CN106000101B - A kind of method of separation of phenylalanine, tyrosine and tryptophan raceme - Google Patents
A kind of method of separation of phenylalanine, tyrosine and tryptophan raceme Download PDFInfo
- Publication number
- CN106000101B CN106000101B CN201610421429.7A CN201610421429A CN106000101B CN 106000101 B CN106000101 B CN 106000101B CN 201610421429 A CN201610421429 A CN 201610421429A CN 106000101 B CN106000101 B CN 106000101B
- Authority
- CN
- China
- Prior art keywords
- sample
- separation
- beta
- raceme
- cyclodextrin
- 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.)
- Expired - Fee Related
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D57/00—Separation, other than separation of solids, not fully covered by a single other group or subclass, e.g. B03C
- B01D57/02—Separation, other than separation of solids, not fully covered by a single other group or subclass, e.g. B03C by electrophoresis
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Electrochemistry (AREA)
- Environmental & Geological Engineering (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Indole Compounds (AREA)
Abstract
A kind of separation of phenylalanine,The method of tyrosine and tryptophan raceme,It is related to a kind of separation D,The separation method of l-amino acid raceme,This method makees chiral selector with Cu and L-Histidine,Using beta-cyclodextrin as separating medium,Disodium hydrogen phosphate is buffer solution,With ligand exchange method,Make Cu,L-Histidine and drawing azoles drug left-right rotary are respectively formed ternary complex,Under the effect of separating medium beta-cyclodextrin,Enantiomer is set to realize separation by capillary electrophoresis,D is detached with ligand exchange capillary electrophoresis,L-amino acid raceme,Chiral selector used in it is Cu,L-Histidine and beta-cyclodextrin,Wherein using beta-cyclodextrin as separating medium,The present invention is easy to operate,Chiral selector is cheap and easy to get and sample and reagent dosage are few,It can be used for internal analysis and the optical purity of products detection of this amino acid.
Description
Technical field
The present invention relates to a kind of separation D, the method for l-amino acid raceme, more particularly to a kind of separation of phenylalanine,
The method of tyrosine and tryptophan raceme.
Background technology
Capillary Electrophoresis has many advantages, such as efficient, quick, easy also known as high performance capillary electrophoresis because of it.It is with high pressure
DC electric field is driving force, the new skill of a kind of electrophoresis detached in capillary by the difference of its distribution coefficient with charged particle
Art, splitter more with clastotype with it effect is high, analyze speed is fast, easy to operate, sample and reagent consumption are few, ring
The advantages that border pollution is small, thus favor is received in chiral separation research.
This method chiral selector type is more, and common selective agent has cyclodextrin, chiral crown ether, big cyclohexanol peptides antibiosis
Element, linear polysaccharide, protein, Chiral surfactant and ligand exchange compound etc. can also use multiple chiral choosings simultaneously
Agent is selected, reinforces separating effect using its synergistic effect, and method is simple, quick, it is at low cost.Currently, not using both at home and abroad
Double chiral selector capillary electrophoresis detach D, the report of l-amino acid raceme.
Invention content
The purpose of the present invention is to provide a kind of method of separation of phenylalanine, tyrosine and tryptophan raceme, the party
Method researchs and develops for such single amino acid using double chiral selector capillary electrophoresis and application provides technical support and guarantor
Barrier.
The purpose of the present invention is what is be achieved through the following technical solutions：
A kind of method of separation of phenylalanine, tyrosine and tryptophan raceme, this method make hand with Cu and L-Histidine
Property selective agent, using beta-cyclodextrin as separating medium, disodium hydrogen phosphate is buffer solution, with ligand exchange method, makes Cu, L- group ammonia
The D of acid and amino acid raceme, L-type are respectively formed ternary complex, under the effect of separating medium beta-cyclodextrin, pass through capillary
Electrophoresis apparatus make enantiomer realize separation, with ligand exchange capillary electrophoresis detach draw azoles drug enantiomer, used in hand
Property selective agent is Cu, L-Histidine and beta-cyclodextrin, wherein using beta-cyclodextrin as separating medium,
Specific separating step is as follows：
A. the configuration of sample：The configuration of sample stock solution, respectively precision weigh phenylalanine, tyrosine and tryptophan and disappear
10 mg of body sample powder is revolved, sets in 10 mL brown volumetric flasks, scale is dissolved and be settled to filtered methanol, is shaken up, is obtained
Sample concentration is the storing solution of 1 mg/mL, and -20 oCRefrigerator preserves；The preparation of sample test solution, precision measure sample deposit
1 mL of liquid in 10 mL brown volumetric flasks, with methanol constant volume to scale, shakes up respectively, obtains 100 μ g/mL test liquids, and 4 oCRefrigerator
It preserves；
B. separation condition：Running buffer, 5 mmol/L sodium dihydrogen phosphates, 8 mmol/L beta-cyclodextrins, 13 mmol/L
L-Histidine, 10 mmol/L copper acetates, pH 5.0, then the ultrasound degassing after 0.45 μm of membrane filtration；Capillary column：Always
Long 53 cm, 45 cm of effective length, 50 μm of internal diameter；Wash column mode：Capillary column successively with 0.1 mol/L sodium hydroxide solutions,
Redistilled water, running buffer respectively rinse sample introduction after 10 min；Between sample introduction 5 are respectively rinsed with redistilled water, running buffer
Next sample introduction is carried out after min；Input mode：Siphon sample introduction, positive sample introduction cathode detection；Sample introduction difference in height：10 cm；When sample introduction
Between：10 s；Column temperature：Room temperature.
A kind of method of separation of phenylalanine, tyrosine and tryptophan raceme, sample operation：Runtime buffer
Liquid and sample are through 0.45 μm of filtering with microporous membrane, and ultrasound degassing, sample are dissolved with methanol, and 4 DEG C of refrigerators save backup.
A kind of method of separation of phenylalanine, tyrosine and tryptophan raceme, the electrophoretic separation condition：
Running buffer：5 mmol/L sodium dihydrogen phosphates contain 8 mmol/L β-CD, 13 mmol/L L-Histidines and 10 mmol/L vinegar
Sour copper is adjusted to pH 5.0；Separation voltage：15 kV；Ultraviolet detection wavelength：250 nm.
Azole medicine is drawn in a kind of method of separation of phenylalanine, tyrosine and tryptophan raceme, the separation
Ligand selected by object enantiomer is Cu, and chiral selector is L-Histidine, their coordination ratios are 1：1.3.
A kind of method of the separation of phenylalanine, tyrosine and tryptophan raceme, the separation of phenylalanine,
53 cm of capillary overall length of tyrosine and tryptophan raceme, 45 cm of effective length, internal diameter：50 mm；Every time before analysis successively
10 min are respectively rinsed with 10%HCl, 1mol/LNaOH, redistilled water and running buffer；Running buffer is used between sample introduction successively
5 min of column is respectively rushed with redistilled water, then carries out sample introduction next time.
A kind of method of the separation of phenylalanine, tyrosine and tryptophan raceme, the separation of phenylalanine,
8 mmol/L β-CD are added when tyrosine and tryptophan raceme in running buffer and are used as separating medium.
Advantages of the present invention is with effect：
1. changing capillary electrophoresis the present invention provides separation of phenylalanine, tyrosine and tryptophan raceme, Cu is used
Make chiral selector with L-Histidine, under the effect of separating medium beta-cyclodextrin, drug mapping is realized by capillary electrophoresis
Body detaches, and the present invention uses multiple chiral selectors, and higher separating effect, easy to operate, hand are obtained using its synergistic effect
Property selective agent is cheap and easy to get and sample and reagent dosage are few.
2. the present invention can be used for internal analysis and the optical purity detection of this amino acid, such single right to research and develop
The quality control for reflecting body amino acid provides technical support and ensures.
Description of the drawings
Fig. 1 is the chromatogram of separation of phenylalanine raceme；
Fig. 2 is separation tyrosine raceme chromatogram；
Fig. 3 is separating tryptophane raceme chromatogram.
Specific implementation mode
The present invention is described in detail with reference to the accompanying drawings.
Fig. 1-3 is separation D, and the chromatogram of l-amino acid raceme, separation analysis time, separating degree was more than in 15 minutes
3。
Embodiment：
1. precision weighs phenylalanine, tyrosine and each 10 mg of tryptophan raceme, 10 mL brown volumetric flasks are set respectively
In, scale is dissolved and be settled to methanol, is shaken up.Each sample concentration be 1 mg/mL storing solution, -20 oCRefrigerator preserves.
Precision measures 1 mL of each sample storing solution respectively in 10 mL brown volumetric flasks, with methanol constant volume to scale, shakes up, obtains 100 μ
G/mL test liquids, it is spare after 0.45 μm of filtering with microporous membrane.
2. preparing 5 mmol/LNaH2PO4Containing 8 mmol/L beta-cyclodextrins, 10 mmol/L copper acetates and 13 mmol/LL-
Histidine makees running buffer, uses H3PO4It is adjusted to pH 5.0 with NaOH, through 0.45 μm of filtering with microporous membrane, and ultrasound degassing is standby
With.Capillary column walks base after respectively rinsing 10 min with 0.1 mol/LNaOH, redistilled water and above-mentioned running buffer successively
Line, after baseline is steady（2 min or so）Using siphon sample introduction, 10 cm of sample introduction height, 10 s of sample injection time, positive sample introduction is negative
Pole is detected, and electrophoretogram is recorded.5 min of column is respectively rushed with redistilled water and running buffer successively between sample introduction, it is left to walk 2 min of baseline
The right side, then into next sample.Separation voltage：15 kV, ultraviolet detection wavelength：290 nm.Separation electrophoresis figure such as Fig. 1-3, phenylpropyl alcohol
Propylhomoserin, tyrosine and tryptophan raceme reach baseline separation.
Claims (1)
1. a kind of method of separation of phenylalanine, tyrosine and tryptophan raceme, which is characterized in that this method is with Cu and L- groups
Propylhomoserin makees chiral selector, and using beta-cyclodextrin as separating medium, disodium hydrogen phosphate is buffer solution, with ligand exchange method, is made
The D of Cu, L-Histidine and amino acid raceme, L-type are respectively formed ternary complex, under the effect of separating medium beta-cyclodextrin,
So that enantiomer is realized separation by capillary electrophoresis, detached with ligand exchange capillary electrophoresis and draw azoles drug enantiomer,
Chiral selector used in it is Cu, L-Histidine and beta-cyclodextrin, wherein using beta-cyclodextrin as separating medium；
Specific separating step is as follows：
A. the configuration of sample：The configuration of sample stock solution, respectively precision weigh phenylalanine, tyrosine and tryptophan raceme
10 mg of sample powder is set in 10 mL brown volumetric flasks, scale is dissolved and be settled to filtered methanol, is shaken up, sample is obtained
Concentration is the storing solution of 1 mg/mL, and -20 DEG C of refrigerators preserve；The preparation of sample test solution, precision measure stock sample solution 1
ML in 10 mL brown volumetric flasks, with methanol constant volume to scale, shakes up respectively, obtains 100 μ g/mL test liquids, and 4 DEG C of refrigerators preserve；
B. separation condition：Running buffer, 5mmol/L sodium dihydrogen phosphates, 8 mmol/L beta-cyclodextrins, 13mmol/L L- group ammonia
Acid, 10mmol/L copper acetates, pH5.0, then the ultrasound degassing after 0.45 μm of membrane filtration；Capillary column：Overall length 53cm, has
Imitate length 45cm, 50 μm of internal diameter；Wash column mode：Capillary column successively with 0.1 mol/L sodium hydroxide solutions, redistilled water,
Running buffer respectively rinses sample introduction after 10min；Next time is carried out after respectively rinsing 5min with redistilled water, running buffer between sample introduction
Sample introduction；Input mode：Siphon sample introduction, positive sample introduction cathode detection；Sample introduction difference in height：10cm；Sample injection time：10s；Column temperature：Room
Temperature；Separation voltage：15 kV；Ultraviolet detection wavelength：250 nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610421429.7A CN106000101B (en) | 2016-06-15 | 2016-06-15 | A kind of method of separation of phenylalanine, tyrosine and tryptophan raceme |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610421429.7A CN106000101B (en) | 2016-06-15 | 2016-06-15 | A kind of method of separation of phenylalanine, tyrosine and tryptophan raceme |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106000101A CN106000101A (en) | 2016-10-12 |
| CN106000101B true CN106000101B (en) | 2018-10-26 |
Family
ID=57088245
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610421429.7A Expired - Fee Related CN106000101B (en) | 2016-06-15 | 2016-06-15 | A kind of method of separation of phenylalanine, tyrosine and tryptophan raceme |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106000101B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109342539A (en) * | 2018-08-31 | 2019-02-15 | 安徽省新康饲料有限公司 | One kind extracting tyrosine and its detection method from cow dung |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101913278B (en) * | 2010-06-30 | 2013-07-24 | 中国科学技术大学 | Preparation method of polydopamine-g-polyethylene oxide capillary coating for separating protein by capillary electrophoresis |
| CN201728058U (en) * | 2010-08-09 | 2011-02-02 | 天津富金环境技术研究有限公司 | Dielectrophoresis separation system for heavy metal recovery |
| CN102703923B (en) * | 2012-05-21 | 2014-11-12 | 沈阳化工大学 | Method for separating lansoprazole and omeprazole racemates |
| CN103063756B (en) * | 2012-11-29 | 2014-07-30 | 沈阳化工大学 | Method for separating antiulcer medicament by double-chiral selector capillary electrophoresis method |
-
2016
- 2016-06-15 CN CN201610421429.7A patent/CN106000101B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN106000101A (en) | 2016-10-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Integrated flow-electrode capacitive deionization and microfiltration system for continuous and energy-efficient brackish water desalination | |
| US7981284B2 (en) | Suppressor utilizing micro ion exchange tube and ion chromatograph utilizing the same | |
| JP7398143B2 (en) | Method for measuring binding site occupancy and/or relative concentration of polymer membrane in liquid and electrosorption device and/or electrofiltration device | |
| JP2011504411A5 (en) | ||
| CN104892710B (en) | A kind of method for purifying reduced form β NADHs | |
| Jiang et al. | An excellent method to produce morpholine by bipolar membrane electrodialysis | |
| CN106000101B (en) | A kind of method of separation of phenylalanine, tyrosine and tryptophan raceme | |
| CN101334376A (en) | Four anti-ulcer medicament capillary pipe electrophoresis chiral isolation analysis method | |
| CN103063756B (en) | Method for separating antiulcer medicament by double-chiral selector capillary electrophoresis method | |
| CN103852528A (en) | Method for detecting posaconazole | |
| US20110163029A1 (en) | Method for substance separation using a cellulose hydrate membrane in size exclusion chromatography | |
| Zhang et al. | Tetraalkylammonium-l-tartrate ionic liquids as sole chiral selectors in capillary electrophoresis | |
| Kartsova et al. | Current status of capillary electrophoresis | |
| CN102703922B (en) | Method for separating pantoprazole from tenatoprazole drug raceme | |
| CN102703923B (en) | Method for separating lansoprazole and omeprazole racemates | |
| CN107551821A (en) | A kind of salt-resistant type anion exchange film chromatography medium and its production and use | |
| Li et al. | Exploring the nanofiltration mass transfer characteristic and concentrate process of procyanidins from grape juice | |
| CN106770595B (en) | Electrophoresis tank for Protein Separation and the column gel electrophoresis apparatus using it | |
| WO2024016656A1 (en) | Alumina film, and preparation method therefor and use thereof | |
| CN107118272A (en) | A kind of cromoci and its endotoxic minimizing technology | |
| CN101766993B (en) | Overall chiral stationary phase of silica gel capillary and preparation method thereof | |
| CN106582289B (en) | Filtration system and filtration method | |
| CN103018354A (en) | Chiral separation analyzing method of antiulcer medicament | |
| BR102014001074A2 (en) | carbon nanotubes restricted to macromolecule binding by albumin coating (racnt-bsa) | |
| CN209301846U (en) | A kind of compression type immediate dialysis device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20181026 Termination date: 20210615 |