CN106501396A - A kind of detection method of hemorrhoid medicine index components content - Google Patents
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Abstract
The invention discloses a kind of detection method of hemorrhoid medicine index components content, which belongs to pharmaceutical technology field.The index components include matrine, hydrochloric acid phellodendrine, berberine hydrochloride and scopolamine hydrobromide.In order to evade the content for only determining single component or two kinds of compositions, the present invention adopts high performance liquid chromatography, is successfully achieved while and quickly determining four index components contents in the preparation of preparation, reference substance solution from hemorrhoid medicine need testing solution and chromatography condition.The detection method also has the characteristics of accuracy, precision are high, and stability, repeatability and specificity are good.
Description
Technical field
A kind of the present invention relates to pharmaceutical technology field, more particularly to detection method of hemorrhoid medicine index components content.
Background technology
Hemorrhoid are a kind of common diseases positioned at anus area, and with age, sickness rate gradually increases.For treating
The piles medicine category of hemorrhoid can be divided into three major types：Western medicine, Chinese medicine and Chinese medicine and western medicine compound preparation, Chinese medicine have curing haemorrhoids inflammation granule,
Hemorrhoid health piece and Hemorrhoid hemostatic pill etc., Chinese medicine and western medicine compound preparation have HUAZHI SHUAN etc., compare Western medicine, Chinese medicine and Chinese medicine and western medicine compound preparation
Composition is relative complex.
《Chinese Pharmacopoeia》One prescription for disclosing HUAZHI SHUAN of version in 2015 is：Bismuth subgallate 200g, Radix Sophorae Flavescentiss 370g,
Cortex Phellodendri 92.5g, Flos Daturae 55.5g, Borneolum Syntheticum 30g.The preparation method of HUAZHI SHUAN is：Radix Sophorae Flavescentiss, Cortex Phellodendri, Flos Daturae add water to cook secondary,
4h, second 2h；Collecting decoction, filters and stands 12h, is 1.12 (60～65 DEG C) by supernatant concentration to relative density
Clear paste, crushed after being dried is into most fine powder；The ethyl hydroxybenzoate of 2.6g is dissolved with ethanol in proper amount；Substrate is separately taken appropriate, heating is molten
Change, add bismuth subgallate, above-mentioned most fine powder, Borneolum Syntheticum and 16.8g polyoxyethylene sorbitan monoleates, ethyl hydroxybenzoate ethanol, mix, fill
Note, that is, make 1000.This product heat clearing and damp drying, hemostasis with astringents, for damp-heat in the large intestine caused by mixed hemorrhoidss, mixed hemorrhoids.At present,
《Chinese Pharmacopoeia》The content assaying method of in bismuth subgallate and Borneolum Syntheticum Borneolum Syntheticum has only been recorded.HUAZHI SHUAN is used as Chinese medicine and western medicine compound recipe
Preparation, if not being dry ingredients simply adduction, but multiple active component integrally play curative effect effect.Prior art does not only have to secondary
The matrine in Borneolum Syntheticum and Radix Sophorae Flavescentiss in gallate-based bismuth, Borneolum Syntheticum is detected still have multiple index components to be detected, is lacked
Comprehensive and medicine total quality can not be reflected.
Additionally, pertinent literature was announced：Detection contains containing for the Chinese medicine Related Component of Radix Sophorae Flavescentiss, Cortex Phellodendri or Flos Daturae medical material
Amount, but these methods are still more unilateral, it is difficult to suitable for the quality testing of HUAZHI SHUAN or even hemorrhoid medicine.
Content of the invention
It is an object of the invention to overcome weak point that prior art is present and provide a kind of more comprehensively, reliable hemorrhoid
The detection method of drug effect measurement component content, of the invention by berberine hydrochloride, hydrochloric acid phellodendrine, matrine and hydrobromic acid Rhizoma Scopoliae Japonicae
Alkali is rapidly quantitative determined using high performance liquid chromatography, it is to avoid only simultaneously as HUAZHI SHUAN or even the index components of piles medicine
The quality of medicine is judged by accident by detecting single component.
The technical solution used in the present invention is：A kind of detection method of hemorrhoid medicine index components content, the index into
Dividing includes that matrine, hydrochloric acid phellodendrine, berberine hydrochloride and scopolamine hydrobromide, the detection method are comprised the following steps：
1) preparation of hemorrhoid medicine need testing solution：Hemorrhoid medicine powder is placed in alcoholic solution and is extracted, filtered, concentration filter
Liquid；
The preparation of reference substance solution：Prepare the right of matrine, hydrochloric acid phellodendrine, berberine hydrochloride and scopolamine hydrobromide
According to product solution；
2) precision draws hemorrhoid medicine need testing solution and reference substance solution respectively, injects high performance liquid chromatograph, determines；
Wherein chromatography condition is：Using gradient elution, mobile phase A is acetonitrile, and Mobile phase B is the acid containing dodecyl sodium sulfate
Solution, Detection wavelength are 210～230nm, and column temperature is 20～30 DEG C.
Used as the further improvement to above-mentioned technical proposal, the Detection wavelength is 220nm, and column temperature is 20 DEG C, now respectively
Between index components (matrine, hydrochloric acid phellodendrine, berberine hydrochloride and scopolamine hydrobromide), separating effect is preferable.
Used as the further improvement to above-mentioned technical proposal, the preparation of the hemorrhoid medicine need testing solution is specially：Will
Hemorrhoid medicine powder is placed in alcoholic solution, and relative to every gram of hemorrhoid medicine, the volume of the alcoholic solution is 20～30mL；At ultrasound
30～60min of reason, adds alcoholic solution after cooling, supplies the weight of reduction；Filtered after 30～60min of freezing, filtrate is dense
It is reduced to the 2/25～1/8 of original volume.
It is highly preferred that relative to every gram of hemorrhoid medicine, the volume of the alcoholic solution is 25mL, and the volume of concentrated solution is substance
Long-pending 1/10, the index components content that now alcoholic solution is extracted is more preferably, it is not necessary to excessively concentrate, and is suitable for high performance liquid chromatography
Determine.
It is highly preferred that the sonication treatment time that the hemorrhoid medicine solution is prepared is 45min, cooling time is 45min.
Used as the further improvement to above-mentioned technical proposal, the preparation of the reference substance solution is specially：Weigh matrine
The reference substance of reference substance, hydrochloric acid phellodendrine reference substance, berberine hydrochloride reference substance and scopolamine hydrobromide, adds the alcohol molten
It is 27.4～438.4 μ g/mL, hydrochloric acid that liquid is configured to matrine for 241.3～3860.0 μ g/mL, hydrochloric acid Cortex Phellodendri alkali concn
The mixed solution that Radix Berberidis Amurensis alkali concn is 25.9～414.0 μ g/mL, scopolamine hydrobromide concentration is 10.1～161.8 μ g/mL.
Reference substance mixed liquid concentration within the above range when, the linear relationship for drawing is more excellent, and the index components content of detection is more accurate.
As the further improvement to above-mentioned technical proposal, the alcoholic solution be 50%～100% methanol solution or
50%～100% ethanol solution.Four kinds of index components effects that 50%～100% methanol or ethanol are extracted are more preferable, content detection
Results contrast objective.
Used as the further improvement to above-mentioned technical proposal, the alcoholic solution is 60% methanol solution.
Used as the further improvement to above-mentioned technical proposal, the acid in the Mobile phase B is glacial acetic acid, phosphoric acid or formic acid,
Percentage by volume of the acid in Mobile phase B is 0.05%～0.3%, and the dodecyl sodium sulfate is in Mobile phase B
Mass percent is 0.1%～0.3%.
Used as the further improvement to above-mentioned technical proposal, percentage by volume of the acid in Mobile phase B is
0.1%, mass percent of the dodecyl sodium sulfate in Mobile phase B is 0.2%.
Used as the further improvement to above-mentioned technical proposal, the program of the gradient elution is：
0～9min, mobile phase A are 33% → 35%, and Mobile phase B is 67% → 65%；
9～18min, mobile phase A are 35% → 40%, and Mobile phase B is 65% → 60%；
18～30min, mobile phase A are 40% → 50%, and Mobile phase B is 60% → 50%；
30～50min, mobile phase A are 50% → 60%, and Mobile phase B is 50% → 40%；
The percentage ratio that above is referred to is percent by volume.
When the program of the gradient elution on the premise of ensureing to separate four kinds of index components, can shorten the reservation of each component
Between, detection efficiency is improved, while guaranteeing that Chromatogram Baseline is more steady.
Used as the further improvement to above-mentioned technical proposal, the chromatographic column used during measure is bonded with octadecylsilane
C of the silica gel for filler18Post.
Used as the further improvement to above-mentioned technical proposal, the hemorrhoid medicine is HUAZHI SHUAN.
The beneficial effects of the present invention is：The present invention is by berberine hydrochloride, hydrochloric acid phellodendrine, matrine and hydrobromic acid east Liang
Henbane alkali has been evaded and has only determined asking for single component or two kinds of component contents as HUAZHI SHUAN or even the index components of hemorrhoid medicine
Topic；But, Chinese medicine and Chinese medicine and western medicine compound preparation complicated component easily affect to separate, determine, and the present invention adopts high-efficient liquid phase color for this
Spectrometry, selects by a large amount of wounds from examination drug solution, the preparation of reference substance solution and chromatographic condition and flow phase system is supplied
The work of the property made successfully is achieved while and quickly determining four index components contents.The detection method also has accurately simultaneously
Degree, precision are high, the characteristics of stability, repeatability and specificity are good.
Description of the drawings
Fig. 1 is the HPLC of berberine hydrochloride, hydrochloric acid phellodendrine, matrine and scopolamine hydrobromide mixed reference substance solution
Chromatogram；
Fig. 2 is HPLC chromatogram of the HUAZHI SHUAN for examination drug solution；
Fig. 3 is the HPLC chromatogram of negative sample solution, and wherein negative sample does not include berberine hydrochloride, hydrochloric acid Cortex Phellodendri
Alkali, matrine and scopolamine hydrobromide.
Specific embodiment
For the object, technical solutions and advantages of the present invention are better described, below in conjunction with specific embodiment and subordinate list pair
The present invention is described further.
Embodiment 1
1) preparation of need testing solution：HUAZHI SHUAN powder about 1g is taken, accurately weighed, put in conical flask with cover；To conical flask
60% methanol 25mL of middle accurate addition, close plug, weighed weight, supersound process 45min；Let cool, then weighed weight, use 60% methanol
Supply the weight of less loss；Shake up, freeze 45min；Filter, precision pipettes subsequent filtrate 20mL, is concentrated into 2mL, obtains final product.
The preparation of reference substance solution：Take berberine hydrochloride reference substance, hydrochloric acid phellodendrine reference substance, matrine reference substance and hydrogen
Bromic acid scopolamine reference substance is appropriate, accurately weighed, plus 60% methanol makes matrine for 1930.0 μ g/mL, hydrochloric acid Huang
Cypress alkali concn is 109.6 μ g/mL, berberine hydrochloride concentration is 103.5 μ g/mL, scopolamine hydrobromide concentration is 20.2 μ g/mL
Mixed reference substance solution.
2) condition determination of high performance liquid chromatography：Chromatographic column is the C with octadecylsilane chemically bonded silica as filler18
Post；Mobile phase A is acetonitrile, and Mobile phase B is 0.1% phosphoric acid solution containing 0.2% dodecyl sodium sulfate；Detection wavelength is
220nm, column temperature are 20 DEG C, and sample size is 10 μ L, and the program of gradient elution is shown in Table 1, and (percentage ratio wherein, being related to is volume basis
Than).The index components content (concentration) of need testing solution=(reference substance solution concentration × need testing solution peak area)/reference substance
Peak area, similarly hereinafter.
Table 1 is gradient elution program
Embodiment 2
Detecting step reference embodiment 1, except for the difference that：In process for preparation for examination drug solution and reference substance solution, with
50% methanol is used as Extraction solvent；In need testing solution process for preparation, supersound process and cooling time are 30min；Mixing control
In product solution matrine be 241.3 μ g/mL, hydrochloric acid Cortex Phellodendri alkali concn be 27.4 μ g/mL, berberine hydrochloride concentration be 25.9
μ g/mL, scopolamine hydrobromide concentration are 10.1 μ g/mL；During high-performance liquid chromatogram determination, Mobile phase B is containing 0.3% dodecane
0.05% phosphoric acid solution of base sodium sulfonate, Detection wavelength are 210nm, and column temperature is 25 DEG C.
Embodiment 3
Detecting step reference embodiment 1, except for the difference that：In process for preparation for examination drug solution and reference substance solution, with
Methanol is used as Extraction solvent；In need testing solution process for preparation, supersound process and cooling time are 60min；Mixing reference substance
In solution matrine be 3860.0 μ g/mL, hydrochloric acid Cortex Phellodendri alkali concn be that 438.4 μ g/mL, berberine hydrochloride concentration are
414.0 μ g/mL, scopolamine hydrobromide concentration are 161.8 μ g/mL；During high-performance liquid chromatogram determination, Mobile phase B is containing 0.1%
0.3% phosphoric acid solution of dodecyl sodium sulfate, Detection wavelength are 230nm, and column temperature is 30 DEG C.
Embodiment 4
Detecting step reference embodiment 1, except for the difference that：In the process for preparation of need testing solution and reference substance solution, with
50% ethanol is used as Extraction solvent.
Embodiment 5
Detecting step reference embodiment 1, except for the difference that：In the process for preparation of need testing solution and reference substance solution, with
70% ethanol is used as Extraction solvent.
Embodiment 6
Detecting step reference embodiment 1, except for the difference that：In the process for preparation of need testing solution and reference substance solution, with second
Alcohol is used as Extraction solvent.
Embodiment 7
Detecting step reference embodiment 1, except for the difference that：During high-performance liquid chromatogram determination, Mobile phase B is containing 0.2% 12
0.1% glacial acetic acid solution of sodium alkyl sulfonate.
Embodiment 8
Detecting step reference embodiment 1, except for the difference that：During high-performance liquid chromatogram determination, Mobile phase B is containing 0.1% 12
0.05% glacial acetic acid solution of sodium alkyl sulfonate.
Embodiment 9
Detecting step reference embodiment 1, except for the difference that：During high-performance liquid chromatogram determination, Mobile phase B is containing 0.3% 12
0.3% glacial acetic acid solution of sodium alkyl sulfonate.
Embodiment 10
Detecting step reference embodiment 1, except for the difference that：During high-performance liquid chromatogram determination, Mobile phase B is containing 0.2% 12
0.1% formic acid solution of sodium alkyl sulfonate.
Embodiment 11
Detecting step reference embodiment 1, except for the difference that：During high-performance liquid chromatogram determination, Mobile phase B is containing 0.1% 12
0.05% formic acid solution of sodium alkyl sulfonate.
Embodiment 12
Detecting step reference embodiment 1, except for the difference that：During high-performance liquid chromatogram determination, Mobile phase B is containing 0.3% 12
0.3% formic acid solution of sodium alkyl sulfonate.
Embodiment 13
The checking that index components content is determined
1 test material
In detection method agents useful for same be commercially available, wherein be used for assay acetonitrile be chromatographically pure
(production of LABSCIENCE companies), water is ultra-pure water, and other reagents are pure for analysis.HUAZHI SHUAN is respected by Community in Baiyunshan, Guangzhou and repaiies a Pharmaceutical
Limited company provides；Matrine reference substance is provided by National Institute for Food and Drugs Control, lot number 805-9001, and content is
100%；Hydrochloric acid phellodendrine reference substance is provided by National Institute for Food and Drugs Control, and lot number is 111895-201303, and content is
95.8%；Berberine hydrochloride reference substance is provided by National Institute for Food and Drugs Control, and lot number is 110713-201212, content
For 86.7%；Scopolamine hydrobromide reference substance is provided by National Institute for Food and Drugs Control, lot number 100049-200308,
Content is 100%.
2 instruments
1260 type high performance liquid chromatographs of Agilent, photodiode array detector (DAD), chromatographic column is
Agilent Eclipse Plus C18Post (250mm × 4.6mm, 5 μm), Agilent ZORBAX SB-C18Post (250mm ×
4.6mm, 5 μm), 4 μ Fusion-RP 80A posts (250mm × 4.6mm, 4 μm) of Phenomenex Synergi.KQ-600DE types
Numerical control supersonic cleaning device (Kunshan Ultrasonic Instruments Co., Ltd.), Precisa 404A, ME204T/02 type electronic analysis day
Flat.
The preparation of 3 hemorrhoid medicine need testing solutions
HUAZHI SHUAN powder about 1g is taken, accurately weighed, put in conical flask with cover；60% methanol 25mL of accurate addition；Close plug cone
Shape bottle, weighed weight, supersound process 45min；Let cool, then weighed weight, the weight for supplying less loss with 60% methanol；Shake up, cold
Freeze 45min；Filter, precision pipettes subsequent filtrate 20mL, is concentrated into 2mL, obtains final product.
The preparation of 4 reference substance solution
Take the control of berberine hydrochloride reference substance, hydrochloric acid phellodendrine reference substance, matrine reference substance and scopolamine hydrobromide
Appropriate product, accurately weighed, plus 60% methanol make matrine for 1930.0 μ g/mL, hydrochloric acid Cortex Phellodendri alkali concn be 109.6 μ
The mixing reference substance that g/mL, berberine hydrochloride concentration are 103.5 μ g/mL, scopolamine hydrobromide concentration is 20.2 μ g/mL is molten
Liquid, obtains final product.
The condition determination of 5 high performance liquid chromatography
Chromatographic column is the C with octadecylsilane chemically bonded silica as filler18Post；With acetonitrile as mobile phase A, to contain
0.1% phosphoric acid solution of 0.2% dodecyl sodium sulfate is Mobile phase B；Detection wavelength is 220nm；Column temperature is 20 DEG C, sample size
For 10 μ L；It is shown in Table the program of 1 gradient elution.
6 linear relationships are investigated
Precision weighs reference substance berberine hydrochloride, hydrochloric acid phellodendrine, matrine and appropriate scopolamine hydrobromide, plus 60%
Methanol makes berberine hydrochloride concentration for 414.0 μ g/mL, and hydrochloric acid Cortex Phellodendri alkali concn is 438.4 μ g/mL, and matrine is
3860.0 μ g/mL, scopolamine hydrobromide concentration are the mixed reference substance solution of 161.8 μ g/mL.Mixing is obtained final product by dilution right
25.9,51.8,103.5,207.0,414.0 μ g/mL are respectively according to the berberine hydrochloride concentration in product solution, corresponding hydrochloric acid is yellow
Cypress alkali concn respectively 27.4,54.8,109.6,219.2,438.4 μ g/mL, corresponding matrine respectively 241.3,
482.5th, 965.0,1930.0,3860.0 μ g/mL, corresponding scopolamine hydrobromide concentration be respectively 10.1,20.2,40.4,
80.9、161.8μg/mL.The each 10 μ L of the above-mentioned reference substance mixed solution of accurate absorption, inject chromatograph of liquid, measure by containing respectively
Determine method measure.With peak area (Y) as vertical coordinate, reference substance concentration (X, μ g/mL) is abscissa, draws standard curve, must return
Equation is returned to be shown in Table 2.From table 2, it can be deduced that matrine is 241.3～3860.0 μ g/mL, hydrochloric acid Cortex Phellodendri alkali concn is
27.4～438.4 μ g/mL, berberine hydrochloride concentration be 25.9～414.0 μ g/mL, scopolamine hydrobromide concentration be 10.1～
In the mixed solution of 161.8 μ g/mL, four index components assume good linear relationship.In routine testing, as long as sample
Concentration then without the need for drawing mark song every time, can be detected using one point external standard method in mark song concentration range.
Table 2 is that result investigated by linear relationship
7 stability tests
Take with portion HUAZHI SHUAN medicine, after preparation, by content assaying method, respectively 0,2,4,8,16,24,32h sample introductions
Determine.As a result find that HUAZHI SHUAN solution is determined in 32h from after preparing basicly stable, be shown in Table 3.
Table 3 is stability test result
8 replica tests
Take with 6 parts of a collection of HUAZHI SHUAN, with reference to " for trying the preparation of drug solution ", hydrochloric acid is carried out using high performance liquid chromatography little
The measure of bark of a cork tree alkali, hydrochloric acid phellodendrine, matrine and scopolamine hydrobromide content, the results are shown in Table 4.
Table 4 is repeated result of the test
The comparison of 9 different chromatographic columns
Take with 2 parts of a collection of HUAZHI SHUAN, by content assaying method, use A posts (Agilent Eclipse Plus C respectively18Post,
Specification 250mm × 4.6mm, 5 μm), B posts (Agilent ZORBAX SB-C18Post, specification 250mm × 4.6mm, 5 μm), C posts
The chromatographic column of (4 μ Fusion-RP 80A posts of Phenomenex Synergi, specification 250mm × 4.6mm, 4 μm) three kinds of models is surveyed
Fixed.As a result find to be measured using the chromatographic column of three kinds of models, the content of HUAZHI SHUAN the results are shown in Table 5 without significant difference.
Measurement result of the table 5 for different chromatographic columns
10 average recoveries are tested
6 parts of the HUAZHI SHUAN with a collection of known content is taken, per part takes about 0.5g, accurately weighed, puts in conical flask with cover；Respectively
Accurate addition 1mL berberine hydrochloride reference substance solution (0.1235mg/mL), 1mL hydrochloric acid phellodendrine reference substance solution
(0.1226mg/mL), 1mL matrines reference substance solution (2.821mg/mL) and 1mL scopolamine hydrobromide (0.02674mg/
ML), remainder is processed and is determined by " preparation of need testing solution ", the results are shown in Table 6～table 9.
The response rate of 6 berberine hydrochloride of table
The response rate of the table 7 for hydrochloric acid phellodendrine
The response rate of the table 8 for matrine
The response rate of the table 9 for scopolamine hydrobromide
11 specificities are tested
The medical material in addition to Cortex Phellodendri, Radix Sophorae Flavescentiss and Flos Daturae is weighed by HUAZHI SHUAN prescription, negative sample is made by preparation technology.
Negative sample solution is made again by the preparation method for supplying examination drug solution.Accurate absorption reference substance solution, HUAZHI SHUAN solution and the moon
The property each 10 μ L of sample solution, are injected separately into chromatograph of liquid, are measured by content assaying method.As a result find little with hydrochloric acid
Bark of a cork tree alkali reference substance, hydrochloric acid phellodendrine reference substance, matrine reference substance and the corresponding position of scopolamine hydrobromide reference substance chromatograph
On, negative sample solution is noiseless, illustrates that the assay method has preferable specificity, sees accompanying drawing 1～3.
Summary, above result of the test show：The detection method is easy to operate, workable, result is accurate, exclusive
Property and favorable reproducibility, are suitable as method of quality control, and can determine hydrochloric acid Radix Berberidis Amurensis under the conditions of same HPLC methods simultaneously
Alkali, hydrochloric acid phellodendrine, the content of four index components of matrine and scopolamine hydrobromide.
Last should be noted that above example is only in order to illustrating technical scheme rather than protect to of the invention
The restriction of shield scope, although being explained in detail to the present invention with reference to preferred embodiment, one of ordinary skill in the art should
Understand, technical scheme can be modified or equivalent, without deviating from the essence of technical solution of the present invention.
Claims (10)
1. a kind of detection method of hemorrhoid medicine index components content, it is characterised in that：The index components include matrine, salt
Sour phellodendrine, berberine hydrochloride and scopolamine hydrobromide, the detection method are comprised the following steps：
1) preparation of hemorrhoid medicine need testing solution：Hemorrhoid medicine powder is placed in alcoholic solution and is extracted, filtered, concentrate filtrate；
The preparation of reference substance solution：Prepare the reference substance of matrine, hydrochloric acid phellodendrine, berberine hydrochloride and scopolamine hydrobromide
Solution；
2) precision draws hemorrhoid medicine need testing solution and reference substance solution respectively, injects high performance liquid chromatograph, determines；Wherein
Chromatography condition is：Using gradient elution, mobile phase A is acetonitrile, and Mobile phase B is sour molten containing dodecyl sodium sulfate
Liquid, Detection wavelength are 210～230nm, and column temperature is 20～30 DEG C.
2. detection method according to claim 1, it is characterised in that：The step 1) in, the hemorrhoid medicine test sample
The preparation of solution is specially：Hemorrhoid medicine powder is placed in alcoholic solution, relative to every gram of hemorrhoid medicine, the body of the alcoholic solution
Product is 20～30mL；30～60min of supersound process, adds alcoholic solution after cooling, supplies the weight of reduction；30～60min of freezing
After filtered, concentrate the filtrate to the 2/25～1/8 of original volume.
3. detection method according to claim 1, it is characterised in that：The preparation of the reference substance solution is specially：Weigh
The reference substance of matrine reference substance, hydrochloric acid phellodendrine reference substance, berberine hydrochloride reference substance and scopolamine hydrobromide, adds institute
It is 27.4～438.4 μ g/ to state alcoholic solution and be configured to matrine for 241.3～3860.0 μ g/mL, hydrochloric acid Cortex Phellodendri alkali concn
It is the mixed of 10.1～161.8 μ g/mL that mL, berberine hydrochloride concentration are 25.9～414.0 μ g/mL, scopolamine hydrobromide concentration
Close solution.
4. the detection method according to Claims 2 or 3, it is characterised in that：The alcoholic solution is that 50%～100% methanol is molten
Liquid or 50%～100% ethanol solution.
5. detection method according to claim 4, it is characterised in that：The alcoholic solution is 60% methanol solution.
6. detection method according to claim 1：Acid in the Mobile phase B be glacial acetic acid, phosphoric acid or formic acid, the acid
Percentage by volume in Mobile phase B is 0.05%～0.3%, quality hundred of the dodecyl sodium sulfate in Mobile phase B
Fraction is 0.1%～0.3%.
7. detection method according to claim 6, it is characterised in that：The percentage by volume of the acid in Mobile phase B be
0.1%, mass percent of the dodecyl sodium sulfate in Mobile phase B is 0.2%.
8. detection method according to claim 1, it is characterised in that：The program of the gradient elution is：
0～9min, mobile phase A are 33% → 35%, and Mobile phase B is 67% → 65%；
9～18min, mobile phase A are 35% → 40%, and Mobile phase B is 65% → 60%；
18～30min, mobile phase A are 40% → 50%, and Mobile phase B is 60% → 50%；
30～50min, mobile phase A are 50% → 60%, and Mobile phase B is 50% → 40%.
9. detection method according to claim 1, it is characterised in that：The chromatographic column used during measure is with octadecyl silicon
C of the alkane bonded silica gel for filler18Post.
10. detection method according to claim 1, it is characterised in that：The hemorrhoid medicine is HUAZHI SHUAN.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107389830B (en) * | 2017-09-20 | 2019-09-03 | 江苏省中医院 | A kind of hemorrhoid complicated by anal fistula smokes the quality determining method of lotion |
| CN111337588A (en) * | 2020-03-12 | 2020-06-26 | 烟台荣昌制药股份有限公司 | Method for measuring dissolution rate of organic ingredients in anal suppository |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107389830B (en) * | 2017-09-20 | 2019-09-03 | 江苏省中医院 | A kind of hemorrhoid complicated by anal fistula smokes the quality determining method of lotion |
| CN111337588A (en) * | 2020-03-12 | 2020-06-26 | 烟台荣昌制药股份有限公司 | Method for measuring dissolution rate of organic ingredients in anal suppository |
| CN115420816A (en) * | 2022-07-21 | 2022-12-02 | 广东万年青制药股份有限公司 | Content determination method of traditional Chinese medicine composition for nourishing yin and cooling blood |
| CN115356413A (en) * | 2022-08-17 | 2022-11-18 | 博诺康源(北京)药业科技有限公司 | Method for detecting scopolamine hydrobromide related substances |
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| CN106501396B (en) | 2018-04-20 |
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