CN110988178A - High performance liquid chromatography detection method for phosphorylcholine - Google Patents
High performance liquid chromatography detection method for phosphorylcholine Download PDFInfo
- Publication number
- CN110988178A CN110988178A CN201911299590.1A CN201911299590A CN110988178A CN 110988178 A CN110988178 A CN 110988178A CN 201911299590 A CN201911299590 A CN 201911299590A CN 110988178 A CN110988178 A CN 110988178A
- Authority
- CN
- China
- Prior art keywords
- phosphorylcholine
- high performance
- performance liquid
- liquid chromatography
- detection method
- 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
- YHHSONZFOIEMCP-UHFFFAOYSA-O phosphocholine Chemical compound C[N+](C)(C)CCOP(O)(O)=O YHHSONZFOIEMCP-UHFFFAOYSA-O 0.000 title claims abstract description 17
- 229950004354 phosphorylcholine Drugs 0.000 title claims abstract description 17
- 238000001514 detection method Methods 0.000 title claims abstract description 15
- 238000004128 high performance liquid chromatography Methods 0.000 title claims abstract description 9
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 12
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims abstract description 8
- 125000003368 amide group Chemical group 0.000 claims abstract description 6
- 239000007788 liquid Substances 0.000 claims abstract description 6
- 230000007935 neutral effect Effects 0.000 claims abstract description 6
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims abstract description 4
- 235000019253 formic acid Nutrition 0.000 claims abstract description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 239000000243 solution Substances 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 3
- 238000010812 external standard method Methods 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 238000010829 isocratic elution Methods 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 238000011002 quantification Methods 0.000 claims 1
- 238000000926 separation method Methods 0.000 abstract description 8
- 238000000034 method Methods 0.000 abstract description 5
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 4
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229960001340 histamine Drugs 0.000 description 2
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 230000002194 synthesizing effect Effects 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 208000004930 Fatty Liver Diseases 0.000 description 1
- 206010019708 Hepatic steatosis Diseases 0.000 description 1
- XSDGQYHSDRSTKI-UHFFFAOYSA-N N1CCOCC1.P(=O)#C[N+](CCO)(C)C Chemical compound N1CCOCC1.P(=O)#C[N+](CCO)(C)C XSDGQYHSDRSTKI-UHFFFAOYSA-N 0.000 description 1
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 1
- 229960004373 acetylcholine Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- -1 chlorinated phosphorylcholine hydrochloride Chemical class 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 208000010706 fatty liver disease Diseases 0.000 description 1
- 238000002013 hydrophilic interaction chromatography Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000003607 modifier Substances 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 231100000240 steatosis hepatitis Toxicity 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
-
- 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
- G01N2030/042—Standards
- G01N2030/047—Standards external
Landscapes
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to the technical field of analytical chemistry, and relates to a high performance liquid chromatography detection method of phosphorylcholine. Separating the sample liquid by a neutral amide bonded phase chromatographic column, isocratically eluting acetonitrile and 0.1 percent formic acid solution, and detecting by an evaporative light detector. The method is simple and convenient, can quickly measure the content of the phosphorylcholine, and has higher accuracy, stability and separation degree.
Description
Technical Field
The invention belongs to the technical field of analytical chemistry, and relates to a high performance liquid chromatography detection method of phosphorylcholine, in particular to a high performance liquid chromatography detection method of phosphorylcholine, which has high accuracy, stability and resolution and is used for neutral amide bonded phase chromatographic column separation and evaporative light detector detection.
Background
The phosphorylcholine exists in the form of chlorinated phosphorylcholine hydrochloride, can participate in synthesizing phospholipid, has the effects of protecting liver, strengthening liver, promoting lipid metabolism and resisting fatty liver, can accelerate methyl transfer and promote liver cell regeneration, and can synthesize acetylcholine in vivo to activate nervous system; decompose histamine, enhance the activity of renal histamine enzyme, and has the function of detoxification, and meanwhile, the phosphorylcholine is also a raw material for synthesizing phosphorylcholine morpholine which is an intermediate of choline medicaments. The phosphorylcholine can be used in the field of medicine, can also be used as a quality modifier for brewed products, and can stabilize amino acid and other wine ingredients.
At present, the content of choline phosphate in the beverage is detected by using a liquid chromatography-mass spectrometry method, but the detection cost of the liquid chromatography-mass spectrometry method is higher, and the universality of the liquid chromatography is not strong, so a high performance liquid chromatography detection method for the choline phosphate is required to be established.
The method is simple to operate, can quickly determine the content of the phosphorylcholine, and has higher accuracy, stability and separation degree.
Disclosure of Invention
The invention aims to provide high performance liquid chromatography detection with higher accuracy, stability and separation degree, and the high performance liquid chromatography detection can realize the rapid determination of the content of the phosphorylcholine by the separation of a neutral amido bonded phase chromatographic column and the detection of an evaporative light detector.
In order to realize the purpose of the invention, the technical scheme of the invention is as follows: dissolving a sample by pure water or methanol, fixing the volume, injecting the sample into a high performance liquid chromatograph after passing through a membrane, collecting a chromatogram under the condition of setting an instrument, and quantifying by an external standard method; wherein, the instrument conditions are set as that the neutral amido bonded phase chromatographic column is separated, the column temperature is 35 ℃, acetonitrile with the volume ratio of 20/80 and 0.1 percent formic acid solution are used as mobile phases for isocratic elution, the column flow is 0.5mL/min, the sample injection volume is 10 mu L, and the detection is carried out by an evaporative light detector.
Has the advantages that: the method adopts neutral amide bonded phase chromatographic column separation and an evaporative light detector for detection, has higher accuracy, stability and separation degree, is simple to operate, can quickly determine the content of the phosphorylcholine, and greatly saves the analysis time and cost.
Drawings
FIG. 1 high performance liquid chromatogram of standard solution
Detailed Description
The present invention is further illustrated by the following examples, but is not limited thereto in any way, and any variations or modifications based on the teachings of the present invention are within the scope of the present invention.
Example one
The high performance liquid chromatograph is matched with an evaporation light detector.
0.1g of the phosphorylcholine standard substance (precisely 0.001g) was weighed out accurately, dissolved in methanol and made to 100 mL. The chromatographic conditions employed were: venusil HILIC chromatography column; column temperature of 35 ℃; acetonitrile and 0.1% formic acid solution are eluted with equal degree by mobile phase and have the volume ratio of 20/80; flow rate of 0.5 mL/min; sample introduction volume of 10 μ L; and detecting by an evaporative light detector. And (4) injecting the standard solution into a high performance liquid chromatograph after passing through a membrane, and recording a chromatogram. 3.75min is a phosphorylcholine spectrum peak, the retention time is appropriate, the separation degree is high, and the peak shape symmetry is good.
Claims (1)
1. A high performance liquid chromatography detection method of phosphorylcholine is characterized in that a sample is dissolved by pure water or methanol and subjected to constant volume, the solution is injected into a high performance liquid chromatograph after passing through a membrane, a chromatogram is collected under the condition of setting an instrument, and the quantification is carried out by an external standard method; wherein, the instrument conditions are set as that the neutral amido bonded phase chromatographic column is separated, the column temperature is 35 ℃, acetonitrile with the volume ratio of 20/80 and 0.1 percent formic acid solution are used as mobile phases for isocratic elution, the column flow is 0.5mL/min, the sample injection volume is 10 mu L, and the detection is carried out by an evaporative light detector.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911299590.1A CN110988178A (en) | 2019-12-12 | 2019-12-12 | High performance liquid chromatography detection method for phosphorylcholine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911299590.1A CN110988178A (en) | 2019-12-12 | 2019-12-12 | High performance liquid chromatography detection method for phosphorylcholine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110988178A true CN110988178A (en) | 2020-04-10 |
Family
ID=70094450
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911299590.1A Pending CN110988178A (en) | 2019-12-12 | 2019-12-12 | High performance liquid chromatography detection method for phosphorylcholine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110988178A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116879428A (en) * | 2023-06-29 | 2023-10-13 | 沈阳金久奇科技有限公司 | High performance liquid analysis method for residual content of phosphorylcholine in L-alpha-phosphorylcholine |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5643716A (en) * | 1992-09-08 | 1997-07-01 | Amur Research Corp. | Diagnostic agent and methods for identifying HIV infected individuals and monitoring their therapy |
| CN102138892A (en) * | 2010-02-03 | 2011-08-03 | 广州汉光医药进出口有限公司 | Choline alfoscerate injection preparation as well as preparation method and detection method thereof |
| CN105388225A (en) * | 2015-10-23 | 2016-03-09 | 苏州天马医药集团天吉生物制药有限公司 | Analysis and detection method for UDPC in medicinal preparation containing citicoline sodium |
| CN109709220A (en) * | 2017-10-25 | 2019-05-03 | 中国科学院大连化学物理研究所 | A combined marker and kit and application for diagnosing bladder cancer |
-
2019
- 2019-12-12 CN CN201911299590.1A patent/CN110988178A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5643716A (en) * | 1992-09-08 | 1997-07-01 | Amur Research Corp. | Diagnostic agent and methods for identifying HIV infected individuals and monitoring their therapy |
| CN102138892A (en) * | 2010-02-03 | 2011-08-03 | 广州汉光医药进出口有限公司 | Choline alfoscerate injection preparation as well as preparation method and detection method thereof |
| CN105388225A (en) * | 2015-10-23 | 2016-03-09 | 苏州天马医药集团天吉生物制药有限公司 | Analysis and detection method for UDPC in medicinal preparation containing citicoline sodium |
| CN109709220A (en) * | 2017-10-25 | 2019-05-03 | 中国科学院大连化学物理研究所 | A combined marker and kit and application for diagnosing bladder cancer |
Non-Patent Citations (2)
| Title |
|---|
| 中华人民共和国国家质量监督检验检疫总局: "《中华人民共和国出入境检验检疫行业标准SN/T 3937-2014》", 9 April 2014 * |
| 杜章斌等: "HPLC-ELSD法测定酶解磷脂中甘油磷脂酰胆碱含量的研究", 《粮食与食品工业》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116879428A (en) * | 2023-06-29 | 2023-10-13 | 沈阳金久奇科技有限公司 | High performance liquid analysis method for residual content of phosphorylcholine in L-alpha-phosphorylcholine |
| CN116879428B (en) * | 2023-06-29 | 2024-04-02 | 沈阳金久奇科技有限公司 | High performance liquid analysis method for residual content of phosphorylcholine in L-alpha-phosphorylcholine |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Pesek et al. | Analysis of hydrophilic metabolites by high-performance liquid chromatography–mass spectrometry using a silica hydride-based stationary phase | |
| Callahan et al. | Profiling of polar metabolites in biological extracts using diamond hydride‐based aqueous normal phase chromatography | |
| CN105717207B (en) | Method that is a kind of while detecting a variety of sugar and sugar alcohol in food | |
| JP6223364B2 (en) | An improved analytical method for the analysis and determination of contaminants in dianhydrogalactitol | |
| Hearn et al. | High-pressured liquid chromatography of amino acids, peptides and proteins: V. Separation of thyroidal iodo-amino acids by hydrophilic ion-paired reversed-phase high-performance liquid chromatography | |
| Edwards et al. | Effect of decreasing column inner diameter and use of off-line two-dimensional chromatography on metabolite detection in complex mixtures | |
| Li et al. | Ultrafiltration liquid chromatography combined with high‐speed countercurrent chromatography for screening and isolating potential α‐glucosidase and xanthine oxidase inhibitors from Cortex Phellodendri | |
| CN112526051B (en) | Fmoc-lysine high performance liquid chromatography determination method | |
| US8334110B2 (en) | Detection of blood plasma amygdalin of dissipating blood stasis botanical | |
| CN110988178A (en) | High performance liquid chromatography detection method for phosphorylcholine | |
| Ding et al. | Simultaneous determination of thirteen aminoalcohol-diterpenoid alkaloids in the lateral roots of Aconitum carmichaeli by solid-phase extraction-liquid chromatography–tandem mass spectrometry | |
| Saudan et al. | Carbon isotopic ratio analysis by gas chromatography/combustion/isotope ratio mass spectrometry for the detection of gamma‐hydroxybutyric acid (GHB) administration to humans | |
| t'Kindt et al. | Evaluation of hydrophilic interaction chromatography versus reversed‐phase chromatography in a plant metabolomics perspective | |
| Wang et al. | Screening and quantitative analysis of nine illicit antiallergics in emulsion cosmetics by ultra-high performance liquid chromatography-quadrupole-time-of-flight high-resolution mass spectrometry | |
| Zhao et al. | pH‐Zone‐refining counter‐current chromatography for two new lipo‐alkaloids separated from refined alkaline extraction of Kusnezoff monkshood root | |
| Zheng et al. | Polymer monolith microextraction online coupled to hydrophilic interaction chromatography/mass spectrometry for analysis of β2‐agonist in human urine | |
| Hirota et al. | High-performance liquid chromatographic determination of the enantiomers of carnitine and acetylcarnitine on a chiral stationary phase | |
| McCalley | Quantitative analysis of alkaloids from Cinchona bark by High-Performance Liquid Chromatography | |
| CN103217498A (en) | Method for detecting dicyandiamide in milk powder with LC-MS (liquid chromatography/mass spectrometry) and sample preparation method | |
| CN107014939A (en) | The method that the switching combination of ion exclusion ion exchange column determines the γ hydroxybutyric acids in urine | |
| Yu et al. | Determination of myriocin in natural and cultured Cordyceps cicadae using 9-fluorenylmethyl chloroformate derivatization and high-performance liquid chromatography with UV-detection | |
| Rattenbury et al. | Separation and quantification of urinary di-and polyamines by gas chromatography with electron capture detection | |
| Corlett et al. | High-performance liquid chromatographic determination of the enantiomers of cyclophosphamide in serum | |
| Günther et al. | Semi-preparative liquid-chromatographic separation of all four stereoisomers of α-bisabolol on tribenzoylcellulose | |
| Jagota et al. | Ion chromatography of amylamine and tert.-butylamine in pharmaceuticals |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20200410 |