CN104215708B - For treating the detection method of content of the Chinese native medicine compound prescription pellet of gynecologic blood diseases - Google Patents
For treating the detection method of content of the Chinese native medicine compound prescription pellet of gynecologic blood diseases Download PDFInfo
- Publication number
- CN104215708B CN104215708B CN201410452215.7A CN201410452215A CN104215708B CN 104215708 B CN104215708 B CN 104215708B CN 201410452215 A CN201410452215 A CN 201410452215A CN 104215708 B CN104215708 B CN 104215708B
- Authority
- CN
- China
- Prior art keywords
- methanol
- protocatechuic acid
- content
- protocatechualdehyde
- blood
- 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.)
- Active
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses the detection method of content of a kind of Chinese native medicine compound prescription pellet for treating gynecologic blood diseases, use the content simultaneously measuring protocatechuic acid and protocatechualdehyde under same chromatographic condition, need testing solution extracts through methanol eddy, prepares in acid condition ether extraction.So effectively eliminate negative interference, the extraction ratio of protocatechuic acid and protocatechualdehyde can be improved again.The method good stability of the present invention, favorable reproducibility, specificity are strong and simple to operate, set up the quality control standard that her blood peace granule is new accordingly, can help effectively to control the end product quality in her blood peace particle manufacture and and improve its quality level, ensure that production technology is controlled, guarantee Clinical practice curative effect, definitely provide reliable guarantee for product stable curative effect.
Description
Technical field
The invention belongs to Chinese medicine content detection technical field, particularly relate to a kind of Chinese native medicine compound prescription pellet for treating gynecologic blood diseases
Detection method of content.
Background technology
She pacifies granule by blood, Yi Xuean (Sugarless type) granule is the Chinese native medicine compound prescription pellet for treating gynecologic blood diseases, osmanthus, Yunnan end
The flavour of a drug such as perfume (or spice), Herba Leonuri, Rhizoma Corydalis, Radix Glycyrrhizae of receiving form, and have promoting blood circulation and hemostasis, promoting the circulation of QI to relieve pain effect.For lochia
After without cease, artificial abortion, metrorrhagia is clean, Chinese medical discrimination belongs to syndrome of blood stasis person.The hemorrhage persistent period can be shortened, alleviate lower abdomen
Pain.In State Food and Drug Administration's standard YBZ00292008, her blood peace granule use constant speed containing survey method
Flowing the most only measures the content of protocatechuic acid, and need testing solution is to use water-saturated n-butanol solution to extract preparation.This method is to master
Want effective ingredient protocatechuic acid to there is bigger negative interference, measure inaccurate, it is impossible to effectively control finished product protocatechuic acid
Content.
Summary of the invention
The technical problem to be solved in the present invention is to provide the use that a kind of good stability, favorable reproducibility, specificity are strong and simple to operate
In the detection method of content of the Chinese native medicine compound prescription pellet for the treatment of gynecologic blood diseases, eliminate negative interference, it is achieved principle active component contains
The accurate detection of amount.
For solving above-mentioned technical problem, the present invention adopts the following technical scheme that the Chinese native medicine compound prescription pellet for treating gynecologic blood diseases
Detection method of content, use high-performance liquid chromatogram determination method (one annex VI B of " Chinese Pharmacopoeia " version in 2010),
With methanol and 1% glacial acetic acid mixed liquor, her blood peace granule or her blood peace sugar type granules are carried out gradient elution, detect Chinese medicine simultaneously
Compound granular protocatechuic acid and the content of protocatechualdehyde；Wherein, Chinese native medicine compound prescription pellet need testing solution extracts through methanol eddy,
Prepare in acid condition ether extraction.
The detection method of content of the above-mentioned Chinese native medicine compound prescription pellet for treating gynecologic blood diseases, is carried out by following operation:
A, chromatographic condition and system suitability
It is filler with octadecylsilylated bonded silica gel；Flowing is made up of methanol and 1% glacial acetic acid, carries out gradient elution,
At 0-6min, methanol volume fraction is 10%, 6-10min, methanol volume fraction for be become 22%, 10-20min from 10%,
Methanol volume fraction maintains 22%, 20-25min, and methanol volume fraction is become 55% from 22%, 25-40min, methanol volume integral
Number maintenance 55%；Detection wavelength is 280nm, flow velocity 1.0ml/min, column temperature 25 DEG C；Number of theoretical plate presses protocatechuic acid and former
Catechu aldehyde peak calculates and is not less than 3000 respectively；
B, the preparation of reference substance solution
Take protocatechuic acid and protocatechualdehyde respectively appropriate, accurately weighed, add methanol and make every 1ml containing protocatechuic acid 25 μ g, former
The mixed solution of catechu aldehyde 10 μ g；
C, the preparation method of need testing solution
Take Chinese native medicine compound prescription pellet finely ground, take about 4g, accurately weighed, add methanol 50ml, be heated to reflux 1 hour, filter,
Filtrate is evaporated, and residue adds 1mol/L hydrochloric acid 20ml and dissolves, and filters, and filtrate extracts 3 times with ether, each 15ml, merges
Ether solution, puts in the conical flask filling 4g anhydrous sodium sulfate standing 30min, point takes ether solution and is evaporated, and residue methanol is appropriate
Make dissolving, be transferred in 10ml measuring bottle, add methanol to scale, shake up, filter with the microporous filter membrane of 0.45 μm, to obtain final product；
D, algoscopy
Precision draws reference substance solution and each 10 μ l of need testing solution respectively, injects chromatograph of liquid, measures, to obtain final product.
For the problem of relevant content detection existence, inventor couple at present her blood peace granule (or she blood peace sugar type granules)
It is improved, and uses the content simultaneously measuring protocatechuic acid and protocatechualdehyde under same chromatographic condition, need testing solution warp
Methanol eddy extracts, and prepares in acid condition ether extraction.Wherein, the ether solution anhydrous sodium sulfate after ether extraction is quiet
Put, be the moisture in order to carry during removing ether extraction, to improve the extraction ratio of two kinds of effective ingredient；And with methanol and
1% glacial acetic acid mixed liquor carries out gradient elution mutually as flowing, can effectively divide the material of opposed polarity in test sample
From, thus eliminate negative interference, and previous have little to no effect for a rear pin, actual mass analysis is had the biggest side
Help.The method good stability of the present invention, favorable reproducibility, specificity are strong and simple to operate, set up her blood peace granule accordingly new
Quality control standard, can help effectively to control the end product quality in her blood peace particle manufacture and and improve its quality level, it is ensured that
Production technology is controlled, it is ensured that Clinical practice curative effect, definitely provides reliable guarantee for product stable curative effect.
Accompanying drawing explanation
Fig. 1 is aging method protocatechuic acid contrast color spectrogram.
Fig. 2 is aging method feminine gender chromatogram.
Fig. 3 is for aging method sample chromatogram figure.
Fig. 4 is reference substance mixed solution chromatogram of the present invention, and A, protocatechuic acid comparison in figure, B, protocatechualdehyde compare.
Fig. 5 is feminine gender sample chromatogram of the present invention.
Fig. 6 is the chromatogram of inventive samples, and A, protocatechuic acid comparison in figure, B, protocatechualdehyde compare.
Detailed description of the invention
The instrument used in following example and reagent:
Agilent1100 high performance liquid chromatograph (includes G1311A quaternary pump, G1313A automatic sampler, G1314A VWD
Detector, G1316A column oven, Agilengt1100 chromatographic work station (Agilent Technologies of the U.S.))；Electronic analysis
Balance: BP211D (Germany Sai Duolisi)；PS-60AL ultrasonic washing instrument (40kHz, 360W)；
Protocatechuic acid reference substance (being purchased from Nat'l Pharmaceutical & Biological Products Control Institute) lot number 110809-200604；Protocatechualdehyde pair
According to product (being purchased from Nat'l Pharmaceutical & Biological Products Control Institute) lot number 110810-200506；Methanol, glacial acetic acid are chromatographically pure, its
Its reagent is all analytical pure.
The preparation of embodiment 1 need testing solution
Take Chinese native medicine compound prescription pellet finely ground, take about 4g, accurately weighed, add methanol 50ml, be heated to reflux 1 hour, filter,
Filtrate is evaporated, and residue adds 1mol/L hydrochloric acid 20ml and dissolves, and filters, and filtrate extracts 3 times with ether, each 15ml, merges
Ether solution, puts in the conical flask filling 4g anhydrous sodium sulfate standing 30min, point takes ether solution and is evaporated, and residue methanol is appropriate
Make dissolving, be transferred in 10ml measuring bottle, add methanol to scale, shake up, filter with the microporous filter membrane of 0.45 μm, to obtain final product.
Embodiment 2 screens the testing conditions of the content of Chinese native medicine compound prescription pellet protocatechuic acid, protocatechualdehyde
The screening of chromatographic condition: investigated methanol-glacial acetic acid: water (10:90:1), methanol-1% glacial acetic acid (90:lO),
Methanol-water (20:80 adjusts PH=2.8 with glacial acetic acid), acetonitrile-1% glacial acetic acid and methanol (A), 1% glacial acetic acid (B) ladder
The flow phase system such as degree eluting, compare through test, finally determine that methanol (A), 1% glacial acetic acid (B) gradient elution complete
State the assay of two kinds of compositions.
25 DEG C, 30 DEG C, 40 DEG C of different column temperatures are investigated；And 0.8,1.0,1.2mL/min different in flow rate is to each effective one-tenth
The impact separated, result of the test shows 25 DEG C, and during flow velocity 1.0mL/min, separating effect is best.
The screening of full wavelength scanner
Protocatechuic acid, protocatechualdehyde reference substance solution are carried out full wavelength scanner (200-400nm), finds that protocatechuic acid exists
There is absorption maximum at 256nm, and protocatechualdehyde has absorption maximum at 280nm.Because protocatechualdehyde content is relatively low, and
At 260nm, sample Interference Peaks is the most, therefore determines that detection wavelength is 280nm.
The selection of chromatographic column: investigated PhenomenexC18Post, Yi Lite C18Post, KromasilC18Post, SapphireC18
Post, result test shows, Yi Lite C18Post (4.6 × 250mm, 5 μm) separating effect is optimal.
Embodiment 3 Chinese native medicine compound prescription pellet (her blood peace granule) protocatechuic acid, the content detection of protocatechualdehyde
Chromatographic condition and system suitability
It is filler with octadecylsilylated bonded silica gel；Flowing is made up of methanol and 1% glacial acetic acid, carries out gradient elution,
At 0-6min, methanol volume fraction is 10%, 6-10min, methanol volume fraction for be become 22%, 10-20min from 10%,
Methanol volume fraction maintains 22%, 20-25min, and methanol volume fraction is become 55% from 22%, 25-40min, methanol volume integral
Number maintenance 55%；Detection wavelength is 280nm, flow velocity 1.0ml/min, column temperature 25 DEG C, sample size: 10 μ l number of theoretical plates are pressed
Protocatechuic acid and protocatechualdehyde peak calculate and are not less than 3000 respectively；
The preparation of reference substance solution
Precision weighs protocatechuic acid reference substance 6.26mg respectively, protocatechualdehyde compares 6.68mg and is respectively placed in the volumetric flask of 50ml
In, dissolve with methanol and be diluted to scale, shaking up, as protocatechuic acid, protocatechualdehyde reference substance storing solution；Precision respectively
Measure above-mentioned storing solution protocatechuic acid 5ml, protocatechualdehyde 2ml, put in the volumetric flask of 25ml, with methanol dilution to scale,
Shake up, obtain every 1ml mixing comparison containing protocatechuic acid (25.04 μ g/ml) and protocatechualdehyde (10.688 μ g/ml)
Product solution.
The preparation method of need testing solution
Preparing by method described in embodiment 1, she is respectively 131001,131002,131101 by blood peace granule lot number.Result
As follows:
Table 1
The methodological study of detection method of content of the present invention:
(1) linear relationship test
Respectively accurate measure above-mentioned reference substance storing solution 1.0,2.0,3.0,4.0,5.0mI, put in 25mL measuring bottle,
Add methanol to scale, shake up.Precision draws each 10 μ l of above-mentioned solution respectively, injects in chromatograph of liquid, in above-mentioned chromatograph
Under the conditions of measure protocatechuic acid, the peak area of protocatechualdehyde.With reference substance solution concentration as abscissa, respectively with protocatechuic acid,
Protocatechualdehyde integrating peak areas value is vertical coordinate, prepares standard curve, and regression equation is respectively as follows:
Y=16.897X-2.48, R2=0.9995 (n=5)；Y=51.897X-17.32, R2=0.9996 (n=5), protocatechuic acid and former
There is good linear relationship the catechu aldehyde range of linearity respectively 5.008～25.04 μ g/ml, 5.344～26.72 in μ g/ml.
(2) Precision Experiment
Accurate above-mentioned protocatechuic acid and the mixing reference substance solution 10 μ L of protocatechualdehyde of drawing, in injection chromatograph of liquid, by upper
State chromatographic condition to measure, parallel assay 5 times.Record 5 protocatechuic acid, the meansigma methodss of protocatechualdehyde peak area to be respectively
419.8、547.5；Calculate RSD and be respectively 0.86%, 0.56% (n=5), show that instrument precision is good.
(3) stability experiment
At 0,2,4,8,12 hours, accurate absorption pacified, with batch her blood, the need testing solution 10 that granule (131001) is made
μ l, injects in chromatograph of liquid, measures by above-mentioned chromatographic condition, records 5 protocatechuic acid, protocatechualdehyde peak area
Meansigma methods is respectively 352.7,201.8；Calculate RSD and be respectively 0.73%, 0.64% (n=5), show sample solution
There is in 12h preferable stability.
(4) repeated experiment
Take with criticizing her blood peace granule (131001) 5 parts, every part of about 4g, accurately weighed, respectively according to described in embodiment 1
Method prepares need testing solution, measures protocatechuic acid and the content of protocatechualdehyde by above-mentioned chromatographic condition.Result protocatechuic acid and
The average content of protocatechualdehyde is respectively 0.797mg/15g, 0.0360mg/15g, calculates RSD and is respectively 0.61%, 0.80%
(n=5), show that this method repeatability is preferable.
(5) sample-adding recovery experiment
Take and pacify granule (131001) (protocatechuic acid content 0.789mg/15g, 0.0788mg/15g) about with her blood a batch of
2.0g, totally five parts, accurately weighed, precision adds protocatechuic acid reference substance (25.04 μ g/ml) 5ml, protocatechualdehyde respectively
Comparison (10.688 μ g/ml) 1ml, the method as described in embodiment 1 prepares need testing solution, surveys by above-mentioned chromatographic condition
Determine the content of protocatechuic acid, protocatechualdehyde, calculate average recovery.Result protocatechuic acid, protocatechualdehyde mean sample reclaim
Being respectively 98.24, RSD is 1.68%；98.83%, RSD are 1.85% (n=5).
Her blood peace granule protocatechuic acid of embodiment 4 contains the comparison of survey aging method and the present invention
Aging method:
Chromatographic condition and system suitability are with octadecylsilane chemically bonded silica as filler: methanol-water-glacial acetic acid (10:
89:1) for flowing phase；Detection wavelength is 256nm.Number of theoretical plate is calculated by protocatechuic acid peak should be not less than 3000.
It is appropriate that the preparation precision of reference substance solution weighs protocatechuic acid reference substance, adds methanol and makes molten containing 25 μ g of every 1ml
Liquid, to obtain final product.
The preparation of need testing solution takes the about 4g of this product under content uniformity item, and accurately weighed, the 40ml that adds water makes dissolving, adds
Hydrochloric acid 0.3ml, stirs evenly, and stands 10 minutes, filters, with water 10ml washing container at twice and filtering residue, merging filtrate and
Washing liquid, extracts 5 times with water-saturated n-butanol, each 15ml, merges n-butanol extracting liquid, reclaims n-butyl alcohol, and residue adds first
Alcohol makes dissolving in right amount, moves in 10ml measuring bottle, adds methanol to scale, shakes up, to obtain final product.
Algoscopy precision respectively draws reference substance solution and each 10 μ l of need testing solution, injects chromatograph of liquid, measures,
Obtain.
(1) comparison (her blood peace granule 131001) of content
Table 2
(2) trace analysis is shown in Fig. 1-6.
Claims (1)
1., for treating a detection method of content for the Chinese native medicine compound prescription pellet of gynecologic blood diseases, use high-performance liquid chromatogram determination method,
It is characterized in that carrying out by following operation:
A, chromatographic condition and system suitability
It is filler with octadecylsilylated bonded silica gel；Flowing is made up of methanol and 1% glacial acetic acid, carries out gradient elution,
At 0-6min, methanol volume fraction is 10%, 6-10min, methanol volume fraction for be become 22%, 10-20min from 10%,
Methanol volume fraction maintains 22%, 20-25min, and methanol volume fraction is become 55% from 22%, 25-40min, methanol volume integral
Number maintenance 55%；Detection wavelength is 280nm, flow velocity 1.0ml/min, column temperature 25 DEG C；Number of theoretical plate presses protocatechuic acid and former
Catechu aldehyde peak calculates and is not less than 3000 respectively；
B, the preparation of reference substance solution
Take protocatechuic acid and protocatechualdehyde respectively appropriate, accurately weighed, add methanol and make every 1ml containing protocatechuic acid 25 μ g, former
The mixed solution of catechu aldehyde 10 μ g；
C, the preparation method of need testing solution
Take her blood peace granule or her blood peace sugar type granules is finely ground, take about 4g, accurately weighed, add methanol 50ml, heat back
Flowing 1 hour, filter, filtrate is evaporated, and residue adds 1mol/L hydrochloric acid 20ml and dissolves, and filters, and filtrate extracts 3 times with ether,
Every time 15ml, merges ether solution, puts standing 30min in the conical flask filling 4g anhydrous sodium sulfate, point takes ether solution and is evaporated,
Residue methanol makes dissolving in right amount, is transferred in 10ml measuring bottle, adds methanol to scale, shakes up, with the micropore of 0.45 μm
Filter membrane filters, and to obtain final product；
D, algoscopy
Precision draws reference substance solution and each 10 μ l of need testing solution respectively, injects chromatograph of liquid, measures, to obtain final product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410452215.7A CN104215708B (en) | 2014-09-05 | 2014-09-05 | For treating the detection method of content of the Chinese native medicine compound prescription pellet of gynecologic blood diseases |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410452215.7A CN104215708B (en) | 2014-09-05 | 2014-09-05 | For treating the detection method of content of the Chinese native medicine compound prescription pellet of gynecologic blood diseases |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104215708A CN104215708A (en) | 2014-12-17 |
| CN104215708B true CN104215708B (en) | 2016-08-31 |
Family
ID=52097422
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410452215.7A Active CN104215708B (en) | 2014-09-05 | 2014-09-05 | For treating the detection method of content of the Chinese native medicine compound prescription pellet of gynecologic blood diseases |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104215708B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106053624A (en) * | 2016-05-12 | 2016-10-26 | 广西万寿堂药业有限公司 | Method for measuring contents of protocatechuic acid and protocatechualdehyde in Yi Xue'an particles |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| MY170986A (en) * | 2009-05-26 | 2019-09-23 | Malaysian Palm Oil Board | Composition comprising caffeoylshikimic acids, protocatechuic acid, hydroxytyrosol, hydroxybenzoic acid and their derivatives and method of preparation thereof |
| KR20110018178A (en) * | 2009-08-17 | 2011-02-23 | 대한민국(관리부서 : 산림청 국립산림과학원장) | Compound isolated from the bark of phellodendron amurense, isolating method of thereof and application |
-
2014
- 2014-09-05 CN CN201410452215.7A patent/CN104215708B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN104215708A (en) | 2014-12-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109212083A (en) | The quality determining method of compound endothelium corneum gigeriae galli chewable tablets | |
| CN102221590B (en) | Method for simultaneously determining multi-index ingredients of Simotang preparation and establishing fingerprint chromatogram thereof | |
| CN101766771A (en) | Quality control method of medicine for gynecopathy treatment | |
| CN108802255A (en) | The method for measuring ganoderic acid A and Ganoderma lucidum triterpenes components content in compound preparation | |
| CN104062374B (en) | The detection method of the Chinese medicine composition of invigorating Qi and tonifying kidney | |
| CN103808835B (en) | The method of 10 kinds of chemical composition contents in HPLC-DAD method Simultaneously test Siwu Tang decoction | |
| CN104345110A (en) | Content determination method for seven compositions in traditional Chinese medicine composition preparation | |
| CN103115984A (en) | Quality control method of medicament for treating leukopenia | |
| CN104597139B (en) | Method for simultaneously determining three kinds of phenylethanoid glycoside compositions in callicarpa nudiflora preparation through HPLC | |
| CN106198810B (en) | A kind of quality determining method of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow suppression | |
| CN103822975A (en) | Test method for pulse-activating preparation adopting pilose asiabell root | |
| CN104215708B (en) | For treating the detection method of content of the Chinese native medicine compound prescription pellet of gynecologic blood diseases | |
| CN103389353B (en) | A kind of method of measuring liposoluble ingredient content in blood-nourishing and brain-refreshing granules by Ultra Performance Liquid Chromatography method | |
| CN104483435B (en) | A kind of detection method of gastrodia-glossy ganoderma granule | |
| CN103230453A (en) | Yinchengao Decoction formula particles, and preparation method and detection method thereof | |
| CN102008541B (en) | Method for simultaneously detecting three main active ingredients in sugar-free type compound wintercreeper preparation | |
| CN105510452A (en) | Multiple index component content determination, fingerprint building and preparation methods of liver-tonifying eyesight-improving oral liquid | |
| CN105067725B (en) | It is a kind of at the same determine shenmei stomach nourishing granule in the root of herbaceous peony, radix glycyrrhizae and the red sage root content method | |
| CN108414667A (en) | The detection method of one seed ginseng osmanthus Yixin granule quality standard | |
| CN101028474B (en) | Method for inspecting the quality of Chinese preparation with Yang-and kidney tonifying functions | |
| CN103575823A (en) | Detection method of 8 chemical components in Tangminling preparation | |
| CN103823000B (en) | A kind of quality determining method of root of Japanese banana | |
| CN103592392B (en) | Determination method for tanshinol content in Naozhengning preparation | |
| CN105891371B (en) | Detection method for Maidang lactation promoting particles | |
| CN105823830A (en) | Method for measuring content of salvianolic acid B and schisandrin in heart-benefiting pulse-invigorating granule by quantitative analysis of multi-components by single marker(QAMS) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |