CN106770828B - Method that is a kind of while measuring multicomponent content in ginkgo biloba p.e and its preparation - Google Patents
Method that is a kind of while measuring multicomponent content in ginkgo biloba p.e and its preparation Download PDFInfo
- Publication number
- CN106770828B CN106770828B CN201611217062.3A CN201611217062A CN106770828B CN 106770828 B CN106770828 B CN 106770828B CN 201611217062 A CN201611217062 A CN 201611217062A CN 106770828 B CN106770828 B CN 106770828B
- Authority
- CN
- China
- Prior art keywords
- preparation
- kaempferol
- quercetin
- reference substance
- 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
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
Abstract
The present invention relates to drug tests, it is more particularly related to a kind of high performance liquid chromatography for measuring multicomponent content in ginkgo leaf (or its extract) and its preparation simultaneously.The method that the present invention establishes can simultaneously be measured the content of 14 kinds of flavonoid glycoside compounds and 2 kinds of phenolic acid compounds in ginkgo leaf (or its extract) and its preparation, provide new analysis method and foundation for the quality control of commercially available gingko leaf preparation.
Description
Technical field
The present invention relates to drug tests, it is more particularly related to which a kind of measure ginkgo biloba p.e simultaneously
And its in preparation multicomponent content high-efficiency liquid chromatography method for detecting.
Background technique
Ginkgo is one of plant most ancient on the earth, is Ginkgoaceae ginkgo platymiscium, the tree species vitality is extremely strong, thousands of
The age of tree in year is not uncommon for, and can be rated as " venerable old man or lady " of plant kingdom.Recorded in Compendium of Material Medica, ginkgo leaf can bushing foster the spirit of nobility, kidney-nourishing
Moistening lung, strengthening brain and inducing resuscitation are promoted longevity.Modern pharmacology research shows that ginkgo leaf has scavenging activated oxygen, reducing blood lipid, enhancing
Central nervous system function, adjust neurotransmitter and hormonal readiness, improve hemorheology state, is anti-inflammatory, antiallergy the effects of.Cause
And it is more and more with the preparation variety that ginkgo leaf is developed, clinical application is more and more extensive.That clinically applies at present has
This ACE Semi, Ginkgo Leaf Agent, Tian Bao Ning, Gin Kgo Plus, Floium Ginkgo, network are glad logical, tablet, the granule, injection of a variety of brands such as ketone for curing coronary heart disease
Agent is mainly used for treating various cardiovascular and cerebrovascular diseases, diabetes, the nervous system disease and respiratory disease etc..With to silver
The research of apricot leaf preparation effect, the various preparations of ginkgo leaf have not only obtained everybody on cardiovascular and cerebrovascular diseases and have approved and answer
With, and clinic is also applied in the other diseases such as Chronic Obstructive Pulmonary Disease.With going deep into for research, gingko leaf preparation
It will clinically be more widely used.
Principle active component is flavonoid glycoside and terpene lactone active material in ginkgo biloba p.e and its preparation, at present
Until, it is generally both at home and abroad the content for measuring these two types of ingredients about the method for quality control of ginkgo leaf and its preparation.About silver
Flavonoid glycoside and terpene lactone contents measuring method, have there is high performance liquid chromatography in apricot leaf and its preparation.For example,
CN104034826 provides a kind of side using HPLC ELSD detector detection ginkgo leaf terpene lactone
Method can detect 4 kinds of terpene lactones using this method simultaneously；Liu can wait to disclose and a kind of be mentioned with HPLC-UV method measurement ginkgo leaf
3 flavonol glycosides contents in object are taken, and measure method (the Chinese traditional Chinese medicine magazine of 4 terpene lactone contents with HPLC-ELSD method
(former Chinese Medicine journal) 2 months the 2nd phases of volume 30 in 2015, page 598~601).The above method is using different detection sides
Method detects terpene lactone and flavonol glycosides respectively, the disadvantage is that needing using different instruments, different sample processing conditions difference
Sample introduction, many and diverse so as to cause operating procedure, simple process is insufficient.
Cui great Ming etc. discloses a kind of 3 kinds of GINKGO BILOBA EXTRACTs using in RP-HPLC measurement ginkgo biloba p.e lipoid plastid
Glycosides (Quercetin, Kaempferol, Isorhamnetin) and 4 kinds of terpene lactones (ginkalide A, ginkolide B, ginkalide C, gingkoes
Lactone) content method (Jilin Auto Industry, 2013,35 (4): 446~449,462)；Gorgeous equality is appointed to disclose one kind
Using HPLC-MS simultaneously measure ginkalide A in Shu Xuening injection (GA), ginkolide B (GB), ginkalide C (GC),
Method (the medicine of 7 component contents such as Bilobalide, (BB) and Quercetin (QCT), Isorhamnetin (ISR) and Kaempferol (KAE)
Object analysis magazine, page 2013,33 (2) the 220th~224).The above method can detect flavonoid glycoside and terpene lactone simultaneously, significantly
Simplify operating process.
The reference substance of each ingredient need to be used when measuring Multiple components content simultaneously in above-mentioned numerous detection methods,
It can allow the control of comprehensive multi-index quality that can make quality control because testing cost is excessively high in actual production and detection work in this way
Index is reduced, and is unfavorable for the thoroughly evaluating of quality of medicinal material.Comment method it can be seen that attempting introducing one and surveying to realize each constituents more
While detection be specification and improve gingko leaf preparation quality control indispensability choosing.
It is (" Chinese that side etc. discloses a kind of " one survey comments method while measuring flavones ingredient in 6 in Shu Xuening injection " more
Pharmacist ", the 10th phase of volume 18 in 2015, page 1652~1656) method；Yang Yanmo etc. discloses a survey and comments technology in ginkgo more
Application study (Hunan University of Traditional Chinese Medicine's master thesis, 2014) in leaf extract and its Jinji preparation.But it is above-mentioned
The detectable component of two methods is less, and cannot detect flavonoids and liposoluble ingredient simultaneously, it is therefore necessary to carry out more
Further investigation, develops better detection technique.
Summary of the invention
14 kinds of flavonoid glycoside groups in ginkgo biloba p.e and its preparation are measured simultaneously the purpose of the present invention is to provide a kind of
Divide the detection method with 2 kinds of phenolic acid class constituent contents.In this way, the quality of preparation can be preferably controlled, guarantees to use
The safety of medicine, can preferably Instructing manufacture, make controlling of production process more it is stringent rationally, enable the customer to full appreciation
Product quality.
To achieve the goals above, the technical solution of the present invention is as follows:
It is a kind of using flavonoid glycoside in hplc simultaneous determination ginkgo leaf, ginkgo biloba p.e and its preparation and
The method of phenolic acid class constituent content, wherein the preparation does not include Shu Xuening injection, the chromatographic condition which uses are as follows: fixed
It is mutually the chromatographic column using octadecyl silane as filler；33 DEG C~37 DEG C of column temperature；Elution flow rate is 0.1~0.5mL/min；
Mobile phase A is the aqueous solution for the formic acid for being 0.1% containing mass ratio；Mobile phase B is the mixed solvent of methanol and acetonitrile, and volume ratio is
1:1；Carry out gradient elution, gradient condition are as follows:
；Using diode array detector, phenolic acid class component is detected using the ultraviolet wavelength within the scope of 270nm~290nm,
And flavonoid glycoside ingredient is detected using the ultraviolet wavelength within the scope of 350nm~370nm simultaneously.
In some embodiments, the preferred column temperature of chromatographic column is 35 DEG C.
In some embodiments, the preferred 0.3mL/min of the elution flow rate.
In some embodiments it is preferred that the ultraviolet wavelength using 280nm detects phenolic acid class component.
In some embodiments it is preferred that the ultraviolet wavelength using 360nm detects flavonoid glycoside component.
In some embodiments it is preferred that detecting phenolic acid class component using the ultraviolet wavelength of 280nm, while using 360nm
Ultraviolet wavelength detect flavonoid glycoside component.
In some embodiments it is preferred that calculating the content of the flavonoid glycoside and phenolic acid class component using external standard method.
Comment method to calculate containing for the flavonoid glycoside and phenolic acid class component in some embodiments it is preferred that surveying using one more
Amount, wherein flavonoid glycoside component is using rutin as internal standard, and phenolic acid class component is using Cryptochlorogenic acid as internal standard.
In some embodiments, sampling volume is 1 μ L.
In some embodiments, the chromatographic column is ACQUITY UPLC BEH C18 chromatographic column.
In some embodiments, the chromatographic column specification is 1.7 μm, 2.1 × 150mm.
2 kinds of phenolic acid class components of the present invention are respectively protocatechuic acid and Cryptochlorogenic acid；14 kinds of flavonoids groups
It is respectively dish legumin, rutin, isoquercitrin, Quercetin -3-O- glucosyl group (1 → 2) rhamnoside, Kaempferol -3-O- rue
{ [6-O- is (right by 2-O- by fragrant glucosides, astragalin, cosmosiin, narcissin, isorhamnetin-3-O-β-D-glycopranoside, 3-O-
The trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnopyranosyl }-Quercetin, Quercetin, { [6-O- is (to the trans- perfume (or spice) of hydroxyl-by 2-O- by 3-O-
Beans acyl)-glucosyl group]-(1-2) rhamnopyranosyl }-Kaempferol, Kaempferol, Isorhamnetin.
Method of the invention has the advantages that compared with prior art:
(1) method of the invention can measure simultaneously ginkgo biloba p.e and its system under the same high-efficient liquid phase chromatogram condition
The content of 14 kinds of flavonoid glycoside components and 2 kinds of phenolic acid class components in agent avoids in the detection frequently replacement liquid-phase condition, improves
Working efficiency is suitble to the requirement of industry mass production.
(2) method durability of the invention is good, easy to accomplish, and instrument equipment and reagent are conventional articles, test
Parameter is also conventional parameter, no harsh conditions, at low cost, and the condition in most of laboratories is able to satisfy.
(3) of the invention one survey comments method, choose two it is cheap, be easy to get, stable ingredient as the completion of internal reference object to it
It is measured while his 14 kinds of ingredients, solves other compositions reference substance and be not easy to obtain caused detection method to be difficult to apply to reality
The problem of production.
Attached drawing
Attached drawing 1: chromatogram of the hybrid standard product solution in 280nm absorbing wavelength；
Attached drawing 2: chromatogram of the hybrid standard product solution in 360nm absorbing wavelength；
Attached drawing 3: chromatogram of the ginkgo biloba p.e in 280nm absorbing wavelength；
Attached drawing 4: chromatogram of the ginkgo biloba p.e in 360nm absorbing wavelength；
Attached drawing 1 is equal to attached drawing 4 are as follows: 1: protocatechuic acid；2: Cryptochlorogenic acid；3: dish legumin；4: rutin；5: isoquercitrin；6:
Quercetin -3-O- glucosyl group (1 → 2) rhamnoside；7: kaempferol-3-O-rutinoside；8: astragalin；9: cosmos
Glycosides；10: narcissin；11: isorhamnetin-3-O-β-D-glycopranoside；{ [6-O- is (to the trans- tonka-bean of hydroxyl-by 2-O- by 12:3-O-
Acyl)-glucosyl group]-rhamnopyranosyl }-Quercetin；13: Quercetin；14:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-
Glucosyl group]-(1-2) rhamnopyranosyl }-Kaempferol；15: Kaempferol；16: Isorhamnetin.
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, chromatographic condition
Chromatographic column: ACQUITY UPLC BEH C18 (1.7 μm, 2.1 × 150mm)；
Mobile phase B: methanol/acetonitrile (1:1)；
Mobile phase A: ultrapure water (contains 0.1% formic acid)；
Flow velocity: 0.3mL/min；
Column temperature: 35 DEG C；
Sampling volume: 1 μ L；
Diode array detector: detecting phenolic acid class component using 280nm ultraviolet wavelength, and ultraviolet using 360nm simultaneously
Wavelength detecting flavonoid glycoside component；
Analysis time: 35min；
Gradient elution method:
2. the preparation of solution
(1) preparation of reference substance stock solution
Precision weighs protocatechuic acid, Cryptochlorogenic acid, dish legumin, rutin, isoquercitrin, Quercetin -3-O- glucosyl group (1
→ 2) rhamnoside, kaempferol-3-O-rutinoside, astragalin, cosmosiin, narcissin, Isorhamnetin -3-O- β-D-
Glucoside, 3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnopyranosyl }-Quercetin, Quercetin,
3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-(1-2) rhamnopyranosyl }-Kaempferol, Kaempferol, different mouse
Li Su reference substance 5mg, respectively plus methanol be made into concentration be 1mg/mL single reference substance stock solution, set saved in 4 DEG C of refrigerators it is standby
With.
(2) sample preparation methods
Ginkgo Leaf is taken, precision weighs 1g, is placed in 10ml volumetric flask, adds 70% methanol solution to set graduation mark, ultrasound
Extract 30min.Extracting solution is centrifuged 10min with the speed of 14000r/min, is repeated twice, takes supernatant as ginkgo leaf sample
Stock solution is set in 4 DEG C of refrigerators and is saved backup.
(3) measuring method
External standard method: precision measures reference substance solution and each 1 μ l of test solution, injects liquid chromatograph, records chromatogram.
One survey comments method: precision measures reference substance solution and each 1 μ l of test solution, injects liquid chromatograph, record color
Spectrogram, by the substitute reference substance method of the correction up factor with calculated by peak area to get wherein correction factor is shown in Table 1.
3. methodological study
(1) standard curve
By upper protocatechuic acid, Cryptochlorogenic acid, dish legumin, rutin, isoquercitrin, Quercetin -3-O- glucosyl group (1 → 2)
Rhamnoside, kaempferol-3-O-rutinoside, astragalin, cosmosiin, narcissin, Isorhamnetin -3-O- β-D- grape
Glucosides, 3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnopyranosyl }-Quercetin, Quercetin, 3-O-
{ 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-(1-2) rhamnopyranosyl }-Kaempferol, Kaempferol, Isorhamnetin
Reference substance be configured to concentration be respectively 40ug/ml, 40ug/ml, 150ug/ml, 200ug/ml, 50ug/ml, 100ug/ml,
200ug/ml、50ug/ml、40ug/ml、200ug/ml、40ug/ml、100ug/ml、40ug/ml、100ug/ml、40ug/ml、
The mixing mother liquor of 30ug/ml, and it is diluted to 8 points step by step with 2 times.By above-mentioned mixing mother liquor according to above-mentioned 1 μ of chromatographic condition sample introduction
L records peak area, and spectrogram Fig. 1, spectrogram Fig. 2 under ultraviolet wavelength 360nm are obtained under ultraviolet wavelength 280nm.Under the conditions of ultraviolet, with
Sample introduction concentration is abscissa, peak area is ordinate, draws standard curve, acquires linear regression equation.Respectively with signal-to-noise ratio S/N
3:1 and 10:1 is as detection limit (LOD) and minimum quantitative limit (LOQ).Its standard curve and the range of linearity are shown in Table 1.It can from table
To learn, related coefficient is all larger than 0.999 to 16 compounds in the linear range, shows that 16 compound linear relationships are good.
(2) calculating of relative correction factor
There are two types of the calculation methods of relative correction factor: first is that after each compound standard curve intercept is corrected to zero, tiltedly
The ratio between rate is the correction factor of each compound；Second is that wherein A is according to fa/b=fa/fb=(Aa/Ca)/(Ab/Cb) is defined
The peak area of compound, C are concentration, calculate average value after acquiring each correction factor to obtain the final product.Flavonoid glycoside compound is with rutin
For internal standard, for phenolic acid compound using Cryptochlorogenic acid as internal standard, each compound is suitable for the first calculation method.The phase of each compound
To correction factor in table 1.
1 standard curve of table and the range of linearity, LODs, LOQs and relative correction factor (RCF)
Note: A: protocatechuic acid；B: Cryptochlorogenic acid；C: dish legumin；D: rutin；E: isoquercitrin；F: Quercetin -3-O- grape
Glycosyl (1 → 2) rhamnoside；G: kaempferol-3-O-rutinoside；H: astragalin；I: cosmosiin；J: narcissin；K:
Isorhamnetin-3-O-β-D-glycopranoside；L:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnose
Base }-Quercetin；M: Quercetin；N:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-(1-2) rhamnose
Base }-Kaempferol；O: Kaempferol；P: Isorhamnetin.
(3) precision, stability, repetitive test
The standard solution for preparing basic, normal, high three concentration respectively, to test solution carry out in a few days, day to day precision
With the investigation of stability.Withinday precision is basic, normal, high three horizontal same a test solution continuous sample introductions in one day
6 times, day to day precision is interior continuous sample introduction 6 times for three days on end.Stability experiment is the test solution of basic, normal, high 3 concentration
Respectively 0,2,4,6,8,12, for 24 hours in sample introduction.Repeated experiment is to prepare 6 parts of ginkgo biloba p.e sample solutions in parallel, continuously
Sample introduction.By external standard method and " one surveys comment more ", method acquires the rate of recovery of each compound, and calculates RSD value, repeated experiment knot respectively
Fruit is shown in Table 1, and precision and stability experiment the results are shown in Table 2, table 3.The result shows that this method is accurate, stablizes, repeatability height, it can
To be used for ginkgo biloba p.e assay.
2 external standard method of table calculates precision, stability test result
Note: A: protocatechuic acid；B: Cryptochlorogenic acid；C: dish legumin；D: rutin；E: isoquercitrin；F: Quercetin -3-O- grape
Glycosyl (1 → 2) rhamnoside；G: kaempferol-3-O-rutinoside；H: astragalin；I: cosmosiin；J: narcissin；K:
Isorhamnetin-3-O-β-D-glycopranoside；L:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnose
Base }-Quercetin；M: Quercetin；N:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-(1-2) rhamnose
Base }-Kaempferol；O: Kaempferol；P: Isorhamnetin.
The mark of table 3 one comments method to calculate precision, stability test result
Note: A: protocatechuic acid；B: Cryptochlorogenic acid；C: dish legumin；D: rutin；E: isoquercitrin；F: Quercetin -3-O- grape
Glycosyl (1 → 2) rhamnoside；G: kaempferol-3-O-rutinoside；H: astragalin；I: cosmosiin；J: narcissin；K:
Isorhamnetin-3-O-β-D-glycopranoside；L:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnose
Base }-Quercetin；M: Quercetin；N:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-(1-2) rhamnose
Base }-Kaempferol；O: Kaempferol；P: Isorhamnetin.
(4) sample recovery rate
The ginkgo biloba p.e sample of known content is taken to set in volumetric flask, it is accurate respectively to be added in ginkgo biloba p.e sample
80%, 100%, the 120% of each compounds content, 3 parts of each concentration operation repetitive, according to " preparations of 2 (2) sample solutions "
Lower method is handled, and calculates the rate of recovery with (the original amount of measured amount -)/scalar quantity × 100%, and respectively by external standard method and
" one surveys comment more " method acquires the sample recovery rate of each compound, and calculates RSD value, the results are shown in Table 4 and 5.The results show that each chemical combination
For the sample recovery rate of object between 89.1%-105%, RSD value is respectively less than 5%.
4 external standard method of table calculates sample recovery rate
Note: A: protocatechuic acid；B: Cryptochlorogenic acid；C: dish legumin；D: rutin；E: isoquercitrin；F: Quercetin -3-O- grape
Glycosyl (1 → 2) rhamnoside；G: kaempferol-3-O-rutinoside；H: astragalin；I: cosmosiin；J: narcissin；K:
Isorhamnetin-3-O-β-D-glycopranoside；L:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnose
Base }-Quercetin；M: Quercetin；N:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-(1-2) rhamnose
Base }-Kaempferol；O: Kaempferol；P: Isorhamnetin.
Table 5 one is surveyed comments method to calculate sample recovery rate more
Note: A: protocatechuic acid；B: Cryptochlorogenic acid；C: dish legumin；D: rutin；E: isoquercitrin；F: Quercetin -3-O- grape
Glycosyl (1 → 2) rhamnoside；G: kaempferol-3-O-rutinoside；H: astragalin；I: cosmosiin；J: narcissin；K:
Isorhamnetin-3-O-β-D-glycopranoside；L:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnose
Base }-Quercetin；M: Quercetin；N:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-(1-2) rhamnose
Base }-Kaempferol；O: Kaempferol；P: Isorhamnetin.
4. sample size measures
" one surveys comment more " analysis method is used to measure two kinds of ingredient in ginkgo biloba p.e: phenolic acid class and flavonoids
Ingredient.3 batches of ginkgo biloba p.es sequentially determining, chromatogram under " 1 chromatographic condition " is as shown in Figure 3 and Figure 4.By external standard method and
" one surveys comment more " method result compares, and is as a result listed in table 6.The result shows that " one surveys comment more " analysis method and " external standard method "
Result out is almost the same (RSDs < 5%), illustrates that " one surveys comment more " method can be used to carry out quality control to Shu Xuening injection
System.
The content of table 63 batches of ginkgo biloba p.es, 16 chemical components
Note: A: protocatechuic acid；B: Cryptochlorogenic acid；C: dish legumin；D: rutin；E: isoquercitrin；F: Quercetin -3-O- grape
Glycosyl (1 → 2) rhamnoside；G: kaempferol-3-O-rutinoside；H: astragalin；I: cosmosiin；J: narcissin；K:
Isorhamnetin-3-O-β-D-glycopranoside；L:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnose
Base }-Quercetin；M: Quercetin；N:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-(1-2) rhamnose
Base }-Kaempferol；O: Kaempferol；P: Isorhamnetin；E.M represents the calculated content of external standard method method, and N.M represents " few mark to be commented "
The calculated content of method, R.D are relative deviation.
The verifying of 5 correction factors
In order to further verify the applicability of relative correction factor, by ginkgo biloba p.e according to " 2 (2) sample solutions
Preparation " under item method handled, calculate opposite improvement factor under different wave length, different temperatures and flow velocity respectively, and with
Correction factor obtained by upper experiment is compared analysis.The results are shown in Table 7, and under different temperatures, flow velocity, relative correction factor is steady
It is fixed reliable, difference at different wavelengths, but it is more stable.
7 correction factor verification result of table
Note: A: protocatechuic acid；B: Cryptochlorogenic acid；C: dish legumin；D: rutin；E: isoquercitrin；F: Quercetin -3-O- grape
Glycosyl (1 → 2) rhamnoside；G: kaempferol-3-O-rutinoside；H: astragalin；I: cosmosiin；J: narcissin；K:
Isorhamnetin-3-O-β-D-glycopranoside；L:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnose
Base }-Quercetin；M: Quercetin；N:3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-(1-2) rhamnose
Base }-Kaempferol；O: Kaempferol；P: Isorhamnetin.
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 (1)
1. a kind of using 14 kinds of flavonoid glycosides in hplc simultaneous determination ginkgo leaf, ginkgo biloba p.e and its preparation
The method of component and 2 kinds of phenolic acid class constituent contents, wherein the preparation does not include Shu Xuening injection, it is characterised in that use
Chromatographic condition are as follows: stationary phase is the chromatographic column using octadecyl silane as filler；35 DEG C of column temperature；Elution flow rate is 0.3mL/
min；Mobile phase A is the aqueous solution for the formic acid for being 0.1% containing mass ratio；Mobile phase B is the mixed solvent of methanol and acetonitrile, volume
Than being 1:1；Carry out gradient elution, gradient condition are as follows:
Using diode array detector, phenolic acid class component is detected using 280nm ultraviolet wavelength, is examined using 360nm ultraviolet wavelength
Survey flavonoid glycoside component；Wherein flavonoid glycoside component is using rutin as internal standard, and phenolic acid class component is using Cryptochlorogenic acid as internal standard；
The method, steps are as follows:
(1) preparation of reference substance stock solution
Precision weighs protocatechuic acid, Cryptochlorogenic acid, dish legumin, rutin, isoquercitrin, Quercetin -3-O- glucosyl group (1 → 2)
Rhamnoside, kaempferol-3-O-rutinoside, astragalin, cosmosiin, narcissin, Isorhamnetin -3-O- β-D- grape
Glucosides, 3-O- { 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-rhamnopyranosyl }-Quercetin, Quercetin, 3-O-
{ 2-O- [6-O- (to the trans- coumaric acyl of hydroxyl -)-glucosyl group]-(1-2) rhamnopyranosyl }-Kaempferol, Kaempferol, Isorhamnetin
5 mg of reference substance, respectively plus methanol is made into the single reference substance stock solution that concentration is 1mg/mL；
(2) sample preparation methods
Ginkgo Leaf is taken, precision weighs 1g, is placed in 10ml volumetric flask, adds 70% methanol solution to graduation mark, ultrasonic extraction
Extracting solution is centrifuged 10min with the speed of 14000r/min, is repeated twice, takes supernatant as ginkgo leaf sample by 30min；
(3) measuring method
Reference substance solution and each 1 μ l of test solution are measured, liquid chromatograph is injected, records chromatogram；By the correction up factor
Substitute reference substance method is with calculated by peak area to get the content of each component；Wherein the calculation method of relative correction factor is Jiang Gehua
After conjunction object standard curve intercept is corrected to zero, the ratio between slope is the correction factor of each compound.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611217062.3A CN106770828B (en) | 2016-12-26 | 2016-12-26 | Method that is a kind of while measuring multicomponent content in ginkgo biloba p.e and its preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611217062.3A CN106770828B (en) | 2016-12-26 | 2016-12-26 | Method that is a kind of while measuring multicomponent content in ginkgo biloba p.e and its preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106770828A CN106770828A (en) | 2017-05-31 |
| CN106770828B true CN106770828B (en) | 2019-07-02 |
Family
ID=58924818
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611217062.3A Active CN106770828B (en) | 2016-12-26 | 2016-12-26 | Method that is a kind of while measuring multicomponent content in ginkgo biloba p.e and its preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106770828B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107064397A (en) * | 2017-06-16 | 2017-08-18 | 黑龙江珍宝岛药业股份有限公司 | A kind of Shu Xuening injection method of quality control that many evaluation amount methods and finger-print are surveyed based on one |
| CN107884483B (en) * | 2017-09-27 | 2020-12-29 | 黑龙江珍宝岛药业股份有限公司 | Method for measuring content of flavonoid component in ginkgo leaf and preparation thereof and application |
| CN108279278B (en) * | 2017-12-29 | 2020-09-04 | 广州白云山和记黄埔中药有限公司 | Method for separating flavonoid components and application thereof |
| CN108362809B (en) * | 2018-02-01 | 2020-07-10 | 大连工业大学 | Quality evaluation method of ginkgo leaf and extract and preparation thereof |
| CN112240914A (en) * | 2019-07-18 | 2021-01-19 | 福建中医药大学 | Method for detecting flavone components in anoectochilus formosanus with different appearance phenotypes |
| CN111323500A (en) * | 2019-10-17 | 2020-06-23 | 蚌埠火鹤制药股份有限公司 | Establishment method of HPLC fingerprint spectrum of hypericum japonicum granules |
| CN110824071B (en) * | 2019-12-18 | 2021-09-28 | 神威药业集团有限公司 | Method for detecting lignans and flavonol glycosides in ginkgo leaf extract or preparation thereof |
| CN112697899B (en) * | 2020-12-07 | 2022-04-12 | 中国药科大学 | Detection method of ginkgo flavonol glycosides |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999034810A1 (en) * | 1998-01-12 | 1999-07-15 | Her Majesty In Right Of Canada As Represented By The Minister Of Agriculture And Agri-Food Canada | A process for the isolation, recovery and purification of non-polar extractives |
| CN102539553A (en) * | 2011-12-15 | 2012-07-04 | 石家庄东方药业有限公司 | Method for establishing fingerprint spectrum of liver-enhancing medicine |
| CN103175912A (en) * | 2013-02-06 | 2013-06-26 | 南京中医药大学 | Method for controlling quality of ginkgo leaves and extract thereof |
| CN104458995A (en) * | 2014-12-17 | 2015-03-25 | 通化华夏药业有限责任公司 | Method for rapid analysis of flavonoid glycoside and phenolic acid active ingredients in ixeris sonchifolia injection by virtue of high performance liquid chromatography (HPLC) |
| CN106198782A (en) * | 2016-06-24 | 2016-12-07 | 广州中医药大学 | A kind of can realize the content analysis of 18 components in Folium Ilicis and the method for quality control of similarity evaluation simultaneously |
-
2016
- 2016-12-26 CN CN201611217062.3A patent/CN106770828B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999034810A1 (en) * | 1998-01-12 | 1999-07-15 | Her Majesty In Right Of Canada As Represented By The Minister Of Agriculture And Agri-Food Canada | A process for the isolation, recovery and purification of non-polar extractives |
| CN102539553A (en) * | 2011-12-15 | 2012-07-04 | 石家庄东方药业有限公司 | Method for establishing fingerprint spectrum of liver-enhancing medicine |
| CN103175912A (en) * | 2013-02-06 | 2013-06-26 | 南京中医药大学 | Method for controlling quality of ginkgo leaves and extract thereof |
| CN104458995A (en) * | 2014-12-17 | 2015-03-25 | 通化华夏药业有限责任公司 | Method for rapid analysis of flavonoid glycoside and phenolic acid active ingredients in ixeris sonchifolia injection by virtue of high performance liquid chromatography (HPLC) |
| CN106198782A (en) * | 2016-06-24 | 2016-12-07 | 广州中医药大学 | A kind of can realize the content analysis of 18 components in Folium Ilicis and the method for quality control of similarity evaluation simultaneously |
Non-Patent Citations (3)
| Title |
|---|
| A Screening Method for the Identification of Glycosylated Flavonoids and Other Phenolic Compounds Using a Standard Analytical Approach for All Plant Materials;LONG-ZE LIN 等;《J.Agric.Food Chem.》;20070127;第55卷(第4期);第1084-1096页 |
| RP-HPLC 法测定竹叶提取物中黄酮类和酚酸类成分;龚金焱 等;《中草药》;20100131;第41卷(第1期);第63-65页 |
| 银杏叶提取物的研究进展;徐艳芬 等;《药物评价研究》;20101231;第33卷(第6期);第452-456页 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106770828A (en) | 2017-05-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106770828B (en) | Method that is a kind of while measuring multicomponent content in ginkgo biloba p.e and its preparation | |
| CN105891355B (en) | It is a kind of while determine the detection method of flavone compound and terpene lactones compounds content in ginkgo biloba p.e and its preparation | |
| CN106706809B (en) | Method that is a kind of while measuring multicomponent content in Shu Xuening injection | |
| CN108362809B (en) | Quality evaluation method of ginkgo leaf and extract and preparation thereof | |
| CN107884483B (en) | Method for measuring content of flavonoid component in ginkgo leaf and preparation thereof and application | |
| CN107064397A (en) | A kind of Shu Xuening injection method of quality control that many evaluation amount methods and finger-print are surveyed based on one | |
| CN105467059A (en) | Quality detecting method for traditional Chinese medicine composition for treating hematuresis | |
| CN106124639A (en) | The multicomponent content assaying method of Eucommia ulmoides | |
| CN107037167A (en) | The method of flavones ingredient content in multi-target ingredient quantitative determination ginkgo leaf | |
| CN110927311B (en) | Construction method of UPLC (ultra performance liquid chromatography) characteristic spectrum of dogbane leaf medicinal material and method for measuring content of flavonoid component of dogbane leaf medicinal material | |
| CN109030678A (en) | The content assaying method of 5 kinds of chemical components in a kind of northeast Chinese Ixeris | |
| CN106501393B (en) | A method of detection hickory chick Nucleosides | |
| CN104237416B (en) | The HPLC method of 6 kinds of flavones ingredient content in Simultaneously test poacynum hendersonii woodson leaf | |
| CN104374841B (en) | Tablet of antelope's horn for common cold quality control is with reference to product and purposes | |
| CN108956800A (en) | The measuring method of ellagic acid content in cortex moutan | |
| CN104678004A (en) | Quality control method for kudzuvine root and hawthorn lipid-lowering particles | |
| Wu et al. | Simultaneous determination of nine flavonoids in beagle dog by HPLC with DAD and application of Ginkgo biloba extracts on the pharmacokinetic | |
| CN114034798B (en) | Red water dendrobium stem flower fingerprint construction and content determination method | |
| CN109991327A (en) | One surveys the methods for commenting method evaluation field thistle quality more | |
| CN102078503A (en) | Detection method for pulse-activating decoction traditional Chinese medicine preparation | |
| CN108414642A (en) | The content assaying method and purposes of flavonol glycosides prototype ingredient in a kind of Shu Xuening injection | |
| CN108061763A (en) | Multicomponent content assaying method in longan leaf | |
| CN111896637B (en) | Detection method of Jinqing intermediate and fingerprint spectrum construction method thereof | |
| CN104597168B (en) | Ramulus Mori refines decoction pieces content assaying method | |
| CN101441201B (en) | Method for detecting contents of puerarin and rhodiola rosea glycosides in Paotianhongyangshejiu |
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 |