CN104215703A - Detection method of glycoside macro-molecule substances in qi-tonifying pulse-restoring injection preparation - Google Patents
Detection method of glycoside macro-molecule substances in qi-tonifying pulse-restoring injection preparation Download PDFInfo
- Publication number
- CN104215703A CN104215703A CN201310219306.1A CN201310219306A CN104215703A CN 104215703 A CN104215703 A CN 104215703A CN 201310219306 A CN201310219306 A CN 201310219306A CN 104215703 A CN104215703 A CN 104215703A
- Authority
- CN
- China
- Prior art keywords
- mobile phase
- sample
- chromatographic
- detection method
- water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a detection method of glycoside macro-molecule substances in a qi-tonifying pulse-restoring injection (lyophilized) preparation, and in particular, the method is as follows: high performance gel chromatography and an ELSD (evaporative light-scattering detector) are used for detecting, an Ultrahydrogel 250 chromatographic column (7.8mm * 300mm) is used, and cyclohexanecarboxylic acid-water (1:5000) as a mobile phase is used for detecting, the liquid phase detection method of the macro-molecule substances (impurities) is established, and the method is simple in operation, reliable in results and high in sensitivity.
Description
Technical field:
The present invention relates to a kind of detection method of Chinese medicine composition, particularly the detection method of glucosides class macromolecular substances in a kind of injection Yiqi and vein recovery.
Background technology:
Injection Yiqi and vein recovery (freeze-drying) is the compound Chinese medicinal preparation for drip-feed be made up of red ginseng, the tuber of dwarf lilyturf, the fruit of Chinese magnoliavine, has Yiqi and vein recovery, the curative effect of nourishing Yin and promoting production of body fluid.Be mainly used in coronary heart disease angina pectoris syndrome of deficiency of both qi and yin, disease sees chest impediment and cardialgia, palpitation, tiredness with no desire to speak, have a dizzy spell, lustreless complexion, tongue are light, few tongue or stripping tongue, arteries and veins thin and delicate or knot generation; Chronic left heart insufficiency II, III level syndrome of deficiency of both qi and yin caused by coronary heart disease, disease sees palpitaition, breathe hard very then that One's breath come in heavy gasps, sensation of oppression and faint pain in the chest, time when doing only, lassitude hypodynamia, pale complexion, dynamic then sweating, light, the few tongue of tongue or stripping tongue, arteries and veins is thin and delicate or tie generation.
Injection Yiqi and vein recovery (freeze-drying) proterties: this product is lurid loose block; Have draw moist.Get 1 bottle of content, the 2 ~ 3ml that adds water is brownish red clear liquid after dissolving.Usage and dosage: drip-feed.Every day 1 time, each 8 bottles, instil with 250ml ~ 500ml5% glucose injection or normal saline dilution posterior vein.About 40 per minute.2 weeks courses for the treatment of.
In injection Yiqi and vein recovery (freeze-drying), principal ingredient is carbohydrate, saponins, lignanoids etc.; Wherein main active is saponins and lignanoids, and its molecular weight great majority are no more than 1000.And the material that in prescription medicinal material, relative molecular weight is larger such as protein, polypeptide, polysaccharide etc. are the materials removed as impurity in extraction and preparation technical process.Should not exist in finished product, for ensureing the quality of finished product, be necessary the detection these macromolecular substances being carried out to quantitative and qualitative analysis, this research is just based on above-mentioned situation, establish High Performance Gel Permeation liquid chromatography-evaporative light-scattering detector and the macromolecule material that may exist in injection Yiqi and vein recovery (freeze-drying) is detected, the method is easy, quick and sensitive, can be used for the inspection of macromolecular substances in injection Yiqi and vein recovery (freeze-drying), has Practical significance.
Summary of the invention:
The invention provides the detection method of glucosides class macromolecular substances in a kind of injection Yiqi and vein recovery, described method is for detect injection Yiqi and vein recovery sample with High Performance Gel Permeation liquid chromatography-evaporative light-scattering detector, and wherein, sample detection parameter is:
Chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses; Column temperature 25-35 DEG C; Mobile phase is formic acid-water 1:2500-7500, and mobile phase pH value is 2-7; Flow velocity 0.25-0.75mlmin
-1; Detecting device: drift pipe temperature 85-100 DEG C, air pressure 0.1-0.2MPa.
Wherein sample detection mobile phase is preferably formic acid-water 1:5000, flow velocity 0.5mlmin
-1, mobile phase pH value is 3; Detector parameters is preferred: drift pipe temperature 90 DEG C, air pressure 0.17MPa;
The detection method of glucosides class macromolecular substances in above-mentioned injection Yiqi and vein recovery, preferred detection parameter is: column temperature 30 DEG C; Mobile phase is formic acid-water 1:5000, and mobile phase pH value is 3; Flow velocity 0.5mlmin
-1; Detector parameters: drift pipe temperature 90 DEG C, air pressure 0.17MPa.
The detection method of above-mentioned High Performance Gel Permeation liquid chromatography-evaporative light-scattering detector to injection Yiqi and vein recovery (freeze-drying) sample comprises:
The preparation of detected sample, method is as follows:
Get injection Yiqi and vein recovery (freeze-drying) to be mixed with concentration with ultrapure water and to be about 50-200mgmL
-1solution, shake up, through 0.25-0.65 μm of filtering with microporous membrane, obtain detected sample.
Sample detection, method is as follows:
Get subsequent filtrate, injecting chromatograph, sample size 10-40 μ L,
Described chromatographic chromatographic condition is as follows:
Chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm);
Column temperature (25-35) DEG C;
Mobile phase is formic acid-water (1:2500-7500), and mobile phase pH value is 2-7;
Flow velocity 0.25-0.75mlmin
-1.
Detector parameters: drift pipe temperature 85-100 DEG C, air pressure 0.1-0.2MPa; Sample size 10-40 μ L.
What retention time was greater than 16min is small molecular weight material, and retention time is macromolecular substances in 16min.
The most preferred technical scheme of the present invention is as follows:
Wherein, the compound method of sample:
Get 1 injection Yiqi and vein recovery (freeze-drying) to be mixed with concentration with ultrapure water and to be about 100mgmL
-1solution, shake up, through 0.45 μm of filtering with microporous membrane, get subsequent filtrate, sample introduction 20 μ L,
Chromatographic condition:
Chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm);
Column temperature 30 DEG C;
Mobile phase is formic acid-water 1:5000, and mobile phase pH value is 3;
Flow velocity 0.5mlmin
-1.
Detector parameters: drift pipe temperature 90 DEG C, air pressure 0.17MPa; Sample size 20 μ L.
Detection method of the present invention, be through screening obtain, screening process is as follows:
1 instrument and reagent
Waters2695 high performance liquid chromatograph, Waters2420 type evaporative light-scattering detector, Waters Ultrahydrogel
tM250 gel chromatographic columns (7.8 × 300mm; I.d), XWK-3A pneumatic pump (Tianjin Hua Sheng analytical instrument factory), XS105 plum Teller precise electronic analytical balance (plum Teller-Tuo benefit instrument Shanghai, Shanghai company limited); By Chinese pharmaceutical biological product, dextran molecular weight standards identifies that institute provides (lot number 140640-201002 relative molecular weight divides 180,2500,4600,7100,10000,21400,41100,84400),
Injection Yiqi and vein recovery (freeze-drying) is provided by Tianjin TianShiLi ZhiJiao Medicine Co., Ltd, and ultrapure water is made by oneself.
2 methods and result
2.1 chromatographic condition chromatographic columns are Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm); Column temperature 30 DEG C; Mobile phase is formic acid-water; Mobile phase PH3 flow velocity 0.5mlmin
-1.Detector parameters: drift pipe temperature 90 DEG C, air pressure 0.17MPa; Sample size 20 μ L.
It is 180,2500 that 2.2 molecular weight marker thing retention times investigation precisions take 8 parts of relative molecular weights, and the dextran molecular weight standards of 4600,7100,10000,21400,41100,84400 is appropriate, is mixed with each about 1mgmL with ultrapure water
-1solution, simultaneously water makes placebo solution, and by above-mentioned chromatographic process, sample introduction respectively, measure, Fig. 1 is shown in by collection of illustrative plates.Fig. 1 is the HPGPC figure of 8 kinds of different molecular weight dextran reference substances and blank
The retention time of table 18 kind of different molecular weight dextran reference substance
As can be known from Table 1, what retention time was greater than 16min is small molecular weight material, thus retention time in 16min can initial guess its be macromolecular substances.
2.3 instrument precisions investigate the dextran reference substance that precision weighing relative molecular weight is 2500,4600,10000 and 41100, and dilute with water obtains concentration is respectively 1.18mgmL
-1, 1.24mgmL
-1, 1.32mgmL
-1and 1.36mgmL
-1solution, continuous sample introduction 6 times respectively, measures retention time, calculates RSD, result relative molecular weight be the dextran standard items of 4600,10000 and 41100 and the RSD of retention time be respectively 0.3093%, 0.3077% and 0.2717%(n=6), show that precision is good.
2.4 reference substance minimum detectability precision weighings get the dextran reference substance that relative molecular mass is 2500,4600,10000 and 41100, are diluted with water to scale respectively, and obtaining concentration is 2.16mgmL
-1, 2.25mgmL
-1, 2.08mgmL
-1and 2.44mgmL
-1solution, be diluted to 20 times, 50 times, 100 times, 200 times, 500 times respectively, sample introduction analysis.Result shows, the two is when being diluted to 500 times, and the s/n at sample peak is respectively 7.52,3.13,4.21 and 3.37, therefore can judge that the minimum detectability of dextran reference substance is about 4 μ gmL
-1.
2.5 reference substance study on the stability get above-mentioned two kinds of dextran reference substance solution, place 0 respectively, 2,4, measure by above-mentioned condition sample introduction after 8h, calculate the RSD of retention time, result relative molecular mass be the RSD of the retention time of the dextran reference substance of 2500,4600,10000 and 41100 be respectively 0.1013%, 0.1260%, 0.3514% and 0.2237%(n=4), result shows that reference substance solution is stable in 8h.
2.6 sample determinations are got 10 batches of injection Yiqi and vein recoveries (freeze-drying) (source: Tianjin TianShiLi ZhiJiao Medicine Co., Ltd) respectively and are mixed with concentration with ultrapure water and are about 100mgmL
-1solution, shake up, through 0.45 μm of filtering with microporous membrane, get subsequent filtrate, sample introduction 20 μ L, measures by the method described in step 2.1.Sample HPGPC figure is shown in Fig. 2.Fig. 2 is the HPGPC-ELSD figure of 10 batches of injection Yiqi and vein recoveries (freeze-drying)
Result show survey the appearance time of 10 batches of injection Yiqi and vein recovery (freeze-drying) samples all higher than 16min, can in judgement sample without macromolecular substances or content extremely low.
For proving the accuracy of this method, in above 3 batch samples without large molecular saccharides glycoside material, add dextran reference substance carry out measuring to obtain figure below, Fig. 3 is that to add molecular weight be the HPGPC-ELSD figure molecular weight of the dextran standard items of 10000 and 41100 to 3 batches of injection Yiqi and vein recoveries (freeze-drying) is that the concentration of the dextran standard items of 10000 and 41100 is about 1mgmL
-, other parameters are identical with said method.
2.7 sample stabilities are investigated precisions and are taken injection Yiqi and vein recovery (freeze-drying), are settled to 100mgmL by water-soluble solution
-1.Investigate respectively 0,2,4,8h place after sample stability in aqueous.Gained HPGPC figure is shown in Fig. 4.Fig. 4 sample stability in aqueous
Result shows: sample dissolution in aqueous solution after measure in 8h and do not occur macromolecular substances, therefore 8h internal stability is good in aqueous for sample.
3 analysis and thinking
3.1 at present, and the detection for the macromolecule material in traditional Chinese medicine there is no regulation method can be for referencial use.Method validation in this research carries out with reference to " Chinese Pharmacopoeia " 2010 editions one annex Ⅹ VIII quality standards in Chinese drugs analytical approach checking basic demand.
3.2 in conjunction with the result of study of the ratio shared by all kinds of chemical compositions in injection Yiqi and vein recovery (freeze-drying): wherein total sugar content ratio is in the formulation higher, therefore this research selects dextran molecular weight standards for mark, set up detection method, for detecting the macromolecular substances that may exist in preparation.
The separation principle of 3.3 foundation gel chromatographies: under same experimental conditions, the molecular mass of material is larger, more not easily enters gel voids, and retention time is shorter; Otherwise molecular mass is less, retention time is longer.Set up the method based on this principle to detect the macromolecule material in traditional Chinese medicine production process, simple and easy to do.And by contrasting the applicability of Ultrahydrogel250 chromatographic column (7.8mm × 300mm) and Ultrahydrogel500 chromatographic column (7.8mm × 300mm), select Ultrahydrogel250 chromatographic column (7.8mm × 300mm) to carry out method for building up.Evaporative light-scattering detector is designed for highly effective liquid phase chromatographic system, comparatively before utilize differential refraction detector to set up method advantageously, the method not only stability is higher, and precision is better, and sensitivity is also improved, minimum detectability (in dextran) is about 4 μ gmL
-1.In addition, evaporative light-scattering detector may be used for gradient elution, and comparatively differential refraction detector is more convenient for further Optimal Development new method, and the scope of application is wider.
3.4 the method formic acid-water are that mobile phase pH is about 3.0, can get rid of saponins and lignan component and produce interference because of physics polymerized form aggregation to macromolecular substances measurement result through investigating with this flowing.Sample concentration is selected to be about 100mgmL during sample determination
-1, namely get this product 1 bottle, inject and make dissolving with water 2.6mL; With RID detection method
[4]compare and both significantly increased sample concentration, improve the sensitivity detecting macromolecular substances in sample, consistent with the concentration of defects inspecting in injection Yiqi and vein recovery (freeze-drying) quality standard check item again.
The following drawings is 10 batch products of appeal sugar-free glycoside material, but in mensuration process, mobile phase changes water into, and additive method is identical with the application's assay method: Fig. 5, the HPGPC-ELSD figure of 10 batches of injection Yiqi and vein recoveries (freeze-drying).
Known when mobile phase is water by above accompanying drawing, itself is not containing the sample of large molecular saccharides glycoside material, occur in its collection of illustrative plates that large molecule is mixed peak, illustrated and saponins and lignan component can not be avoided to produce interference because of physics polymerized form aggregation to macromolecular substances measurement result when taking water as mobile phase.
This research adopts HPGPC-ELSD to detect glucosides class macromolecular substances in injection Yiqi and vein recovery (freeze-drying), easy and simple to handle, result prepares reliable and sensitivity is higher, could measure after original assay method detecting device will balance 12h, each sample determination needs 1h, and sensitivity is not high.New method: the short about 15-30min of detecting device equilibration time, each sample determination needs 30min, highly sensitive.。Therefore, this method can be used as the method for quality control of Yiqi and vein recovery (freeze-drying).Also the detection that can be macromolecular substances in other Chinese medicine preparations provides reference.
Assay method of the present invention is effective, simple to operate, is easy to promote.
Accompanying drawing explanation
Figure 18 plants the HPGPC figure of different molecular weight dextran reference substance and blank
The HPGPC-ELSD figure of Figure 21 0 batch of injection Yiqi and vein recovery (freeze-drying)
Figure 33 criticizes injection Yiqi and vein recovery (freeze-drying), and to add molecular weight be the dextran standard items of 10000 and 41100
Fig. 4 sample stability in aqueous
The HPGPC-ELSD figure of Figure 51 0 batch of injection Yiqi and vein recovery (freeze-drying)
Embodiment:
By the following examples, further illustrate the present invention, but not as limitation of the present invention.
Embodiment 1
Get injection Yiqi and vein recovery (freeze-drying) (source: Tianjin TianShiLi ZhiJiao Medicine Co., Ltd) to be mixed with concentration with ultrapure water and to be about 100mgmL
-1solution, shake up, through 0.45 μm of filtering with microporous membrane, get subsequent filtrate, sample introduction 20 μ L, chromatographic condition: chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm); Column temperature 30 DEG C; Mobile phase is formic acid-water (1:5000); Mobile phase PH3, flow velocity 0.5mlmin
-1.Detector parameters: drift pipe temperature 90 DEG C, air pressure 0.17MPa; Sample introduction measures.
Testing result: retention time is greater than 16min
Embodiment 2
Get injection Yiqi and vein recovery (freeze-drying) (source: Tianjin TianShiLi ZhiJiao Medicine Co., Ltd) to be mixed with concentration with ultrapure water and to be about 50mgmL
-1solution, shake up, through 0.25 μm of filtering with microporous membrane, get subsequent filtrate, sample introduction 10 μ L, chromatographic condition: chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm); Column temperature 25 DEG C; Mobile phase is formic acid-water (1:2500); Mobile phase PH2, flow velocity 0.25mlmin
-1.Detector parameters: drift pipe temperature 85 DEG C, air pressure 0.1MPa; Sample introduction measures.
Embodiment 3
Get injection Yiqi and vein recovery (freeze-drying) (source: Tianjin TianShiLi ZhiJiao Medicine Co., Ltd) to be mixed with concentration with ultrapure water and to be about 200mgmL
-1solution, shake up, through 0.65 μm of filtering with microporous membrane, get subsequent filtrate, sample introduction 40 μ L, chromatographic condition: chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm); Column temperature 35 DEG C; Mobile phase is formic acid-water (1:7500); Mobile phase PH7, flow velocity 0.75mlmin
-1.Detector parameters: drift pipe temperature 100 DEG C, air pressure 0.2MPa; Sample introduction measures.
Embodiment 4
Get injection Yiqi and vein recovery (freeze-drying) (source: Tianjin TianShiLi ZhiJiao Medicine Co., Ltd) to be mixed with concentration with ultrapure water and to be about 150mgmL
-1solution, shake up, through 0.5 μm of filtering with microporous membrane, get subsequent filtrate, sample introduction 30 μ L, chromatographic condition: chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm); Column temperature 35 DEG C; Mobile phase is formic acid-water (1:6000); Mobile phase PH5, flow velocity 0.65mlmin
-1.Detector parameters: drift pipe temperature 85 DEG C, air pressure 0.17MPa; Sample introduction measures.
Embodiment 5
Get injection Yiqi and vein recovery (freeze-drying) (source: Tianjin TianShiLi ZhiJiao Medicine Co., Ltd) to be mixed with concentration with ultrapure water and to be about 80mgmL
-1solution, shake up, through 0.45 μm of filtering with microporous membrane, get subsequent filtrate, sample introduction 20 μ L, chromatographic condition: chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm); Column temperature 30 DEG C; Mobile phase is formic acid-water (1:4000); Mobile phase PH6, flow velocity 0.7mlmin
-1.Detector parameters: drift pipe temperature 95 DEG C, air pressure 0.17MPa; Sample introduction measures.
Embodiment 6
Detection method comprises:
The preparation of detected sample, method is as follows:
Get injection Yiqi and vein recovery (freeze-drying) to be mixed with concentration with ultrapure water and to be about 50mgmL
-1solution, shake up, through 0.5 μm of filtering with microporous membrane, obtain detected sample.
Sample detection, method is as follows:
Get subsequent filtrate, injecting chromatograph, sample size 30 μ L,
Described chromatographic chromatographic condition is as follows:
Chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm);
Column temperature (25) DEG C;
Mobile phase is formic acid-water (1:2500), and mobile phase pH value is 2;
Flow velocity 0.25mlmin
-1.
Detector parameters: drift pipe temperature 85 DEG C, air pressure 0.1MPa; Sample size 20 μ L.
Testing result: retention time is greater than 16min.
Embodiment 7
Detection method comprises:
The preparation of detected sample, method is as follows:
Get injection Yiqi and vein recovery (freeze-drying) to be mixed with concentration with ultrapure water and to be about 200mgmL
-1solution, shake up, through 0.65 μm of filtering with microporous membrane, obtain detected sample.
Sample detection, method is as follows:
Get subsequent filtrate, injecting chromatograph, sample size 40 μ L,
Described chromatographic chromatographic condition is as follows:
Chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm);
Column temperature (35) DEG C;
Mobile phase is formic acid-water (1:7500), and mobile phase pH value is 7;
Flow velocity 0.75mlmin
-1.
Detector parameters: drift pipe temperature 100 DEG C, air pressure 0.2MPa; Sample size 40 μ L.
Testing result: retention time is greater than 16min.
Embodiment 8
Detection method comprises:
The preparation of detected sample, method is as follows:
Get injection Yiqi and vein recovery (freeze-drying) to be mixed with concentration with ultrapure water and to be about 150mgmL
-1solution, shake up, through 0.5 μm of filtering with microporous membrane, obtain detected sample.
Sample detection, method is as follows:
Get subsequent filtrate, injecting chromatograph, sample size 30 μ L,
Described chromatographic chromatographic condition is as follows:
Chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm);
Column temperature (25) DEG C;
Mobile phase is formic acid-water (1:2500), and mobile phase pH value is 2;
Flow velocity 0.25mlmin
-1.
Detector parameters: drift pipe temperature 85 DEG C, air pressure 0.1MPa; Sample size 10 μ L.
Testing result: retention time is greater than 16min.
Claims (6)
1. the detection method of glucosides class macromolecular substances in injection Yiqi and vein recovery, described method, for detect injection Yiqi and vein recovery sample with High Performance Gel Permeation liquid chromatography-evaporative light-scattering detector, is characterized in that, sample detection parameter is:
Chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses;
Column temperature 25-35 DEG C;
Mobile phase is formic acid-water 1:2500-7500, and mobile phase pH value is 2-7;
Flow velocity 0.25-0.75mlmin
-1;
Detecting device: drift pipe temperature 85-100 DEG C, air pressure 0.1-0.2MPa.
2. detection method as claimed in claim 1, it is characterized in that, sample detection mobile phase is formic acid-water 1:5000, flow velocity 0.5mlmin
-1, mobile phase pH value is 3.
3. detection method as claimed in claim 1, is characterized in that, detector parameters: drift pipe temperature 90 DEG C, air pressure 0.17MPa.
4. detection method as claimed in claim 1, it is characterized in that, sample detection parameter is:
Column temperature 30 DEG C;
Mobile phase is formic acid-water 1:5000, and mobile phase pH value is 3;
Flow velocity 0.5mlmin
-1;
Detector parameters: drift pipe temperature 90 DEG C, air pressure 0.17MPa.
5. detection method as claimed in claim 1, is characterized in that, said method comprising the steps of:
Detect injection Yiqi and vein recovery (freeze-drying) sample with High Performance Gel Permeation liquid chromatography-evaporative light-scattering detector, detection method comprises:
The preparation of detected sample, method is as follows:
Get injection Yiqi and vein recovery (freeze-drying) to be mixed with concentration with ultrapure water and to be about 50-200mgmL
-1solution, shake up, through 0.25-0.65 μm of filtering with microporous membrane, obtain detected sample;
Described chromatographic chromatographic condition is as follows:
Chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses (7.8 × 300mm);
Column temperature 25-35 DEG C;
Mobile phase is formic acid-water 1:2500-7500, and mobile phase pH value is 2-7;
Flow velocity 0.25-0.75mlmin
-1;
Detector parameters: drift pipe temperature 85-100 DEG C, air pressure 0.1-0.2MPa.
6. detection method as claimed in claim 5, is characterized in that, said method comprising the steps of:
Detect injection Yiqi and vein recovery (freeze-drying) sample with High Performance Gel Permeation liquid chromatography-evaporative light-scattering detector, detection method comprises:
The preparation of detected sample, method is as follows:
Get injection Yiqi and vein recovery (freeze-drying) to be mixed with concentration with ultrapure water and to be about 100mgmL
-1solution, shake up, through 0.45 μm of filtering with microporous membrane, obtain detected sample;
Described chromatographic chromatographic condition is as follows:
Chromatographic column is Waters Ultrahydrogel
tM250 gel chromatographic columnses;
Column temperature 30 DEG C;
Mobile phase is formic acid-water 1:5000, and mobile phase pH value is 3;
Flow velocity 0.5mlmin
-1;
Detector parameters: drift pipe temperature 90 DEG C, air pressure 0.17MPa.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310219306.1A CN104215703B (en) | 2013-06-04 | 2013-06-04 | The detection method of glycoside macromolecular substances in a kind of injection Yiqi and vein recovery |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310219306.1A CN104215703B (en) | 2013-06-04 | 2013-06-04 | The detection method of glycoside macromolecular substances in a kind of injection Yiqi and vein recovery |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104215703A true CN104215703A (en) | 2014-12-17 |
| CN104215703B CN104215703B (en) | 2017-06-23 |
Family
ID=52097417
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310219306.1A Active CN104215703B (en) | 2013-06-04 | 2013-06-04 | The detection method of glycoside macromolecular substances in a kind of injection Yiqi and vein recovery |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104215703B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104634913A (en) * | 2015-01-30 | 2015-05-20 | 北京赛升药业股份有限公司 | Quality control method of bozhi glycopeptide injection liquid |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004323687A (en) * | 2003-04-25 | 2004-11-18 | Nippon Synthetic Chem Ind Co Ltd:The | Ethylene-vinyl acetate copolymer saponification product and its application |
| JP2005106591A (en) * | 2003-09-30 | 2005-04-21 | Sumitomo Chemical Co Ltd | Analytical method for structural unit in acrylic resin |
| CN101539550A (en) * | 2009-04-28 | 2009-09-23 | 李绍平 | Qualitative and quantitative analysis method for polyoses |
| CN102243215A (en) * | 2010-05-10 | 2011-11-16 | 辽宁诺康医药有限公司 | Detection method for water-soluble glucomannan |
| CN102445500A (en) * | 2010-10-14 | 2012-05-09 | 天津天士力之骄药业有限公司 | Method for detecting macromolecular substances in Yiqifumai injection |
-
2013
- 2013-06-04 CN CN201310219306.1A patent/CN104215703B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004323687A (en) * | 2003-04-25 | 2004-11-18 | Nippon Synthetic Chem Ind Co Ltd:The | Ethylene-vinyl acetate copolymer saponification product and its application |
| JP2005106591A (en) * | 2003-09-30 | 2005-04-21 | Sumitomo Chemical Co Ltd | Analytical method for structural unit in acrylic resin |
| CN101539550A (en) * | 2009-04-28 | 2009-09-23 | 李绍平 | Qualitative and quantitative analysis method for polyoses |
| CN102243215A (en) * | 2010-05-10 | 2011-11-16 | 辽宁诺康医药有限公司 | Detection method for water-soluble glucomannan |
| CN102445500A (en) * | 2010-10-14 | 2012-05-09 | 天津天士力之骄药业有限公司 | Method for detecting macromolecular substances in Yiqifumai injection |
Non-Patent Citations (3)
| Title |
|---|
| 周如真: "GPC-LALLS联机测定右旋糖酐", 《中国药学杂志》 * |
| 林晓等: "凝胶过滤色谱-蒸发光散射法检测麦冬多糖色谱分离的研究", 《中草药》 * |
| 韩晓萍等: "HPGPC法检测注射用益气复脉（冻干）中的糖苷类大分子物质", 《中成药》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104634913A (en) * | 2015-01-30 | 2015-05-20 | 北京赛升药业股份有限公司 | Quality control method of bozhi glycopeptide injection liquid |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104215703B (en) | 2017-06-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102466658B (en) | Measurement method of content of 5-hydroxymethylfurfural (5-HMF) in injection | |
| CN102445500B (en) | Method for detecting macromolecular substances in Yiqifumai injection | |
| CN105092741B (en) | A kind of method that high performance liquid chromatography detects the caprolactam of 3 amino 2 | |
| CN101900712B (en) | Pharmacokinetic testing method of Exendin-4 | |
| CN103808835B (en) | The method of 10 kinds of chemical composition contents in HPLC-DAD method Simultaneously test Siwu Tang decoction | |
| CN103926332B (en) | A kind of Ultra Performance Liquid Chromatography method of uridine, guanosine and adenosine content in Simultaneously test pinellia tuber extract | |
| CN104678031B (en) | High performance liquid chromatography detects the method for atractyloside and/or earboxyatractylosida | |
| CN103604879B (en) | Detection method of polystictus versicolor and preparation containing polystictus versicolor | |
| CN106324167B (en) | Method for determining flavonoid components in astragalus extract by UPLC | |
| CN106124682B (en) | A kind of composition method of inspection of Radix Et Caulis Acanthopanacis Senticosi injection | |
| CN105954432B (en) | A kind of detection method of ginkolide B content | |
| CN105510452B (en) | Multi-target ingredient assay, fingerprint map construction and the preparation method of liver-benefiting eye-brightening oral liquid | |
| CN104215703B (en) | The detection method of glycoside macromolecular substances in a kind of injection Yiqi and vein recovery | |
| CN102068553B (en) | Method for constructing high performance liquid chromatographic (HPLC) fingerprint of Mammary lump preparation arising from qi stagnation and blood stasis | |
| CN104101657A (en) | Method for determining content of multiple components in Chinese medicinal composition preparation | |
| CN103115985B (en) | Method for synchronously detecting six flavonoid constituents in sowthistle-leaf ixeris seedling injection in HPLC-DAD (High Performance Liquid Chromatography-Diode Array detector) method | |
| CN111896637B (en) | Detection method of Jinqing intermediate and fingerprint spectrum construction method thereof | |
| CN113092640B (en) | Method for detecting benzyl alcohol and benzaldehyde in heparin sodium injection | |
| WO2009155756A1 (en) | Method for determining the contents of oligosaccharides in morinda officinalis chinese medicine or extraction thereof | |
| CN107655995A (en) | Method that is a kind of while detecting three kinds of flavone compounds in ginkgo leaf | |
| CN102590408A (en) | Method for detecting macromolecular substances in perhexiline injection | |
| CN107976494B (en) | Construction of standard characteristic spectrum of Kangfu tincture and quality detection method thereof | |
| CN109709241A (en) | The measuring method of high molecular weight substance in Chinese medicine injection containing liposoluble ingredient | |
| CN102520081B (en) | Detection method for traditional Chinese preparation capable of activating blood circulation to dissipate blood stasis | |
| CN103487517A (en) | Determination method for theophylline in doxofylline injection preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |