CN110231417A - A kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content - Google Patents
A kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content Download PDFInfo
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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
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Abstract
The invention discloses a kind of measuring methods of SHEXIANG ZHUANGGU GAO multicomponent content, include following steps: 1) preparing inner mark solution；2) it takes menthol reference substance, camphor reference substance, gaultherolin reference substance and borneol reference substance to be placed in same volumetric flask, inner mark solution is added, is diluted to scale with ethyl acetate, reference substance solution can be obtained；3) SHEXIANG ZHUANGGU GAO is taken, obtains sample to be tested after shearing；4) ethyl acetate solution is used, dissolution extracts sample to be tested, obtains test solution；5) using gas chromatography measurement reference substance solution each component content and test solution each component content；Gas chromatographic detection condition are as follows: chromatographic column selects the capillary column with PEG 20000 (PEG20M) for stationary phase；Sample volume 1uL；140 DEG C of column temperature；The problem of the technical solution used in the present invention solves the measuring method that can measure SHEXIANG ZHUANGGU GAO Multicomponent content directly under same chromatographic condition currently without one kind, leads to content detection low efficiency.
Description
Technical field
The present invention relates to medication components content detection fields, and in particular to a kind of measurement of SHEXIANG ZHUANGGU GAO multicomponent content
Method.
Background technique
SHEXIANG ZHUANGGU GAO is for easing pain, anti-inflammatory, is used for rheumatalgia, arthralgia, pain in the loins, neuralgia, and DOMS is sprained,
It dampens；SHEXIANG ZHUANGGU GAO is a kind of Chinese patent drug, wherein plurality of Chinese component and Western medicine component are contained, in order to effectively monitor musk deer
The quality of fragrant bone-strengthening plaster needs to carry out assay to components various in SHEXIANG ZHUANGGU GAO, but in order to avoid between component
Interference, at present to the measuring method of SHEXIANG ZHUANGGU GAO content, can only select using liquid chromatograph or gas chromatograph respectively,
Each component is detected respectively, the survey of Multicomponent content can be measured directly under same chromatographic condition without one kind
The problem of determining method, leading to content detection low efficiency.
Summary of the invention
The purpose of the present invention is to provide a kind of measuring methods of SHEXIANG ZHUANGGU GAO multicomponent content, solve to not have at present
There is a kind of measuring method that can measure SHEXIANG ZHUANGGU GAO Multicomponent content directly under same chromatographic condition, content is caused to be examined
The problem of surveying low efficiency.
In order to solve the above technical problems, the invention adopts the following technical scheme: a kind of SHEXIANG ZHUANGGU GAO multicomponent content
Measuring method, include following steps:
1) inner mark solution is prepared；
2) menthol reference substance, camphor reference substance, gaultherolin reference substance and borneol reference substance is taken to be placed in same volumetric flask,
Inner mark solution is added, is diluted to scale with ethyl acetate, reference substance solution can be obtained；
3) SHEXIANG ZHUANGGU GAO is taken, obtains sample to be tested after shearing；
4) ethyl acetate solution is used, dissolution extracts sample to be tested, obtains test solution；
5) using gas chromatography measurement reference substance solution each component content and test solution each component content；
Gas chromatographic detection condition are as follows: chromatographic column selects the capillary column with PEG 20000 (PEG20M) for stationary phase；Into
Sample amount 1uL；140 DEG C of column temperature；
Further, step 1) prepares the concrete operations of inner mark solution are as follows: it is appropriate that precision weighs naphthalene, and ethyl acetate is added, is made
The solution of every 1mL ethyl acetate naphthalene containing 10mg to get arrive inner mark solution；
Further, step 2 preparation reference substance solution concrete operations are as follows: take respectively menthol reference substance, camphor reference substance,
Salicylic acid reference substance, each 10mg of borneol reference substance, are placed in together in 10mL volumetric flask, and inner mark solution 1mL is added in precision, use acetic acid
Ethyl ester is diluted to scale, shakes up, and reference substance solution can be obtained；
Further, step 3) prepares the concrete operations of sample to be tested are as follows: takes SHEXIANG ZHUANGGU GAO 210cm2, is cut into fillet, removes
Cover lining obtains sample to be tested；
Further, step 4) includes:
Step S1: sample to be tested is placed in conical flask, and ethyl acetate 50mL, weighed weight is added in precision；
Step S2: after heating and refluxing extraction, letting cool, then weighed weight；
Step S3: less loss weight is supplied with ethyl acetate；
Step S4: filtering, precision draw subsequent filtrate 20mL in 25mL volumetric flask, and 2.5mL inner mark solution is added in precision, uses acetic acid
Ethyl ester is diluted to scale, shakes up, and test solution can be obtained；
Further, time 55min~65min that step S2 is heated to reflux, 75 DEG C~80 DEG C of temperature holding are slightly boiled；
Further, step S2 is let cool to 20 DEG C~30 DEG C of temperature；
Further, the time 60min that step S2 is heated to reflux, temperature are kept for 80 DEG C；
Further, the detection temperature of step 5) gas chromatograph is
Further technical scheme steps 5) in, theoretical cam curve calculates >=5000 by naphthalene peak.
Compared with prior art, the beneficial effects of the present invention are:
1, detect that menthol in SHEXIANG ZHUANGGU GAO, camphor, gaultherolin, borneol are respective to be contained simultaneously using same chromatographic condition
Amount, it is simple and efficient to handle, it is at low cost；
2, 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；
3, by selecting suitable testing conditions, so that the HPLC measures separating degree, the standard of each component content in SHEXIANG ZHUANGGU GAO
True property, reproducibility, stability etc. all meet standard requirements.
Detailed description of the invention
Fig. 1 is reference substance solution spectrogram.
Fig. 2 is test solution spectrogram.
Fig. 3 is menthol canonical plotting.
Fig. 4 is camphor canonical plotting.
Fig. 5 is gaultherolin canonical plotting.
Fig. 6 is borneol 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: a kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content, includes following steps:
1) inner mark solution is prepared；
2) menthol reference substance, camphor reference substance, gaultherolin reference substance and borneol reference substance is taken to be placed in same volumetric flask,
Inner mark solution is added, is diluted to scale with ethyl acetate, reference substance solution can be obtained；
3) SHEXIANG ZHUANGGU GAO is taken, obtains sample to be tested after shearing；
4) ethyl acetate solution is used, dissolution extracts sample to be tested, obtains test solution；
5) using gas chromatography measurement reference substance solution each component content and test solution each component content；
Gas chromatographic detection condition are as follows: chromatographic column selects the capillary column with PEG 20000 (PEG20M) for stationary phase；Into
Sample amount 1uL；140 DEG C of column temperature.
Wherein, the column length of capillary column is 30m, internal diameter 0.32mm, film thickness 0.25um.
This method SHEXIANG ZHUANGGU GAO detected, by weight score calculate, and include following composition: medicinal material medicinal extract 95~
110 parts, 0.30~0.50 part of Moschus, 30~50 parts of menthol, 20~35 parts of gaultherolin, 2~5 parts of chondroitin sulfate, ice
25~35 parts of piece, 6~8 parts of bagodryl hydrochloride, 28~38 parts of camphor.
The SHEXIANG ZHUANGGU GAO the preparation method comprises the following steps: take 100 parts of medicinal material medicinal extract, 0.24 part of Moschus, 40 parts of menthols,
25.5 parts of gaultherolins, 4.8 parts of chondroitin sulfates, 30 parts of borneols, 6 parts of bagodryl hydrochlorides and 30 parts of camphors, separately plus 5.3~
The matrix of 5.7 times of weight after coating is made, carries out painting cream, dissection, and cover lining is cut into small pieces, and SHEXIANG ZHUANGGU GAO can be obtained.
Wherein, matrix is made of rubber, rosin.
Wherein, step 1) prepares the concrete operations of inner mark solution are as follows: it is appropriate that precision weighs naphthalene, and ethyl acetate is added, is made
The solution of every 1mL ethyl acetate naphthalene containing 10mg to get arrive inner mark solution.
Wherein, step 2 preparation reference substance solution concrete operations are as follows: take respectively menthol reference substance, camphor reference substance,
Salicylic acid reference substance, each 10mg of borneol reference substance, are placed in together in 10mL volumetric flask, and inner mark solution 1mL is added in precision, use acetic acid
Ethyl ester is diluted to scale, shakes up, and reference substance solution can be obtained.
Wherein, step 3) prepares the concrete operations of sample to be tested are as follows: takes SHEXIANG ZHUANGGU GAO 210cm2, is cut into fillet, removes
Cover lining obtains sample to be tested.
Wherein, step 4) includes:
Step S1: sample to be tested is placed in conical flask, and ethyl acetate 50mL, weighed weight is added in precision；
Step S2: after heating and refluxing extraction, letting cool, then weighed weight；
Step S3: less loss weight is supplied with ethyl acetate；
Step S4: filtering, precision draw subsequent filtrate 20mL in 25mL volumetric flask, and 2.5mL inner mark solution is added in precision, uses acetic acid
Ethyl ester is diluted to scale, shakes up, and 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.
Time 55min~65min that step S2 is heated to reflux, 75 DEG C~80 DEG C of temperature holding being heated to reflux are slightly boiled.
Step S2 is let cool to 20 DEG C~30 DEG C of temperature.
The time that step S2 is heated to reflux is 60min；The temperature being heated to reflux is kept for 80 DEG C.
The detection temperature of gas chromatograph is 260 DEG C in step 5).
In step 5), theoretical cam curve calculates >=5000 by naphthalene peak.
Testing result is as shown in Fig. 1 Fig. 2, wherein peak sequence is followed successively by camphor, menthol, borneol, naphthalene, salicylic acid first
Ester.
Method validation:
(1) linear to investigate
It is accurate respectively to draw menthol (3.537mg/ml), camphor (3.514mg/ml), gaultherolin (3.514mg/ml), dragon
Brain (3.555mg/ml) reference substance mixed solution 1,2,3,5,8ml to 10ml measuring bottle, are separately added into internal standard naphthalene (10.33mg/ml)
Each 1ml adds ethyl acetate to scale, and precision draws 1 μ l sample introduction, records chromatography, to compare peak area (A to) with internal standard peak area
The ratio between (in A) (A right/A in) is abscissa, is vertical to compare the ratio between same internal standard (in C) of sample volume (C to) (C right/C in) (μ g)
Coordinate mapping, calculate regression equation are as follows: A menthol=0.8037B-0.0689 (r=0.9990), menthol 0.3514 μ g~
2.8112 μ g, A camphor=0.7709B-0.0894 (r=0.9992), camphor is in 0.3514 μ g of μ g~2.8112, A gaultherolin
=0.5381B-0.0205 (r=0.9995), gaultherolin is in 0.3514 μ g of μ g~2.8112, A borneol=0.6661B-
0.0468 (r=0.9990), borneol linear relationship between 0.3555 μ of μ g~2.8440 g are good.
1 menthol linear relationship of table
2 camphor linear relationship of table
3 gaultherolin linear relationship of table
4 borneol linear relationship of table
(2) precision test
Accurate draw contains menthol (1.0611mg/ml), camphor (1.0542mg/ml), gaultherolin (1.0542mg/ respectively
Ml), borneol (1.0665mg/ml), internal standard naphthalene (1.033mg/ml) 1 μ l of mixed solution repeat sample introduction 6 times, record chromatography, knot
It is respectively 0.40%, 1.21%, 3.22%, 1.27% that fruit, which is shown in Table 5, RSD, illustrates there is good precision.
5 Precision test result of table
(3) repetitive test
Take a sample to measure replication 6 times by content assaying method described in text, calculate, the results are shown in Table 6, menthol, camphor,
Gaultherolin, borneol RSD are respectively 0.66% or more, illustrate there is good repeatability.
6 repetitive test result of table
(4) recovery test
It is recycled using sample-adding, precision takes sample (the menthol 26.04mg/100cm2, camphor 19.84mg/ for having measured content
100cm2, gaultherolin 13.96mg/100cm2, borneol 14.47mg/100cm2) 50cm2, add menthol, camphor, water
Poplar acid methyl esters, borneol reference substance are appropriate, measure by chromatographic condition described in text, calculate the rate of recovery, the results are shown in Table 7, table 8,9 and of table
Table 10, menthol, camphor, gaultherolin, borneol the rate of recovery be 97.45%, 97.58%, 100.75%, 99.06%, RSD be
0.66%, 0.69%, 0.77%, 0.34%, illustrate that this law has the good rate of recovery.
7 menthol recovery test result of table
8 camphor recovery test of table
9 gaultherolin recovery test result of table
10 borneol recovery test result of table
(5) specificity
SHEXIANG ZHUANGGU GAO and negative sample is taken to prepare sample solution and negative sample solution by sample solution preparation method, by upper
Chromatographic condition is stated, takes reference substance solution, sample solution, negative sample solution injection liquid chromatograph, sample solution chromatography respectively
It is equipped with corresponding chromatographic peak in reference substance solution chromatography corresponding positions, and negative noiseless.
(6) durability
The selection of Extraction solvent: taking 5 parts of SHEXIANG ZHUANGGU GAO, every part of 210cm2, is cut into fillet, removes cover lining, splits in conical flask,
Ethyl alcohol, methanol, each 50ml of ethyl acetate is added in precision, and weighed weight heating and refluxing extraction 1 hour, lets cool, with ethyl alcohol, methanol,
Ethyl acetate respectively supplies less loss weight, and filtration, precision is drawn subsequent filtrate 20ml and set in 25ml measuring bottle, and inner mark solution is added in precision
2.5ml adds ethyl alcohol, methanol, ethyl acetate to be diluted to scale, shake up to get.Each 1 μ l of test solution is drawn, gas phase color is injected
Spectrometer measures, and calculates, the results are shown in Table 10, illustrates that SHEXIANG ZHUANGGU GAO uses ethyl acetate as Extraction solvent, content is extracted completeer
Entirely.
The selection of 11 Extraction solvent of table
The selection of sample extraction method: taking 2 parts of SHEXIANG ZHUANGGU GAO, every part of 210cm2, is cut into fillet, removes cover lining, sets tool respectively
It filling in conical flask, ethyl acetate 50ml is added in precision, and a copy of it is ultrasonically treated 60 minutes, another refluxing extraction 60 minutes,
It takes out, lets cool, supply less loss weight with ethyl acetate, shake up, filter, precision is drawn subsequent filtrate 20ml and set in 25ml measuring bottle, essence
Close addition inner mark solution 2.5ml, adds ethyl acetate to be diluted to scale, shake up to get.Each 1 μ l of draw solution injects gas-chromatography
Instrument measures, and calculates, the results are shown in Table 12, and refluxing extraction content is obviously higher than ultrasonic extraction content, therefore selects refluxing extraction.
The selection of 12 extracting method of table
The selection of sample extraction time: taking 3 parts of SHEXIANG ZHUANGGU GAO, every part of 210cm2, is cut into fillet, removes cover lining, sets tool respectively
It fills in conical flask, ethyl acetate 50ml is added in precision, refluxing extraction 15,30,60,90 minutes respectively, takes out, lets cool, use acetic acid
Ethyl ester supplies less loss weight, shakes up, and filtration, precision is drawn subsequent filtrate 20ml and set in 25ml measuring bottle, and inner mark solution is added in precision
2.5ml adds ethyl acetate to be diluted to scale, shake up to get.Each 1 μ l of draw solution injects gas chromatograph, measures, and calculates,
It the results are shown in Table 13, just extract completely within refluxing extraction 60 minutes, therefore the selective extraction time is 60 minutes.
The selection of 13 extraction time of table
(7) sample stability is tested
Take SHEXIANG ZHUANGGU GAO, by embodiment 1 step 4) prepare, by step 5) chromatographic condition measure A it is right/A in, as a result see
Table 14.
14 sample stability test result of table
Three batches of sample assay results: see and measure menthol, camphor, bigcatkin willow in three batches of bone-strengthening musk cream drugs according to this method
The content of sour methyl esters, borneol, the results are shown in Table 15.
Although reference be made herein to invention has been described for multiple explanatory embodiments of the invention, however, it is to be understood that
Those skilled in the art can be designed that a lot of other modification and implementations, these modifications and implementations will fall in this Shen
It please be within disclosed scope and spirit.It more specifically, within the scope of the present disclosure and claims, can be to master
The building block and/or layout for inscribing composite configuration carry out a variety of variations and modifications.In addition to what is carried out to building block and/or layout
Outside modification and improvement, to those skilled in the art, other purposes also be will be apparent.
Claims (10)
1. a kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content, which is characterized in that include following steps:
1) inner mark solution is prepared；
2) menthol reference substance, camphor reference substance, gaultherolin reference substance and borneol reference substance is taken to be placed in same volumetric flask,
Inner mark solution is added, is diluted to scale with ethyl acetate, reference substance solution can be obtained；
3) SHEXIANG ZHUANGGU GAO is taken, obtains sample to be tested after shearing；
4) ethyl acetate solution is used, dissolution extracts sample to be tested, obtains test solution；
5) using gas chromatography measurement reference substance solution each component content and test solution each component content；
Gas chromatographic detection condition are as follows: chromatographic column selects the capillary column with PEG 20000 (PEG20M) for stationary phase；Into
Sample amount 1uL；140 DEG C of column temperature.
2. a kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content according to claim 1, it is characterised in that: step 1)
Prepare the concrete operations of inner mark solution are as follows: it is appropriate that precision weighs naphthalene, and ethyl acetate is added, and every 1mL ethyl acetate is made containing 10mg
The solution of naphthalene to get arrive inner mark solution.
3. a kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content according to claim 1, it is characterised in that: step 2
Prepare the concrete operations of reference substance solution are as follows: take menthol reference substance, camphor reference substance, salicylic acid reference substance, borneol pair respectively
It according to each 10mg of product, is placed in 10mL volumetric flask together, inner mark solution 1mL is added in precision, is diluted to scale with ethyl acetate, shakes
It is even, reference substance solution can be obtained.
4. a kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content according to claim 1, it is characterised in that: step 3)
Prepare the concrete operations of sample to be tested are as follows: take SHEXIANG ZHUANGGU GAO 210cm2, it is cut into fillet, cover lining is removed, obtains sample to be tested.
5. a kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content according to claim 1, it is characterised in that: step 4)
Include:
Step S1: sample to be tested is placed in conical flask, and ethyl acetate 50mL, weighed weight is added in precision；
Step S2: after heating and refluxing extraction, letting cool, then weighed weight；
Step S3: less loss weight is supplied with ethyl acetate；
Step S4: filtering, precision draw subsequent filtrate 20mL in 25mL volumetric flask, and 2.5mL inner mark solution is added in precision, uses acetic acid
Ethyl ester is diluted to scale, shakes up, and test solution can be obtained.
6. a kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content according to claim 5, it is characterised in that: step S2
The time being heated to reflux is 55min~65min；75 DEG C~85 DEG C of temperature holding being heated to reflux is slightly boiled.
7. a kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content according to claim 5, it is characterised in that: step S2
It lets cool to 20 DEG C~30 DEG C of temperature.
8. a kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content according to claim 6, it is characterised in that: step S2
The time being heated to reflux is 60min；The temperature being heated to reflux is kept for 80 DEG C.
9. a kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content according to claim 1, it is characterised in that: step 5)
The detection temperature of gas chromatograph is 260 DEG C.
10. a kind of measuring method of SHEXIANG ZHUANGGU GAO multicomponent content according to claim 1, it is characterised in that: step
5) in, chromatographic column theoretical cam curve calculates >=5000 by naphthalene peak.
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Application publication date: 20190913 |