CN110308220A - The orange red sufficient flesh Xi Shi Bao carotenoid identification of one kind and content assaying method - Google Patents
The orange red sufficient flesh Xi Shi Bao carotenoid identification of one kind and content assaying method Download PDFInfo
- Publication number
- CN110308220A CN110308220A CN201910535437.8A CN201910535437A CN110308220A CN 110308220 A CN110308220 A CN 110308220A CN 201910535437 A CN201910535437 A CN 201910535437A CN 110308220 A CN110308220 A CN 110308220A
- Authority
- CN
- China
- Prior art keywords
- carotenoid
- orange red
- ethyl acetate
- identification
- flesh
- 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.)
- Pending
Links
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
-
- 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
- G01N30/89—Inverse chromatography
Abstract
The orange red sufficient flesh Xi Shi Bao carotenoid identification of one kind and content assaying method, are related to nature strength, identification and assay.1) carotenoid is extracted；2) concentrated extracting solution；3) major carotenoids are separated；4) major carotenoids are identified；5) content of major carotenoids is measured.Using carotenoid in the orange red sufficient flesh Xi Shi Bao body of ethyl acetate soak extraction, major carotenoids type is analyzed and identified using ultraviolet specrophotometer and liquid chromatograph-mass spectrometer after evaporation and concentration, on the basis of high performance liquid chromatography, standard curve is established with major carotenoids content in computation organization's sample with standard items.Strong operability, quantitative accurate, repeated effect are good.Not only there is strong specific aim to the pigment parsing of orange red sufficient flesh Xi Shi Bao, but also extends in other shellfishes and uses.
Description
Technical field
The present invention relates to nature strength, identification and assays, more particularly, to a kind of orange red sufficient flesh Xi Shi Bao
Carotenoid identification and content assaying method.
Background technique
Carotenoid is that a kind of fat-soluble colored compound can be passive by higher plant, algae and Microbe synthesis
Object conversion absorbs that bright-coloured body colour is presented.Carotenoid is distributed widely in the marine animals body such as fish, shellfish, shrimp, crab,
Its content height has become important economic characters.At least more than 60 kinds of carotenoid in known shellfish according to statistics, more
Plate guiding principle, Gastropoda, Bivalvia and Cephalopoda are distributed.The current research in relation to seashells color focuses primarily upon shell
Not only polymorphism is presented in shell color for color, actually shellfish, and soft tissue also has different colours.As the mature ovarian of Bivalve is mostly
Red, crocus, mature spermary are mostly white；Adductor muscle in addition to white, may be there are also the colors such as red, orange.Shellfish software
In tissue it is this it is orange, red may be because it is rich in caused by carotenoid.
Abalone belongs to Mollusca (Mollusca), Gastropoda (Gastropoda), Bao Ke (Haliotidae), Bao category
(Haliotis).It is known as first of " Hai Bazhen ", is one of highest shellfish of China's economic value.Xi Shi Bao (Haliotis
Gigantea) NATURAL DISTRIBUTION was introduced a fine variety in 2003 in Japan, South Korea's littoral sea from China, and individual is larger, was resisted in Bao class
Sick power is most strong, has very high cultivation potentiality.In general, Xi Shi Bao foot is in light brown yellow more, but find in a group less
The mutated individual of bright orange-red is presented in the sufficient flesh of ratio (about 2%).Compared with common sufficient flesh, this orange red sufficient flesh is not only
It is in beautiful color, and it is rich in carotenoid, it is absorbed for research marine animal carotenoid, transhipment and metabolic mechanism provide
Cast material.However, the research at present about carotenoid Identification of Species in shellfish soft tissue is very few, only Li Ning ([1] Lee
Peaceful Chinese Marine University Ph.D. Dissertation .2010) reported that Patinopecten yessoensis (Patinopecten yessoensis) was orange
Adductor muscle is rich in two Carotenoids of scallop alcohol ketone and pectenoxanthin.The research precise Identification goes out the orange adductor muscle of Patinopecten yessoensis
Carotenoid type, but method and step is complicated.([2] Zhang Qiqing, Guo Wei, Cai Zhixing blue-green pigments of abalone shell mentions Zhang Qiqing
Take method application publication number CN 101717590A) etc. invent a kind of extraction method of blue-green pigments of abalone shell, the technical method
Simply, remaining shell is complete after pigment extracts, but cannot identify its type.It yet there are no orange red mutant class in the prior art recklessly
Radish element Identification of Species and assay are reported.
Summary of the invention
The object of the present invention is to provide a kind of orange red foots for the body colour parsing that can solve the problems, such as orange red sufficient flesh Xi Shi Bao
The identification of flesh Xi Shi Bao carotenoid and content assaying method.
The present invention the following steps are included:
1) carotenoid is extracted；
In step 1), the specific method for extracting carotenoid can are as follows: tissue sample is put into vacuum refrigeration and is done
The dry 48h of dry machine, is clayed into power with grinder or mortar, is dissolved in 20ml ethyl acetate after weighing 0.7~1.5g foot flesh powder, adds
Enter 0.02gBHT (2,6- di-tert-butyl-4-methy phenol), 4 DEG C of centrifugations 5 of homogenate 5~10min, 4000r/min freezing~
10min takes supernatant；It is homogenized above, is centrifuged, the step of supernatant is taken to be repeated 3 times, sufficiently to extract the carotenoids in sample
Element, by supernatant collection in pear shape bottle, extraction process whole process is protected from light, to avoid carotenoid illumination effect under degradation and
Isomerization.
2) concentrated extracting solution；
In step 2), the specific method of the concentrated extracting solution can are as follows: rotate by supernatant obtained by step 1) dense
Contracting, is dried to powder for concentrate using solvent work station, is stored in -20 DEG C of refrigerators；The condition of the concentrated by rotary evaporation can are as follows:
10 DEG C of refrigerator temps <, 33 DEG C of water bath temperature <；The solvent work station can be GecVacEZ-2plus, program: LOW
BP；33 DEG C of temperature <；Time to final stage:30~40min；Final stage:30~40min.
3) major carotenoids are separated；
In step 3), the specific method of the separation major carotenoids can are as follows: (is worn using high performance liquid chromatograph
Pacify Ultimate3000), access reverse-phase chromatographic column ZorbaxSB-C18 (5 μ m 4.6 × 250mm, Agilent
Technologies, Santa Clara, CA, USA) carotenoid type in orange red Xi Shi Bao body is separated, condition is column
Temperature, 25 DEG C；Sampling volume, 20 μ l；Mobile phase, by volume, Yi Jing ︰ Jia Chun ︰ dichloromethane solution=40 ︰, 56 ︰ 4；Flow velocity
1.0ml/min；To advanced optimize separating effect, using forward chromatographic column YMC-Pack SIL column (250 × 4.6mm, 5
μm, 12nm, YMC CO., Ltd., Kyoto, Japan) optimization separating effect, mobile phase is that normal hexane and ethyl acetate carry out ladder
Degree elution, using linear gradient elution: 1) Zheng Yi Wan ︰ ethyl acetate=40 ︰ 60,10min；2) normal hexane=100,5min；3)
Zheng Yi Wan ︰ ethyl acetate=40 ︰ 60 balance 5min, column temperature, 25 DEG C；Flow velocity 1.5ml/min.
4) major carotenoids are identified；
In step 4), the specific method of the identification major carotenoids can are as follows: to identify isolated main peak,
Using liquid chromatograph-mass spectrometer (kyoto, Japan Shimadzu LCMS-8050), it is true that ESI-MS analysis is carried out using positive ion mode
Its fixed molecular weight, mass spectral analysis condition and step 3) reverse-phase chromatography are consistent；Infer what step 3) obtained according to its molecular weight
Main peak is zeaxanthin or lutein, confirms orange red Xi Shi by comparing zeaxanthin, lutein, beta carotene standard items
Bao major carotenoids type.
5) content of major carotenoids is measured.
In step 5), the specific method of the content of the measurement major carotenoids can are as follows: by step 3) and 4)
Obtaining major carotenoids type in orange red sufficient flesh Xi Shi Bao body is zeaxanthin and beta carotene, will be obtained by step 2)
Carotenoid powder is dissolved in 2ml ethyl acetate, and sample to be tested is made with the membrane filtration in 0.22 μm of aperture；Use efficient liquid phase
Chromatograph (wears peace Ultimate 3000), access forward chromatographic column YMC-PackSIL (250 × 4.6mm, 5 μm, 12nm,
YMCCO., Ltd., Kyoto, Japan), 25 DEG C of column temperature, mobile phase is that normal hexane and ethyl acetate carry out gradient elution；Using line
Property gradient elution: 1) Zheng Yi Wan ︰ ethyl acetate=40 ︰ 60,10min；2) normal hexane=100,5min；3) Zheng Yi Wan ︰ acetic acid second
Ester=40 ︰ 60 balance 5min, flow rate of mobile phase, and: 1.5ml/min measures light absorption value at 446nm；By 1.0mg zeaxanthin mark
Quasi- product are dissolved in 5.0ml ethyl acetate respectively, and the standard items stoste of 200 μ g/ml is made, and dilute stoste with ethyl acetate, are prepared into
Concentration is respectively the working solution of 1.25,2.5,5,10,25,50,100 μ g/ml；Each concentration work is measured with high performance liquid chromatograph
Liquid light absorption value at 446nm, obtains standard curve, obtains zeaxanthin concentration in sample using standard curve, uses step 1)
Middle tissue powder weight data calculates zeaxanthin concentration in basic stitch sample；The measuring method and zeaxanthin of beta carotene
It is similar, working solution concentration need to be only changed to 1.25,2.5,5.0,10.0,15.0 μ g/ml in step.
The present invention uses carotenoid in the orange red sufficient flesh Xi Shi Bao body of ethyl acetate soak extraction, makes after evaporation and concentration
Major carotenoids type is analyzed and identified with ultraviolet specrophotometer and liquid chromatograph-mass spectrometer, in high-efficient liquid phase color
On the basis of spectrometry, standard curve is established with major carotenoids content in computation organization's sample with standard items.
Remarkable advantage of the invention is: strong operability, quantitative accurate, and repeated effect is good.Both to orange red sufficient flesh west
The pigment parsing of family name Bao has strong specific aim, and extends in other shellfishes and use.
Detailed description of the invention
Fig. 1 is carotenoid high-efficient liquid phase chromatogram in orange red sufficient flesh Xi Shi Bao body.
Fig. 2 is carotenoid high-efficient liquid phase chromatogram in common sufficient flesh Xi Shi Bao body.
Fig. 3 is carotenoid mass spectrogram in orange red sufficient flesh Xi Shi Bao body.
Specific embodiment
Following embodiment will the present invention is further illustrated in conjunction with attached drawing.
Tissue raw material in the embodiment of the present invention are as follows: the soft tissues such as sufficient flesh, sexual gland or gill of orange red foot flesh Xi Shi Bao.
The embodiment of the present invention includes following steps:
1) it extracts carotenoid: tissue sample being put into the dry 48h of vacuum freeze drier, is ground with grinder or mortar
At powder.0.70~1.50g foot flesh powder is weighed, weight is recorded.Be dissolved in 20ml ethyl acetate, be added 0.02gBHT (2,
6- di-tert-butyl-4-methy phenol), it is homogenized 5~10min, 4000r/min freezes (4 DEG C) 5~10min of centrifugation, takes supernatant.
It is homogenized, is centrifuged above, taking the step of supernatant in triplicate, sufficiently to extract the carotenoid in sample, by supernatant collection
In pear shape bottle.Extraction process whole process is protected from light, to avoid carotenoid degradation and isomerization under illumination effect.
2) concentrated extracting solution: by supernatant obtained by step 1) carry out concentrated by rotary evaporation (10 DEG C of refrigerator temps <, water-bath pot temperature
Spend 33 DEG C of <).Use solvent work station (GecVacEZ-2plus, program: LOW BP；33 DEG C of temperature <；time to final
stage30-40min；Final stage30-40min) concentrate is dried to powder, it is stored in -20 DEG C of refrigerators.
3) major carotenoids are separated: using high performance liquid chromatograph (wearing peace Ultimate3000), accessing reverse phase color
Compose column ZorbaxSB-C18 (5 μ m 4.6 × 250mm, Agilent Technologies, Santa Clara, CA, USA) separation
Carotenoid type in orange red Xi Shi Bao body, condition are column temperature, 25 DEG C；Sampling volume, 20 μ l；Mobile phase, Yi Jing ︰ first
56 ︰ 4, v/v of Chun ︰ dichloromethane solution=40 ︰；Flow velocity 1.0ml/min.To advanced optimize separating effect, normal-phase chromatography is used
Column YMC-Pack SIL column (250 × 4.6mm, 5 μm, 12nm, YMC CO., Ltd., Kyoto, Japan) optimization separation effect
Fruit.Mobile phase is normal hexane (A) and ethyl acetate (B) carries out gradient elution.Using linear gradient elution: 1) A ︰ B=40 ︰ 60,
10min；2) A=100,5min；3) A ︰ B=40 ︰ 60 balances 5min.Column temperature, 25 DEG C；Flow velocity 1.5ml/min.
4) major carotenoids are identified: to identify isolated main peak, utilizing liquid chromatograph-mass spectrometer (day
This capital of a country Shimadzu LCMS-8050), using positive ion mode carry out ESI-MS analyze determine its molecular weight, mass spectral analysis condition with
Step 3) reverse-phase chromatography is consistent.Infer that the main peak that step 3) obtains is zeaxanthin or lutein according to its molecular weight, is
It further confirms that, confirms the orange red main class Hu trailing plants of Xi Shi Bao by comparing zeaxanthin, lutein, beta carotene standard items
Bu Su type.
By step 3) and 4) 5) measure the content of major carotenoids: obtaining main in orange red sufficient flesh Xi Shi Bao body
Carotenoid type is zeaxanthin and beta carotene.Carotenoid powder obtained by step 2) is dissolved in 2ml acetic acid second
Sample to be tested is made with the membrane filtration in 0.22 μm of aperture in ester.It (wears using high performance liquid chromatograph and pacifies Ultimate 3000),
It accesses forward chromatographic column YMC-PackSIL (250 × 4.6mm, 5 μm, 12nm, YMCCO., Ltd., Kyoto, Japan).Column temperature 25
DEG C, mobile phase is normal hexane (A) and ethyl acetate (B) carries out gradient elution.Using linear gradient elution: 1) A ︰ B=40 ︰ 60,
10min；2) A=100,5min；3) A ︰ B=40 ︰ 60 balances 5min.Flow rate of mobile phase: 1.5ml/min.It measures and inhales at 446nm
Light value.1.0mg zeaxanthin standard items are dissolved in 5.0ml ethyl acetate respectively, the standard items stoste of 200 μ g/ml is made.Use second
Acetoacetic ester dilutes stoste, is prepared into the working solution that concentration is respectively 1.25,2.5,5,10,25,50,100 μ g/ml.With efficient liquid
Chromatography measures each concentration working solution light absorption value 446nm at, obtains standard curve, by the light absorption value of sample to be tested and the ratio between
To zeaxanthin concentration in acquisition sample.Using powder qualitative data is organized in step 1), zeaxanthin in basic stitch sample is calculated
Concentration.The measuring method of beta carotene is similar, working solution concentration only need to be changed to 1.25 in step, 2.5,5.0,
10.0,15.0 μ g/ml.
Specific embodiment is given below.
Fig. 1 provides carotenoid high-efficient liquid phase chromatogram in orange red sufficient flesh Xi Shi Bao body, and Fig. 2 provides common sufficient flesh west
Carotenoid high-efficient liquid phase chromatogram in family name's Bao body, Fig. 3 provide carotenoid mass spectrogram in orange red sufficient flesh Xi Shi Bao body.
Embodiment 1
The sufficient flesh powder for weighing the orange red sufficient flesh Xi Shi Bao of 1.0g, is dissolved in 20ml ethyl acetate, 0.02gBHT (2,6- is added
Di-tert-butyl-4-methy phenol), it is homogenized 3min, 4000r/min freezes (4 DEG C) centrifugation 5min, takes supernatant.The above homogenate, from
The heart takes the step of supernatant to be repeated 3 times, by supernatant collection in pear shape bottle.With Rotary Evaporators (- 10 DEG C of refrigerator temps, water
32 DEG C of bath temperature) supernatant flashed into dope, reuse solvent work station (GecVacEZ-2plus, program: LOW BP；
33 DEG C of temperature；time to final stage 40min；Final stage40min) in be dried to powder.Powder is dissolved in
Extracting solution to be measured is made with 0.22 μm of membrane filtration in 2ml ethyl acetate.Extracting solution is dissolved in acetonitrile, methanol and methylene chloride
Mobile phase solution in (40 ︰, 56 ︰ 4, v/v).Using liquid chromatograph-mass spectrometer (kyoto, Japan Shimadzu LCMS-8050), adopt
ESI-MS analysis is carried out with positive ion mode, by comparison zeaxanthin, lutein, beta carotene standard items confirmatory sample
Contained carotenoid type is zeaxanthin and beta carotene.
Embodiment 2
The orange red sufficient flesh Xi Shi Bao tissue powder of 1.0g is weighed, 20ml ethyl acetate is dissolved in, 0.02gBHT (2,6- bis- is added
Tert-butyl-4-methyl-Phenol), it is homogenized 5min, 4000r/min freezes (4 DEG C) centrifugation 5min, takes supernatant.The above homogenate, from
The heart takes the step of supernatant to be repeated 3 times, by supernatant collection in pear shape bottle.With Rotary Evaporators (- 10 DEG C of refrigerator temps, water
32 DEG C of bath temperature) supernatant flashed into dope, reuse solvent work station (GecVacEZ-2plus, program: LOW BP；
33 DEG C of temperature；time to final stage 40min；Final stage 40min) in be dried to powder.Powder is dissolved in
Sample to be tested is made with 0.22 μm of membrane filtration in 2ml ethyl acetate.(peace Ultimate is worn using high performance liquid chromatograph
3000), forward chromatographic column YMC-PackSIL (250 × 4.6mm, 5 μm, 12nm, YMCCO., Ltd., Kyoto, Japan) is accessed.
25 DEG C of column temperature, mobile phase is normal hexane (A) and ethyl acetate (B) carries out gradient elution.Using linear gradient elution: 1) A ︰ B=
40 ︰ 60,10min；2) A=100,5min；3) A ︰ B=40 ︰ 60 balances 5min；Flow velocity 1.5ml/min.Preparing concentration is respectively
1.25, the zeaxanthin standard items working solution of 2.5,5,10,25,50,100 μ g/ml, concentration is respectively 1.25,2.5,5,10,
25, the beta carotene working solution of 50,100 μ g/ml makes the standard curve that concentration corresponds to light absorption value respectively.By sample extinction
Value is compared with standard curve, obtains zeaxanthin concentration and beta carotene concentration in sample.
Claims (6)
1. a kind of orange red sufficient flesh Xi Shi Bao carotenoid identification and content assaying method, it is characterised in that including following step
It is rapid:
1) carotenoid is extracted；
2) concentrated extracting solution；
3) major carotenoids are separated；
4) major carotenoids are identified；
5) content of major carotenoids is measured.
2. a kind of orange red sufficient flesh Xi Shi Bao carotenoid identification and content assaying method as described in claim 1, feature
It is in step 1), the extraction carotenoid method particularly includes: tissue sample is put into vacuum freeze drier and is done
Dry 48h, is clayed into power with grinder or mortar, is dissolved in 20ml ethyl acetate after weighing 0.7~1.5g foot flesh powder, is added
0.02g2,6- di-tert-butyl-4-methy phenol, are homogenized 5~10min, and 4000r/min freezes 4 DEG C of 5~10min of centrifugation, takes supernatant
Liquid；It is homogenized above, is centrifuged, the step of supernatant is taken to be repeated 3 times, sufficiently to extract the carotenoid in sample, supernatant is received
It combines in pear shape bottle, extraction process whole process is protected from light, to avoid carotenoid degradation and isomerization under illumination effect.
3. a kind of orange red sufficient flesh Xi Shi Bao carotenoid identification and content assaying method as described in claim 1, feature
It is in step 2), the concentrated extracting solution method particularly includes: supernatant obtained by step 1) is subjected to concentrated by rotary evaporation, is used
Concentrate is dried to powder by solvent work station, is stored in -20 DEG C of refrigerators；The condition of the concentrated by rotary evaporation can are as follows: refrigerator
10 DEG C of temperature <, 33 DEG C of water bath temperature <；The solvent work station can be GecVacEZ-2plus, program: LOW BP；Temperature
33 DEG C of <；Time to final stage:30~40min；Final stage:30~40min.
4. a kind of orange red sufficient flesh Xi Shi Bao carotenoid identification and content assaying method as described in claim 1, feature
It is in step 3), the separation major carotenoids method particularly includes: use high performance liquid chromatograph, access reverse phase
Chromatographic column ZorbaxSB-C18 separates carotenoid type in orange red Xi Shi Bao body, and condition is column temperature, and 25 DEG C；Sample introduction body
Product, 20 μ l；Mobile phase, by volume, Yi Jing ︰ Jia Chun ︰ dichloromethane solution=40 ︰, 56 ︰ 4；Flow velocity 1.0ml/min；For into one
Step optimization separating effect, using forward chromatographic column YMC-Pack SIL column optimize separating effect, mobile phase be normal hexane and
Ethyl acetate carries out gradient elution, using linear gradient elution: 1) Zheng Yi Wan ︰ ethyl acetate=40 ︰ 60,10min；2) normal hexane
=100,5min；3) Zheng Yi Wan ︰ ethyl acetate=40 ︰ 60 balance 5min, column temperature, 25 DEG C；Flow velocity 1.5ml/min.
5. a kind of orange red sufficient flesh Xi Shi Bao carotenoid identification and content assaying method as described in claim 1, feature
It is in step 4), the identification major carotenoids method particularly includes: to identify isolated main peak, utilize liquid
Phase chromatograph-mas spectrometer carries out ESI-MS using positive ion mode and analyzes its determining molecular weight, mass spectral analysis condition and step
3) reverse-phase chromatography is consistent；Infer that the main peak that step 3) obtains is zeaxanthin or lutein according to its molecular weight, passes through ratio
Orange red Xi Shi Bao major carotenoids type is confirmed to zeaxanthin, lutein, beta carotene standard items.
6. a kind of orange red sufficient flesh Xi Shi Bao carotenoid identification and content assaying method as described in claim 1, feature
It is in step 5), the content of the measurement major carotenoids method particularly includes: obtain by step 3) and 4) orange
Major carotenoids type is zeaxanthin and beta carotene in red foot flesh Xi Shi Bao body, by class Hu trailing plants obtained by step 2)
Bu Su powder is dissolved in 2ml ethyl acetate, and sample to be tested is made with the membrane filtration in 0.22 μm of aperture；Use high performance liquid chromatography
Instrument accesses forward chromatographic column YMC-PackSIL, and 25 DEG C of column temperature, mobile phase is that normal hexane and ethyl acetate carry out gradient elution；It adopts
With linear gradient elution: 1) Zheng Yi Wan ︰ ethyl acetate=40 ︰ 60,10min；2) normal hexane=100,5min；3) Zheng Yi Wan ︰ second
Acetoacetic ester=40 ︰ 60 balance 5min, flow rate of mobile phase, and: 1.5ml/min measures light absorption value at 446nm；By 1.0mg maize
Matter standard items are dissolved in 5.0ml ethyl acetate respectively, and the standard items stoste of 200 μ g/ml is made, and dilute stoste, system with ethyl acetate
For the working solution at concentration being respectively 1.25,2.5,5,10,25,50,100 μ g/ml；Each concentration is measured with high performance liquid chromatograph
Working solution light absorption value at 446nm, obtains standard curve, obtains zeaxanthin concentration in sample using standard curve, uses step
Rapid 1) middle tissue powder weight data, calculates zeaxanthin concentration in basic stitch sample；The measuring method and corn of beta carotene
Yellow matter is similar, and difference is in step for working solution concentration to be changed to 1.25,2.5,5.0,10.0,15.0 μ g/ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910535437.8A CN110308220A (en) | 2019-06-20 | 2019-06-20 | The orange red sufficient flesh Xi Shi Bao carotenoid identification of one kind and content assaying method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910535437.8A CN110308220A (en) | 2019-06-20 | 2019-06-20 | The orange red sufficient flesh Xi Shi Bao carotenoid identification of one kind and content assaying method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110308220A true CN110308220A (en) | 2019-10-08 |
Family
ID=68076260
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910535437.8A Pending CN110308220A (en) | 2019-06-20 | 2019-06-20 | The orange red sufficient flesh Xi Shi Bao carotenoid identification of one kind and content assaying method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110308220A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111307976A (en) * | 2020-03-12 | 2020-06-19 | 宁波大学 | High-flux detection method for carotenoid in aquatic product |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102735767A (en) * | 2012-06-04 | 2012-10-17 | 华中科技大学 | Method for detecting content of carotenoids having cis-trans isomers through high-polarity mobile phase |
| CN104090062A (en) * | 2014-07-30 | 2014-10-08 | 江苏省农业科学院 | Detection method for single-cis and double-cis lutein isomers |
| CN104297407A (en) * | 2014-10-23 | 2015-01-21 | 中国农业科学院作物科学研究所 | Ultra-high performance liquid chromatographic determination method for content of carotenoid in wheat |
| CN105693583A (en) * | 2015-12-30 | 2016-06-22 | 晨光生物科技集团股份有限公司 | Method for removing harmful ingredients in carotenoid |
-
2019
- 2019-06-20 CN CN201910535437.8A patent/CN110308220A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102735767A (en) * | 2012-06-04 | 2012-10-17 | 华中科技大学 | Method for detecting content of carotenoids having cis-trans isomers through high-polarity mobile phase |
| CN104090062A (en) * | 2014-07-30 | 2014-10-08 | 江苏省农业科学院 | Detection method for single-cis and double-cis lutein isomers |
| CN104297407A (en) * | 2014-10-23 | 2015-01-21 | 中国农业科学院作物科学研究所 | Ultra-high performance liquid chromatographic determination method for content of carotenoid in wheat |
| CN105693583A (en) * | 2015-12-30 | 2016-06-22 | 晨光生物科技集团股份有限公司 | Method for removing harmful ingredients in carotenoid |
Non-Patent Citations (5)
| Title |
|---|
| R. B. MCGEACHIN 等: "Determination of Carotenoid Pigments, Retinol, and α-Tocopherol in Feeds, Tissues, and Blood Serum by Normal Phase High Performance Liquid Chromatography", 《POULTRY SCIENCE》 * |
| 何卿 等: "植物类胡萝卜素提取与分析技术研究进展", 《植物学报》 * |
| 刘洋 等: "鲑鱼肉和饲料中类胡萝卜素的分析方法", 《水产科学》 * |
| 李宁: "虾夷扇贝橘红色闭壳肌产生的原因及其在育种中的应用", 《中国博士学位论文全文数据库 农业科技辑》 * |
| 石婧 等: "水产动物组织中类胡萝卜素的研究进展", 《食品工业科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111307976A (en) * | 2020-03-12 | 2020-06-19 | 宁波大学 | High-flux detection method for carotenoid in aquatic product |
| CN111307976B (en) * | 2020-03-12 | 2022-06-28 | 宁波大学 | High-throughput detection method for carotenoid in aquatic product |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sawicki et al. | Betalain profile, content and antioxidant capacity of red beetroot dependent on the genotype and root part | |
| Li et al. | Characterization of phenolics, betacyanins and antioxidant activities of the seed, leaf, sprout, flower and stalk extracts of three Amaranthus species | |
| Rebecca et al. | Pigment identification and antioxidant properties of red dragon fruit (Hylocereus polyrhizus) | |
| Roriz et al. | Floral parts of Gomphrena globosa L. as a novel alternative source of betacyanins: Optimization of the extraction using response surface methodology | |
| Çekiç et al. | Comparison of antioxidant capacity and phytochemical properties of wild and cultivated red raspberries (Rubus idaeus L.) | |
| Sass-Kiss et al. | Differences in anthocyanin and carotenoid content of fruits and vegetables | |
| Lee et al. | Characterization and quantitation of anthocyanins in purple-fleshed sweet potatoes cultivated in Korea by HPLC-DAD and HPLC-ESI-QTOF-MS/MS | |
| CN103919712B (en) | Cordyceps militaris extract, and preparation method and application thereof | |
| Yao et al. | Free radical scavenging capability, antioxidant activity and chemical constituents of Pyrola incarnata Fisch. leaves | |
| Fernández-Marín et al. | Isorhapontigenin: a novel bioactive stilbene from wine grapes | |
| Fu et al. | Chemical composition and antioxidant activity of Chinese wild raspberry (Rubus hirsutus Thunb.) | |
| Luo et al. | Preparative separation of grape skin polyphenols by high-speed counter-current chromatography | |
| Feng et al. | Polyphenol profile and antioxidant activity of the fruit and leaf of Vaccinium glaucoalbum from the Tibetan Himalayas | |
| Wang et al. | Resveratrol synthesis under natural conditions and after UV-C irradiation in berry skin is associated with berry development stages in ‘Beihong’(V. vinifera× V. amurensis) | |
| Ji et al. | Investigation of the distribution and season regularity of resveratrol in Vitis amurensis via HPLC–DAD–MS/MS | |
| Li et al. | Polyphenol stilbenes from fenugreek (Trigonella foenum-graecum L.) seeds improve insulin sensitivity and mitochondrial function in 3T3-L1 adipocytes | |
| CN105717209A (en) | Method for extracting carotenoid components in pepper leaves and quantitatively detecting content of carotenoid components | |
| CN105241965B (en) | A kind of method of on-line quick detection sample total economic value | |
| Zhu et al. | Composition and contents of anthocyanins in different pomegranate cultivars | |
| CN103033578B (en) | Method for quickly and preliminarily screening excellent tea tree resource based on contained contents | |
| Nie et al. | Preparative separation of three terpenoids from edible brown algae Sargassum fusiforme by high-speed countercurrent chromatography combined with preparative high-performance liquid chromatography | |
| CN110308220A (en) | The orange red sufficient flesh Xi Shi Bao carotenoid identification of one kind and content assaying method | |
| CN107698458A (en) | A kind of extracting method of double (to the coumaric acyl) spermidines of N1, N5 | |
| CN101967302A (en) | Extraction method of wild chrysanthemum carotenoid pigment and perpetration method thereof | |
| Chen et al. | Identification of key taste components in Baccaurea ramiflora Lour. fruit using non-targeted metabolomics |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191008 |