CN108020616A - The content assaying method of Astragaloside IV in the anti-Party A in Five Sacred Mountins - Google Patents
The content assaying method of Astragaloside IV in the anti-Party A in Five Sacred Mountins Download PDFInfo
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Abstract
The present invention relates to a kind of content assaying method of Astragaloside IV, and in particular to the content assaying method of Astragaloside IV in a kind of anti-Party A in Five Sacred Mountins.Using the method for high performance liquid chromatography evaporative photodetector, include the following steps：1) chromatographic condition is determined；2) preparation of reference substance solution；3) preparation of test solution：A, precision weighs test sample 5g, adds methanol and is heated to reflux, obtains extracting solution；B, extracting solution filters, and filter residue, merging filtrate are washed with methanol；C, the filtrate that is concentrated under reduced pressure is closely dry, adds aqueous alkali dissolving, is then extracted with water-saturated n-butanol, obtain extract；D, extract is washed with ammonia solution, is concentrated to dryness, and is added methanol constant volume to 10mL, is obtained test solution；4) measure.Negative control of the present invention does not interfere with, and has stronger specificity and good reappearance, and obtained chromatogram baseline is steady, and chromatographic peak peak shape is good, reaches baseline separation, is a kind of detection effective analysis method of Astragaloside IV component.
Description
Technical field
The present invention relates to a kind of content assaying method of Astragaloside IV, and in particular to Astragaloside IV in a kind of anti-Party A in Five Sacred Mountins
Content assaying method.
Background technology
The anti-Party A in Five Sacred Mountins is the clinical proved recipe of Shandong Prov. Hospital Zhao Jiajun president exploitation, mainly by radix astragali, ginseng etc. ten
Taste Chinese medicine forms, supplementing qi and nourishing yin, promoting blood circulation and removing blood stasis, softening and resolving hard mass, adjusts immune function.It is high to be clinically mainly used for thyroid function
Into, tubercle and simple goiter, endocrine exophthalmos etc..The medicine prescription is unique, meets the modern times to thyroid disease
Study of pathogenesis, not only Endocrine have dual regulation, also have certain effect to hemopoietic system and immune system, can rise
High leukocytic, elimination thyroid hormone secretion is excessive and uses the side effect caused by antithyroid skin medicine.
Using radix astragali as main ingredient in the anti-Party A in Five Sacred Mountins, sweet, tepor.Return lung, the spleen channel；Major function：Tonifying Qi and lifting yang, Gu table stops
Sweat, inducing diuresis for removing edema, blood-nourishing of promoting the production of body fluid, the stagnant logical numbness of row, pus draining and toxin expelling, expelling pus and promoting granulation；For treatment deficiency of vital energy weak, anorexia and loose stool, under middle gas
Fall into, rush down prolapse of the anus for a long time, uterine bleeding of having blood in stool, exterior deficiency spontaneous perspiration, deficiency of vital energy oedema, Heat Diabetes, blood deficiency chlorosis, hemiplegia, numbness pain is numb, carbuncle
Subcutaneous ulcer is difficult to burst, burst for a long time and does not holds back.Modern research shows that：Radix astragali contains a variety of chemical compositions, such as saponins, lignin, polysaccharide, Huang
Ketone etc..Astragaloside IV contained by it is triterpenoid saponins, is the principle active component of radix astragali, and the anti-first capsule in Five Sacred Mountins
Principle active component.Therefore, the quality standard evaluation means as the anti-first capsule in Five Sacred Mountins, Determination of Astragaloside it is accurate
Whether be ensure product curative effect key problem in technology.
Tlc scanning determination Astragaloside content is used in party's proper mass standard, sample treatment is complicated, and the rate of recovery is low,
Measurement result is not accurate enough, it is impossible to meets the requirement of its content of Accurate Determining.Meanwhile because in Astragaloside IV molecular structure without conjugation
Double bond, no UV absorption can not Accurate Determining its content using conventional UV detector detection.Therefore selection evaporative light detection
Device is detected it.Moreover, Astragaloside IV content in Milkvetch Root is extremely low, need to be enriched with sample and located before detection
Reason.If processing method is not thorough, Astragaloside content testing result can produce larger fluctuation, cause testing result not accurate enough
Really.At the same time in the anti-Party A in Five Sacred Mountins except radix astragali in addition to the also medicinal material such as ginseng, radix glycyrrhizae, its contain substantial amounts of triterpenoid saponin and sapogenin into
Divide, easily disturb the assay of Astragaloside IV.
Astragaloside IV analog is as follows in Radix Astragali：
Table 1：Astragaloside IV analogue
| Title | R1 | R2 | R3 |
| Astragaloside I | Ac | Ac | H |
| Different astragaloside I | Ac | H | Ac |
| Acetyl astragaloside I | Ac | Ac | Ac |
| Astragaloside II | Ac | H | H |
| Different astragaloside II | H | Ac | H |
| Astragaloside IV (Astragaloside IV) | H | H | H |
The content of Astragaloside IV from Radix Astragali is extremely low,《Chinese Pharmacopoeia》Pointed out in versions first in 2015：Milkvetch Root
The content of middle Astragaloside IV must not be less than 0.040%.The aglycon of Astragaloside IV is cycloastragenol, its aglycon three is connected with 1 point
Sub- xylose, 6 are connected with 1 molecule glucose.Its analogue is often connected with acetyl group on each hydroxyl of 3 xyloses.Ammonia tries
In liquid washing process while acid interfering material is removed, above-mentioned acetyl group can be also promoted to hydrolyze.Ammonia solution washing process is anti-
Should be incomplete, Determination of Astragaloside can be caused to produce larger fluctuation.
The content of the invention
The object of the present invention is to provide the content assaying method of Astragaloside IV in a kind of anti-Party A in Five Sacred Mountins, negative control does not have
Interference, has stronger specificity and good reappearance, obtained chromatogram baseline is steady, and chromatographic peak peak shape is good, reaches
Baseline separation, is a kind of detection effective analysis method of Astragaloside IV component.
The content assaying method of Astragaloside IV in the anti-Party A in Five Sacred Mountins of the present invention, using high performance liquid chromatography-evaporation
The method of photodetector, includes the following steps：
1) chromatographic condition is determined：
Chromatography column packing：Octadecyl silane；
Mobile phase：Acetonitrile-water, volume ratio 31:69；
Flow velocity：1.0mL/min；
Column temperature：40℃；
Theoretical cam curve：Calculated by Astragaloside IV peak, not less than 3000 (i.e. more than or equal to 3000)；
Evaporate the condition of photodetector：
Drift tube temperature：101.1℃；
Gas flow rate：2.7L/min；
2) preparation of reference substance solution：
Accurately weighed Astragaloside IV reference substance, adds methanol that solution of every 1mL containing 0.5mg is made；
3) preparation of test solution：
A, precision weighs test sample 5g, adds methanol and is heated to reflux, obtains extracting solution；
B, extracting solution filters, and filter residue, merging filtrate are washed with methanol；
C, the filtrate that is concentrated under reduced pressure is closely dry, adds aqueous alkali dissolving, is then extracted with water-saturated n-butanol, obtain extract；
D, extract is washed with ammonia solution, is concentrated to dryness, and is added methanol constant volume to 10mL, is obtained test solution；
4) measure：
It is accurate respectively to draw 10 μ L of reference substance solution, 20 μ L of 20 μ L and test solution, liquid chromatograph is injected, using outer
Two-point method measure is marked, up to measurement result.
Preparing for the test solution is specific as follows：
Precision weighs test sample 5g, adds methanol 100mL and is heated to reflux 1h, extracting solution filtering, filter is washed with 40mL methanol
Slag, merging filtrate, is concentrated under reduced pressure near dry, and adding 20~50mL aqueous alkalis to shake 5~60min makes its dissolving, with the positive fourth of water saturation
Alcohol extracts 4 times, each 40mL, merges butanol extraction liquid, is washed 3 times with ammonia solution, each 50mL, shakes 1min, just every time
Butanol, before immunoassay liquid is concentrated to dryness, and adds methanol constant volume to 10mL, up to test solution.
Wherein：
Aqueous alkali is ammonium hydroxide, mass concentration is 2~20% sodium hydrate aqueous solution, mass concentration are 2~20%
Potassium hydroxide aqueous solution or the triethylamine aqueous solution that mass concentration is 2~20%.
Preferably, the test solution prepare it is specific as follows：
Precision weighs test sample 5g, adds methanol 100mL to be heated to reflux 1h, and extracting solution filtering, filter residue is washed with 40mL methanol,
Merging filtrate, is concentrated under reduced pressure near dry, adds 20mL ammonium hydroxide shaking 5min to make its dissolving, extracts 4 times with water-saturated n-butanol, every time
40mL, merges butanol extraction liquid, is washed 3 times, each 50mL with ammonia solution, shakes 1min, butanol extraction liquid concentration every time
To doing, add methanol constant volume to 10mL, up to test solution.
In order to verify stability, accuracy, specificity and the system suitability of measure content method, side has been carried out to method
Science of law checking test, to ensure that the content assaying method can be as the method for quality control of the anti-first capsule in Five Sacred Mountins.
1st, instrument and reagent, reagent
1.1 instrument
High performance liquid chromatograph：Shimadzu 20A；Chromatographic column：Wondasil C18 5μm 4.6×250mm；Evaporative light detects
Device： ELSD 2000ES；Electronic balance：CPA225D.
1.2 reagents and reagent
Reference substance：Astragaloside IV (lot number：110781-201515, content 93.0%) ground purchased from Chinese food drug assay
Study carefully institute；
Reagent：Methanol, acetonitrile (chromatographically pure, Beijing lark prestige Science and Technology Ltd.)；Water is ultra-pure water；
Sample：Five Sacred Mountins anti-first capsule (Qidu Pharmaceutical Co., Ltd., Shandong Prov., lot number：170302)；
Negative control sample：(Qidu Pharmaceutical Co., Ltd., Shandong Prov., lot number 160904).
2nd, sample preparation prescription and preparation method
2.1 sample preparation prescriptions
Prepared fleece flower root 20g, fruit of glossy privet 10g, coix seed 12g, radix astragali 30g, curcuma zedoary 20g, stir-baked RHIZOMA SPARGANII with vinegar 13g, radix glycyrrhizae 3g, white flower
HERBA HEDYOTIS DIFFUSAE 10g, rhizoma dioscoreae nipponicae 10g, ginseng 30g.
2.2 preparation method
Curcuma zedoary, stir-baked RHIZOMA SPARGANII with vinegar, oldenlandia diffusa are weighed into raw material in proportion, are put into volatile oil extracting equipment, extraction volatilization
Oil；When the dregs of a decoction and prepared fleece flower root, the fruit of glossy privet, radix astragali, radix glycyrrhizae, rhizoma dioscoreae nipponicae add 10 times of 80% ethanol solution infiltration 24 small, 3 are decocted
It is secondary, every time 2 it is small when, decoct temperature control at 75~85 DEG C, collecting decoction, is concentrated under reduced pressure into relative density 1.30；Gained medicinal extract
It is dried with the mode of being dried under reduced pressure；Gained dry extract is crushed with pulverizer and crosses 100 mesh sieves；Remaining coix seed, ginseng powder
Broken machine crushes and crosses 100 mesh sieves；Extract powder and ginseng pulverate are mixed, the volatile oil of extraction is added and adds suitable amount of sucrose, is made
Particle.Add proper lubrication agent and load softgel shell, to obtain the final product.
2.3 negative control sample preparation prescriptions
Prepared fleece flower root 20g, fruit of glossy privet 10g, coix seed 12g, curcuma zedoary 20g, stir-baked RHIZOMA SPARGANII with vinegar 13g, radix glycyrrhizae 3g, oldenlandia diffusa
10g, rhizoma dioscoreae nipponicae 10g, ginseng 30g.
The preparation method of 2.4 negative control samples
Curcuma zedoary, stir-baked RHIZOMA SPARGANII with vinegar, oldenlandia diffusa are weighed into raw material in proportion, are put into volatile oil extracting equipment, extraction volatilization
Oil；When the dregs of a decoction and prepared fleece flower root, the fruit of glossy privet, radix glycyrrhizae, rhizoma dioscoreae nipponicae add 10 times of 80% ethanol solution infiltration 24 small, decoction 3 times, every time
2 it is small when, decoct temperature control at 75~85 DEG C, collecting decoction, is concentrated under reduced pressure into relative density 1.30；Gained medicinal extract is dry with decompression
Dry mode is dried；Gained dry extract is crushed with pulverizer and crosses 100 mesh sieves；Remaining coix seed, ginseng are crushed with pulverizer
And cross 100 mesh sieves；Extract powder and ginseng pulverate are mixed, the volatile oil of extraction is added and adds suitable amount of sucrose, particle is made.Again plus
Enter proper lubrication agent and load softgel shell, to obtain the final product.
3rd, method screening process
Each stage sample is detected during to Determination of Astragaloside, as a result as follows：
Table 2：Each stage sample testing result of Determination of Astragaloside
Table 3：Insoluble solvent dissolving result measure
As can be seen from Table 3, test solution is after methanol eddy extracts, using purified water sample dissolution, through just
After butanol, before immunoassay, ammonia solution washing times rise to 5 times from 3 times, survey Astragaloside content still fluctuate it is larger, it is impossible to reach accurate
Really measure.And after purified water dissolving is changed to buck dissolving, shaking 5min is fully hydrolyzed Astragaloside IV analog.Again through just
Butanol, before immunoassay, after ammonia solution washing, Astragaloside content can be determined accurately in test solution.Meanwhile the result listed by table 2
Show, after extracting n-butyl alcohol, the operation for retaining ammonia solution washing is still needed to, to remove pigment impurity.
4th, methodological study
4.1 specificity
The anti-first capsule sample (lot number 170302) in Five Sacred Mountins and negative control are prepared respectively according to the preparation method under 2.2 and 2.4
Sample (lot number 160904).According to the preparation method of test solution, test solution and negative control solution are prepared respectively.
Respectively draw reference substance solution, test solution, negative control solution injection liquid chromatograph, according to set chromatographic condition into
Row measure, the results showed that, in the corresponding position of Astragaloside IV chromatographic peak, noiseless peak occurs in negative control chromatographic peak, chromatography
Figure is shown in Fig. 1-Fig. 3.
4.2 linear relationships are investigated
Compound concentration is the radix astragali first of 0.04mg/mL, 0.05mg/mL, 0.1mg/mL, 0.2mg/mL, 0.5mg/mL respectively
Glycosides reference substance solution, precision are drawn above-mentioned each 20 μ L of concentrations control product solution injections high performance liquid chromatograph, are measured.Measure
It the results are shown in Table 4.
Table 4：Linear relationship measurement result
| Numbering | Concentration (mg/mL) | Sample size μ L | Sample size μ g | Lg (sample size) | Peak area | Lg (peak area) |
| 1 | 0.04 | 20 | 0.80 | -0.0969 | 93028 | 4.9686 |
| 2 | 0.05 | 20 | 1.00 | 0.0000 | 148904 | 5.1729 |
| 3 | 0.10 | 20 | 2.00 | 0.3010 | 408038 | 5.6107 |
| 4 | 0.20 | 20 | 4.00 | 0.6021 | 1280073 | 6.1072 |
| 5 | 0.50 | 20 | 10.00 | 1.0000 | 5612867 | 6.7492 |
The logarithmic regression equation of sample size and peak area is：Lgy=1.6038lgx+5.1424 (R2=0.9993, x=into
Sample amount μ g, y=peak area).
The result shows that：Sample size linear relationship in 0.80~10.00 μ g ranges is good.
4.3 accuracy (recovery test)
Precision weighs the anti-first capsule sample 2.5g in 170302 batches of Five Sacred Mountins, totally 9 parts, is separately added into 1mL, 2mL, 3mL
The reference substance solution of 0.5mg/mL, handles sample according to the processing mode of test solution, obtains basic, normal, high by three by each parallel three parts
Totally 9 parts of the test solution of a concentration.Reference substance solution, 9 parts of test solution injection liquid chromatographs are drawn respectively, according to both
Fixed chromatographic condition is measured, as a result as follows：
Table 5：Accuracy test measurement result
Result above shows that the average recovery rate of Astragaloside IV is 100.57%, RSD 0.84%, illustrates that accuracy is good
It is good.
4.4 precision (repetitive test)
Precision weighs the anti-first capsule test sample 5g in 170302 batches of Five Sacred Mountins, and sample is handled according to the processing mode of test solution
Product.6 parts of test samples of parallel preparation.Reference substance solution, 6 parts of test solution injection liquid chromatographs are drawn respectively, according to set
Chromatographic condition be measured, it is as a result as follows：
Table 6：Astragaloside IV repeatability measurement result
Result above shows, 6 parts of parallel test samples of same batch of sample preparation, and the Astragaloside IV average content measured is
0.28mg/g, RSD 1.68%, the reappearance of illustration method are good.
4.5 solution stability testing
Precision weighs the anti-first capsule test sample 5.0g in 170302 batches of Five Sacred Mountins, and sample is handled according to the processing mode of test solution
Product.Reference substance solution, test solution are drawn respectively, inject liquid chromatograph respectively at 0h, 2h, 4h, 8h, 12h, 24h, according to
Set chromatographic condition is measured, as a result as follows：
Table 7：Reference substance solution, test solution Stability Determination result
The result shows that reference substance and test sample peak area are respectively 0.21% and 0.14% to the RSD of numerical value, show that 24 is small
When it is interior, reference substance and test solution are stablized.
Beneficial effects of the present invention are as follows：
1st, the present invention provides one kind to utilize HPLC ELSD detector Accurate Determining treatment thyroid gland
The assay method of Astragaloside content in the compound Chinese medicinal preparation of disease.
2nd, method of quality control of the invention, adds the operation of buck dissolving, sample before extracting n-butyl alcohol through making radix astragali
First glycosides analogue is fully hydrolyzed, and is made Astragaloside IV extraction more complete, be ensure that the steady of final assay result
It is fixed.Extraction times, solvent dosage and the extraction time of n-butanol are controlled at the same time, make extraction complete, stringent control ammonia solution
Dosage and washing times, eliminate the interference of impurity in detection sample to greatest extent.
3rd, testing result shows do not have in the anti-Party A's negative control sample in Five Sacred Mountins on the corresponding position of Astragaloside IV chromatographic peak
There is Interference Peaks appearance, which has stronger specificity and good reappearance, and obtained chromatogram baseline is steady, peak
Shape is good, reaches baseline separation, is a kind of detection Astragaloside IV component accurately and effectively analysis method.
Brief description of the drawings
Fig. 1 is Astragaloside IV reference substance solution chromatogram of the present invention；
Fig. 2 is the anti-first capsule test solution chromatogram in Five Sacred Mountins of the present invention
Fig. 3 is the anti-first capsule negative control solution chromatogram in Five Sacred Mountins of the present invention.
Embodiment
The present invention is described further with reference to embodiments.
Embodiment 1
The content of Astragaloside IV in the anti-first capsule in Five Sacred Mountins is measured, is comprised the following steps that：
1) chromatographic condition is determined
Chromatography column packing：Octadecyl silane；
Mobile phase：Acetonitrile-water, volume ratio 31:69；
Flow velocity：1.0mL/min；
Column temperature：40℃；
Theoretical cam curve：Calculated by Astragaloside IV peak, more than or equal to 3000；
Evaporate the condition of photodetector：
Drift tube temperature：101.1℃；
Gas flow rate：2.7L/min.
2) preparation of reference substance solution
Take Astragaloside IV reference substance appropriate, it is accurately weighed, add methanol that solution of every 1mL containing 0.5mg is made, to obtain the final product.
3) preparation of test solution
Precision weighs capsule 's content 5g, adds methanol 100mL to be heated to reflux 1h, extracting solution filtering, is washed with 40mL methanol
Filter residue, merging filtrate, is concentrated under reduced pressure near dry, adds 20mL ammonium hydroxide shaking 5min to make its dissolving, is extracted 4 times with water-saturated n-butanol,
Each 40mL, merges butanol extraction liquid, is washed 3 times with ammonia solution, each 50mL, shakes 1min, butanol extraction liquid every time
Be concentrated to dryness, add methanol constant volume to 10mL, to obtain the final product.
4) measure
Precision draws 10 μ L of reference substance solution, 20 μ L of 20 μ L and test solution, liquid chromatograph is injected, using external standard two
Point method measure, to obtain the final product.
The content for measuring Astragaloside IV in the anti-first capsule in every Five Sacred Mountins is 0.03mg.
Embodiment 2
The content of Astragaloside IV in the anti-first piece in Five Sacred Mountins is measured, is comprised the following steps that：
1) chromatographic condition is determined
Chromatography column packing：Octadecyl silane；
Mobile phase：Acetonitrile-water, volume ratio 31:69；
Flow velocity：1.0mL/min；
Column temperature：40℃；
Theoretical cam curve：Calculated by Astragaloside IV peak, more than or equal to 3000；
Evaporate the condition of photodetector：
Drift tube temperature：101.1℃；
Gas flow rate：2.7L/min.
2) preparation of reference substance solution
Take Astragaloside IV reference substance appropriate, it is accurately weighed, add methanol that solution of every 1mL containing 0.5mg is made, to obtain the final product.
3) preparation of test solution
Take test sample appropriate, finely ground, precision weighs 5g, adds methanol 100mL to be heated to reflux 1h, extracting solution filtering, uses 40mL
Methanol washs filter residue, and merging filtrate, is concentrated under reduced pressure near dry, adds 20mL ammonium hydroxide shaking 5min to make dissolving, is extracted with water-saturated n-butanol
Take 4 times, each 40mL, merge butanol extraction liquid, washed 3 times with ammonia solution, each 50mL, shakes 1min, n-butanol every time
Extract is concentrated to dryness, and adds methanol constant volume to 10mL, to obtain the final product.
4) measure
Precision draws 10 μ L of reference substance solution, 20 μ L of 20 μ L and test solution, liquid chromatograph is injected, using external standard two
Point method measure, to obtain the final product.
Measure the anti-first piece in every Five Sacred Mountins and 0.03mg is calculated as with Astragaloside IV containing radix astragali.
Claims (8)
- A kind of 1. content assaying method of Astragaloside IV in anti-Party A in Five Sacred Mountins, using high performance liquid chromatography-evaporation photodetector Method, it is characterised in that：Include the following steps：1) chromatographic condition is determined：Chromatography column packing：Octadecyl silane；Mobile phase：Acetonitrile-water, volume ratio 31:69；Flow velocity：1.0mL/min；Column temperature：40℃；Theoretical cam curve：Calculated by Astragaloside IV peak, not less than 3000；Evaporate the condition of photodetector：Drift tube temperature：101.1℃；Gas flow rate：2.7L/min；2) preparation of reference substance solution：Accurately weighed Astragaloside IV reference substance, adds methanol that solution of every 1mL containing 0.5mg is made；3) preparation of test solution：A, precision weighs test sample 5g, adds methanol and is heated to reflux, obtains extracting solution；B, extracting solution filters, and filter residue, merging filtrate are washed with methanol；C, the filtrate that is concentrated under reduced pressure is closely dry, adds aqueous alkali dissolving, is then extracted with water-saturated n-butanol, obtain extract；D, extract is washed with ammonia solution, is concentrated to dryness, and is added methanol constant volume to 10mL, is obtained test solution；4) measure：It is accurate respectively to draw 10 μ L of reference substance solution, 20 μ L of 20 μ L and test solution, liquid chromatograph is injected, using external standard two Point method measure, up to measurement result.
- 2. the content assaying method of Astragaloside IV in the anti-Party A in Five Sacred Mountins according to claim 1, it is characterised in that：Step a In, the dosage of methanol is 100mL, be heated to reflux the time for 1 it is small when.
- 3. the content assaying method of Astragaloside IV in the anti-Party A in Five Sacred Mountins according to claim 1, it is characterised in that：Step b In, the dosage of methanol is 40mL.
- 4. the content assaying method of Astragaloside IV in the anti-Party A in Five Sacred Mountins according to claim 1, it is characterised in that：Step c In, adding 20~50mL aqueous alkalis and shaking 5~60 minutes makes its dissolving.
- 5. the content assaying method of Astragaloside IV in the anti-Party A in Five Sacred Mountins according to claim 4, it is characterised in that：Step c In, adding 20mL aqueous alkalis and shaking 5 minutes makes its dissolving.
- 6. the content assaying method of Astragaloside IV in the anti-Party A in Five Sacred Mountins according to claim 1, it is characterised in that：Step c In, hydroxide that aqueous alkali is ammonium hydroxide, mass concentration is 2~20% sodium hydrate aqueous solution, mass concentration are 2~20% Aqueous solutions of potassium or the triethylamine aqueous solution that mass concentration is 2~20%.
- 7. the content assaying method of Astragaloside IV in the anti-Party A in Five Sacred Mountins according to claim 1, it is characterised in that：Step c In, extracted 4 times with water-saturated n-butanol, each 40mL.
- 8. the content assaying method of Astragaloside IV in the anti-Party A in Five Sacred Mountins according to claim 1, it is characterised in that：Step d In, extract is washed 3 times, each 50mL with ammonia solution, every time shaking 1 minute.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109342619A (en) * | 2018-11-14 | 2019-02-15 | 神威药业集团有限公司 | Stilbene Huang leads to the measuring method of Astragaloside content in secret soft capsule |
| CN110967422A (en) * | 2019-11-21 | 2020-04-07 | 吉林师范大学 | Method for determining content of astragaloside in qi-tonifying and blood-nourishing tablet |
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| CN111337595A (en) * | 2020-04-13 | 2020-06-26 | 内蒙古天创药业科技股份有限公司 | Preparation method of test solution for determining content of astragaloside in astragalus medicinal material |
| CN115792017A (en) * | 2022-12-06 | 2023-03-14 | 中国中医科学院眼科医院 | Quality detection method of traditional Chinese medicine composition for reducing recurrence of herpes simplex viral keratitis |
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