CN105954430A - Detection method for serum cholesterol oxide - Google Patents

Detection method for serum cholesterol oxide Download PDF

Info

Publication number
CN105954430A
CN105954430A CN201610537108.3A CN201610537108A CN105954430A CN 105954430 A CN105954430 A CN 105954430A CN 201610537108 A CN201610537108 A CN 201610537108A CN 105954430 A CN105954430 A CN 105954430A
Authority
CN
China
Prior art keywords
cholesterol
serum
solution
detection method
oxide
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201610537108.3A
Other languages
Chinese (zh)
Other versions
CN105954430B (en
Inventor
韩晓菲
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Pingdingshan huixinyuan Biotechnology Co.,Ltd.
Original Assignee
Dalian University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Dalian University filed Critical Dalian University
Priority to CN201610537108.3A priority Critical patent/CN105954430B/en
Publication of CN105954430A publication Critical patent/CN105954430A/en
Application granted granted Critical
Publication of CN105954430B publication Critical patent/CN105954430B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/04Preparation or injection of sample to be analysed
    • G01N30/06Preparation
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N2030/022Column chromatography characterised by the kind of separation mechanism
    • G01N2030/027Liquid chromatography
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/04Preparation or injection of sample to be analysed
    • G01N30/06Preparation
    • G01N2030/062Preparation extracting sample from raw material
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/04Preparation or injection of sample to be analysed
    • G01N30/06Preparation
    • G01N2030/067Preparation by reaction, e.g. derivatising the sample

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention provides a detection method for serum cholesterol oxide. The method comprises: sample treatment, saponification, solid-phase extraction, silylation derivatization and HPLC-MS/MS (High Performance Liquid Chromatography-Tandem Mass Spectrometry) analysis. The method can be used for quantitatively detecting automatically oxidized representative products in the serum cholesterol oxide at the same time and has a good application prospect.

Description

A kind of detection method of serum cholesterol oxide
Technical field
The invention belongs to biomarker technical field of analysis and detection, be specifically related to a kind of cholesterol in serum oxygen The detection method of compound.
Background technology
Cholesterol has an important meaning to the physiological function maintaining human normal, but too much cholesterol heap in vivo The long-pending health that can affect again human body self.Numerous studies have shown that: the danger of cardiovascular disease (CVD) because of Element is relevant with oxidative stress increase.Wherein, cholesterol oxidation product COPs is the non-of body oxidative stress status One of mark of Chang Teshu.
In 1966, Brooks etc. detected that oxidized gallbladder is solid first in human atherosclerosis organizes Alcohol, 26-oxycholesterol and 7-ketone group cholesterol.But, people think atherosclerotic important wind always The danger factor is hypercholesterolemia, so concentrate in hypercholesterolemia the most always, right Cholesterol oxidized forms does not draw attention rare report.In recent years, deep along with to low density lipoprotein, LDL (LDL) Entering research, research finds first, and one of risks and assumptions that cardiovascular is the most serious is probably oxidized cholesterol, I.e. cholesterol oxide (cholesterol oxidation products), this compounds is more solid than inoxidized gallbladder Alcohol is more easily caused atherosclerosis, increases the risk of heart disease and stroke outbreak.Cholesterol oxide is a lot Aspect shows its toxic action, such as, can increase carcinogenic probability, Cytotoxic, promote cell to occur Sudden change, makes steroid compound metabolism disorder, substantially can work atherosclerosis, and harmful to human is good for Health.At present, there is the cholesterol oxide in numerous food studied, including egg products, Animal fat Deng.Scientific research result demonstrates, cholesterol and oxidation product thereof are the risk of cardiovascular disease to a certain extent The factor, but cholesterol oxide and the research of cardiovascular disease generation potential mechanism indefinite.
The a series of of A ring, B ring or side chain are caused under the environmental conditions such as cholesterol illumination in vitro, heating Automatic oxidation reaction, thus produce multiple oxidation product.Cholesterol can also be produced by autoxidation mode in vivo Raw.In theory, cholesterol oxide can have more than 70 to plant, and some of them are not autoxidizable generation in cholesterol body Table product, and some other cholesterol oxide is autoxidizable representative product in cholesterol body, can be interior Source property produces or obtains from diet.
Summary of the invention
For prior art thinks little of the detection of cholesterol in serum oxide, the easy external autoxidation of cholesterol The cholesterol oxide that internal autoxidation forms cholesterol oxide, cannot produce carries out classification and Detection, place During reason, the loss of oxygen sterin causes defects such as being difficult to the most quantitatively, serum composition complex jamming is many, the present invention The detection method of a kind of serum cholesterol oxide is provided.
Specifically, the application provides the detection method of a kind of serum cholesterol oxide, it is characterised in that bag Include following steps:
(1) sample treatment: extract serum with chloroform-methanol
(2) saponification: with KOH solution, dehydrated alcohol, the concussion saponification of BHT room temperature;
(3) Solid-Phase Extraction: the solution after saponification crosses SPE solid-phase extraction column;
(4) derivatization: the product of step (3) is derived with Tri-Sil reagent;
(5) LC-MS analysis: liquid chromatograph uses RP-18e cation seperation column, mass spectrum uses API4000 Triple quadrupole mass spectrograph.
The detection method of serum cholesterol oxide of the present invention, it is characterised in that described step (1) sample Process includes: target step in first adding quantitatively in serum before extraction, described quantitative in be designated as non-internal The cholesterol oxidation species that autoxidation produces, such as 19-oxycholesterol etc..
The detection method of serum cholesterol oxide of the present invention, it is characterised in that described step (1) includes Configure chloroform-methanol mixture by the volume ratio of 1:2, serum is diluted 10 with described chloroform-methanol mixed liquor Times, vortex concussion 1min, stand extraction 3min, 4000g and be centrifuged 10min, draw chloroform layer.
The detection method of serum cholesterol oxide of the present invention, it is characterised in that described step (2) saponification Including: add the KOH solution of 60% (w/v), dehydrated alcohol, BHT room temperature concussion saponification 24h, its The addition of middle KOH solution and dehydrated alcohol is 10 times of serum volume, and BHT's is final concentration of 5-50ng/mL。
The detection method of serum cholesterol oxide of the present invention, it is characterised in that described step (3) solid phase Extraction includes: by the solution ether extraction after saponification 3 times, then with distilled water wash 2 times, anhydrous slufuric acid The dried 30 DEG C of rotations of sodium are evaporated off the ether of residual;After adding the n-hexane dissolution of serum volume 5 times, cross SPE Solid-phase extraction column, through normal hexane: diethyl ether (95:5, v/v), normal hexane: diethyl ether (90:10, v/v), After diethyl ether (80:20, v/v) washing, with acetone: methanol (60:20v/v) eluting.
The detection method of serum cholesterol oxide of the present invention, it is characterised in that described step (4) derives Change includes: taking the solution 1ml after saponification, nitrogen dries up, and adds 400 μ LTri-Sil reagent, 60 DEG C of insulations 45min, rotation steaming adds methanol after drying: in the solution of water (3:1, v/v).
The detection method of serum cholesterol oxide of the present invention, it is characterised in that described step (5) liquid matter In combination analysis, HPLC parameter is as follows: solution A is methanol: water (3:1, v/v), and solution B is isopropanol; The solution A of 0-0.5min 60%, the solution B of 40%;0.5-3.5min linear elution, solution B is carried by 40% High to 70%;> 3.5min, the solution B of 100%;Flow velocity is 1ml/min.
The detection method of serum cholesterol oxide of the present invention, it is characterised in that described step (5) liquid matter In combination analysis, mass spectrometry parameters is: source temperature 500, atomization electric current 3 μ A, gas curtain air pressure 25psi, atomization gas Pressure 25psi.
The detection method of serum cholesterol oxide of the present invention, it is characterised in that described serum cholesterol oxygen Compound choosing freely 7 α-OH cholesterol, 7 β-OH cholesterol, 5 β, 6 beta epoxide cholesterol, 5 α, 6 α- The component of epoxycholesterol, 7 ketone group cholesterol, 25-OH cholesterol composition.
What detection method of the present invention reached has the beneficial effect that:
(1) by sample treatment, saponification, Solid-Phase Extraction, cholesterol in serum class component is isolated;Pass through The cholesterol oxide that derivatization makes structure similar more adapts to follow-up HPLC-MS/MS detection, it is to avoid blood The interference of complicated ingredient in clear, improve accuracy of detection.
(2) by the internal non-existent cholesterol oxide of addition as quantitative internal standard, itself and internal cholesterol Oxide physicochemical character, chemical constitution are similar, and the loss in reason step is similar, using it as internal standard throughout It is obtained in that in described extracorporeal treatment step the cholesterol oxide ratio of loss, thus to the cholesterol recorded Oxide content is proofreaded, and improves the accuracy that serum cholesterol oxide is quantitative.
(3) the method for the invention one-time detection i.e. can be simultaneously to generation autoxidizable in cholesterol in serum body Table product, 7 α-OH cholesterol, 7 β-OH cholesterol, 5 β, 6 beta epoxide cholesterol, 5 α, 6 α- Epoxycholesterol, 7 ketone group cholesterol, 25-OH cholesterol carry out detection by quantitative respectively.
Accompanying drawing explanation
Fig. 1 is cholesterol oxide HPLC-MS/MS collection of illustrative plates.
Wherein: 1:7 α-OH cholesterol, 2:7 β-OH cholesterol, 3:7-ketone group cholesterol, 4:5 β, 6 Beta epoxide cholesterol, 5:5 α, 6 α-epoxycholesterol, 6: cholesterol, 7:19-OH cholesterol, 8: 25-OH cholesterol.
Detailed description of the invention
Embodiment 1: the extraction of cholesterol oxide
1. take normal individual in blood bank, serum 1mL to be checked, be added thereto to 19-OH cholesterol to final concentration 10 μ g/mL, add 10mL chloroform-methanol mixture (chloroform: methanol=1:2, v/v).Vortex concussion 1min, Room temperature stands extraction 3min, 4000g and is centrifuged 10min, draws chloroform layer.
2. add the KOH solution of 10mL 60% (w/v), 10mL dehydrated alcohol, 0.5 μ g BHT, room Temperature concussion 24h, makes the oil component saponification in serum, prevents cholesterol oxidation simultaneously.
3., by non-saponification organic facies ether extraction 3 times, then with distilled water wash 2 times, anhydrous sodium sulfate is dried Rear 30 DEG C of rotations are evaporated off the solvent of residual.Use 5mL n-hexane dissolution, cross SPE solid-phase extraction column, Jing Guozheng Hexane: diethyl ether (95:5, v/v), normal hexane: diethyl ether (90:10, v/v), diethyl ether (80:20, v/v) After washing, with acetone: methanol (60:20v/v) eluting.
4. taking the solution 1ml after Solid-Phase Extraction, nitrogen dries up, and adds 400 μ LTri-Sil reagent, 60 DEG C of guarantors Temperature 45min, rotation steaming adds methanol after drying: in the solution of water (3:1, v/v).
Embodiment 2:HPLC-MS/MS is analyzed
Liquid chromatograph uses RP-18e cation seperation column (50 × 4.6cm).275 series injector automatically, 275 Series high-resolution binary pump.HPLC parameter is as follows: solution A is methanol: water (3:1, v/v), solution B For isopropanol;The solution A of 0-0.5min 60%, the solution B of 40%, 0.5-3.5;0.5-3.5min linear elution, Solution B is brought up to 70% by 40%;> 3.5min, the solution B of 100%;Flow velocity is 1ml/min.
Mass spectrum uses API4000 triple quadrupole mass spectrograph to carry out ACPI mass spectrum.Mass Spectrometry Conditions is set to: source temperature Degree 500, atomization electric current 3 μ A, gas curtain air pressure 25psi, atomization air pressure 25psi.Mass Spectrometry Conditions such as table 1:
Table 1: Mass Spectrometry Conditions
Parameter index Numerical value
Detector 385.3/367.3
Solve bunch voltage 70V
Entrance potential 10V
Exit potential 8V
Impact energy 28eV
Above-mentioned HPLC-MS/MS method can complete the detection to six kinds of cholesterol oxides, institute in 6 minutes Obtain collection of illustrative plates as it is shown in figure 1, measure its peak area.Addition according to 19-OH cholesterol obtains after proofreading Obtain various cholesterol oxide content such as table 2 in serum.
Table 2: in serum, various cholesterol oxide content are as follows:
Numbering Cholesterol oxide Ng/mL serum
1 7 α-OH cholesterol 9.5
2 7 β-OH cholesterol 22.0
3 7-ketone group cholesterol 7.3
4 5 β, 6 beta epoxide cholesterol 5.8
5 5 α, 6 α-epoxycholesterol 5.6
6 25-OH cholesterol 8.2
Should be appreciated that the above-described embodiment technology only for the explanation present invention is conceived and feature, its object is to allow Person skilled in the art will appreciate that present disclosure and is carried out, and can not limit this with this Bright protection domain, all equivalence changes made according to spirit of the invention or modification, all should contain at this In the protection domain of invention.

Claims (10)

1. the detection method of a serum cholesterol oxide, it is characterised in that comprise the following steps:
(1) sample treatment: extract serum with chloroform-methanol;
(2) saponification: with KOH solution, dehydrated alcohol, the concussion saponification of BHT room temperature;
(3) Solid-Phase Extraction: the solution after saponification crosses SPE solid-phase extraction column;
(4) derivatization: the product of step (3) is derived with Tri-Sil reagent;
(5) LC-MS analysis: liquid chromatograph uses RP-18e cation seperation column, mass spectrum uses API4000 triple Quadrupole mass spectrometer.
2. the detection method of serum cholesterol oxide as claimed in claim 1, it is characterised in that described step (1) Sample treatment includes: target step in first adding quantitatively in serum before extraction, described quantitative in be designated as The cholesterol oxidation species that non-internal autoxidation produces.
3. the detection method of serum cholesterol oxide as claimed in claim 2, it is characterised in that described interior It is designated as 19-OH cholesterol.
4. the detection method of serum cholesterol oxide as claimed in claim 1, it is characterised in that described step (1) Chloroform-methanol mixture is configured including by the volume ratio of 1:2, serum is dilute with described chloroform-methanol mixed liquor Release 10 times, vortex concussion 1min, stand extraction 3min, 4000g and be centrifuged 10min, draw chloroform layer.
5. the detection method of serum cholesterol oxide as claimed in claim 1, it is characterised in that described step (2) Saponification includes: add the KOH solution of 60% (w/v), dehydrated alcohol, the concussion saponification of BHT room temperature 24h, wherein the addition of KOH solution and dehydrated alcohol is 10 times of serum volume, the end of BHT Concentration is 5-50ng/mL.
6. the detection method of serum cholesterol oxide as claimed in claim 1, it is characterised in that described step (3) Solid-Phase Extraction includes: by the solution ether extraction after saponification 3 times, then by distilled water wash 2 times, nothing The dried 30 DEG C of rotations of aqueous sodium persulfate are evaporated off the solvent of residual;Add the n-hexane dissolution of serum volume 5 times After, cross SPE solid-phase extraction column, through normal hexane: diethyl ether (95:5, v/v), normal hexane: diethyl After the washing of ether (90:10, v/v), diethyl ether (80:20, v/v), with acetone: methanol (60:20v/v) Eluting.
7. the detection method of serum cholesterol oxide as claimed in claim 1, it is characterised in that described step (4) Derivatization includes: taking the solution 1ml after Solid-Phase Extraction, nitrogen dries up, and adds 400 μ LTri-Sil reagent, 60 DEG C of insulation 45min, rotation steaming adds methanol after drying: in the solution of water (3:1, v/v).
8. the detection method of serum cholesterol oxide as claimed in claim 1, it is characterised in that described step (5) During LC-MS is analyzed, HPLC parameter is as follows: solution A is methanol: water (3:1, v/v), and solution B is Isopropanol;The solution A of 0-0.5min 60%, the solution B of 40%;0.5-3.5min linear elution, solution B is brought up to 70% by 40%;> 3.5min, the solution B of 100%;Flow velocity is 1ml/min.
9. the detection method of serum cholesterol oxide as described in claim 1 or 8, it is characterised in that described step (5) in LC-MS analysis, mass spectrometry parameters is: source temperature 500, atomization electric current 3 μ A, gas curtain air pressure 25psi, atomization air pressure 25psi.
10. the detection method of serum cholesterol oxide as described in claim 1-6, it is characterised in that described serum gallbladder The choosing of sterin oxide freely 7 α-OH-cholesterol, 7 β-OH cholesterol, 5 β, 6 beta epoxide cholesterol, One group of component group of 5 α, 6 α-epoxycholesterol, 7 ketone group cholesterol, 25-OH cholesterol composition.
CN201610537108.3A 2016-07-08 2016-07-08 A kind of detection method of serum cholesterol oxide Active CN105954430B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610537108.3A CN105954430B (en) 2016-07-08 2016-07-08 A kind of detection method of serum cholesterol oxide

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610537108.3A CN105954430B (en) 2016-07-08 2016-07-08 A kind of detection method of serum cholesterol oxide

Publications (2)

Publication Number Publication Date
CN105954430A true CN105954430A (en) 2016-09-21
CN105954430B CN105954430B (en) 2018-10-16

Family

ID=56899624

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610537108.3A Active CN105954430B (en) 2016-07-08 2016-07-08 A kind of detection method of serum cholesterol oxide

Country Status (1)

Country Link
CN (1) CN105954430B (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107748099A (en) * 2017-10-20 2018-03-02 西北大学 A kind of self-heating solid phase saponifiable extraction column preparation method for blood sample pre-treatment
CN108776185A (en) * 2018-07-24 2018-11-09 曲阜师范大学 A kind of cholesterol of hydroxyl and its determination method of metabolin
CN109507351A (en) * 2018-12-07 2019-03-22 大连大学 A kind of derivatization method and its analyzing detecting method of oxysterol substance
CN114487185A (en) * 2022-01-21 2022-05-13 武汉迈特维尔生物科技有限公司 Separation and identification method of cholesterol pathway

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101999674A (en) * 2010-11-08 2011-04-06 文利新 Pork cooking method capable of reducing cholesterol oxide

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101999674A (en) * 2010-11-08 2011-04-06 文利新 Pork cooking method capable of reducing cholesterol oxide

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
AKIRA HONDA等: "Highly sensitive quantification of 7a-hydroxy-4-cholesten-3-one in human serum by LC-ESI-MS/MS", 《JOURNAL OF LIPID RESEARCH》 *
ALEX SEVANIAN等: "ANALYSIS OF PLASMA CHOLESTEROL OXIDATION PRODUCTS USING GAS- AND HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY/MASS SPECTROMETRY", 《FREE RADICAL BIOLOGY & MEDICINE》 *
B.H. CHEN等: "Evaluation of the analysis of cholesterol oxides by LC", 《JOURNAL OF CHROMATOGRAPHY A》 *
E. RAZZAZI-FAZELI等: "Determination of cholesterol oxides in processed food using highperformance", 《JOURNAL OF CHROMATOGRAPHY A》 *
MICHALIS S等: "Development of a Reliable Analytical Protocol for the Isolation of Cholesterol Oxidation Products—a Comparison of Different", 《FOOD ANAL. METHODS》 *
T.P. BUSCH等: "Artifact generation and monitoring in analysis of cholesterol oxide products", 《ANALYTICAL BIOCHEMISTRY》 *
柯星等: "酶解-UPLC-MS/MS法同时测定咸鸭蛋黄中的11种胆固醇氧化物", 《分析化学》 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107748099A (en) * 2017-10-20 2018-03-02 西北大学 A kind of self-heating solid phase saponifiable extraction column preparation method for blood sample pre-treatment
CN107748099B (en) * 2017-10-20 2020-09-01 西北大学 Preparation method of self-heating solid-phase saponification extraction column for pretreatment of blood sample
CN108776185A (en) * 2018-07-24 2018-11-09 曲阜师范大学 A kind of cholesterol of hydroxyl and its determination method of metabolin
CN109507351A (en) * 2018-12-07 2019-03-22 大连大学 A kind of derivatization method and its analyzing detecting method of oxysterol substance
CN114487185A (en) * 2022-01-21 2022-05-13 武汉迈特维尔生物科技有限公司 Separation and identification method of cholesterol pathway
CN114487185B (en) * 2022-01-21 2024-05-28 武汉迈特维尔生物科技有限公司 Separation and identification method of cholesterol pathway

Also Published As

Publication number Publication date
CN105954430B (en) 2018-10-16

Similar Documents

Publication Publication Date Title
Alara et al. Optimization of microwave-assisted extraction of flavonoids and antioxidants from Vernonia amygdalina leaf using response surface methodology
CN105954430A (en) Detection method for serum cholesterol oxide
Di Donna et al. Rapid assay of resveratrol in red wine by paper spray tandem mass spectrometry and isotope dilution
Li et al. Screening and characterization of natural antioxidants in four Glycyrrhiza species by liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry
Zheng et al. Simultaneous characterization and quantitation of 11 coumarins in Radix Angelicae Dahuricae by high performance liquid chromatography with electrospray tandem mass spectrometry
Suárez et al. Improved method for identifying and quantifying olive oil phenolic compounds and their metabolites in human plasma by microelution solid-phase extraction plate and liquid chromatography–tandem mass spectrometry
Jin et al. A new strategy for the discovery of epimedium metabolites using high-performance liquid chromatography with high resolution mass spectrometry
Ávila et al. Ultrasound-assisted extraction and silylation prior to gas chromatography–mass spectrometry for the characterization of the triterpenic fraction in olive leaves
Giménez et al. Pentacyclic triterpene in Olea europaea L: A simultaneous determination by high-performance liquid chromatography coupled to mass spectrometry
De Combarieu et al. Identification of Ruscus steroidal saponins by HPLC-MS analysis
Zălaru et al. Polyphenols in Coreopsis tinctoria Nutt. fruits and the plant extracts antioxidant capacity evaluation
Li et al. The strategy for establishment of the multiple reaction monitoring based characteristic chemical profile of triterpenes in Alismatis rhizoma using two combined tandem mass spectrometers
Condrat et al. Qualitative and quantitative analysis of gallic acid in Alchemilla vulgaris, Allium ursinum, Acorus calamus and Solidago virga-aurea by chip-electrospray ionization mass spectrometry and high performance liquid chromatography
Siani et al. Chemical composition of South American Burseraceae non‐volatile oleoresins and preliminary solubility assessment of their commercial blend
El-Haci et al. Phenolics content and antioxidant activity of some organic extracts of endemic medicinal plant Anabasis aretioides Coss. & Moq. from Algerian Sahara
CN106338552A (en) Detection method for 3-MCPD ester in oil product
Liu et al. Metabolism profile of timosaponin B‐II in urine after oral administration to rats by ultrahigh‐performance liquid chromatography/quadrupole‐time‐of‐flight mass spectrometry
Carretero et al. A simplified method for HPLC‐MS analysis of sterols in vegetable oil
CN111249321A (en) Method for simultaneously preparing black currant polyphenol and polysaccharide
Farheen et al. Triterpenoids and triterpenoid saponins from the aerial parts of Fagonia indica Burm
CN107389847A (en) A kind of method of lipid component in quick analysis Bee Pollen
Pérez-Arantegui et al. Colorants and oils in Roman make-ups–an eye witness account
Sánchez-González et al. Liquid chromatography–mass spectrometry determination in plasma of maslinic acid, a bioactive compound from Olea europaea L.
da Silva Antonio et al. Phytochemistry by design: a case study of the chemical composition of Ocotea guianensis optimized extracts focused on untargeted metabolomics analysis
Yu et al. Nontargeted metabolomics and enzyme inhibitory and antioxidant activities for chemical and biological characterization of jujube (Ziziphus jujuba) extracts

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
TR01 Transfer of patent right

Effective date of registration: 20211210

Address after: 467000 Xia Zhuang Cun Bei, Shilong District, Pingdingshan City, Henan Province

Patentee after: Pingdingshan huixinyuan Biotechnology Co.,Ltd.

Address before: 230051 room 1414, building D, Yinhe happiness Plaza, intersection of Luzhou Avenue and Fuzhou Road, Baohe District, Hefei City, Anhui Province

Patentee before: Hefei keyiguo Information Technology Co.,Ltd.

Effective date of registration: 20211210

Address after: 230051 room 1414, building D, Yinhe happiness Plaza, intersection of Luzhou Avenue and Fuzhou Road, Baohe District, Hefei City, Anhui Province

Patentee after: Hefei keyiguo Information Technology Co.,Ltd.

Address before: 116622 No. 10, Xuefu Avenue, Dalian economic and Technological Development Zone, Liaoning

Patentee before: DALIAN University

TR01 Transfer of patent right