CN105929066A - Method for determining andrographolide and dehydroandrographoline in andrographis tablet by using HPLC - Google Patents
Method for determining andrographolide and dehydroandrographoline in andrographis tablet by using HPLC Download PDFInfo
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- CN105929066A CN105929066A CN201610259545.3A CN201610259545A CN105929066A CN 105929066 A CN105929066 A CN 105929066A CN 201610259545 A CN201610259545 A CN 201610259545A CN 105929066 A CN105929066 A CN 105929066A
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- andrographolide
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- dehydrorographolide
- andrographis
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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
- 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
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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- Chemical & Material Sciences (AREA)
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- General Health & Medical Sciences (AREA)
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- Treatment Of Liquids With Adsorbents In General (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a method for determining andrographolide and dehydroandrographoline in an andrographis tablet by using HPLC. The objective of the invention is to provide the determination method with the advantages of high detection sensitivity, simple operation and good repeatability. The method comprises the following steps: weighing 1 g of sample powder, placing the sample powder into an extraction pool with a volume of 10 mL and subjecting the sample to static extraction with methanol at 60 DEG C twice; after completion of extraction, fixing the volume of obtained extract to a constant volume of 20 mL, sucking up 1 mL of the extract into a centrifuge tube, carrying out vortex treatment for 2 min and then carrying out centrifugation at a speed of 10000 r/min for 3 min; and collecting supernatant, carrying out uniform mixing, carrying out filtering with a filter membrane with a size of 0.22 [mu]m and injecting filtrate into a high-performance liquid chromatograph for detection. The method belongs to the technical field of chemical detection.
Description
Technical field
The present invention provides a kind of method measuring andrographolide and dehydrorographolide, tool
Say body, be that a kind of HPLC measures in andrographis tablet in andrographolide and dehydration Herba Andrographis
The method of ester；Belong to technical field of chemical detection.
Background technology
Andrographis tablet records in " Chinese Pharmacopoeia 2005 version one, its check item is tablet
The content of regulation in rules of preparations.Chinese medicine oral administration solid preparations is many with drug powder or extract
For material, effective ingredient must be discharged by the process of disintegrate and dissolution, be absorbed into blood
And play therapeutical effect.
According to high performance liquid chromatography (one annex VI D of version pharmacopeia in 2010), assay method is such as
Under.
1, chromatographic condition and system suitability
With octadecylsilane chemically bonded silica as filler；With methanol-water (60:40) it is
Flowing phase；Detection wavelength is 254nm.Number of theoretical plate presses the calculating of dehydrorographolide peak should
It is not less than 2000.
2, the preparation of reference substance solution
Take dehydrorographolide reference substance appropriate, accurately weighed, add methanol and make every 1ml
Solution containing 0.1mg, to obtain final product.
3, the preparation of need testing solution
Take this product 20 (small pieces) or 10 (large stretch of), remove coating, accurate title
Fixed, finely ground, take 0.5g, accurately weighed, put in tool plug conical flask, accurate addition methanol 25
Ml, close plug, weighed weight, soak 1 hour, supersound process (power 250W, frequency 3
3kHz) 30 minutes, let cool, more weighed weight, supply the weight of less loss with methanol, shake
Even, filter, precision measures subsequent filtrate 10ml (remaining subsequent filtrate is standby), is added in neutrality
On alumina column (internal diameter is 1.5cm for 200～300 mesh, 5g), wash with methanol 20ml
De-, collect eluent, put in 50ml measuring bottle, add methanol to scale, shake up, to obtain final product.
4, measure
Precision draws reference substance solution and each 10 μ l of need testing solution respectively, injects liquid chromatograph
Instrument, measures, to obtain final product.
This product every (C20H28O4) Han dehydrorographolide, small pieces must not be less than 4.0
Mg, sheet must not be less than 8.0mg
Summary of the invention
For above-mentioned deficiency, it is an object of the invention to provide a kind of detection sensitivity high, operation
Simply, reproducible HPLC measures andrographolide and dehydration Herba Andrographis in andrographis tablet
The method of lactone.
For solving above-mentioned technical problem, the technical scheme that the present invention provides is as follows:
A kind of HPLC measures andrographolide and the side of dehydrorographolide in andrographis tablet
Method, the method comprises the steps: successively to take sample powder 1g, is placed in 10mL abstraction pool
In, with methanol to sample at temperature 60 C, static extraction 5min, repeats to extract 2 times, carries
By extracting solution methanol constant volume to 20mL after taking into, 1mL is in centrifuge tube in absorption, whirlpool
Whirlpool 2min, 10000r/min are centrifuged 3min, collect supernatant, mix 0.22 micron
Filter membrane, takes filtrate injection high performance liquid chromatograph and detects.
Further, above-mentioned HPLC measures andrographolide and dehydration in andrographis tablet and wears
The method of heart lotus lactone, described high performance liquid chromatograph is Agilent Agilent-1200.
Further, above-mentioned HPLC measures andrographolide and dehydration in andrographis tablet and wears
The method of heart lotus lactone, described chromatographic condition is as follows:
Chromatographic column: StableBond SB-phenyl post；
Flowing phase: acetonitrile A-0.3% aqueous formic acid B；
Mobile phase A is acetonitrile, and Mobile phase B is 0.5% formic acid；Gradient elution program: 0～
1min, 80%A, 20%B；1～12min, 85%～15%A, 15%～85%B；12～
15min, 50%A, 50%B；3min is balanced under 50%A, 50%B.
Flow velocity 1mL/min；
Wavelength: 273nm, 287nm；
Column temperature: 35 DEG C.
Sample size: 5 μ l.
Further, above-mentioned HPLC measures andrographolide and dehydration in andrographis tablet
The method of andrographolide, described chromatographic column specification: 250mm × 4.6mm, 5 μm.
Compared with prior art, the technical scheme that the present invention provides establishes HPLC mensuration and wears
Andrographolide and the method for dehydrorographolide in heart lotus sheet, the method is easy and simple to handle,
Accurately and reliably, can be used for the quality control of andrographis tablet.
Detailed description of the invention
Below in conjunction with detailed description of the invention, the claim of the present invention is done the most in detail
Illustrate, but do not constitute any limitation of the invention, any in the claims in the present invention protection
The amendment of the limited number of time that scope is made, still within the claims of the present invention.
Involved in the present invention to percentage composition concentration, in addition to specified otherwise, solute is liquid
Be volumetric concentration, solute be solid be mass concentration.
Embodiment 1
A kind of HPLC measures andrographolide and the side of dehydrorographolide in andrographis tablet
Method, the method comprises the steps: successively to take sample powder 1g, is placed in 10mL abstraction pool
In, with methanol to sample at temperature 60 C, static extraction 5min, repeats to extract 2 times, carries
By extracting solution methanol constant volume to 20mL after taking into, 1mL is in centrifuge tube in absorption, whirlpool
Whirlpool 2min, 10000r/min are centrifuged 3min, collect supernatant, mix 0.22 micron
Filter membrane, takes filtrate injection Agilent Agilent-1200 high performance liquid chromatograph and detects.
Described chromatographic condition is as follows:
Chromatographic column: StableBond SB-phenyl post (250mm × 4.6mm, 5 μm)；
Flowing phase: acetonitrile A-0.3% aqueous formic acid B；
Mobile phase A is acetonitrile, and Mobile phase B is 0.5% formic acid；Gradient elution program: 0～
1min, 80%A, 20%B；1～12min, 85%～15%A, 15%～85%B；12～
15min, 50%A, 50%B；3min is balanced under 50%A, 50%B.
Flow velocity 1mL/min；
Wavelength: 273nm, 287nm；
Column temperature: 35 DEG C.
Sample size: 5 μ l.
Take with a sample solution, continuous sample introduction 6 times, calculate andrographolide peak area
RSD=0.5%, the peak area RSD=0.3% of dehydrorographolide, show that the method is accurate
Degree is good
Taking need testing solution, in 0,2,4,8, l2,24h measures.Wear in result 24h
The peak area RSD=0.48% (n=6) of heart lotus lactone, the peak area of dehydrorographolide
RSD=0.3% (n=6), shows that this product has good stability.
Taking 5 parts of same batch sample powder, the method provided by the application measures, and records dehydration
Andrographolide, the average content of andrographolide are respectively 49.6mg.g-1、23.3mg.g- 1, RSD is respectively 1.5%, 2.1%.
Reference substance solution: respectively take andrographolide reference substance, dehydrorographolide reference substance is fitted
Amount, adds methanol and makes containing andrographolide 16.6g.mL-1And dehydrorographolide
10.2g.mL-The reference substance mixed solution of 1, to obtain final product
Taking the sample powder about 0.042g of above-mentioned known content, accurately weighed, accurate addition wears
Heart lotus lactone reference substance solution (0.83mg.mL-1) 1mL, dehydrorographolide reference substance
2mL(1.02mg.mL-1), same treatment, by above-mentioned chromatographic condition sample introduction, respectively at
Mensuration trap at 273nm, 287nm, andrographolide average recovery rate 99.8%,
RSD1.12%, dehydrorographolide average recovery rate 99.6%, RSD1.2%.
Claims (4)
1. andrographolide and dehydrorographolide during a HPLC measures andrographis tablet
Method, it is characterised in that the method comprises the steps: to take sample powder 1g successively, puts
In 10mL abstraction pool, with methanol to sample at temperature 60 C, the static 5min that extracts, weight
Extract 2 times again, by extracting solution methanol constant volume to 20mL after having extracted, draw 1mL
In centrifuge tube, whirlpool 2min, 10000r/min are centrifuged 3min, collect supernatant, mixing
Cross the filter membrane of 0.22 micron, take filtrate injection high performance liquid chromatograph and detect.
HPLC the most according to claim 1 measure in andrographis tablet andrographolide and
The method of dehydrorographolide, it is characterised in that described high performance liquid chromatograph is peace
Prompt human relations Agilent-1200.
In HPLC the most according to claim 1 and 2 mensuration andrographis tablet in Herba Andrographis
Ester and the method for dehydrorographolide, it is characterised in that described chromatographic condition is as follows:
Chromatographic column: Stab l eBond SB-phenyl post；
Flowing phase: acetonitrile A-0.3% aqueous formic acid B；
Mobile phase A is acetonitrile, and Mobile phase B is 0.5% formic acid；Gradient elution program: 0～
1mi n, 80%A, 20%B；1～12mi n, 85%～15%A, 15%～85%B；12～
15mi n, 50%A, 50%B；3mi n is balanced under 50%A, 50%B；
Flow velocity 1mL/mi n；
Wavelength: 273nm, 287nm；
Column temperature: 35 DEG C；
Sample size: 5 μ l.
HPLC the most according to claim 3 measures andrographolide in andrographis tablet
Method with dehydrorographolide, it is characterised in that described chromatographic column specification: 250
Mm × 4.6mm, 5 μm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610259545.3A CN105929066A (en) | 2016-04-25 | 2016-04-25 | Method for determining andrographolide and dehydroandrographoline in andrographis tablet by using HPLC |
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|---|---|---|---|
| CN201610259545.3A CN105929066A (en) | 2016-04-25 | 2016-04-25 | Method for determining andrographolide and dehydroandrographoline in andrographis tablet by using HPLC |
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| CN201610259545.3A Pending CN105929066A (en) | 2016-04-25 | 2016-04-25 | Method for determining andrographolide and dehydroandrographoline in andrographis tablet by using HPLC |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106501407A (en) * | 2016-11-02 | 2017-03-15 | 百奥森（江苏）食品安全科技有限公司 | The detection method of active constituent content in a kind of chuanxinlian tablet |
| CN106950294A (en) * | 2017-02-22 | 2017-07-14 | 江西普正植物药科技协同创新有限公司 | A kind of antiphlogistic Chunwang capsule quality determining method |
| CN109142588A (en) * | 2018-10-25 | 2019-01-04 | 广州白云山光华制药股份有限公司 | A kind of LianZhixiaoyan Capsule HPLC characteristic spectrum and its construction method and application |
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| CN105021723A (en) * | 2015-07-06 | 2015-11-04 | 广西壮族自治区药用植物园 | Method for simultaneous determination of content of geniposide, andrographolide and dehydroandrographolide in Zhimai tablets for clearing heat |
| CN105467023A (en) * | 2014-09-05 | 2016-04-06 | 天士力制药集团股份有限公司 | Method for determining andrographis paniculata herb fingerprint and four diterpene lactones through UPLC |
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2016
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Patent Citations (2)
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| CN105467023A (en) * | 2014-09-05 | 2016-04-06 | 天士力制药集团股份有限公司 | Method for determining andrographis paniculata herb fingerprint and four diterpene lactones through UPLC |
| CN105021723A (en) * | 2015-07-06 | 2015-11-04 | 广西壮族自治区药用植物园 | Method for simultaneous determination of content of geniposide, andrographolide and dehydroandrographolide in Zhimai tablets for clearing heat |
Non-Patent Citations (3)
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| 徐晓琴 等: "反相高效液相色谱法测定穿心莲药材及其制剂中的穿心莲内酯和脱水穿心莲内酯", 《色谱》 * |
| 王贞媛 等: "穿心莲片中内酯类成分的"一测多评"", 《中华中医药学刊》 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106501407A (en) * | 2016-11-02 | 2017-03-15 | 百奥森（江苏）食品安全科技有限公司 | The detection method of active constituent content in a kind of chuanxinlian tablet |
| CN106950294A (en) * | 2017-02-22 | 2017-07-14 | 江西普正植物药科技协同创新有限公司 | A kind of antiphlogistic Chunwang capsule quality determining method |
| CN109142588A (en) * | 2018-10-25 | 2019-01-04 | 广州白云山光华制药股份有限公司 | A kind of LianZhixiaoyan Capsule HPLC characteristic spectrum and its construction method and application |
| CN109142588B (en) * | 2018-10-25 | 2021-07-09 | 广州白云山光华制药股份有限公司 | HPLC (high performance liquid chromatography) characteristic spectrum of Lianzhi anti-inflammation capsules as well as construction method and application thereof |
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Application publication date: 20160907 |