CN110187031A - The measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment - Google Patents
The measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment Download PDFInfo
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- CN110187031A CN110187031A CN201910507252.6A CN201910507252A CN110187031A CN 110187031 A CN110187031 A CN 110187031A CN 201910507252 A CN201910507252 A CN 201910507252A CN 110187031 A CN110187031 A CN 110187031A
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- G—PHYSICS
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- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
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Abstract
The invention discloses a kind of measuring methods of urea content in Moschus blood-circulation and removing blood stasis ointment, which is characterized in that includes following steps: 1) preparing the reference substance solution of urea using methanol solution as solvent；2) Moschus blood-circulation and removing blood stasis ointment is taken, obtains sample to be tested after shearing；3) with alcohol solvent, dissolution extracts sample to be tested, obtains test solution；4) using the content of urea in high effective liquid chromatography for measuring reference substance solution and test solution；Chromatographic test strip part are as follows: using amino bonded silica gel as filler, mobile phase is the acetonitrile-aqueous solution of Volume fraction 92:8；Detection wavelength is 190nm；Sample volume 10uL；The method that technical characteristic adopted by the present invention solves measurement urea content at present is relatively complicated, and due in Moschus blood-circulation and removing blood stasis ointment interfering components it is more, lead to be difficult to easy the problem of accurately detecting urea content in Moschus blood-circulation and removing blood stasis ointment.
Description
Technical field
The present invention relates to medication components content detection fields, and in particular to urea content in a kind of Moschus blood-circulation and removing blood stasis ointment
Measuring method.
Background technique
Moschus blood-circulation and removing blood stasis ointment has the effect of activating microcirculation and removing stasis medicinal, anti-inflammatory analgetic；For joint sprain, soft tissue bruise is anxious
Property lumbar sprain lumbar muscle strain, scapulohumeral periarthritis, pernio of not bursting, nodositas is red；Moschus blood-circulation and removing blood stasis ointment is as a kind of Chinese patent drug, composition
In do not comprise only several taste Chinese materia medica preparations, further include having western medicine composition；In order to examine and determine in Moschus blood-circulation and removing blood stasis ointment it is each effectively at
Part, there is the measuring method to wherein various component contents, but the rare measuring method to wherein urea content；Dermatology with
Certain medicaments containing urea improve the humidity of skin, thus content of the urea in Moschus blood-circulation and removing blood stasis ointment to drug effect have compared with
Big influence；At present measurement urea content method it is relatively complicated, and due in Moschus blood-circulation and removing blood stasis ointment interfering components it is more, lead
Cause is difficult to easy the problem of accurately detecting urea content in Moschus blood-circulation and removing blood stasis ointment.
Summary of the invention
The purpose of the present invention is to provide a kind of measuring methods of urea content in Moschus blood-circulation and removing blood stasis ointment, solve measurement urine
The method of cellulose content is relatively complicated, and due in Moschus blood-circulation and removing blood stasis ointment interfering components it is more, cause to be difficult to easy accurately to examine
The problem of measuring urea content in Moschus blood-circulation and removing blood stasis ointment.
In order to solve the above technical problems, the invention adopts the following technical scheme: urea in a kind of Moschus blood-circulation and removing blood stasis ointment
The measuring method of content, which is characterized in that include following steps:
1) reference substance solution of urea is prepared using methanol solution as solvent；
2) Moschus blood-circulation and removing blood stasis ointment is taken, obtains sample to be tested after shearing；
3) with alcohol solvent, dissolution extracts sample to be tested, obtains test solution；
4) using the content of urea in high effective liquid chromatography for measuring reference substance solution and test solution；
Chromatographic test strip part are as follows: using amino bonded silica gel as filler, mobile phase is that the acetonitrile-water of Volume fraction 93:7 is molten
Liquid；Detection wavelength is 200nm；Sample volume 20uL；
Further, the Moschus blood-circulation and removing blood stasis ointment includes following composition: Moschus, Radix Notoginseng, safflower, Radix Salviae Miltiorrhizae, boric acid, camphor,
Dragon's blood, urea, belladonna liquid extract, bagodryl hydrochloride and procaine hydrochloride；
Further, the Moschus blood-circulation and removing blood stasis ointment is made using following preparation method:
Step S1: being ground into coarse powder for Radix Notoginseng, Radix Salviae Miltiorrhizae, and safflower is added and is uniformly mixed, and the ethyl alcohol of 6 times of amounts is added as solvent, shines
Percolation under liquid extract and extract item carries out diacolation after impregnating 24 hours, collects liquid of filtering, and the liquid that will filter obtains after being concentrated under reduced pressure
To concentrate；
Step S2: after boric acid, camphor are dissolved with ethyl alcohol, being added in the concentrate that step S1 is obtained, and adjusts total volume with ethyl alcohol, obtains
To lotion；
Step S3: 2.8 times of amounts are added in Moschus, dragon's blood, urea, belladonna liquid extract, bagodryl hydrochloride and procaine hydrochloride
After matrix, coating is made, is coated on lotion, cutting, cover lining are cut into small pieces to get the Moschus blood-circulation and removing blood stasis ointment is arrived；
Further, the concrete operations of the reference substance solution of urea are prepared in step 1) are as follows: take urea reference substance appropriate, precision claims
After fixed, add methanol that the solution of every 1mL methanol reference substance of urea containing 0.025mg is made, reference substance solution can be obtained；
Further, the concrete operations of sample to be tested are prepared in step 2 are as follows: take Moschus blood-circulation and removing blood stasis ointment 100cm2, be cut into narrow
Item removes cover lining, obtains sample to be tested；
Further, step 3) includes:
Step A: sample to be tested is placed in stuffed conical flask, and the accurate 50mL ethyl alcohol measured, weighed weight is added；
Step B: after heating and refluxing extraction, letting cool, then weighed weight；
Step C: supplying the weight of less loss with ethyl alcohol, shake up, and filtering takes subsequent filtrate, test solution can be obtained；
Further, the time being heated to reflux in step B is 55min~65min, and keeping temperature is 78 DEG C~83 DEG C slightly boiled；
Further, it after being heated to reflux, lets cool to temperature and reaches 20 DEG C~30 DEG C；
Further, the temperature for the chromatographic column that chromatography detects in step 4) is 20 DEG C -40 DEG C；
Further technical solution is that the type of elution that chromatography detects in step 4) is isocratic elution.
Compared with prior art, the beneficial effects of the present invention are:
1, the release of active principle can more be allowed by being heated to reflux processing sample, big to effective component solubility and dissolve to other compositions
Small solvent is spent, effective component is dissolved out from sample, guarantees that material liquid component is uniform, improves yield when extracting；
2, using the acetonitrile-aqueous solution of Volume fraction 93:7 as mobile phase, can smoothly appearance improve detection efficiency, peak compared with
It is good, no hangover and miscellaneous peak is few；Liquid chromatography for measuring is directlyed adopt without performing the derivatization to urea, it is simple and convenient；
3, by selecting suitable testing conditions, so that the separating degree of urea content in HPLC measurement Moschus blood-circulation and removing blood stasis ointment,
Accuracy, reproducibility, stability etc. all meet standard requirements.
Detailed description of the invention
Fig. 1 is reference substance solution chromatogram.
Fig. 2 is test solution chromatogram.
Fig. 3 is urea canonical plotting.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
Embodiment 1: the measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment, includes following steps:
1) reference substance solution of urea is prepared using methanol solution as solvent；
2) Moschus blood-circulation and removing blood stasis ointment is taken, obtains sample to be tested after shearing；
3) with alcohol solvent, dissolution extracts sample to be tested, obtains test solution；
4) using the content of urea in high effective liquid chromatography for measuring reference substance solution and test solution；
Chromatographic test strip part are as follows: using amino bonded silica gel as filler, mobile phase is that the acetonitrile-water of Volume fraction 93:7 is molten
Liquid；Detection wavelength is 200nm；Sample volume 20uL.
The Moschus blood-circulation and removing blood stasis ointment includes following composition: Moschus, Radix Notoginseng, safflower, Radix Salviae Miltiorrhizae, boric acid, camphor, dragon's blood,
Urea, belladonna liquid extract, bagodryl hydrochloride and procaine hydrochloride.
The Moschus blood-circulation and removing blood stasis ointment is made using following preparation method:
Step S1: being ground into coarse powder for Radix Notoginseng, Radix Salviae Miltiorrhizae, and safflower is added and is uniformly mixed, and the ethyl alcohol of 6 times of amounts is added as solvent, shines
Percolation under liquid extract and extract item carries out diacolation after impregnating 24 hours, collects liquid of filtering, and the liquid that will filter obtains after being concentrated under reduced pressure
To concentrate；
Step S2: after boric acid, camphor are dissolved with ethyl alcohol, being added in the concentrate that step S1 is obtained, and adjusts total volume with ethyl alcohol, obtains
To lotion；
Step S3: 2.8 times of amounts are added in Moschus, dragon's blood, urea, belladonna liquid extract, bagodryl hydrochloride and procaine hydrochloride
After matrix, coating is made, is coated on lotion, cutting, cover lining are cut into small pieces to get the Moschus blood-circulation and removing blood stasis ointment is arrived.
Wherein, matrix is made of rubber and rosin, is prepared using pure plant, green and healthy, and has good
Structural strength.
The concrete operations of the reference substance solution of urea are prepared in step 1) are as follows: take urea reference substance appropriate, after accurately weighed,
Add methanol that the solution of every 1mL methanol reference substance of urea containing 0.025mg is made, reference substance solution can be obtained.
The concrete operations of sample to be tested are prepared in step 2 are as follows: take Moschus blood-circulation and removing blood stasis ointment 100cm2, be cut into fillet, remove
Cover lining obtains sample to be tested.
Step 3) includes:
Step A: sample to be tested is placed in stuffed conical flask, and the accurate 50mL ethyl alcohol measured, weighed weight is added；
Step B: after heating and refluxing extraction, letting cool, then weighed weight；
Step C: supplying the weight of less loss with ethyl alcohol, shake up, and filtering takes subsequent filtrate, test solution can be obtained.
The release of active principle can more be allowed by being heated to reflux processing sample, big and molten to other compositions to effective component solubility
Effective component is dissolved out by Xie Du small solvent from sample, is guaranteed that material liquid component is uniform, is improved yield when extracting.
Wherein, the time being heated to reflux in step B is 55min~65min, and keeping temperature is 78 DEG C~83 DEG C slightly boiled；Add
After heat reflux, lets cool to temperature and reach 20 DEG C~30 DEG C.
Wherein, the temperature of the chromatographic column of middle chromatography detection is 20 DEG C~40 DEG C.
Wherein, the type of elution that chromatography detects in step 4) is isocratic elution.
(1) linear to investigate
Precision draws urea reference substance solution (0.02499mg/ml) 2,6,10,14,18,22 μ l sample introduction, chromatography is recorded, with peak face
Product is abscissa, is ordinate mapping with sample volume (μ g), calculates regression equation are as follows: A=34815.0689C-136.0095 (r=
0.9999), regression equation was fitted the linear equation of origin are as follows: A=34471.9966C；By a test solution, sample introduction is analyzed,
Gained peak area substitutes into upper two formula respectively and calculates, relative deviation 0.46%, therefore is believed that standard curve crosses origin, regression equation
No intercept.Thus one point external standard method calculating can be used in assay.Linear relationship is good between 0.05 μ of μ g~0.5498 g.
1 urea linear relationship of table is investigated
(2) precision test
Reference substance solution 0.02499mg/ml is taken, is repeated sample introduction 6 times, chromatography is recorded, the results are shown in Table 2, RSD 0.77%, explanation has
Good precision.
2 precision test of table
(3) repetitive test
It takes a sample to prepare 6 parts of test solutions by step 3), respectively sample introduction, records chromatography, calculate, the results are shown in Table 3, RSD is
0.66%, illustrate there is good repeatability.
3 repetitive test of table
(4) recovery test
Recycled using sample-adding, precision draws the sample 50cm2(1.2352mg/100cm2 for having measured content), add urea pair
It is appropriate according to product, it is measured by chromatographic condition described in step 1), calculates the rate of recovery, the results are shown in Table 4, the rate of recovery of urea is 101.0%,
RSD is 1.06%, illustrates that this law has the good rate of recovery.
4 recovery test of table
(5) specificity
Moschus blood-circulation and removing blood stasis ointment and negative sample is taken to prepare sample solution and negative sample solution by sample solution preparation method,
By step 4) chromatographic condition, reference substance solution, sample solution, negative sample solution injection liquid chromatograph are taken respectively, and sample is molten
Liquid chromatography is equipped with corresponding chromatographic peak in reference substance solution chromatography corresponding positions, and negative noiseless.
(6) durability
The selection of sample extraction solvent: taking 5 parts of Moschus blood-circulation and removing blood stasis ointment, and every part of 100cm2 is set in stuffed conical flask, accurate respectively
Ethyl alcohol, methanol, 70% ethyl alcohol, 80% ethyl alcohol, each 50ml of 90% ethyl alcohol is added, weighed weight heating and refluxing extraction 60 minutes, is let cool,
Weighed weight again is supplied less loss weight with ethyl alcohol, methanol, 70% methanol, 80% methanol, 90% methanol respectively, is shaken up, and filtration takes continuous
Filtrate is as test solution.Each 10 μ l injection liquid chromatograph of above-mentioned solution is taken respectively, is calculated, be the results are shown in Table 5, illustrate this product
Using ethyl alcohol as Extraction solvent, content is extracted more complete.
The selection of 5 Extraction solvent of table
The selection of sample extraction method: taking 2 parts of Moschus blood-circulation and removing blood stasis ointment, and every part of 100cm2 is set in stuffed conical flask respectively, accurate
Ethyl alcohol 50ml is added, a copy of it is ultrasonically treated 60 minutes, another refluxing extraction 60 minutes is taken out, lets cool, supplied with ethyl alcohol
Less loss weight, shakes up, and filtration takes 10 μ l subsequent filtrates to inject liquid chromatograph respectively, calculates, the results are shown in Table 6, refluxing extraction content
It is obviously higher than ultrasonic extraction content, therefore select refluxing extraction.
The selection of 6 extracting method of table
Sample extraction time effects: taking 4 parts of Moschus blood-circulation and removing blood stasis ointment, and every part of 100cm2 is set in stuffed conical flask respectively, and precision adds
Enter ethyl alcohol 50ml, refluxing extraction 15,30,60,90 minutes respectively, takes out, let cool, supply less loss weight with ethyl alcohol, shake up, filter
It crosses, 10 μ l subsequent filtrates is taken to inject liquid chromatograph respectively, calculate content, the results are shown in Table 7, just mention completely within refluxing extraction 60 minutes
It takes, therefore the selective extraction time is 60 minutes.
7 extraction time of table influences
(7) three batches of sample assays
Although reference be made herein to invention has been described for multiple explanatory embodiments of the invention, however, it is to be understood that ability
Field technique personnel can be designed that a lot of other modification and implementations, these modifications and implementations will fall in the application public affairs
Within the scope and spirit opened.It more specifically, within the scope of the present disclosure and claims, can be to theme group
The building block and/or layout for closing layout carry out a variety of variations and modifications.In addition to the deformation carried out to building block and/or layout
Outer with improving, to those skilled in the art, other purposes also will be apparent.
Claims (10)
1. the measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment, which is characterized in that include following steps:
1) reference substance solution of urea is prepared using methanol solution as solvent；
2) Moschus blood-circulation and removing blood stasis ointment is taken, obtains sample to be tested after shearing；
3) with alcohol solvent, dissolution extracts sample to be tested, obtains test solution；
4) using the content of urea in high effective liquid chromatography for measuring reference substance solution and test solution；
Chromatographic test strip part are as follows: using amino bonded silica gel as filler, mobile phase is that the acetonitrile-water of Volume fraction 93:7 is molten
Liquid；Detection wavelength is 200nm；Sample volume 20uL.
2. the measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment according to claim 1, it is characterised in that: institute
Stating Moschus blood-circulation and removing blood stasis ointment includes following composition: Moschus, Radix Notoginseng, safflower, Radix Salviae Miltiorrhizae, boric acid, camphor, dragon's blood, urea, belladonna stream
Medicinal extract, bagodryl hydrochloride and procaine hydrochloride.
3. the measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment according to claim 2, it is characterised in that: institute
Moschus blood-circulation and removing blood stasis ointment is stated to be made using following preparation method:
Step S1: being ground into coarse powder for Radix Notoginseng, Radix Salviae Miltiorrhizae, and safflower is added and is uniformly mixed, and the ethyl alcohol of 6 times of amounts is added as solvent, shines
Percolation under liquid extract and extract item carries out diacolation after impregnating 24 hours, collects liquid of filtering, and the liquid that will filter obtains after being concentrated under reduced pressure
To concentrate；
Step S2: after boric acid, camphor are dissolved with ethyl alcohol, being added in the concentrate that step S1 is obtained, and adjusts total volume with ethyl alcohol, obtains
To lotion；
Step S3: 2.8 times of amounts are added in Moschus, dragon's blood, urea, belladonna liquid extract, bagodryl hydrochloride and procaine hydrochloride
After matrix, coating is made, is coated on lotion, cutting, cover lining are cut into small pieces to get the Moschus blood-circulation and removing blood stasis ointment is arrived.
4. the measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment according to claim 1, it is characterised in that: step
It is rapid 1) in prepare urea reference substance solution concrete operations are as follows: take urea reference substance appropriate, after accurately weighed, methanol added to be made
The solution of every 1mL methanol reference substance of urea containing 0.025mg, can be obtained reference substance solution.
5. the measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment according to claim 1, it is characterised in that: step
It is rapid 2) in prepare the concrete operations of sample to be tested are as follows: take Moschus blood-circulation and removing blood stasis ointment 100cm2, it is cut into fillet, cover lining is removed, obtains
Sample to be tested.
6. the measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment according to claim 1, it is characterised in that: step
It is rapid 3) to include:
Step A: sample to be tested is placed in stuffed conical flask, and the accurate 50mL ethyl alcohol measured, weighed weight is added；
Step B: after heating and refluxing extraction, letting cool, then weighed weight；
Step C: supplying the weight of less loss with ethyl alcohol, shake up, and filtering takes subsequent filtrate, test solution can be obtained.
7. the measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment according to claim 6, it is characterised in that: step
The time being heated to reflux in rapid B is 55min~65min, and keeping temperature is 78 DEG C~83 DEG C slightly boiled.
8. the measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment according to claim 6, it is characterised in that: add
After heat reflux, lets cool to temperature and reach 20 DEG C~30 DEG C.
9. the measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment according to claim 1, it is characterised in that: step
It is rapid 4) in chromatography detection chromatographic column temperature be 20 DEG C -40 DEG C.
10. the measuring method of urea content in a kind of Moschus blood-circulation and removing blood stasis ointment according to claim 1, it is characterised in that:
The type of elution that chromatography detects in step 4) is isocratic elution.
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Cited By (1)
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| KR101286225B1 (en) * | 2011-06-09 | 2013-07-23 | 윤근수 | Multi-transformation folding bicycle |
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