase is
0.3-0.35ml/min。
Advantages of the present invention essentially consists in：Minute is short, and high performance liquid chromatography continuous mode is no more than 23min；Flowing
The scientific matching of phase constituent makes mobile phase good fluidity, and sample will not be attached on performance liquid chromatographic column during the test,
Improve the accuracy and repeatability of class measure.
Embodiment
Below in conjunction with specific embodiment, the present invention will be described in detail, herein with the illustrative examples and explanation of the present invention
For explaining the present invention, but it is not as a limitation of the invention.
Embodiment one
A kind of method for measuring Flos Tagetis Erectae extract Lutein content, comprises the following steps：
(1) preparation of standard curve；
Lutein standard items are prepared into series of standards liquid with organic solvent, carry out high performance liquid chromatography test：
C30Performance liquid chromatographic column；
Mobile phase：
For A phases by acetonitrile, ethanol, ethyl acetate mixing gained, weight ratio is 1000 successively：500：300；
For B phases by water, acetone, triethylamine mixing gained, weight ratio is 800 successively：450：3；
Gradient elution：
0-1min：The volume fraction of mobile phase A is 100%, and the volume fraction of Mobile phase B is 0；
1-10min：The volume fraction of mobile phase A is 70%, and the volume fraction of Mobile phase B is 30%；
10-17min：The volume fraction of mobile phase A is 50%, and the volume fraction of Mobile phase B is 50%；
17-20min：The volume fraction of mobile phase A is 0%, and the volume fraction of Mobile phase B is 100%；
20-23min：The volume fraction of mobile phase A is 100%, and the volume fraction of Mobile phase B is 0；
(2) peak area of each concentration of lutein standard solution is recorded, it is yellow with leaf using the peak area of lutein as ordinate
The concentration of element is abscissa, makes the equation of linear regression of lutein；
(3) Flos Tagetis Erectae extract Lutein assay：
Flos Tagetis Erectae extract is dissolved in organic solvent, membrane filtration, obtains lutein sample to be tested, according to step (1)
Chromatographic condition lutein sample to be tested is measured, record lutein sample to be tested peak area, the peak area substitute into
In the equation of linear regression of lutein, the content of lutein sample to be tested Lutein is calculated.
The organic solvent of the step (1) and step (3) is that the ratio between acetone and methanol, both volumes are 4：1.
The membrane filtration of the step (3) is to use fenestra as 0.30 micron of film.
Flos Tagetis Erectae extract is dissolved in organic solvent in the step (3), by ultrasonic vibration, then is carried out at film
Reason.
The ultrasonic power is 100W, and supersonic frequency is 15KHz, concussion duration 15min.
The performance liquid chromatographic column specification used in high performance liquid chromatography test is 4.6mm*100mm, filler diameter
It is 2.0um.
The column temperature of the performance liquid chromatographic column is 32 DEG C, and sampling volume is 6ul, and flow rate of mobile phase is 0.3ml/min.
Embodiment two
A kind of method for measuring Flos Tagetis Erectae extract Lutein content, comprises the following steps：
(1) preparation of standard curve；
Lutein standard items are prepared into series of standards liquid with organic solvent, carry out high performance liquid chromatography test：
C30Performance liquid chromatographic column；
Mobile phase：
For A phases by acetonitrile, ethanol, ethyl acetate mixing gained, weight ratio is 900 successively：450：150；
For B phases by water, acetone, triethylamine mixing gained, weight ratio is 750 successively：300：2；
Gradient elution：
0-1min：The volume fraction of mobile phase A is 100%, and the volume fraction of Mobile phase B is 0；
1-10min：The volume fraction of mobile phase A is 70%, and the volume fraction of Mobile phase B is 30%；
10-17min：The volume fraction of mobile phase A is 50%, and the volume fraction of Mobile phase B is 50%；
17-20min：The volume fraction of mobile phase A is 0%, and the volume fraction of Mobile phase B is 100%；
20-23min：The volume fraction of mobile phase A is 100%, and the volume fraction of Mobile phase B is 0；
(2) peak area of each concentration of lutein standard solution is recorded, it is yellow with leaf using the peak area of lutein as ordinate
The concentration of element is abscissa, makes the equation of linear regression of lutein；
(3) Flos Tagetis Erectae extract Lutein assay：
Flos Tagetis Erectae extract is dissolved in organic solvent, membrane filtration, obtains lutein sample to be tested, according to step (1)
Chromatographic condition lutein sample to be tested is measured, record lutein sample to be tested peak area, the peak area substitute into
In the equation of linear regression of lutein, the content of lutein sample to be tested Lutein is calculated.
The organic solvent of the step (1) and step (3) is that the ratio between acetone and methanol, both volumes are 3：2.
The membrane filtration of the step (3) is to use fenestra as 0.35 micron of film.
Flos Tagetis Erectae extract is dissolved in organic solvent in the step (3), by ultrasonic vibration, then is carried out at film
Reason.
The ultrasonic power is 150W, and supersonic frequency is 30KHz, concussion duration 20min.
The performance liquid chromatographic column specification used in high performance liquid chromatography test is 4.6mm*100mm, filler diameter
It is 2.0um.
The column temperature of the performance liquid chromatographic column is 32 DEG C, and sampling volume is 6ul, and flow rate of mobile phase is 0.35ml/min.
Embodiment three
A kind of method for measuring Flos Tagetis Erectae extract Lutein content, comprises the following steps：
(1) preparation of standard curve；
Lutein standard items are prepared into series of standards liquid with organic solvent, carry out high performance liquid chromatography test：
C30Performance liquid chromatographic column；
Mobile phase：
For A phases by acetonitrile, ethanol, ethyl acetate mixing gained, weight ratio is 800 successively：450：100；
For B phases by water, acetone, triethylamine mixing gained, weight ratio is 650 successively：100：0.5；
Gradient elution：
0-1min：The volume fraction of mobile phase A is 100%, and the volume fraction of Mobile phase B is 0；
1-10min：The volume fraction of mobile phase A is 70%, and the volume fraction of Mobile phase B is 30%；
10-17min：The volume fraction of mobile phase A is 50%, and the volume fraction of Mobile phase B is 50%；
17-20min：The volume fraction of mobile phase A is 0%, and the volume fraction of Mobile phase B is 100%；
20-23min：The volume fraction of mobile phase A is 100%, and the volume fraction of Mobile phase B is 0；
(2) peak area of each concentration of lutein standard solution is recorded, it is yellow with leaf using the peak area of lutein as ordinate
The concentration of element is abscissa, makes the equation of linear regression of lutein；
(3) Flos Tagetis Erectae extract Lutein assay：
Flos Tagetis Erectae extract is dissolved in organic solvent, membrane filtration, obtains lutein sample to be tested, according to step (1)
Chromatographic condition lutein sample to be tested is measured, record lutein sample to be tested peak area, the peak area substitute into
In the equation of linear regression of lutein, the content of lutein sample to be tested Lutein is calculated.
The organic solvent of the step (1) and step (3) is that the ratio between acetone and methanol, both volumes are 4-2：1-3.
The membrane filtration of the step (3) is to use fenestra as 0.40 micron of film.
Flos Tagetis Erectae extract is dissolved in organic solvent in the step (3), by ultrasonic vibration, then is carried out at film
Reason.
The ultrasonic power is 200W, and supersonic frequency is 35KHz, concussion duration 15-20min.
The performance liquid chromatographic column specification used in high performance liquid chromatography test is 4.6mm*100mm, filler diameter
It is 2.0um.
The column temperature of the performance liquid chromatographic column is 32 DEG C, and sampling volume is 6ul, and flow rate of mobile phase is 0.35ml/min.
The technical solution provided above the embodiment of the present invention is described in detail, specific case used herein
The principle and embodiment of the embodiment of the present invention are set forth, the explanation of above example is only applicable to help and understands this
The principle of inventive embodiments；Meanwhile for those of ordinary skill in the art, according to the embodiment of the present invention, in specific embodiment party
There will be changes in formula and application range, in conclusion this specification content should not be construed as limiting the invention.
Claims (7)
- A kind of 1. method for measuring Flos Tagetis Erectae extract Lutein content, it is characterised in that：Comprise the following steps：(1) preparation of standard curve；Lutein standard items are prepared into series of standards liquid with organic solvent, carry out high performance liquid chromatography test：C30Performance liquid chromatographic column；Mobile phase：For A phases by acetonitrile, ethanol, ethyl acetate mixing gained, weight ratio is 1000-800 successively：500-450：300-100；For B phases by water, acetone, triethylamine mixing gained, weight ratio is 800-650 successively：450-100：3-0.5；Gradient elution：0-1min：The volume fraction of mobile phase A is 100%, and the volume fraction of Mobile phase B is 0；1-10min：The volume fraction of mobile phase A is 70%, and the volume fraction of Mobile phase B is 30%；10-17min：The volume fraction of mobile phase A is 50%, and the volume fraction of Mobile phase B is 50%；17-20min：The volume fraction of mobile phase A is 0%, and the volume fraction of Mobile phase B is 100%；20-23min：The volume fraction of mobile phase A is 100%, and the volume fraction of Mobile phase B is 0；(2) peak area of each concentration of lutein standard solution is recorded, using the peak area of lutein as ordinate, with lutein Concentration is abscissa, makes the equation of linear regression of lutein；(3) Flos Tagetis Erectae extract Lutein assay：Flos Tagetis Erectae extract is dissolved in organic solvent, membrane filtration, obtains lutein sample to be tested, according to the color of step (1) Spectral condition is measured lutein sample to be tested, records the peak area of lutein sample to be tested, and it is yellow which is substituted into leaf In the equation of linear regression of element, the content of lutein sample to be tested Lutein is calculated.
- A kind of 2. method for measuring Flos Tagetis Erectae extract Lutein content according to claim 1, it is characterised in that：The organic solvent of the step (1) and step (3) is that the ratio between acetone and methanol, both volumes are 4-2：1-3.
- A kind of 3. method for measuring Flos Tagetis Erectae extract Lutein content according to claim 2, it is characterised in that：The membrane filtration of the step (3) is to use fenestra as 0.30-0.40 microns of film.
- A kind of 4. method for measuring Flos Tagetis Erectae extract Lutein content according to claim 1, it is characterised in that：Flos Tagetis Erectae extract is dissolved in organic solvent in the step (3), by ultrasonic vibration, then carries out film process.
- A kind of 5. method for measuring Flos Tagetis Erectae extract Lutein content according to claim 4, it is characterised in that：The ultrasonic power is 100-200W, and supersonic frequency is 15-35KHz, concussion duration 15-20min.
- A kind of 6. method for measuring Flos Tagetis Erectae extract Lutein content according to claim 1, it is characterised in that：The performance liquid chromatographic column specification used in the high performance liquid chromatography test is 4.6mm*100mm, and filler diameter is 2.0um。
- A kind of 7. method for measuring Flos Tagetis Erectae extract Lutein content according to claim 1, it is characterised in that：The column temperature of the performance liquid chromatographic column is 32 DEG C, and sampling volume is 6ul, and flow rate of mobile phase is 0.3-0.35ml/min.
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| CN112920101A (en) * | 2019-12-06 | 2021-06-08 | 中国科学院大连化学物理研究所 | Method for purifying and separating lutein in marigold |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109212094A (en) * | 2018-10-30 | 2019-01-15 | 晨光生物科技集团股份有限公司 | The detection method of quercetagetin in marigold residue |
| CN109212094B (en) * | 2018-10-30 | 2021-03-30 | 晨光生物科技集团股份有限公司 | Method for detecting quercitin in marigold dregs |
| CN112920101A (en) * | 2019-12-06 | 2021-06-08 | 中国科学院大连化学物理研究所 | Method for purifying and separating lutein in marigold |
| CN112920101B (en) * | 2019-12-06 | 2022-05-10 | 中国科学院大连化学物理研究所 | Method for purifying and separating lutein in marigold |
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Application publication date: 20180508 |