CN110007033A - Synchronous detection qinghaosu, dihydroartemisinine, Artesunate content method - Google Patents
Synchronous detection qinghaosu, dihydroartemisinine, Artesunate content method Download PDFInfo
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- CN110007033A CN110007033A CN201910397238.5A CN201910397238A CN110007033A CN 110007033 A CN110007033 A CN 110007033A CN 201910397238 A CN201910397238 A CN 201910397238A CN 110007033 A CN110007033 A CN 110007033A
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- G—PHYSICS
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- 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
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- 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
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- 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
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Abstract
The present invention provide it is a kind of it is synchronous detection qinghaosu, dihydroartemisinine, Artesunate content method, to sweet wormwood sample without pre-treatment derivatization, the content of qinghaosu, dihydroartemisinine, Artesunate sample is directly detected using UPLC method, using Shimadzu Shim-pack XR-ODS II as chromatographic column, 0.1% acetum-acetonitrile is that mobile phase carries out isocratic elution, and method specificity is strong, precision is high, stability is good, accuracy is high.The present invention is obviously shortened detection time, simplifies experimental procedure, reduces test error, the intact isolated qualitative and quantitative analysis of three kinds of samples can be realized in 15 minutes, greatly improve working efficiency.Reagent of the present invention and consumptive material are cheap and easy to get, and operation is quick and easy, practical, are suitable for the content detection of qinghaosu, dihydroartemisinine, Artesunate sample.
Description
Technical field
The invention belongs to analytical chemistry fields, and in particular to a kind of to utilize qinghaosu, double hydrogen in the synchronous test sample of UPLC
The method of qinghaosu, Artesunate content.
Background technique
Sweet wormwood is the dry aerial parts of compositae plant artemisia annua ArtemisiaannuaL., is that drug effect high price is low simply
The honest and clean Chinese medicine being easy to get, property bitter, acrid, cold, returns liver, gallbladder channel, has effects that clearing away summerheat, except steaming, preventing malaria, generates heat for summer-heat evil,
Fever due to yin deficiency, night fever abating at dawn, hectic fever due to yin labor heat, malaria fever and chills, the diseases such as jaundice with damp-heat pathogen mainly contain sequiterpene, flavones, cumarin etc.
Ingredient, wherein qinghaosu (Artemisinin) is the principle active component of sweet wormwood, and artemisinine is times in sweet wormwood containing peroxy-radical
Sesquiterpene lactone drug has the effects that antipyretic, immune, antimalarial, antibacterial, anti parasitic, and being that China is only gains international recognition
Antimalarial agent.Dihydroartemisinine, Artesunate are artemisinin derivative, are effective antimalarial monomers, are had antiviral, anti-
Bacterium, anti-inflammatory, immunological regulation and other effects.
In recent years, qinghaosu and derivative have caused international great attention, and raw medicinal material has realized standardized cultivation,
Therefore in Accurate Determining sweet wormwood the content of artemisinine and derivative the seed selection cultivation of sweet wormwood, qinghaosu related drugs are produced and its
Pharmacological effect research is all of great significance.Currently, the measuring method of qinghaosu and derivative specifically includes that gravimetric method, ultraviolet
Spectrophotometry, fluorescence analysis, gas chromatography, Supercritical fluid chromatography analytic approach, high performance liquid chromatography etc..Due to qinghaosu
And its derivative dihydroartemisinine, Artesunate are the sesquiterpenoids (compound structure is as follows) containing peroxide bridge, three
Compound UV absorption is weaker, reacts insensitive in UV detector, therefore the detection to sweet wormwood sample, generally requires to sample
Carry out pre-treatment derivatization, cause the operating procedure of sample detection to increase, test it is more cumbersome, and experimental results error compared with
Greatly, repeatability is bad, is that the quality control of sweet wormwood sample increases difficulty.Double hydrogen of qinghaosu and its derivative in the prior art simultaneously
The measurement of qinghaosu, Artesunate is all individually to detect, and not only needs higher cost, it is also necessary to which longer time is unfavorable for
Industrial production.
Summary of the invention
In view of this, utilizing qinghaosu, double hydrogen blueness in the synchronous test sample of UPLC the purpose of the present invention is to provide a kind of
The method of artemisin, Artesunate content, this method can be widely used for the quality control of Chinese medicine sweet wormwood Related product, can be effectively
Simplify experimental implementation, saves testing cost and time.Unless otherwise specified, percentage of the present invention is mass percent.
To achieve the above object, the present invention provides the following technical solution:
Qinghaosu, dihydroartemisinine, Artesunate contain in quantifier elimination in using the synchronous test sample of UPLC, inventor
It was found that structure is very approximate since qinghaosu, dihydroartemisinine, Artesunate are all the sesquiterpenoids containing peroxide bridge,
And dihydroartemisinine, there is also epimer, the sample peak of each drug ingedient is easy to be overlapped, while solvent peak there is also
Apparent interference.It is found after numerous studies, chromatographic column, mobile phase, type of elution, flow velocity, Detection wavelength, column temperature etc. are some
The control of column parameter can efficiently separate qinghaosu in sample, dihydroartemisinine, Artesunate, method stability and repeatability
It is good, it is suitble to industrial application.The present invention utilizes qinghaosu in the synchronous test sample of UPLC, dihydroartemisinine, Artesunate content
Method, which is characterized in that use following chromatographic condition: chromatographic column: Shimadzu Shim-pack XR-ODS II；Mobile phase and elution
Mode: with 0.1% acetum: acetonitrile (50:50) is mobile phase isocratic elution；Flow velocity: 0.1~1.0ml/min；Detect wave
It is long: 190~210nm；Column temperature: 25-35 DEG C；Sample volume: 1-20 μ l.
Further, the present invention using qinghaosu in the synchronous test sample of UPLC, dihydroartemisinine, Artesunate content side
Method, including following experimental procedure:
(1) chromatographic condition
Chromatographic column: Shimadzu Shim-pack XR-ODS II
Mobile phase and type of elution: with 0.1% acetum: acetonitrile (50:50) is mobile phase isocratic elution
Flow velocity: 0.1~1.0ml/min
Detection wavelength: 190~210nm
Column temperature: 25-35 DEG C
Sample volume: 1-20 μ l
(2) prepared by test solution
Precision measurement Artemisia annua extract is appropriate, adds methanol that test solution is made.
(3) prepared by reference substance solution
Qinghaosu, dihydroartemisinine, Artesunate reference substance are taken, accurately weighed appropriate, methanol, which is added, to be made to dissolve and dilute
Constant volume, as reference substance solution.
(4) it measures
Precision measures test solution and injects Ultra Performance Liquid Chromatography instrument, records chromatogram；Another accurate measure mixes control
Product solution injects high performance liquid chromatograph, is measured in the same method；By external standard method with qinghaosu, double hydrogen sweet wormwoods in calculated by peak area test sample
The content of element, Artesunate.
In the present invention, the preparation of the test solution uses following methods:
A. test sample is solid sample: test sample is taken, it is accurately weighed, and it sets in 100ml measuring bottle, adds 50% methanol 20ml, surpass
Sound 30min adds methanol solution to be diluted to scale constant volume, shakes up, and filtration takes subsequent filtrate to make test solution.
B. test sample is fluid sample: appropriate test sample is taken with liquid-transfering gun, is set in 100ml measuring bottle, methanol 20ml is added, ultrasound
10min adds methanol solution to be diluted to scale constant volume, shakes up, and filtration takes subsequent filtrate to make test solution.
In the present invention, the preparation of the reference substance solution uses following methods:
It is appropriate that qinghaosu, dihydroartemisinine, Artesunate reference substance are weighed respectively, it is accurately weighed, it sets in 50ml measuring bottle, adds
Methanol 15ml, ultrasonic 20min add methanol solution to be diluted to scale constant volume, shake up, and compare product solution.
Specifically, it is a kind of using qinghaosu in the synchronous test sample of UPLC, dihydroartemisinine, Artesunate content side
Method, including following experimental procedure:
(1) chromatographic condition
Chromatographic column: Shimadzu Shim-pack XR-ODS II
Mobile phase and type of elution: with 0.1% acetum: acetonitrile (50:50) is mobile phase isocratic elution
Flow velocity: 0.3ml/min
Detection wavelength: 194nm
Column temperature: 30 DEG C
Sample volume: 5 μ l
(2) prepared by test solution
A. test sample is solid sample: test sample is taken, it is accurately weighed, and it sets in 100ml measuring bottle, adds 50% methanol 20ml, surpass
Sound 30min adds methanol solution to be diluted to scale constant volume, shakes up, and filtration takes subsequent filtrate to make test solution；
B. test sample is fluid sample: appropriate test sample is taken with liquid-transfering gun, is set in 100ml measuring bottle, methanol 20ml is added, ultrasound
10min adds methanol solution to be diluted to scale constant volume, shakes up, and filtration takes subsequent filtrate to make test solution；
(3) prepared by reference substance solution
It is appropriate that qinghaosu, dihydroartemisinine, Artesunate reference substance are weighed respectively, it is accurately weighed, it sets in 50ml measuring bottle and adds
Methanol 15ml, ultrasonic 20min add methanol solution to be diluted to scale constant volume, shake up, and compare product solution；
(4) it measures
Precision measures test solution and injects Ultra Performance Liquid Chromatography instrument, records chromatogram；Another accurate measure mixes control
Product solution injects high performance liquid chromatograph, is measured in the same method；By external standard method with qinghaosu, double hydrogen sweet wormwoods in calculated by peak area test sample
The content of element, Artesunate.
The utility model has the advantages that the present invention provides and a kind of utilizes qinghaosu, dihydroartemisinine, sweet wormwood amber in the synchronous test sample of UPLC
The method of ester content, to sweet wormwood sample be not necessarily to derivatization treatment, using UPLC method directly detect qinghaosu, dihydroartemisinine,
The content of Artesunate sample, using Shimadzu Shim-pack XR-ODS II as chromatographic column, 0.1% acetum-acetonitrile is flowing
Isocratic elution is mutually carried out, method specificity is strong, precision is high, stability is good, accuracy is high.When the present invention is obviously shortened detection
Between, simplify experimental procedure, reduce test error, the intact isolated qualitative, quantitative point of three kinds of samples can be realized in 15 minutes
Analysis, greatly improves working efficiency.Reagent of the present invention and consumptive material are cheap and easy to get, and operation is quick and easy, practical, are suitable for
Content detection containing qinghaosu, dihydroartemisinine, Artesunate sample.
Detailed description of the invention:
Fig. 1: reference substance mixed solution map；
Fig. 2: Artemisinin solid paste map；
Fig. 3: sweet wormwood extracting solution map.
Specific embodiment:
In order to keep the purpose of the present invention, technical scheme and beneficial effects clearer, in conjunction with attached drawing to of the invention preferred
Embodiment is described in detail:
One, detection method of content
1, chromatographic condition
Chromatographic column: Shimadzu Shim-pack XR-ODS II
Mobile phase and type of elution: with 0.1% acetum: acetonitrile (50:50) is mobile phase isocratic elution
Flow velocity: 0.3ml/min
Detection wavelength: 194nm
Column temperature: 30 DEG C
Sample volume: 5 μ l
2, prepared by test solution
Test sample is solid sample: test sample is taken, it is accurately weighed, and it sets in 100ml measuring bottle, adds 50% methanol 20ml, ultrasound
30min adds methanol solution to be diluted to scale constant volume, shakes up, and filtration takes subsequent filtrate to make test solution.
Test sample is fluid sample: appropriate test sample taken with liquid-transfering gun, is set in 100ml measuring bottle, methanol 20ml is added, ultrasound
10min adds methanol solution to be diluted to scale constant volume, shakes up, and filtration takes subsequent filtrate to make test solution.
3, prepared by reference substance solution
It is appropriate that qinghaosu, dihydroartemisinine, Artesunate reference substance are weighed respectively, it is accurately weighed, it sets in 50ml measuring bottle, adds
Methanol 15ml, ultrasonic 20min add methanol solution to be diluted to scale constant volume, shake up, and compare product solution.
4, it measures
Precision measures test solution and injects Ultra Performance Liquid Chromatography instrument, records chromatogram；Another accurate measure mixes control
Product solution injects high performance liquid chromatograph, is measured in the same method；By external standard method with qinghaosu, double hydrogen sweet wormwoods in calculated by peak area test sample
The content of element, Artesunate, calculation formula are as follows:
A is supplied: test sample peak area；A pairs: reference substance peak area；C is supplied: test sample concentration；C pairs: reference substance concentration
M: sample relative molecular mass number；Test sample is solid, and unit mg/g, test sample is liquid, unit mg/ml.
Two, the verifying of content assaying method
1, specificity is investigated
By the above chromatographic condition, precision measures reference substance mixed solution, Qinghaosu solution, dihydroartemisinine pair
Sample introduction is distinguished according to product solution, Artesunate reference substance solution, test solution, the negative sample solution of non-dosing material, mobile phase
5.0 μ L acquire chromatographic data, record chromatogram.The chromatogram of reference substance mixed solution is shown in Fig. 1, determines qinghaosu, double hydrogen sweet wormwoods
The chromatography Average residence time of element (for epimer, therefore two chromatographic peaks), Artesunate, three compounds separation are good,
And separating degree is all larger than 1.5, while theoretical cam curve is all larger than 5000, meets content detection and requires (being shown in Table 1), from chromatogram
As can be seen that the retention time of each monomeric compound chromatographic peak is consistent, mobile phase and extraction reagent do not significantly interfere with sample,
It further illustrates under chromatographic condition of the present invention, three compounds can realize intact separation detection.
1 qinghaosu of table, dihydroartemisinine, Artesunate Average residence time, separating degree, theoretical cam curve
| Compound | Average residence time (min) | Separating degree (R) | Theoretical cam curve (USP) |
| Qinghaosu | 10.409 | -- | 5799 |
| Dihydroartemisinine | 4.221 | 11.118 | 9145 |
| Dihydroartemisinine | 7.105 | 5.807 | 8243 |
| Artesunate | 9.137 | 2.95 | 8202 |
2, linear relationship is investigated
The reference substance of 0.5mg/mL is configured as stock solution, precision pipette reference substance stock solution 0.05,0.1,0.2,1,
2,5,10mL are respectively placed in 10mL volumetric flask, with methanol dilution to scale.Sample introduction is distinguished by the above chromatographic condition, records color
Spectrogram carries out linear regression to sample introduction concentration (x) with peak area (y), obtains the equation of linear regression and correlation of three compounds
Coefficient r and the range of linearity (being shown in Table 2), show three kinds of compounds in the linear range, and peak area and concentration presentation are good linear
Relationship can be used for content detection.
2 linear relationship of table investigates result
| Compound | Regression equation | r | The range of linearity (μ g/mL) |
| Qinghaosu | Y=61004x-128.03 | 0.998 | 1.2~450 |
| Dihydroartemisinine | Y=81004x-114.03 | 0.997 | 2.4~392 |
| Artesunate | Y=120004x-241.03 | 0.996 | 2.1~750 |
3, precision is investigated
Precision weighs that each reference substance is appropriate, the solution that concentration is 0.096mg/mL is configured to, by the above chromatographic condition sample introduction 5
μ L is continuously repeated sample introduction 6 times, is recorded chromatographic peak area, and calculate relative standard deviation (RSD), be the results are shown in Table 3.It can from table 3
To find out, each Compound RS D is to meet regulation, shows that the method for the present invention precision is good.
3 precision of table investigates result
| Compound | RSD (%) |
| Qinghaosu | 1.23 |
| Dihydroartemisinine | 1.55 |
| Artesunate | 0.96 |
4, study on the stability
Precision weighs that each reference substance is appropriate, is configured to the solution that concentration is 0.096mg/mL, by the operation of the above chromatographic condition,
Respectively 0,1,2,4,6,12, sample introduction for 24 hours, measure chromatographic peak area, calculate the relative standard deviation of each compound peaks area
RSD the results are shown in Table 4.As can be seen from Table 4, each Compound RS D is to meet regulation, shows that the method for the present invention stability is good
It is good.
4 study on the stability result of table
| Compound | RSD (%) |
| Qinghaosu | 1.59 |
| Dihydroartemisinine | 1.98 |
| Artesunate | 1.45 |
5, accuracy is investigated
Sample recovery rate experiment: precision weighs 9 parts of extracting solution test solution, and every part of 1mL is equally divided into 3 groups and sets respectively
In 50mL volumetric flask, the accurate reference substance that corresponding amount is added, is prepared into test solution to be measured, by the above chromatographic condition respectively
Operation calculates sample recovery rate and relative standard deviation RSD, as a result see the table below 5.According to the experimental results, this method result is quasi-
It is really reliable.
Result is investigated in 5 accuracy of table
| Compound | RSD (%) | It is loaded average recovery rate (%) |
| Qinghaosu | 1.92 | 99.86 |
| Dihydroartemisinine | 1.56 | 100.85 |
| Artesunate | 1.09 | 101.23 |
6, quantitative limit and detection limit are investigated
It is appropriate that precision weighs reference substance, is diluted step by step with methanol solution.It is measured by the above liquid phase chromatogram condition sample introduction, record
Chromatogram.It is detection limit with signal-to-noise ratio 3:1, using signal-to-noise ratio 10:1 as quantitative limit, testing result is shown in Table 6.
6 quantitative limit of table and detection limit investigate result
| Compound | Quantitative limit (μ g/mL) | Detection limit (μ g/mL) |
| Qinghaosu | 0.085 | 0.28 |
| Dihydroartemisinine | 0.095 | 0.32 |
| Artesunate | 0.056 | 0.19 |
Three, sample detection
1, solid sample
Test sample preparation: taking Chinese medicine sweet wormwood 5g, by decocting extraction, extracts medical fluid 100mL, is condensed into cream, spare.Essence
The close paste 1g that weighs is placed in 100mL measuring bottle, and methanol 20ml, ultrasonic 10min is added to add methanol solution to be diluted to scale constant volume, shake
Even, filtration takes subsequent filtrate to make test solution.
Reference substance preparation: qinghaosu, dihydroartemisinine, Artesunate reference substance about 10mg, accurately weighed, addition are taken respectively
Methanol makes dissolved dilution constant volume, as reference substance solution；
By aforementioned chromatographic condition measurement qinghaosu, dihydroartemisinine, Artesunate content, test sample result see Fig. 2 and
Table 7.As shown in Figure 2, qinghaosu, dihydroartemisinine, Artesunate content be respectively 0.32mg/g, 0.16mg/g, 0.36mg/
g。
7 qinghaosu of table, dihydroartemisinine, Artesunate Average residence time, separating degree, theoretical cam curve
2, fluid sample
Test sample preparation: taking Chinese medicine sweet wormwood 5g, and by alcohol extraction, it is spare to extract medical fluid 50mL.Precision weighs extraction
Medical fluid 1mL is placed in 100mL measuring bottle, and methanol 20ml, ultrasonic 10min is added to add methanol solution to be diluted to scale constant volume, shake up, filter
It crosses, subsequent filtrate is taken to make test solution.
Reference substance preparation: qinghaosu, dihydroartemisinine, Artesunate reference substance about 10mg, accurately weighed, addition are taken respectively
Methanol makes dissolved dilution constant volume, as reference substance solution；
By aforementioned chromatographic condition measurement qinghaosu, dihydroartemisinine, Artesunate content, test sample result see Fig. 3 and
Table 8.From the figure 3, it may be seen that the content of qinghaosu, dihydroartemisinine, Artesunate is respectively 0.40mg/g, 0.06mg/g, 0.16mg/
g。
8 qinghaosu of table, dihydroartemisinine, Artesunate Average residence time, separating degree, theoretical cam curve
| Compound | Average residence time (min) | Separating degree (R) | Theoretical cam curve (USP) |
| Qinghaosu | 10.409 | -- | 5655 |
| Dihydroartemisinine | 4.061 | 11.465 | 7842 |
| Dihydroartemisinine | 7.150 | 7.304 | 9853 |
| Artesunate | 9.767 | 1.625 | 11049 |
Finally illustrate: preferred embodiment above is only used to illustrate the technical scheme of the present invention and not to limit it, although by upper
It states preferred embodiment the present invention is described in detail, however, those skilled in the art should understand that, can be in form
Various changes are made to it in upper and details, without departing from claims of the present invention limited range.
Claims (5)
1. it is a kind of using qinghaosu in the synchronous test sample of UPLC, dihydroartemisinine, Artesunate content method, feature exists
In using following chromatographic condition: chromatographic column: Shimadzu Shim-pack XR-ODS II；Mobile phase and type of elution: with 0.1% vinegar
Acid solution: acetonitrile (50:50) is mobile phase isocratic elution；Flow velocity: 0.1 ~ 1.0ml/min；Detection wavelength: 190 ~ 210nm；Column
Temperature: 25-35 DEG C；Sample volume: 1-20 μ l.
2. the method as described in claim 1, including following experimental procedure:
(1) chromatographic condition
Chromatographic column: Shimadzu Shim-pack XR-ODS II
Mobile phase and type of elution: with 0.1% acetum: acetonitrile (50:50) is mobile phase isocratic elution
Flow velocity: 0.1 ~ 1.0ml/min
Detection wavelength: 190 ~ 210nm
Column temperature: 25-35 DEG C
Sample volume: 1-20 μ l；
(2) prepared by test solution
Precision measurement Artemisia annua extract is appropriate, adds methanol that test solution is made；
(3) prepared by reference substance solution
Qinghaosu, dihydroartemisinine, Artesunate reference substance are taken, accurately weighed appropriate, methanol, which is added, to be made to dissolve and dilute constant volume,
As reference substance solution；
(4) it measures
Precision measures test solution and injects Ultra Performance Liquid Chromatography instrument, records chromatogram；Another accurate measurement mixing reference substance is molten
Liquid injects high performance liquid chromatograph, is measured in the same method；By external standard method with qinghaosu in calculated by peak area test sample, dihydroartemisinine,
The content of Artesunate.
3. method according to claim 2, which is characterized in that the preparation of the test solution uses following methods:
A. test sample is solid sample: test sample is taken, it is accurately weighed, and it sets in 100ml measuring bottle, adds 50% methanol 20ml, ultrasound
30min adds methanol solution to be diluted to scale constant volume, shakes up, and filtration takes subsequent filtrate to make test solution；
B. test sample is fluid sample: appropriate test sample is taken with liquid-transfering gun, is set in 100ml measuring bottle, methanol 20ml is added, ultrasound
10min adds methanol solution to be diluted to scale constant volume, shakes up, and filtration takes subsequent filtrate to make test solution.
4. method according to claim 2, which is characterized in that the preparation of the reference substance solution uses following methods:
It is appropriate that qinghaosu, dihydroartemisinine, Artesunate reference substance are weighed respectively, it is accurately weighed, it sets in 50ml measuring bottle, adds methanol
15ml, ultrasonic 20min add methanol solution to be diluted to scale constant volume, shake up, and compare product solution.
5. it is a kind of using qinghaosu in the synchronous test sample of UPLC, dihydroartemisinine, Artesunate content method, including it is following
Experimental procedure:
(1) chromatographic condition
Chromatographic column: Shimadzu Shim-pack XR-ODS II
Mobile phase and type of elution: with 0.1% acetum: acetonitrile (50:50) is mobile phase isocratic elution
Flow velocity: 0.3ml/min
Detection wavelength: 194nm
Column temperature: 30 DEG C
Sample volume: 5 μ l；
(2) prepared by test solution
A. test sample is solid sample: test sample is taken, it is accurately weighed, and it sets in 100ml measuring bottle, adds 50% methanol 20ml, ultrasound
30min adds methanol solution to be diluted to scale constant volume, shakes up, and filtration takes subsequent filtrate to make test solution；
B. test sample is fluid sample: appropriate test sample is taken with liquid-transfering gun, is set in 100ml measuring bottle, methanol 20ml is added, ultrasound
10min adds methanol solution to be diluted to scale constant volume, shakes up, and filtration takes subsequent filtrate to make test solution；
(3) prepared by reference substance solution
It is appropriate that qinghaosu, dihydroartemisinine, Artesunate reference substance are weighed respectively, it is accurately weighed, it sets in 50ml measuring bottle, adds methanol
15ml, ultrasonic 20min add methanol solution to be diluted to scale constant volume, shake up, and compare product solution；
(4) it measures
Precision measures test solution and injects Ultra Performance Liquid Chromatography instrument, records chromatogram；Another accurate measurement mixing reference substance is molten
Liquid injects Ultra Performance Liquid Chromatography instrument, is measured in the same method；By external standard method with qinghaosu, double hydrogen sweet wormwoods in calculated by peak area test sample
The content of element, Artesunate.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114200024A (en) * | 2020-09-17 | 2022-03-18 | 昆药集团股份有限公司 | Method for determining dissolution and detection of dihydroartemisinin tablet |
| CN117007733A (en) * | 2023-09-28 | 2023-11-07 | 威胜生物医药(苏州)股份有限公司 | High performance liquid chromatography determination method for five components in sweet wormwood herb paste |
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