CN107144623B - A method of oxidation-resistant active ingredient in online quickly screening and quantitative Chinese medicine - Google Patents
A method of oxidation-resistant active ingredient in online quickly screening and quantitative Chinese medicine Download PDFInfo
- Publication number
- CN107144623B CN107144623B CN201710513206.8A CN201710513206A CN107144623B CN 107144623 B CN107144623 B CN 107144623B CN 201710513206 A CN201710513206 A CN 201710513206A CN 107144623 B CN107144623 B CN 107144623B
- Authority
- CN
- China
- Prior art keywords
- abts
- capillary electrophoresis
- water phase
- chinese medicine
- buffer solution
- 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.)
- Active
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
Abstract
The invention belongs to field of medicaments, and in particular to one kind can screen but also the ABTS of the oxidation-resistant active ingredient in quantitative Chinese medicine+- CE-DAD method.In order to verify the feasibility of this method, for having selected Floium Ginkgo, screens and 9 kinds of oxidation-resistant active ingredients present in it have been determined.The pH value and phosphate concn of buffer, the concentration of lauryl sodium sulfate and beta-cyclodextrin, the reaction conditions such as the content and temperature of acetonitrile and voltage are optimized in experimentation.At identical conditions, ABTS+With the successive sample introduction of Shu Xuening injection sample, then there is the ingredient and ABTS of antioxidant activity+It reacts and by separation detection.Meanwhile the these types of oxidation-resistant active ingredient detected can also be quantified using determining condition, and find the total content of these active constituents and the wired sexual intercourse of total antioxidant activity in sample.Methodology the results show that the present invention be can be applied to reactive compound in screening Chinese medicine in line method, and easy, reliable, low consumption, environmental protection and be easy to automate.
Description
Technical field
The invention belongs to pharmaceutical technology fields, and in particular to the antioxidant activity in Chinese medicine can be screened but also be quantified to one kind
The method of ingredient.
Background technique
In recent years, the primary structure especially lipid and DNA of scientists there's a growing conviction that active oxygen can destroy cell,
Then cause meronecrosis, promote aging even carcinogenic.And antioxidant activity can scavenging activated oxygen, to inhibit disease of old people,
Various cardiovascular and cerebrovascular diseases and neurodegenerative disease, and the generation of the pre- anti-cancer of energy, delay senescence.Natural products, especially
It is to be proved in Chinese medicine containing a large amount of oxidation-resistant active ingredient.Therefore, we should be anti-oxidant as finding using Chinese medicine
The emphasis resource of object.
Ginkgo leaf is a kind of Chinese medicine with antioxidant activity, is widely used in treating various diseases of old people.It is mentioned with ginkgo leaf
Take object be primary raw material made of Shu Xuening injection similarly there is antioxidant activity, due to it is almost without side-effects often by with
In treatment coronary heart disease, the cardiovascular diseases such as angina pectoris and cerebral angiospasm.In view of the safety of medication, it should which control is relaxed comprehensively
The quality of Xuening injection.Although having had many methods for the oxidation-resistant active ingredient in Study of Traditional Chinese Medicine, as LC (S.Lin,
Studies on quality control of Shuxuening injections.TCM Univ.of Fujian
Master's (2014)), LC-MS (J.Liang, A dynamic multiple reaction monitoring method
for the multiple components quantification of complex traditional Chinese
medicine preparations:Niuhuang Shangqing pill as an example,J.Chromatogr.A,
1294 (2013) 58-69) and GC-MS (Y.Hu, W.Kong, X.Yang, L.Xie, J.Wen, M.Yang, GC-MS combined
with chemometric techniques for the quality control and original
discrimination of Curcumae longae rhizome:analysis of essential oils,
J.Sep.Sci.37 (2014) 404-411), but the method for analyzing Shu Xuening injection is seldom.Someone is studied with LC-MS
Floium Ginkgo, but also only surveyed wherein various active constituents content (J.Zaiyou, W.Wenquan, X.Guifang, M.Li,
H.Junling,Comprehensive quality evaluation of Chishao by HPLC,Nutr Hosp.28
(2013)1681-1687).Another there are the problem of be exactly that present existing method cannot be simultaneously from pharmacological activity and content
Two aspects synthetically carry out quality evaluation to Floium Ginkgo.Xiao-Ping Ding, Jin Qi and Yan-Xu Chang et al. are established
A kind of method combination antioxidant activity of online HPLC-DAD-CL to ginkgo leaf carry out quality control (X.P.Ding, Q.Jin,
Y.X.Chang,L.L.Mu,D.N.Zhu,B.Y.Yu,P.S.Xie,T.A.Van Beek,Quality control of
flavonoids in Ginkgo biloba leaves by high-performance liquid chromatography
with diode array detection and on-line radical scavenging activity detection,
J.Chromatogr.A,1216(2009)2204-2210).If using for reference the method to analyze Shu Xuening injection, there can be consumption
The problems such as duration and big organic solvent consumption.
As a kind of modern emerging analysis means, Capillary Electrophoresis (capillary electrophoresis, CE) because
For with easy to operate, analysis time section is efficiently widely used in the advantages of low consumption including Pharmaceutical Analysis (D.S.El, H.D.A.Elhady,d.G.C.G.K.Scriba,Recent advances in capillary
Electrophoretic migration techniques for pharmaceutical analysis (2013-2015),
Electrophoresis, 37 (2016) 1591-1608), many sides of the quality control of bioactivity screening and natural products
Face.For example, Dubber and Kanfer application inverts flow direction-micellar electrokinetic chromatography to determine the flavonoids and terpene lactones in ginkgo leaf
Class compound (M.J.Dubber, Application of reverse-flow micellar electrokinetic
chromatography for the simultaneous determination of flavonols and terpene
Trilactones in Ginkgo biloba dosage forms, J.Chromatogr.A, 1122 (2006) 266-274),
The method can reduce the use of organic reagent but not can reflect the antioxidant activity of ginkgo leaf.In addition to this, Yan-Xu
Chang etc. establishes a kind of online DPPH-CE-DAD (DAD: diode array detector) method comprehensively to evaluate Redujing Granules
Injection, this method DPPH and CE are combined the total antioxidant activity for having measured Reduning injection and every kind have it is anti-
The ingredient of oxidation activity content (Y.X.Chang, J.Liu, Y.Bai, J.Li, E.W.Liu, J.He, X.C.Jiao,
Z.Z.Wang,X.M.Gao,B.L.Zhang,The activity-integrated method for quality
assessment of reduning injection by on-line DPPH-CE-DAD,PloS one,9(2014)
e106254).This method can solve the problem of conventional method cannot reflect sample activity, but in this method isolating active at
The condition difference divided and survey total antioxidant activity.In view of above-mentioned various problems, it is necessary to establish it is a kind of simple, quickly, can
The method leaned on comprehensively evaluates the pharmacological action and quality of traditional Chinese medicine injection.
It is the method for being used primarily for measurement Chinese medicine and its preparation antioxidant activity, most of the method that experiment, which is added, in free radical
It is realized by microplate reader, oxygen radical mainly uses 1,1- diphenyl -2- trinitrophenyl-hydrazine (DPPH) and 2, and 2- joins nitrogen-two
(3- ethyl-benzothiazole -6- sulfonic acid) di-ammonium salts (ABTS+).Using ABTS+To measure the antioxidant activity of Chinese medicine or its preparation
Method be typically all combine ultraviolet specrophotometer off-line mode (S.H.Hwang, Z.Wang, H.N.Yoon, S.S.Lim,
Xanthium strumarium as an Inhibitor ofα-Glucosidase,Protein Tyrosine
Phosphatase 1β,Protein Glycation and ABTS+for Diabetic and Its Complication,
Molecules,21(2016)；X.Wang,X.Li,H.Li,Reassessment of Antioxidant Activity of
Baicalein in vitro, Asian J.Pharm.Biomed.Research, 1 (2011) 186-194), it is evident that it is this
The operation of method needs progress step by step to react offline, separation detection, more complicated.In order to simplify operation, online LC-ABTS+Method measurement Antioxidant Capacity of Chinese Herb be successively reported (K.M.Kalili, S.S.De, H.T.Van, F.Lynen, V.A.De,
Comprehensive two-dimensional liquid chromatography coupled to the ABTS
radical scavenging assay:A powerful method for the analysis of phenolic
antioxidants,Anal.and Bioanal.Chem.406(2014)4233-4242；K.J.Lee,N.Y.Song,
Y.C.Oh,W.K.Cho,J.Y.Ma,Isolation and Bioactivity Analysis of Ethyl Acetate
Extract from Acer tegmentosum Using In Vitro Assay and On-Line Screening
HPLC-ABTS(+)System,J.Anal.Methods in Chem.2014(2014)150509；A.A.
Antioxidant components of Viburnum opulus L.determined by on-line HPLC-UV-
ABTS radical scavenging and LC-UV-ESI-MS methods,Food Chem.175(2015)106-114)。
But since analysis instrument is still liquid phase, the usage amount of organic reagent is still very big in whole experiment process.In order to solve exist
The problem of, we by integrating CE and ABTS online+A kind of method that oxidation-resistant active ingredient is quickly screened from Chinese medicine is established,
And it selects for Floium Ginkgo and is specifically described this method.
Summary of the invention
The purpose of the present invention is to provide the oxidation-resistant active ingredients in a kind of online quickly screening or/and quantitative Chinese medicine
ABTS+- CE-DAD method, the method are applicable to separation, screening Chinese medicine and its oxidation-resistant active ingredient in injection and to samples
Product carry out quality control.
To achieve the goals above, the invention adopts the following technical scheme:
A kind of ABTS of online quickly screening or/and the oxidation-resistant active ingredient in quantitative Chinese medicine+- CE-DAD method, institute
Stating ABTS is that 2,2- joins nitrogen-two (3- ethyl-benzothiazole -6- sulfonic acid) di-ammonium salts, and the CE is capillary electrophoresis, described
DAD is diode array detector, the composition of the Capillary Electrophoresis water phase buffer solution of the method be for 10mM~30mM phosphate,
The beta-cyclodextrin of 0mM~10mM, the second that the lauryl sodium sulfate and percent by volume of 20mM~60mM is 2.5%~10%
Nitrile, wherein the phosphate is to pass through NaH2PO4And Na2HPO4It is mixed to get；The Capillary Electrophoresis water phase buffer solution
PH value is 6.5~7.5；In whole experiment process, voltage is 18kV~22kV, and the temperature of cartridge is controlled at 19 DEG C~25 DEG C；In
The Detection wavelength of medicine sample is 360nm, ABTS+Detection wavelength be 405nm.
In some embodiments, the phosphate in the composition of the Capillary Electrophoresis water phase buffer solution is selected from 10mM
~20mM or 20mM~30mM, preferably 20mM.
In some embodiments, the phosphate in the composition of the Capillary Electrophoresis water phase buffer solution is to pass through
The NaH of mole2PO4And Na2HPO4It is mixed to get.
In some embodiments, the beta-cyclodextrin in the composition of the Capillary Electrophoresis water phase buffer solution is selected from
0mM~5mM or 5mM~10mM, preferably 5mM.
In some embodiments, the lauryl sodium sulfate in the composition of the Capillary Electrophoresis water phase buffer solution
Selected from 20mM~40mM or 40mM~60mM, preferably 40mM.
In some embodiments, the acetonitrile percent by volume in the composition of the Capillary Electrophoresis water phase buffer solution
Selected from 2.5%~5%, 5%~7.5%, 7.5%~10%, preferably 7.5.
In some embodiments, the pH value of the Capillary Electrophoresis water phase buffer solution be selected from 6.5~7.0 or 7.0~
7.5, preferably 7.0.
In some embodiments, the voltage is selected from 18kV~20kV or 20kV~22kV, preferably 20kV；The cartridge
Temperature control be selected from 19 DEG C~22 DEG C or 22 DEG C~25 DEG C, preferably 22 DEG C.
In some embodiments, the present invention screens or/and measures online the Capillary Electrophoresis water of oxidation-resistant active ingredient
The composition of phase buffer is for 20mM phosphate, the beta-cyclodextrin of 50mM, the lauryl sodium sulfate and percent by volume of 40mM
For 7.5% acetonitrile, wherein the phosphate is by equimolar amounts NaH2PO4And Na2HPO4It is mixed to get；The hair
The pH value of cons electrophoresis water phase buffer solution is 7.0；In whole experiment process, voltage is 20kV, and the temperature of cartridge is controlled at 22 DEG C；
The Detection wavelength of traditional Chinese medicine sample is 360nm, ABTS+Detection wavelength be 405nm；Traditional Chinese medicine sample and ABTS+Sample introduction be all
50mbar sample introduction under pressure 5s.
In some embodiments, the Chinese medicine is Shu Xuening injection.
The beneficial effects of the present invention are:
(1) the online ABTS that the present invention establishes+- CE-DAD method can be used to the screening from traditional Chinese medicine injection and quantitatively resist
Oxidative active ingredients, and quality control is carried out to corresponding traditional Chinese medicine injection.Be not found in pertinent literature library it is this
Line ABTS+- CE-DAD method analyzes traditional Chinese medicine injection, and is also for the first time by ABTS+It is combined online with CE.
(2) the method for the present invention is used, can quickly measure the total antioxidant activity of Shu Xuening injection simultaneously under identical conditions
And it is successfully determined the ingredient with antioxidant activity, the content of these subsequent active constituents can also be measured.
(3) CE is not only a separating tool in this method and is the container that can be reacted.In this hair
In bright research process, to the online ABTS+As a result the methodology validation of-CE-DAD method proves the activity that this task is established
Integration method is applicable to separation, screening Chinese medicine and its oxidation-resistant active ingredient in injection and carries out quality control to sample
System.
Attached drawing
Fig. 1: each parameter is to the transit time of 14 compound peaks (including ABTS+) and the effect of separating degree
Wherein: Fig. 1 (A) is the effect of Capillary Electrophoresis water phase buffer solution pH value；
Fig. 1 (B) is the effect of buffer salinity；
Fig. 1 (C) is the effect of lauryl sodium sulfate (SDS) concentration；
Fig. 1 (D) is the effect of beta-cyclodextrin (β-CD) concentration；
Fig. 1 (E) is the effect of acetonitrile (ACN) concentration；
Fig. 1 (F) is the effect of voltage；
Fig. 1 (G) is the effect of temperature；
Wherein abscissa in figure 1 is butterfly legumin, 2 be rutin, 3 be isoquercitrin, 4 be Quercetin -3-O- glucosyl group
(1-2) rhamnoside, 5 be kaempferol-3-O-rutinoside, 6 be narcissin, 7 be Kaempferol -7-O- β-D-Glucose glycosides, 8 be
Cosmosiin, 9 be 3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnopyranosyl } Quercetin, 10 be
Quercetin, 11 be 3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnopyranosyl } Kaempferol, 12 be mountain
How phenol, 13 be Isorhamnetin.
Fig. 2: " ABTS+" and " ABTS+The electrophorogram of solution on-line mixing Shu Xuening injection "
Wherein: that high is " ABTS+" electrophorogram, that low is " ABTS+ and Shu Xuening injection on-line mixing after "
Electrophorogram.
Fig. 3: Floium Ginkgo electrophorogram
Wherein: being the electrophorogram of Floium Ginkgo Yu water on-line mixing above, here is Floium Ginkgo and ABTS+On-line mixing
Electrophorogram.
Fig. 4: ten three hybrid standard product of compound and the electrophorogram of Shu Xuening injection
Wherein: being the electrophorogram of standard items mixture above, here is the electrophorogram of Shu Xuening injection.
Fig. 5: the relationship of Shu Xuening injection total content and total antioxidant activity.
Specific embodiment
Following is that the present invention is further explained in conjunction with specific embodiments.But these embodiments be only limitted to illustrate the present invention without
It is for limiting the scope of the invention.The experimental method of specific experiment condition is not specified in the following example, usually according to routine
Condition.
Embodiment:
1 instrument and material
1.1 instrument
Capillary electrophoresis: Agilent 7100, diode array detector DAD；Non- coating quartz capillary column: Hebei
The sharp Feng chromatography device Co., Ltd of Yongnian；Superpure water machine: Millipore company, II type of Mill-Q；Supercentrifuge: the U.S.
Sigma company, 3K15；Ultrasonic cleaner: Kunshan Ultrasonic Instruments Co., Ltd., KQ-250E；A ten thousandth balance: Germany
Sartorius company, BP121S；Ten a ten thousandth balances: Mettler Toledo company, Switzerland, AX205；Vortex mixer:
Shanghai Hu Xi analysis instrument factory, XW-80A；Ultrafiltration membrane: Tianjin Jin Teng experimental facilities Co., Ltd
1.2 reagent
Acetonitrile: chromatographically pure, Merck KGaA company；Methanol: chromatographically pure, Merck KGaA company；Ultrapure water: Millipore is super
Purification of water system is made；Sodium dihydrogen phosphate: pure, Tianjin Kermel Chemical Reagent Co., Ltd. is analyzed；Disodium hydrogen phosphate: point
Analyse pure, Tianjin Kermel Chemical Reagent Co., Ltd.；Sodium hydroxide: pure, the limited public affairs of Tianjin Ke Miou chemical reagent are analyzed
Department；Lauryl sodium sulfate (SDS): pure, Beijing Suo Laibao Science and Technology Ltd is analyzed；Beta-cyclodextrin (β-CD): analysis is pure, north
Jing Suolaibao Science and Technology Ltd.；2,2- joins nitrogen-two (3- ethyl-benzothiazole -6- sulfonic acid) di-ammonium salts (ABTS): the U.S.
Sigma-aldrich company；Potassium peroxydisulfate: pure, Tianjin Kermel Chemical Reagent Co., Ltd. is analyzed.
1.3 standard items
Butterfly legumin, rutin, isoquercitrin, Quercetin -3-o- glucosyl group (1-2) rhamnoside, Kaempferol -3-o- rue
Glucosides, narcissin, Kaempferol -7-o- β-D-Glucose glycosides, cosmosiin, { [6-O- is (to the trans- tonka-bean of hydroxyl-by 2-O- by 3-O-
Acyl)-glucosyl group]-rhamnopyranosyl } Quercetin, 3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-sandlwood
Glycosyl } Kaempferol is purchased from Chengdu Man Site Biotechnology Co., Ltd, and purity is greater than 98.0%.
1.4 sample
20 batches of Shu Xuening injections are provided by martial prowess medicine company.All samples are through Tianjin University Of Traditional Chinese Medicine Chang Yanxu researcher
Audit confirmation, deposits in Chinese medicine study institute, Tianjin University Of Traditional Chinese Medicine.
2 experiment conditions
2.1 instrument condition
All experiments are real in the Agilent capillary electrophoresis equipped with UV detector (Waldbronn, Germany)
Existing, instrumentation and data analysis are realized on Agilent chem workstation.The vitreous silica that CE separation is 50 μm in internal diameter
It is carried out in capillary, overall length 60.2cm (effective length 52cm) (Hebei, sharp Feng).New capillary should be under 50mbar pressure
Successively 10min is rushed with 1.0M NaOH, 0.1M NaOH and deionized water to activate inner wall.Capillary should be before each run
2min is rushed with 0.1M NaOH under 50mbar pressure, respective background electrolyte solution rushes 3min.After last time separates daily,
Capillary should rush respectively 10min with 0.1M NaOH and deionized water under 50mbar pressure.In order to guarantee that it is good that transit time has
Good repeatability, background buffer in bottle every should be run more to be renewed twice.
2.2 experiment condition
The composition of online screening antioxidant activity and the Capillary Electrophoresis water phase buffer solution for measuring antioxidant activity is 20mM
Phosphate, the beta-cyclodextrin of 5mM, the lauryl sodium sulfate of 40mM and 7.5% acetonitrile；Capillary Electrophoresis water phase buffer solution
PH value be 7.Wherein, phosphate is NaH by equimolar amounts in Capillary Electrophoresis water phase buffer solution2PO4And Na2HPO4It is mixed
What conjunction obtained.In whole experiment process, voltage application is 20kV, and the temperature of cartridge is controlled at 22 DEG C.All sample feedings
It is all in 50mbar sample introduction under pressure 5s.Test sample and ABTS+Wavelength be 360nm and 405nm respectively.
The preparation of 3 standard items, quality-control sample and sample
All flavonoids standard items use methanol to be dissolved into 2mg/mL as stock solution, and then stand-by storage liquid mixes simultaneously again
Be diluted to be using 50% methanol as each constituent concentration of solvent 0.1mg/mL mixed standard solution.ABTS and potassium peroxydisulfate difference
Being dissolved into concentration with deionized water is 9mg/mL and 1.516mg/mL, then by ABTS and potassium persulfate solution with the ratio of 1:2
20-24h is kept in dark place at room temperature to aoxidize sufficiently up to ABTS in mixing+.Obtained ABTS+Free-atom aqueous solution is protected from light in room temperature
Under the conditions of can stablize preservation 3-4 days, with being diluted to 1.5mg/mL using deionized water before.All mark product with all protecting before
There are 4 DEG C, in addition to ABTS+All solution outside free-atom aqueous solution are all filtered with 0.22 μm of nylon filter using preceding.
Butterfly legumin, rutin, isoquercitrin, narcissin, Quercetin -3-o- glucosyl group (1-2) rhamnoside, Kaempferol -
3-o- rutinoside, Kaempferol -7-o- β-D-Glucose glycosides, cosmosiin, { [6-O- is (to the trans- tonka-bean of hydroxyl-by 2-O- by 3-O-
Acyl)-glucosyl group]-rhamnopyranosyl } Quercetin, 3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-sandlwood
Glycosyl } quality-control sample of Kaempferol includes basic, normal, high three concentration, and which is the mixed mark by diluting certain concentration
Matrix obtain to respective concentration and final is 50% methanol.
The foundation of 4 methods
Firstly, the pH value and phosphate, beta-cyclodextrin of optimization Capillary Electrophoresis water phase buffer solution, the concentration of SDS and acetonitrile
And the deposition conditions such as voltage and temperature come detect separation Shu Xuening injection in ingredient and ABTS+.By comparing online mixed
Close sample and water and sample solution and ABTS+The electrophorogram of solution screens oxidation-resistant active ingredient, has antioxidant activity
Reduced in the latter case at sub-summit.By comparing ABTS+On-line mixing water is obtained with the sample after dilution respectively
Electrophoretogram determines total antioxidant activity.In experimentation, sample and ABTS+In 50mbar sample introduction under pressure 5s.
About methodology, precision, linear, the stability and the rate of recovery for 24 hours of this method have been separately verified.It is linearly with six
For a point come what is established, precision and for 24 hours stability are the (n realized by measuring the quality-control sample of basic, normal, high three concentration
=6).The rate of recovery of 9 ingredients is realized by the way that the mixed mark solution of isoconcentration is added in the sample of known concentration.It is above-mentioned
The sample of primary sample and mark-on is detected with the condition optimized, and the rate of recovery is calculated with following formula: the rate of recovery
(%)=(the original amount of measured amount -)/additional amount × 100%.
5 results and discussion
The effect of 5.1pH value
For transit time and separating degree, the pH value of Capillary Electrophoresis water phase buffer solution is vital, because
The change of pH will lead to the migration rate of electrolyte and electroosmotic flow changes.In order to determine optimal pH value, in 20mM phosphoric acid
Under conditions of salt, 5mM β-CD, 60mM SDS and 7.5% acetonitrile, optimizing pH is 6.5,7.0,7.5.As institute in Fig. 1 (A)
Show, total transit time is gradually decreased as pH rises to 7.5 from 6.5, and walking always from line, the chemical combination when pH is 6.5
The separating degree of object is not satisfactory.Total transit time is little in the difference of pH7 and 7.5, but before comparing the Trendline of pH7 and 7.5
Several points can be seen that preceding several compounds the pH separating degree for being 7.5 and not as good as pH be 7 it is good.Therefore, select pH for 7 into
The further research of row.
The effect of 5.2 buffer salinities
FOR ALL WE KNOW, buffer salt is indispensable a part in Capillary Electrophoresis water phase buffer solution, because in electricity
It is the substance for generating electric current under the action of pressure.The buffer salt of various concentration can make in sample in Capillary Electrophoresis water phase buffer solution
Ingredient generate different migratory behaviours, therefore, the concentration of buffer salt needs to optimize.The phosphate of pH7 is investigated respectively herein
The concentration of 10mM, 20mM and 30mM.Fig. 1 (B) shows the increase with phosphate concn, and total transit time can also extend.?
When 10mM and 30mM, ABTS+Cannot be completely separable with rutin, and separating degree makes moderate progress when 20mM.Therefore 20mM is most suitable
Phosphate concn.
The effect of 5.3 lauryl sodium sulfate (SDS) concentration
In micellar electrokinetic capillary chromatography (MEKC), SDS is a kind of common additive, and concentration is just more than its CMC value
Micella can be formed, therefore, good separating degree is highly desirable the concentration of optimization SDS in order to obtain.We have studied different dense
The effect that the SDS (20mM, 40mM and 60mM) of degree generates the Multiple components separation in Floium Ginkgo.Pass through Fig. 1 (C), Ke Yiguan
Observe from 20mM to 60mM that analysis time is increasingly longer, and the separating degree between most of peak is also increasing.In 20mM,
The ordinate of each point does not have too many differences on figure, that is to say, that the separating degree of each compound is less desirable.However, in 60mM
When narcissin and Kaempferol -7-O- glucoside separating degree compared with 40mM when declined.Finally, 40mM is by as optimal dense
Degree carries out the experiment of next step.
The effect of 5.4 beta-cyclodextrins (β-CD) concentration
Cyclodextrin has conical cavity special construction, is that one kind is added in Capillary Electrophoresis water phase buffer solution and can pass through
The substance for changing the migratory behaviour of similar molecules to separate them.In order to improve peak shape and improve the separation of part of compounds
Degree, β-CD have been added into Capillary Electrophoresis water phase buffer solution.Fixed other conditions are constant, investigated 0mM, 5mM and 10mM
Effect of the β-CD of concentration to ingredient each in Shu Xuening injection.With the increase of β-CD concentration, analysis time shorten and
Peak shape narrows.However, can be seen that the increase with β-CD concentration, separating degree by comparing three Trendline in Fig. 1 (D)
Also declined.After considering various factors, it is believed that the β-CD of 5mM is the best concentration of isolating target compound.
The effect of 5.5 acetonitriles (ACN) concentration
Most of Capillary Electrophoresis water phase buffer solution is water-soluble, but the compound in quite a few sample exists
Dissolubility in water is simultaneously bad.ACN is as a kind of common organic additive, after being added in Capillary Electrophoresis water phase buffer solution
It can prevent analyte from generating precipitating in electrophoresis process.ACN does not have UV absorption, and viscosity is not also high, therefore will not be to detection
Compound generate interference.We select 2.5%, 5%, 7.5% and 10% ACN to be investigated.We can from Fig. 1 (E)
To find out, at 2.5%, the separating degree of the ingredient in sample is not satisfactory, in addition, the increase although as ACN concentration separates
Degree makes moderate progress, but analysis time also extends.And peak shape is preferable when 7.5%, therefore, 7.5% be confirmed as it is optimal
Condition.
The effect of 5.6 voltages and temperature
In addition to factor mentioned above, voltage and temperature also can separating degree to analyte and transit time have an impact.
Voltage provides driving force, and therefore, disengaging time can be reduced by improving voltage, but excessively high voltage can also generate more coke
It has burning ears.Temperature mainly influences the viscosity of Capillary Electrophoresis water phase buffer solution.Voltage (18kV, 20kV and 22kV) and cartridge are investigated
Isolated influence (respectively see Fig. 1 (F) Fig. 1 (G)) of the temperature (19 DEG C, 22 DEG C and 25 DEG C) to compound, when voltage 20kV and temperature
When spending 22 DEG C, separating degree is best.
In conclusion what is finally determined is to the optimal conditions of each target compound in separation detection Floium Ginkgo: 20mM
Phosphate (pH7.5), 40mM SDS, 5mM β-CD, 7.5%ACN, voltage 22kV, 22 DEG C of temperature.
5.7 methodology validation
Butterfly legumin, rutin, isoquercitrin, Quercetin -3-o- glucosyl group (1-2) rhamnoside, Kaempferol -3-o- rue
Glucosides, narcissin, Kaempferol -7-o- β-D-Glucose glycosides, cosmosiin, { [6-O- is (to the trans- tonka-bean of hydroxyl-by 2-O- by 3-O-
Acyl)-glucosyl group]-rhamnopyranosyl } Quercetin, 3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-sandlwood
Glycosyl } Kaempferol standard curve and the range of linearity it is as listed in Table 1.LOD and LOQ is by 3 times of signal-to-noise ratio and 10 respectively
Times signal-to-noise ratio computation is got, and the range of the LOD of these compounds is in 0.3 μ gmL-1-1.8μg·mL-1Between, the range of LOQ
It is 1 μ gmL-1-6μg·mL-1。
1. linear equation of table, the range of linearity, LODs, the rate of recovery (n=6) of LOQs and nine compound
Rate of recovery test result is listed in table 1, and 9 average recovery rates (n=6) with antioxidative activity compound exist
Between 96.9-104.5%, and RSD% value < 4.49%.In conclusion this method is used to separate and quantify in traditional Chinese medicine injection
Oxidation-resistant active ingredient be it is stable, accurately.
In order to determine the foundation method reliability, investigated precision (including in a few days and day to day precision) and accurate
Degree, the results are shown in Table 2.All compounds include ABTS+Precision RSD % value 7.5% hereinafter, accuracy exists
Between 97.1-106.3%.The result shows that this method is reliable, it is accurately, reproducible.
In order to evaluate the stability of each ingredient in the analysis process in sample, the factor that another needs is investigated is
Stability for 24 hours.The results are shown in Table 2, it can be seen that all compounds be stored at 4 DEG C for 24 hours after three concentration levels
Retention is respectively within the scope of 95.9-101.7%, 97.3-100.3% and 99.6-100.9% and ABTS+Retention be
99.6%, including ABTS+The RSD value of all compounds inside is respectively less than 6.48%.The result shows that the target chemical combination in sample
Object is stable in storage and analytic process.
In table 2. days, accuracy in the daytime and precision, nine compounds and ABTS+(n=6) stability
The total antioxidant activity of 5.8 on-line determination samples
Using this foundation in line method, the total antioxidant activity of sample can be also measured.Shu Xuening injection is due to clear
Except the ability of sample free radical is stronger, thus sample with ABTS+32 times are diluted with deionized water before online reaction.It uses respectively
ABTS+It is reacted online with the deionized water of sample and equivalent after dilution, ABTS in the case of two kinds of measurement+Peak area (n=3).
Compare obtained electrophorogram (as shown in Figure 2), it can be seen that ABTS+Postpeak is reacted online with the Shu Xuening injection diluted
Area decreases.Relative inhibition is calculated using following formula: inhibiting rate (%)=(P0-P1)/P0× 100%), wherein P0
It is ABTS+With the area of deionized water on-line mixing, P1It is ABTS+With the face after the Shu Xuening injection on-line mixing after dilution
Product.The results are shown in Table 3 for the total antioxidant activity inhibiting rate of 20 batches of Shu Xuening injections, and Shu Xuening injection has anti-as the result is shown
Oxidation activity, and somewhat difference between the Shu Xuening injection of different batches.Therefore, the online ABTS of the foundation+-CE-
DAD method can be used to measure the total antioxidant activity for the traditional Chinese medicine injection for having complex matrices.
The content and total inhibiting rate (n=3) of nine compounds in 3. sample of table
Oxidation-resistant active ingredient in 5.9 online screening samples
In order to screen the oxidation-resistant active ingredient in Shu Xuening injection, ABTS+It (1.5mg/mL) and dilutes twice
Shu Xuening injection successive sample introduction 5s (n=3) under 50mbar pressure.By with not and ABTS+The sample electrophoresis figure pair of reaction
Than nine ingredients (as shown in Figure 3) with antioxidant activity have been filtered out from Floium Ginkgo sample according to the reduction of peak height.With
Afterwards, by sample electrophoresis figure and reference substance compare it is (as shown in Figure 4) known to this nine compounds be respectively butterfly legumin, reed
Fourth, isoquercitrin, Quercetin -3-O- glucosyl group (1-2) rhamnoside, kaempferol-3-O-rutinoside, Kaempferol -7-O-
β-D-Glucose glycosides, cosmosiin, 3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnopyranosyl }
Quercetin, 3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnopyranosyl } Kaempferol.Pass through above-mentioned knot
Fruit judges it is found that the these types of ingredient filtered out is the main antioxidant active constituent in Shu Xuening injection.
The assay of oxidation-resistant active ingredient in 5.10 samples
Measure the content of nine oxidation-resistant active ingredients in 20 batches of samples respectively with the method for foundation, the results are shown in Table 3.
As can be seen from the table, butterfly legumin, rutin, isoquercitrin, Quercetin -3-o- glucosyl group (1-2) rhamnoside, Kaempferol -
3-o- rutinoside, narcissin, Kaempferol -7-o- β-D-Glucose glycosides, cosmosiin, { [6-O- is (to hydroxyl-by 2-O- by 3-O-
Trans- coumaric acyl)-glucosyl group]-rhamnopyranosyl } Quercetin, 3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucose
Base]-rhamnopyranosyl } content range of Kaempferol is 0.0517-0.0669mgmL respectively-1, 0.0586-0.0767mgmL-1, 0.0122-0.0181mgmL-1, 0.0357-0.0578mgmL-1, 0.0579-0.0925mgmL-1, 0.0070-
0.0103mg·mL-1, 0.0305-0.0423mgmL-1, 0.0655-0.0869mgmL-1And 0.0452-0.0831mg
mL-1.The average value of the total content of 20 batches of samples is 0.453mgmL-1And RSD% is 7.21%, illustrates 20 batches of sample rooms
There is fraction of difference.This phenomenon has the result of some differences corresponding with the antioxidant activity of above-mentioned different batches sample room.
In order to evaluate the quality of Shu Xuening injection, the oxidation-resistant active ingredient that this experiment is investigated different samples always contains
Relationship between amount and antioxidant activity.As shown in figure 5, having between total antioxidant activity and nine oxidation-resistant active ingredient total contents
Apparent linear relationship (R=0.9456).The result shows that the antioxidant activity of Shu Xuening injection depends on these activity
The content of ingredient, therefore the quality of Shu Xuening injection can be evaluated by measuring the content of these ingredients.That is, this
The secondary online ABTS established by selecting nine oxidation-resistant active ingredients as target compound+- CE-DAD method can be used
To evaluate the quality of different batches Shu Xuening injection.
6 conclusions
It is well known that ABTS+Be another other than DPPH be often used in the oxygen of measurement sample antioxidant activity from
By base, but it is used in line ABTS+- CE-DAD method has not been reported to screen oxidation-resistant active ingredient.It is first in this task
It is secondary by ABTS+Foundation screening antioxidative activity compound is combined with CE-DAD instrument, wherein selecting Shu Xuening injection
Method to verify the foundation as an example.Nine antioxidant activities are screened from Shu Xuening injection using this method
Object is closed, and successfully they are quantified.It can be seen that with the online ABTS of the foundation+- CE-DAD method is from complicated base
It is simple, quick, accurate, reliable and environmentally friendly when screening oxidation-resistant active ingredient in the Chinese medicine of matter or its preparation and being quantified,
And the sample room antioxidant activity ability of different batches can be evaluated.It, should compared with the method for tradition evaluation Chinese materia medica preparation quality
Online ABTS+- CE-DAD method combines sample pharmacological action to evaluate its quality, and organic examination in the entire experiment process
The consumption of agent is all lower.To sum up, the active integration method of the foundation is a kind of novel, reliable and powerful point
Analysis method, can not only screen and quantify the oxidation-resistant active ingredient in Chinese medicine and its injection, and can to analyzed sample into
The control of row quality.In following research, it is contemplated that by this method in conjunction with mass spectrum, for determining work unknown in natural products
Property compound, or with improve sensitivity technology in conjunction with come detect the low content in screening sample even trace compound.
It should be understood that after reading the above teachings of the present invention, those skilled in the art can make the present invention
Various changes or modification, these equivalent forms also fall within the scope of the appended claims of the present application.
Claims (15)
1. the ABTS of a kind of online quickly screening or/and the oxidation-resistant active ingredient in quantitative Chinese medicine Shu Xuening injection+-CE-
DAD method, the oxidation-resistant active ingredient are butterfly legumin, rutin, isoquercitrin, Quercetin -3-O- glucosyl group (1-2) sandlwood
Glucosides, kaempferol-3-O-rutinoside, Kaempferol -7-O- β-D-Glucose glycosides, cosmosiin, { [6-O- is (right by 2-O- by 3-O-
The trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnopyranosyl } Quercetin, { [6-O- (to the trans- coumaric acyl of hydroxyl -)-Portugal 2-O- 3-O-
Grape glycosyl]-rhamnopyranosyl } Kaempferol, the ABTS is that 2,2- joins nitrogen-two (3- ethyl-benzothiazole -6- sulfonic acid) di-ammonium salts,
The CE is capillary electrophoresis, and the DAD is diode array detector, which is characterized in that online screening or/and measurement are anti-
The composition of the Capillary Electrophoresis water phase buffer solution of oxidative active ingredients is for 10mM~30mM phosphate, β-ring of 0mM~10mM
Dextrin, the acetonitrile that the lauryl sodium sulfate and percent by volume of 20mM~60mM is 2.5%~10%, wherein the phosphate
It is to pass through NaH2PO4And Na2HPO4It is mixed to get；The pH value of the Capillary Electrophoresis water phase buffer solution is 6.5~7.5；It is whole
In a experimentation, voltage is 18kV~22kV, and the temperature of cartridge is controlled at 19 DEG C~25 DEG C；The Detection wavelength of traditional Chinese medicine sample is
360nm, ABTS+Detection wavelength be 405nm.
2. ABTS according to claim 1+- CE-DAD method, which is characterized in that the Capillary Electrophoresis water phase buffer solution
Composition in the phosphate be selected from 10mM~20mM or 20mM~30mM.
3. ABTS according to claim 2+- CE-DAD method, which is characterized in that the Capillary Electrophoresis water phase buffer solution
Composition in the phosphate be 20mM.
4. ABTS according to claim 1+- CE-DAD method, which is characterized in that the Capillary Electrophoresis water phase buffer solution
Composition in the phosphate be NaH by equimolar amounts2PO4And Na2HPO4It is mixed to get.
5. according to claim 1, ABTS described in 2 or 3+- CE-DAD method, which is characterized in that the Capillary Electrophoresis water phase
The beta-cyclodextrin in the composition of buffer is selected from 0mM~5mM or 5mM~10mM.
6. the ABTS according to claim 5+- CE-DAD method, which is characterized in that the Capillary Electrophoresis water phase buffer solution
Composition in the beta-cyclodextrin be 5mM.
7. ABTS according to claim 1,2 or 3+- CE-DAD method, which is characterized in that the Capillary Electrophoresis water phase is slow
The lauryl sodium sulfate in the composition of fliud flushing is selected from 20mM~40mM or 40mM~60mM.
8. ABTS according to claim 7+- CE-DAD method, which is characterized in that the Capillary Electrophoresis water phase buffer solution
Composition in the lauryl sodium sulfate be 40mM.
9. ABTS according to claim 1,2 or 3+- CE-DAD method, which is characterized in that the Capillary Electrophoresis water phase is slow
The acetonitrile percent by volume in the composition of fliud flushing is selected from 2.5%~5%, 5%~7.5%, 7.5%~10%.
10. ABTS+-CE-DAD method according to claim 9, which is characterized in that the Capillary Electrophoresis water phase buffering
The acetonitrile percent by volume in the composition of liquid is 7.5.
11. ABTS+-CE-DAD method according to claim 1,2 or 3, which is characterized in that the Capillary Electrophoresis water phase
The pH value of buffer is selected from 6.5~7.0 or 7.0~7.5.
12. ABTS+-CE-DAD method according to claim 11, which is characterized in that the Capillary Electrophoresis water phase buffering
The pH value of liquid is 7.0.
13. ABTS+-CE-DAD method according to claim 1,2 or 3, which is characterized in that the voltage be selected from 18kV~
20kV or 20kV~22kV；The temperature control of the cartridge is selected from 19 DEG C~22 DEG C or 22 DEG C~25 DEG C.
14. ABTS+-CE-DAD method according to claim 13, which is characterized in that the voltage is 20 kV；The card
The temperature control of box is 22 DEG C.
15. ABTS+-CE-DAD method according to claim 1, which is characterized in that screen or/and measure online and is anti-oxidant
The composition of the Capillary Electrophoresis water phase buffer solution of active constituent is the beta-cyclodextrin of 50mM for 20mM phosphate, the 12 of 40mM
The acetonitrile that sodium alkyl sulfate and percent by volume are 7.5%, wherein the phosphate be by equimolar amounts NaH2PO4 and
What Na2HPO4 was mixed to get；The pH value of the Capillary Electrophoresis water phase buffer solution is 7.0；In whole experiment process, voltage is
The temperature of 20kV, cartridge are controlled at 22 DEG C；The Detection wavelength of traditional Chinese medicine sample is 360nm, and the Detection wavelength of ABTS+ is 405nm；
The sample introduction of traditional Chinese medicine sample and ABTS+ are all in 50mbar sample introduction under pressure 5s.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710513206.8A CN107144623B (en) | 2017-06-29 | 2017-06-29 | A method of oxidation-resistant active ingredient in online quickly screening and quantitative Chinese medicine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710513206.8A CN107144623B (en) | 2017-06-29 | 2017-06-29 | A method of oxidation-resistant active ingredient in online quickly screening and quantitative Chinese medicine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107144623A CN107144623A (en) | 2017-09-08 |
| CN107144623B true CN107144623B (en) | 2019-05-24 |
Family
ID=59785761
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710513206.8A Active CN107144623B (en) | 2017-06-29 | 2017-06-29 | A method of oxidation-resistant active ingredient in online quickly screening and quantitative Chinese medicine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107144623B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108693287B (en) * | 2018-05-23 | 2019-10-08 | 安徽瑞思威尔科技有限公司 | A kind of method of antioxidant content in detection white wine |
| CN112114022A (en) * | 2020-09-24 | 2020-12-22 | 中国科学院兰州化学物理研究所 | Method for precisely screening Chinese herbal medicine active molecules by using capillary electrophoresis enzyme inhibitor |
| CN114935622B (en) * | 2022-05-05 | 2023-06-16 | 杭州师范大学 | Method for simultaneously and online enriching and separating phenolic compounds in red sage root by adopting amphiphilic affinity preconcentration method |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5898493A (en) * | 1996-06-07 | 1999-04-27 | Iowa State University Research Foundation, Inc. | Capillary electrophoresis-fluorescence line narrowing system (CE-FLNS) for on-line structural characterization |
| CN102841152A (en) * | 2011-12-31 | 2012-12-26 | 山西振东泰盛制药有限公司 | Detection method of finger-print of Shuxuening injection |
| CN105241965A (en) * | 2014-07-09 | 2016-01-13 | 西南大学 | Method of on-line quickly detecting total anti-oxidizing property of sample |
| CN105891355A (en) * | 2016-03-31 | 2016-08-24 | 河北神威药业有限公司 | Detecting method for measuring content of flavonoid compounds and terpene lactone compounds in ginkgo leaf extract or preparations of ginkgo leaf extract at same time |
| CN105891356A (en) * | 2016-03-31 | 2016-08-24 | 河北神威药业有限公司 | Detecting method for measuring content of flavonoid compounds and terpene lactone compounds in ginkgo bioba leaf extract at same time |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH11230935A (en) * | 1998-02-18 | 1999-08-27 | Yokogawa Analytical Systems Inc | Method and device for analyzing saccharide by capillary electrophoresis |
| US9316629B2 (en) * | 2013-09-19 | 2016-04-19 | King Fahd University Of Petroleum And Minerals | Method of determining phenoxy herbicides in water samples by phase transfer microextraction with simultaneous derivatization and gas-chromatography mass-spectrometry analysis |
-
2017
- 2017-06-29 CN CN201710513206.8A patent/CN107144623B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5898493A (en) * | 1996-06-07 | 1999-04-27 | Iowa State University Research Foundation, Inc. | Capillary electrophoresis-fluorescence line narrowing system (CE-FLNS) for on-line structural characterization |
| CN102841152A (en) * | 2011-12-31 | 2012-12-26 | 山西振东泰盛制药有限公司 | Detection method of finger-print of Shuxuening injection |
| CN105241965A (en) * | 2014-07-09 | 2016-01-13 | 西南大学 | Method of on-line quickly detecting total anti-oxidizing property of sample |
| CN105891355A (en) * | 2016-03-31 | 2016-08-24 | 河北神威药业有限公司 | Detecting method for measuring content of flavonoid compounds and terpene lactone compounds in ginkgo leaf extract or preparations of ginkgo leaf extract at same time |
| CN105891356A (en) * | 2016-03-31 | 2016-08-24 | 河北神威药业有限公司 | Detecting method for measuring content of flavonoid compounds and terpene lactone compounds in ginkgo bioba leaf extract at same time |
Non-Patent Citations (2)
| Title |
|---|
| Antioxidant, haemolytic activities and HPLC–DAD–ESI–MSn characterization of phenolic compounds from root bark of Juniperus oxycedrus subsp.oxycedrus;Tarik Mohammed Chaouche 等;《Industrial Crops and Products》;20141112;第64卷;第182-187页 |
| 毛细管电泳法测定螺内酯片剂中螺内酯的含量;于海霞 等;《山西医科大学学报》;20151031;第46卷(第10期);第995-998页 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107144623A (en) | 2017-09-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Martins et al. | Determination of parabens in shampoo using high performance liquid chromatography with amperometric detection on a boron-doped diamond electrode | |
| Huang et al. | Studies on principal components and antioxidant activity of different Radix Astragali samples using high-performance liquid chromatography/electrospray ionization multiple-stage tandem mass spectrometry | |
| CN107144623B (en) | A method of oxidation-resistant active ingredient in online quickly screening and quantitative Chinese medicine | |
| Liu et al. | Sensitive and robust UPLC–MS/MS method to determine the gender-dependent pharmacokinetics in rats of emodin and its glucuronide | |
| Memon et al. | Simultaneous determination of quercetin, rutin, naringin, and naringenin in different fruits by capillary zone electrophoresis | |
| Duan et al. | LC–MS/MS determination and pharmacokinetic study of five flavone components after solvent extraction/acid hydrolysis in rat plasma after oral administration of Verbena officinalis L. extract | |
| Liu et al. | Simultaneous determination of icariin, icariside II and osthole in rat plasma after oral administration of the extract of Gushudan (a Chinese compound formulation) by LC–MS/MS | |
| CN105675779A (en) | Quantitative detection method of polysaccharide containing uronic acid | |
| Maier et al. | Study on the use of boromycin as a chiral selector in capillary electrophoresis | |
| Sun et al. | Simultaneous determination of four flavonoids and one phenolic acid in rat plasma by LC–MS/MS and its application to a pharmacokinetic study after oral administration of the Herba Desmodii Styracifolii extract | |
| Xia et al. | A new method for quantitative determination of two uronic acids by CZE with direct UV detection | |
| Foubert et al. | Rapid quantification of 14 saponins of Maesa lanceolata by UPLC–MS/MS | |
| Zheng et al. | A new combined method of stable isotope-labeling derivatization-ultrasound-assisted dispersive liquid–liquid microextraction for the determination of neurotransmitters in rat brain microdialysates by ultra high performance liquid chromatography tandem mass spectrometry | |
| Peng et al. | Simultaneous separation and concentration of neutral analytes by cyclodextrin assisted sweeping-micellar electrokinetic chromatography | |
| CN106814153A (en) | The liquid chromatography mass screening technique of the double target spot anticoagulating active materials based on fibrin ferment and clotting factor | |
| Jiang et al. | Analysis of flavonoids in propolis and Ginkgo biloba by micellar electrokinetic capillary chromatography | |
| Qiu et al. | LC–MS/MS method for the simultaneous quantification of 11 compounds of Ginkgo biloba extract in lysates of mesangial cell cultured by high glucose | |
| Luo et al. | Optimization of ultrasound-assisted extraction (UAE) of phenolic compounds from Crataegus pinnatifida leaves and evaluation of antioxidant activities of extracts | |
| Wu et al. | Direct and simultaneous recognition of the positional isomers of aminobenzenesulfonic acid by TIMS-TOF-MS | |
| Agrawal et al. | Simultaneous pharmacokinetic assessment of phytopharmaceuticals in fenugreek extract using LC–MS/MS in Sprague–Dawley rats | |
| CN103210308B (en) | Method of determining quantity of herbal medicine-derived components | |
| Guo et al. | Separation and detection of tropane alkaloids in Anisodus tanguticus by capillary electrophoresis-electrochemiluminescence | |
| Zhou et al. | Screening and mechanism study of components targeting DNA from the Chinese herb Lonicera japonica by liquid chromatography/mass spectrometry and fluorescence spectroscopy | |
| Xu et al. | Improvement of sugar analysis sensitivity using anion-exchange chromatography–electrospray ionization mass spectrometry with sheath liquid interface | |
| Dong et al. | Screening of lignan patterns in Schisandra species using ultrasonic assisted temperature switch ionic liquid microextraction followed by UPLC-MS/MS analysis |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |