CN103134867A - Stable method for measuring calycosin heteroside content in Astragalus mongholicus submicron powder - Google Patents
Stable method for measuring calycosin heteroside content in Astragalus mongholicus submicron powder Download PDFInfo
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- CN103134867A CN103134867A CN2012105287329A CN201210528732A CN103134867A CN 103134867 A CN103134867 A CN 103134867A CN 2012105287329 A CN2012105287329 A CN 2012105287329A CN 201210528732 A CN201210528732 A CN 201210528732A CN 103134867 A CN103134867 A CN 103134867A
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- radix astragali
- methanol
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- ZZAJQOPSWWVMBI-UHFFFAOYSA-N calycosin Chemical compound C1=C(O)C(OC)=CC=C1C1=COC2=CC(O)=CC=C2C1=O ZZAJQOPSWWVMBI-UHFFFAOYSA-N 0.000 title claims abstract description 47
- QUQPHWDTPGMPEX-UHFFFAOYSA-N Hesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(COC4C(C(O)C(O)C(C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-UHFFFAOYSA-N 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 31
- 239000000843 powder Substances 0.000 title claims abstract description 31
- 241000045403 Astragalus propinquus Species 0.000 title abstract 4
- 235000006533 astragalus Nutrition 0.000 title abstract 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 89
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 18
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims abstract description 12
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims abstract description 6
- 235000019253 formic acid Nutrition 0.000 claims abstract description 6
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 4
- 229930182478 glucoside Natural products 0.000 claims description 39
- -1 calycosin glucoside Chemical class 0.000 claims description 37
- 239000009636 Huang Qi Substances 0.000 claims description 27
- 238000012360 testing method Methods 0.000 claims description 14
- 239000013558 reference substance Substances 0.000 claims description 12
- 238000002360 preparation method Methods 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 8
- 238000001514 detection method Methods 0.000 claims description 7
- 238000013461 design Methods 0.000 claims description 5
- 239000000706 filtrate Substances 0.000 claims description 5
- 238000004064 recycling Methods 0.000 claims description 5
- 238000010992 reflux Methods 0.000 claims description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- 239000000945 filler Substances 0.000 claims description 4
- 238000004811 liquid chromatography Methods 0.000 claims description 2
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 2
- 239000000047 product Substances 0.000 claims description 2
- 239000000377 silicon dioxide Substances 0.000 claims description 2
- 230000006641 stabilisation Effects 0.000 claims description 2
- 238000011105 stabilization Methods 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims 1
- 238000001914 filtration Methods 0.000 abstract description 2
- 238000001704 evaporation Methods 0.000 abstract 1
- 230000008020 evaporation Effects 0.000 abstract 1
- 239000003814 drug Substances 0.000 description 17
- 239000000243 solution Substances 0.000 description 17
- 238000003556 assay Methods 0.000 description 6
- 239000012086 standard solution Substances 0.000 description 6
- 238000000227 grinding Methods 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 238000004090 dissolution Methods 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 239000007791 liquid phase Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 238000004891 communication Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 230000000857 drug effect Effects 0.000 description 2
- 150000008131 glucosides Chemical class 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 241000599985 Beijerinckia mobilis Species 0.000 description 1
- 240000004670 Glycyrrhiza echinata Species 0.000 description 1
- 235000001453 Glycyrrhiza echinata Nutrition 0.000 description 1
- 235000006200 Glycyrrhiza glabra Nutrition 0.000 description 1
- 235000017382 Glycyrrhiza lepidota Nutrition 0.000 description 1
- VTAJIXDZFCRWBR-UHFFFAOYSA-N Licoricesaponin B2 Natural products C1C(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2)C(O)=O)C)(C)CC2)(C)C2C(C)(C)CC1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O VTAJIXDZFCRWBR-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940126678 chinese medicines Drugs 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- LPLVUJXQOOQHMX-UHFFFAOYSA-N glycyrrhetinic acid glycoside Natural products C1CC(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2=O)C(O)=O)C)(C)CC2)(C)C2C(C)(C)C1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O LPLVUJXQOOQHMX-UHFFFAOYSA-N 0.000 description 1
- 229960004949 glycyrrhizic acid Drugs 0.000 description 1
- UYRUBYNTXSDKQT-UHFFFAOYSA-N glycyrrhizic acid Natural products CC1(C)C(CCC2(C)C1CCC3(C)C2C(=O)C=C4C5CC(C)(CCC5(C)CCC34C)C(=O)O)OC6OC(C(O)C(O)C6OC7OC(O)C(O)C(O)C7C(=O)O)C(=O)O UYRUBYNTXSDKQT-UHFFFAOYSA-N 0.000 description 1
- 239000001685 glycyrrhizic acid Substances 0.000 description 1
- 235000019410 glycyrrhizin Nutrition 0.000 description 1
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 229940010454 licorice Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000003595 mist Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229940126701 oral medication Drugs 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
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- Medicines Containing Plant Substances (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a stable method for measuring calycosin heteroside content in Astragalus mongholicus submicron powder. Acetonitrile and 0.2% formic acid solution (20 to 80) serve as a moving phase, detecting wavelength is 260nm, calycosin heterosid serves as a comparing object, and a sample is processed through back flow of carbinol, filtering, evaporation to dryness and dissolving. The calycosin heteroside content in the Astragalus mongholicus submicron powde is detected through high performance liquid chromatography and an ultraviolet detector. The method is good in linear relation, high in degree of precision and good in reproducibility, and can serve as a method for measuring the calycosin heteroside content in the Astragalus mongholicus submicron powder, and the method is simple, rapid and easy to operate.
Description
Technical field
The present invention relates to a kind of efficient liquid-phase chromatography method, the assay technology of more particularly to a kind of traditional Chinese medicine ingredients.
Background technology
[0002] ultramicro grinding is nearly new and high technology developed rapidly for 20 years, and micron even nano level micro mist raw material can be processed into for the field of Chinese medicines.In view of crushing the basic process technology in being Chinese medicine production and applying, ultramicro grinding also more and more causes the concern of people, although being started late in herbal pharmaceutical industry, the kind developed is relatively fewer, but distinctive advantage and wide application prospect have been manifested, and has turned into the study hotspot of Chinese medicine circle in recent years.
The maximum advantage of ultramicro grinding Chinese medicine is the bioavilability for effectively increasing medicine.For pharmaceutical principles, the dissolution rate of medicine and the specific grain surface product of medicine are proportionate, and specific surface area is inversely proportional with grain diameter.Therefore, the particle diameter of medicine is smaller, then its surface area is bigger, more contributes to the dissolution of effective ingredient.Ultramicro grinding Chinese medicine and its particle reach the level of superfines, and its specific surface area is significantly increased, and solubility of the medicine in intestines and stomach substantially increases, so as to increase the bioavilability of medicine, and accelerate the drug effect time.Further, since nanometer or the adhesiveness and small particle diameter of micron grain, the increase of anelasticity, is also beneficial to extension medicine and intestinal wall time of contact, enlarged contact areas, so as to improve the bioavilability of Oral drug absorption when not only improving extension local application.Dissolution such as glycyrrhizic acid after licorice piece ultramicro grinding is dramatically increased.Traditional Chinese medicine medicine materical crude slice is often using the method decocted, though having been carried out the improvement of Chinese medicine preparation at present, simply extracts the fraction composition contained by Chinese medicine, accounts for the 10%~30% of total composition, drug effect is greatly affected.Traditional Chinese medicine ingredients can be fully extracted using superfine communication technique, have the advantages that to absorb fast and easy to use.
Superfine communication technique is applied to the Radix Astragali, Radix Astragali Ultramicro-powder is made, but wherein active ingredient calycosin glucoside is not specified by corresponding content assaying method, the present invention carries out content assaying method to the composition calycosin glucoside in Radix Astragali Ultramicro-powder and further explored, and determines calycosin glucoside content assaying method in a kind of Radix Astragali Ultramicro-powder of more stabilization simple to operation.Reliable active constituent content measuring method is provided for the quality control of Radix Astragali Ultramicro-powder.
The content of the invention
Present invention seek to address that calycosin glucoside assay problem in Radix Astragali Ultramicro-powder, to solve the problem of calycosin glucoside is without corresponding content assaying method in Radix Astragali Ultramicro-powder.
Specific method technical scheme is as follows：
A kind of method of calycosin glucoside content in high effective liquid chromatography for measuring Radix Astragali Ultramicro-powder, its chromatographic condition is：
A. filler is octadecylsilane chemically bonded silica；
B. mobile phase is that mixed proportion is 20:80 acetonitrile and 0.2% formic acid solution；
C. the Detection wavelength of UV-detector is 260nm；
D. flow rate of mobile phase used is 1ml/min；
E. sampling volume is 10 μ l.
1. the drafting of calycosin glucoside standard curve
A. the preparation of standard solution：Precision weighs calycosin glucoside standard items 0.01042g, is placed in 200ml volumetric flasks, is dissolved with methanol, is settled to 200ml.
B. the standard solution of methanol dilution various concentrations is used：Precision measures original content standard solution 5ml, plus methanol constant volume is to 10ml, after mixing, precision measures 5ml into another volumetric flask again, plus methanol constant volume is to 10ml, doubling dilution, is made the calycosin glucoside standard solution that concentration is respectively 52.1 μ g/ml, 26.05 μ g/ml, 13.03 μ g/ml, 6.51 μ g/ml, 3.26 μ g/ml in this way.
C. the peak area responded with the calycosin glucoside standard solution of various concentrations is mapped to concentration, obtains the standard curve of calycosin glucoside.
2. the preparation of test sample
Radix Astragali Ultramicro-powder powder about 1g is taken, it is accurately weighed, put in round-bottomed flask, precision adds methanol 50ml, and weighed weight is heated to reflux 4 hours, let cool, then weighed weight, the weight of less loss is supplied with methanol, shake up, filter, precision measures subsequent filtrate 25ml, recycling design is to doing, and residue adds methanol to dissolve, and is transferred in 5ml measuring bottles, plus methanol is to scale, shake up, produce.
3. precision test
Need testing solution is taken, precision is drawn 10 μ l sample introductions, is repeated 5 times, and records peak area, calculates this method precision.Result of the test is shown in Table.
Table calycosin glucoside content assaying method Precision test result
From result, the relative standard deviation of calycosin glucoside is 0.89%, shows that the sample introduction precision of this method is good.
4. recovery test
The preparation of need testing solution：Precision weighs Radix Astragali Ultramicro-powder about 1g, 6 parts, puts in round-bottomed flask, is separately added into calycosin glucoside standard solution 1ml, precision adds methanol 49ml, and weighed weight is heated to reflux 4 hours, let cool, then weighed weight, the weight of less loss is supplied with methanol, shake up, filter, precision measures subsequent filtrate 25ml, recycling design is to doing, and residue adds methanol to dissolve, and is transferred in 5ml measuring bottles, plus methanol is to scale, shake up, produce.
Reference substance solution：Take calycosin glucoside reference substance appropriate, it is accurately weighed, plus the solution that every 1ml contains 50 μ g is made in methanol, produces.
Table calycosin glucoside recovery test result
Brief description of the drawings
Fig. 1 calycosin glucoside standard curves
Fig. 2 calycosin glucoside standard items liquid chromatograms
First Radix Astragali Ultramicro-powder liquid chromatogram of Fig. 3 different sources
Fig. 4 different sources second batch Radix Astragali Ultramicro-powder liquid chromatograms
Embodiment
With reference to embodiment, the invention will be further described, but protection scope of the present invention is not limited to embodiment.
Content detection is carried out to the calycosin glucoside in two batches Radix Astragali Ultramicro-powder by following detection method.
Embodiment oneCalycosin glucoside assay in first Radix Astragali Ultramicro-powder of different sources
According to high performance liquid chromatography（Chinese veterinary pharmacopoeia）Determine.
Detecting instrument Yi Lite high performance liquid chromatographs, EC2000 UV-detectors.
The general C18 of chromatographic column（200mm × 4.6mm, 5 μm）Liquid-phase chromatographic column.Filler is SinoChrom ODS-BP.
Chromatographic condition is with system suitability with the formic acid solution of acetonitrile -2%（20:80）For mobile phase；Detection wavelength is 260nm.Number of theoretical plate is calculated by calycosin glucoside peak should be not less than 3000.
The preparation of reference substance solution takes calycosin glucoside reference substance appropriate, accurately weighed, plus the solution that every 1ml contains 51.0059 μ g is made in methanol, produces.
The preparation of need testing solution takes first Radix Astragali Ultramicro-powder to crush No. four sieves, weighs three parts of 1.0053g, 1.0014g, 1.006g, puts respectively in round-bottomed flask, precision adds methanol 50ml, and weighed weight is heated to reflux 4 hours, let cool, then weighed weight, the weight of less loss is supplied with methanol, shake up, filter, precision measures subsequent filtrate 25ml, recycling design is to doing, and residue adds methanol to dissolve, and is transferred in 5ml measuring bottles, plus methanol is to scale, shake up, produce.
Determination method is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, injects liquid chromatograph, determines, produce.
Calycosin glucoside content results are computed to be shown in Table.
Calycosin glucoside assay result in first Radix Astragali Ultramicro-powder of table
Embodiment twoCalycosin glucoside assay in different sources second batch Radix Astragali Ultramicro-powder
According to high performance liquid chromatography（Chinese veterinary pharmacopoeia）Determine.
Detecting instrument Yi Lite high performance liquid chromatographs, EC2000 UV-detectors.
The general C18 of chromatographic column（200mm × 4.6mm, 5 μm）Liquid-phase chromatographic column.Filler is SinoChrom ODS-BP.
Chromatographic condition is with system suitability with the formic acid solution of acetonitrile -2%（20:80）For mobile phase；Detection wavelength is 260nm.Number of theoretical plate is calculated by calycosin glucoside peak should be not less than 3000.
The preparation of reference substance solution takes calycosin glucoside reference substance appropriate, accurately weighed, plus the solution that every 1ml contains 51.0059 μ g is made in methanol, produces.
The preparation precision of need testing solution weighs second batch Radix Astragali Ultramicro-powder about tri- parts of 1.0023g, 1.0005g, 1.0002g, puts respectively in round-bottomed flask, and precision adds methanol 50ml, weighed weight, is heated to reflux 4 hours, lets cool, weighed weight, the weight of less loss is supplied with methanol, is shaken up again, filtration, precision measures subsequent filtrate 25ml, and recycling design is to dry, residue adds methanol to dissolve, and is transferred in 5ml measuring bottles, plus methanol is to scale, shake up, produce.
Determination method is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, injects liquid chromatograph, determines, produce.Measurement result is shown in Table.
Calycosin glucoside assay result in table second batch Radix Astragali Ultramicro-powder
Claims (7)
1. the content assaying method of calycosin glucoside in a kind of Radix Astragali Ultramicro-powder of stabilization, it is characterised in that step is as follows：
（1）Using high effective liquid chromatography for measuring;
（2）Chromatographic condition is with system suitability using octadecylsilane chemically bonded silica as filler；With the formic acid solution of acetonitrile -0.2%（20:80）For mobile phase；Detection wavelength is 260nm, and number of theoretical plate is calculated by calycosin glucoside peak should be not less than 3000;
（3）The preparation of reference substance solution takes calycosin glucoside reference substance appropriate, accurately weighed, plus the solution that every 1ml contains 50 μ g is made in methanol, produces;
（4）The preparation of need testing solution takes this product powder about 1g, accurately weighed, puts in round-bottomed flask, precision adds methanol 50ml, and weighed weight is heated to reflux 4 hours, let cool, then weighed weight, the weight of less loss is supplied with methanol, shake up, filter, precision measures subsequent filtrate 25ml, recycling design is to doing, and residue adds methanol to dissolve, and is transferred in 5ml measuring bottles, plus methanol is to scale, shake up, produce;
（5）Determination method is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, injects liquid chromatograph, determines, produce.
2. as claimed in claim 1, the content assaying method of calycosin glucoside in stable Radix Astragali Ultramicro-powder, it is characterised in that：Using high performance liquid chromatography.
3. as claimed in claim 1, the content assaying method of calycosin glucoside in Radix Astragali Ultramicro-powder, it is characterised in that：With the formic acid solution of acetonitrile -0.2%（20:80）For mobile phase.
4. as claimed in claim 1, the content assaying method of calycosin glucoside in stable Radix Astragali Ultramicro-powder, it is characterised in that：Using calycosin glucoside as reference substance.
5. as claimed in claim 1, the content assaying method of calycosin glucoside in stable Radix Astragali Ultramicro-powder, it is characterised in that：Detector used is UV-detector.
6. the content assaying method of calycosin glucoside in stable Radix Astragali Ultramicro-powder, it is characterised in that：Detection wavelength is 260nm.
7. the content assaying method of calycosin glucoside in stable Radix Astragali Ultramicro-powder, it is characterised in that：Solvent for use is methanol.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107045030A (en) * | 2017-04-17 | 2017-08-15 | 广西壮族自治区梧州食品药品检验所 | A kind of method of calycosin glucoside in ASE methods extraction Radix Astragali |
| CN107064386A (en) * | 2017-04-17 | 2017-08-18 | 广西壮族自治区梧州食品药品检验所 | A kind of method that ASE HPLC methods determine calycosin glucoside content in the Radix Astragali |
| CN107929420A (en) * | 2017-11-23 | 2018-04-20 | 江西天元药业有限公司 | Improve energy and blood of human body function and improve the Chinese medicine composition and detection method of immunity |
| CN116754667A (en) * | 2023-05-29 | 2023-09-15 | 江苏中兴药业有限公司 | A method for determining the increased content of Shenqi Jianwei granules quality standard |
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- 2012-12-11 CN CN2012105287329A patent/CN103134867A/en active Pending
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107045030A (en) * | 2017-04-17 | 2017-08-15 | 广西壮族自治区梧州食品药品检验所 | A kind of method of calycosin glucoside in ASE methods extraction Radix Astragali |
| CN107064386A (en) * | 2017-04-17 | 2017-08-18 | 广西壮族自治区梧州食品药品检验所 | A kind of method that ASE HPLC methods determine calycosin glucoside content in the Radix Astragali |
| CN107929420A (en) * | 2017-11-23 | 2018-04-20 | 江西天元药业有限公司 | Improve energy and blood of human body function and improve the Chinese medicine composition and detection method of immunity |
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| CN116754667A (en) * | 2023-05-29 | 2023-09-15 | 江苏中兴药业有限公司 | A method for determining the increased content of Shenqi Jianwei granules quality standard |
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Application publication date: 20130605 |