CN106501396B - A kind of detection method of hemorrhoid medicine index components content - Google Patents

A kind of detection method of hemorrhoid medicine index components content Download PDF

Info

Publication number
CN106501396B
CN106501396B CN201610910516.9A CN201610910516A CN106501396B CN 106501396 B CN106501396 B CN 106501396B CN 201610910516 A CN201610910516 A CN 201610910516A CN 106501396 B CN106501396 B CN 106501396B
Authority
CN
China
Prior art keywords
solution
mobile phase
reference substance
detection method
hemorrhoid medicine
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
Application number
CN201610910516.9A
Other languages
Chinese (zh)
Other versions
CN106501396A (en
Inventor
彭红英
江涛
陈晓坚
朱少璇
程君璐
贾佳良
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
GUANGZHOU BAIYUNSHAN JINGXIUTANG PHARMACEUTICAL Co Ltd
Original Assignee
GUANGZHOU BAIYUNSHAN JINGXIUTANG PHARMACEUTICAL Co Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by GUANGZHOU BAIYUNSHAN JINGXIUTANG PHARMACEUTICAL Co Ltd filed Critical GUANGZHOU BAIYUNSHAN JINGXIUTANG PHARMACEUTICAL Co Ltd
Priority to CN201610910516.9A priority Critical patent/CN106501396B/en
Publication of CN106501396A publication Critical patent/CN106501396A/en
Application granted granted Critical
Publication of CN106501396B publication Critical patent/CN106501396B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)

Abstract

The invention discloses a kind of detection method of hemorrhoid medicine index components content, it 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 measuring single component or two kinds of components, the present invention use high performance liquid chromatography, is successfully realized in the preparation of preparation, reference substance solution from hemorrhoid medicine test solution and chromatography condition at the same time and quick measures four index components contents.The detection method also has the characteristics of accuracy, precision are high, and stability, repeatability and specificity are good.

Description

A kind of detection method of hemorrhoid medicine index components content
Technical field
The present invention relates to pharmaceutical technology field, more particularly to a kind of detection method of hemorrhoid medicine index components content.
Background technology
Hemorrhoid are a kind of common diseases positioned at anus area, and with age, incidence gradually increases.For treating The piles medicine category of hemorrhoid can be divided into three categories：Western medicine, Chinese medicine and Chinese and Western medicine compound preparation, Chinese medicine have curing haemorrhoids inflammation particle, Hemorrhoid health piece and Hemorrhoid hemostatic pill etc., Chinese and Western medicine compound preparation have HUAZHI SHUAN etc., compared to Western medicine, Chinese medicine and Chinese and Western medicine compound preparation Component is relative complex.
《Chinese Pharmacopoeia》One prescription for disclosing HUAZHI SHUAN of version in 2015 is：Bismuth subgallate 200g, kuh-seng 370g, Cortex Phellodendri 92.5g, datura flower 55.5g, borneol 30g.The preparation method of HUAZHI SHUAN is：Kuh-seng, Cortex Phellodendri, datura flower add water to cook it is secondary, 4h, second of 2h；Collecting decoction, filters and stands 12h, is 1.12 (60~65 DEG C) by supernatant concentration to relative density Clear cream, crushed after being dried is into most fine powder；The ethyl hydroxy benzoate of 2.6g is dissolved with ethanol in proper amount；Separately take matrix appropriate, heating is molten Change, add bismuth subgallate, above-mentioned most fine powder, borneol and 16.8g polyoxyethylene sorbitan monoleates, ethyl hydroxy benzoate ethanol, mix, fill Note, that is, be made 1000.This product heat-clearing and damp-drying drug, controlling of bleeding with astringents, for the mixed hemorrhoids caused by large intestinal damp-heat, mixed hemorrhoids.At present, 《Chinese Pharmacopoeia》The content assaying method of borneol in bismuth subgallate and borneol is only recorded.HUAZHI SHUAN is as Chinese and Western medicine compound Preparation, if not being dry ingredients simply adduction, but multiple active ingredients integrally play curative effect effect.The prior art does not only have to secondary The matrine in borneol and kuh-seng in gallate-based bismuth, borneol is detected, and still has multiple index components not detected, is lacked It is comprehensive and cannot reflect medicine total quality.
In addition, pertinent literature was announced：Chinese medicine Related Component of the detection containing kuh-seng, Cortex Phellodendri or datura flower medicinal material contains Amount, but these methods are still more unilateral, it is difficult to suitable for the quality testing of HUAZHI SHUAN or even hemorrhoid medicine.
The content of the invention
There is provided it is an object of the invention to overcome in place of the shortcomings of the prior art it is a kind of more comprehensively, reliable hemorrhoid The detection method of drug effect measurement component content, it is of the invention by Berberine hydrochloride, hydrochloric acid phellodendrine, matrine and hydrobromic acid Anisodus luridus Index components of the alkali as HUAZHI SHUAN or even piles medicine, are rapidly quantitative determined, avoided only at the same time using high performance liquid chromatography The quality of medicine is judged by accident by detection single component.
The technical solution adopted by the present invention is：A kind of detection method of hemorrhoid medicine index components content, the index into Dividing includes matrine, hydrochloric acid phellodendrine, Berberine hydrochloride and scopolamine hydrobromide, and the detection method comprises the following steps：
1) preparation of hemorrhoid medicine test solution：Hemorrhoid medicine powder is placed in alcoholic solution and is extracted, is filtered, concentration filter Liquid；
The preparation of reference substance solution：Prepare pair of matrine, hydrochloric acid phellodendrine, Berberine hydrochloride and scopolamine hydrobromide According to product solution；
2) it is accurate respectively to draw hemorrhoid medicine test solution and reference substance solution, inject high performance liquid chromatograph, measure； 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.
Improved as to the further of above-mentioned technical proposal, the Detection wavelength is 220nm, and column temperature is 20 DEG C, at this time respectively Separating effect is preferable between index components (matrine, hydrochloric acid phellodendrine, Berberine hydrochloride and scopolamine hydrobromide).
Improved as to the further of above-mentioned technical proposal, the preparation of the hemorrhoid medicine test 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 is managed, alcoholic solution is added 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 concentrate is substance Long-pending 1/10, the index components content of alcoholic solution extraction at this time is more preferably, it is not necessary to excessively concentration, and it is adapted to high performance liquid chromatography Measure.
It is highly preferred that the sonication treatment time that the hemorrhoid medicine solution is prepared is 45min, cooling time 45min.
Improved as to the further of above-mentioned technical proposal, the preparation of the reference substance solution is specially：Weigh matrine Reference substance, hydrochloric acid phellodendrine reference substance, the reference substance of Berberine hydrochloride reference substance and scopolamine hydrobromide, it is molten to add the alcohol Liquid be configured to matrine be 241.3~3860.0 μ g/mL, hydrochloric acid Cortex Phellodendri alkali concn be 27.4~438.4 μ g/mL, hydrochloric acid The mixed solution that barberry 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 drawn is more excellent, and the index components content of detection is more accurate.
Improved as to the further of above-mentioned technical proposal, the alcoholic solution for 50%~100% methanol solution or 50%~100% ethanol solution.50%~100% methanol or four kinds of index components effects of ethanol extraction are more preferable, content detection Results contrast it is objective.
Improved as to the further of above-mentioned technical proposal, the alcoholic solution is 60% methanol solution.
Being improved as to the further of 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%.
Improved as to the further of 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%.
Improved as to the further of 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%；
Above is referred to percentage be percent by volume.
The program of the gradient elution can be on the premise of ensureing to separate four kinds of index components, when shortening the reservation of each component Between, detection efficiency is improved, while ensure that Chromatogram Baseline is more steady.
Improved as to the further of above-mentioned technical proposal, the chromatographic column that when measure uses is bonded with octadecylsilane Silica gel is the C of filler18Column.
Improved as to the further of above-mentioned technical proposal, the hemorrhoid medicine is HUAZHI SHUAN.
The beneficial effects of the present invention are：The present invention is by Berberine hydrochloride, hydrochloric acid phellodendrine, matrine and hydrobromic acid east Liang Index components of the henbane alkali as HUAZHI SHUAN or even hemorrhoid medicine, have evaded and have only measured asking for single component or two kinds of component contents Topic；But Chinese medicine and Chinese and Western medicine compound preparation complicated component, separation, measure are easily influenced, the present invention uses high-efficient liquid phase color for this Spectrometry, by largely creating from for examination drug solution, the preparation of reference substance solution and chromatographic condition and flow phase system selection The work for the property made successfully realizes while and four index components contents of quick measure.It is accurate that the detection method also has at the same time The characteristics of degree, precision are high, and stability, repeatability and specificity are good.
Brief 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.
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 test solution：HUAZHI SHUAN powder about 1g is taken, it is accurately weighed, put in conical flask with cover；To conical flask 60% methanol 25mL of middle accurate addition, close plug, weighed weight, is ultrasonically treated 45min；Let cool, then weighed weight, with 60% methanol Supply the weight of less loss；Shake up, freeze 45min；Filtering, precision pipette subsequent filtrate 20mL, are concentrated into 2mL, to obtain the 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 hyoscine reference substance is appropriate, accurately weighed, adds 60% methanol that matrine is made yellow for 1930.0 μ g/mL, hydrochloric acid 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) determination condition of high performance liquid chromatography：Chromatographic column is the C using octadecylsilane chemically bonded silica as filler18 Column；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 wherein, being related to is volume basis Than).The index components content (concentration) of test solution=(reference substance solution concentration × test solution peak area)/reference substance Peak area, similarly hereinafter.
Table 1 is gradient elution program
Embodiment 2
Detecting step reference embodiment 1, unlike：In process for preparation for trying drug solution and reference substance solution, with 50% methanol is as Extraction solvent；In test solution process for preparation, it is ultrasonically treated and cooling time is 30min；Mixing control Matrine is 241.3 μ g/mL in product solution, hydrochloric acid Cortex Phellodendri alkali concn is 27.4 μ g/mL, Berberine hydrochloride concentration is 25.9 μ g/mL, scopolamine hydrobromide concentration are 10.1 μ g/mL；When high performance liquid chromatography measures, Mobile phase B is containing 0.3% dodecane 0.05% phosphoric acid solution of base sodium sulfonate, Detection wavelength 210nm, column temperature are 25 DEG C.
Embodiment 3
Detecting step reference embodiment 1, unlike：In process for preparation for trying drug solution and reference substance solution, with Methanol is Extraction solvent；In test solution process for preparation, it is ultrasonically treated and cooling time is 60min；Mix reference substance Matrine is 3860.0 μ g/mL in solution, hydrochloric acid Cortex Phellodendri alkali concn is 438.4 μ g/mL, Berberine hydrochloride concentration is 414.0 μ g/mL, scopolamine hydrobromide concentration are 161.8 μ g/mL；When high performance liquid chromatography measures, Mobile phase B is containing 0.1% 0.3% phosphoric acid solution of dodecyl sodium sulfate, Detection wavelength 230nm, column temperature are 30 DEG C.
Embodiment 4
Detecting step reference embodiment 1, unlike：In test solution and the process for preparation of reference substance solution, with 50% ethanol is as Extraction solvent.
Embodiment 5
Detecting step reference embodiment 1, unlike：In test solution and the process for preparation of reference substance solution, with 70% ethanol is as Extraction solvent.
Embodiment 6
Detecting step reference embodiment 1, unlike：In test solution and the process for preparation of reference substance solution, with second Alcohol is as Extraction solvent.
Embodiment 7
Detecting step reference embodiment 1, unlike：When high performance liquid chromatography measures, 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, unlike：When high performance liquid chromatography measures, 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, unlike：When high performance liquid chromatography measures, 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, unlike：When high performance liquid chromatography measures, Mobile phase B is containing 0.2% 12 0.1% formic acid solution of sodium alkyl sulfonate.
Embodiment 11
Detecting step reference embodiment 1, unlike：When high performance liquid chromatography measures, Mobile phase B is containing 0.1% 12 0.05% formic acid solution of sodium alkyl sulfonate.
Embodiment 12
Detecting step reference embodiment 1, unlike：When high performance liquid chromatography measures, Mobile phase B is containing 0.3% 12 0.3% formic acid solution of sodium alkyl sulfonate.
Embodiment 13
The verification of index components content measure
1 test material
Agents useful for same is commercially available in detection method, wherein the acetonitrile for assay is chromatographically pure (production of LABSCIENCE companies), water is ultra-pure water, and other reagents are pure to analyze.HUAZHI SHUAN is respected by Community in Baiyunshan, Guangzhou repaiies a medicine company 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, lot number 111895-201303, and content is 95.8%；Berberine hydrochloride reference substance is provided by National Institute for Food and Drugs Control, lot number 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 are Agilent Eclipse Plus C18Column (250mm × 4.6mm, 5 μm), Agilent ZORBAX SB-C18Column (250mm × 4.6mm, 5 μm), 4 μ Fusion-RP 80A columns (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 It is flat.
The preparation of 3 hemorrhoid medicine test solutions
HUAZHI SHUAN powder about 1g is taken, it is accurately weighed, put in conical flask with cover；Precision adds 60% methanol 25mL；Close plug cone Shape bottle, weighed weight, is ultrasonically treated 45min；Let cool, then weighed weight, the weight of less loss is supplied with 60% methanol；Shake up, it is cold Freeze 45min；Filtering, precision pipette subsequent filtrate 20mL, are concentrated into 2mL, to obtain the final product.
The preparation of 4 reference substance solutions
Take Berberine hydrochloride reference substance, hydrochloric acid phellodendrine reference substance, matrine reference substance and scopolamine hydrobromide control Appropriate product, it is accurately weighed, add 60% methanol be made matrine be 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, to obtain the final product.
The determination condition of 5 high performance liquid chromatography
Chromatographic column is the C using octadecylsilane chemically bonded silica as filler18Column；Using acetonitrile as mobile phase A, with containing 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, adds 60% It is 414.0 μ g/mL that Berberine hydrochloride concentration, which is made, in methanol, 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.By diluting up to mixing pair It is respectively 25.9,51.8,103.5,207.0,414.0 μ g/mL according to the Berberine hydrochloride concentration in product solution, corresponding hydrochloric acid is yellow Cypress alkali concn is respectively 27.4,54.8,109.6,219.2,438.4 μ g/mL, corresponding matrine is respectively 241.3, 482.5th, 965.0,1930.0,3860.0 μ g/mL, corresponding scopolamine hydrobromide concentration is respectively 10.1,20.2,40.4, 80.9、161.8μg/mL.It is accurate respectively to draw above-mentioned each 10 μ L of reference substance mixed solution, liquid chromatograph is injected, by containing measurement Determine method measure.With peak area (Y) for ordinate, 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 are 25.9~414.0 μ g/mL, scopolamine hydrobromide concentration be 10.1~ In the mixed solution of 161.8 μ g/mL, good linear relationship is presented in four index components.In routine testing, as long as sample Concentration, then without drawing mark song every time, can be detected in mark song concentration range using one point external standard method.
Table 2 investigates result for linear relationship
7 stability tests
Take with a HUAZHI SHUAN medicine, after preparation, by content assaying method, respectively 0,2,4,8,16,24,32h sample introductions Measure.It turns out that measure is basicly stable in 32h from after preparing for HUAZHI SHUAN solution, 3 are shown in Table.
Table 3 is stability test result
8 repetitive tests
Take with a collection of 6 parts of HUAZHI SHUAN, with reference to " for the preparation of examination drug solution ", it is small that hydrochloric acid is carried out using high performance liquid chromatography Bark of a cork tree alkali, hydrochloric acid phellodendrine, the measure of 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 a collection of 2 parts of HUAZHI SHUAN, by content assaying method, respectively with A columns (Agilent Eclipse Plus C18Column, Specification 250mm × 4.6mm, 5 μm), B columns (Agilent ZORBAX SB-C18Column, specification 250mm × 4.6mm, 5 μm), C columns The chromatographic column of (4 μ Fusion-RP 80A columns of Phenomenex Synergi, specification 250mm × 4.6mm, 4 μm) three kinds of models is surveyed It is fixed.It turns out that being 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.
Table 5 is the measurement result of different chromatographic columns
10 sample recovery rates are tested
6 parts of the HUAZHI SHUAN with a collection of known content is taken, every part takes about 0.5g, accurately weighed, puts in conical flask with cover；Respectively Precision adds 1mL Berberine hydrochlorides reference substance solution (0.1235mg/mL), 1mL hydrochloric acid phellodendrine reference substance solutions (0.1226mg/mL), 1mL matrines reference substance solution (2.821mg/mL) and 1mL scopolamine hydrobromides (0.02674mg/ ML), it is remaining to be handled and measured by " preparation of test solution ", it the results are shown in Table 6~table 9.
The rate of recovery of 6 Berberine hydrochloride of table
Table 7 is the rate of recovery of hydrochloric acid phellodendrine
Table 8 is the rate of recovery of matrine
Table 9 is the rate of recovery of scopolamine hydrobromide
11 specificities are tested
The medicinal material in addition to Cortex Phellodendri, kuh-seng and datura flower is weighed by HUAZHI SHUAN prescription, negative sample is made by preparation process. Negative sample solution is made by the preparation method for examination drug solution again.Precision draws reference substance solution, HUAZHI SHUAN solution and the moon Property each 10 μ L of sample solution, are injected separately into liquid chromatograph, are measured by content assaying method.It turns out that small 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 chromatography On, negative sample solution is noiseless, illustrates that the assay method has preferable specificity, sees attached 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 measure hydrochloric acid barberry at the same time under the conditions of same HPLC methods Alkali, hydrochloric acid phellodendrine, the content of four index components of matrine and scopolamine hydrobromide.
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than the present invention is protected The limitation of scope is protected, although being explained in detail with reference to preferred embodiment to the present invention, those of ordinary skill in the art should Understand, can be to technical scheme technical scheme is modified or replaced equivalently, without departing from the essence of technical solution of the present invention.

Claims (5)

  1. A kind of 1. 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 comprise the following steps：
    1) preparation of hemorrhoid medicine test solution：Hemorrhoid medicine powder is placed in alcoholic solution and is extracted, is filtered, concentrates filtrate；
    The preparation of reference substance solution：Prepare the reference substance of matrine, hydrochloric acid phellodendrine, Berberine hydrochloride and scopolamine hydrobromide Solution；
    The solvent that hemorrhoid medicine test solution and reference substance solution use when preparing is 50%~100% methanol solution or 50% ~100% ethanol solution；
    The hemorrhoid medicine is HUAZHI SHUAN；
    2) it is accurate respectively to draw hemorrhoid medicine test solution and reference substance solution, inject high performance liquid chromatograph, measure；Wherein Chromatography condition is：Using gradient elution, mobile phase A is acetonitrile, and Mobile phase B is sour molten containing dodecyl sodium sulfate Liquid, the acid in the Mobile phase B are glacial acetic acid, phosphoric acid or formic acid, and percentage by volume of the acid in Mobile phase B is 0.05% ~0.3%, mass percent of the dodecyl sodium sulfate in Mobile phase B is 0.1%~0.3%；Detection wavelength is 210 ~230nm, column temperature are 20~30 DEG C, and the chromatographic column that when measure uses is the C using octadecylsilane chemically bonded silica as filler18 Column；
    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%.
  2. 2. detection method according to claim 1, it is characterised in that：In the step 1), 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 is ultrasonically treated, alcoholic solution is added after cooling, supplies the weight of reduction；Freeze 30~60min After filtered, concentrate the filtrate to the 2/25~1/8 of original volume.
  3. 3. detection method according to claim 1, it is characterised in that：The preparation of the reference substance solution is specially：Weigh Matrine reference substance, hydrochloric acid phellodendrine reference substance, the reference substance of Berberine hydrochloride reference substance and scopolamine hydrobromide, add institute State alcoholic solution be configured to matrine be 241.3~3860.0 μ g/mL, hydrochloric acid Cortex Phellodendri alkali concn be 27.4~438.4 μ g/ ML, Berberine hydrochloride concentration are 25.9~414.0 μ g/mL, scopolamine hydrobromide concentration is the mixed of 10.1~161.8 μ g/mL Close solution.
  4. 4. detection method according to claim 1, it is characterised in that：The alcoholic solution is 60% methanol solution.
  5. 5. detection method according to claim 1, 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%.
CN201610910516.9A 2016-10-19 2016-10-19 A kind of detection method of hemorrhoid medicine index components content Active CN106501396B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610910516.9A CN106501396B (en) 2016-10-19 2016-10-19 A kind of detection method of hemorrhoid medicine index components content

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610910516.9A CN106501396B (en) 2016-10-19 2016-10-19 A kind of detection method of hemorrhoid medicine index components content

Publications (2)

Publication Number Publication Date
CN106501396A CN106501396A (en) 2017-03-15
CN106501396B true CN106501396B (en) 2018-04-20

Family

ID=58294315

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610910516.9A Active CN106501396B (en) 2016-10-19 2016-10-19 A kind of detection method of hemorrhoid medicine index components content

Country Status (1)

Country Link
CN (1) CN106501396B (en)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
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
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

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102552478A (en) * 2010-12-16 2012-07-11 贵州万胜药业有限责任公司 Quality detection method of Nine Ingredient Hemorrhoid Capsules
CN103316102A (en) * 2013-06-09 2013-09-25 荣昌制药(淄博)有限公司 Detection method for external traditional Chinese medicine preparation for treating haemorrhoids
CN103884815A (en) * 2014-03-13 2014-06-25 贵阳学院 Method for inspecting quality of haemorrhoids suppository

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102552478A (en) * 2010-12-16 2012-07-11 贵州万胜药业有限责任公司 Quality detection method of Nine Ingredient Hemorrhoid Capsules
CN103316102A (en) * 2013-06-09 2013-09-25 荣昌制药(淄博)有限公司 Detection method for external traditional Chinese medicine preparation for treating haemorrhoids
CN103884815A (en) * 2014-03-13 2014-06-25 贵阳学院 Method for inspecting quality of haemorrhoids suppository

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
HPLC法测定化痔栓中人参皂苷Rg1、三七皂苷R1的含量;王桂香;《辽宁中医杂志》;20110331;第38卷(第3期);519-520 *
HPLC法测定复方痔疮栓中小檗碱型生物碱的含量;丁华;《药物分析杂志》;20040331;第24卷(第3期);337-338 *
高效液相色谱法测定克痔栓中盐酸小檗碱的含量;王振军;《中国医院药学杂志》;20050731;第25卷(第7期);678-679 *

Also Published As

Publication number Publication date
CN106501396A (en) 2017-03-15

Similar Documents

Publication Publication Date Title
CN106501396B (en) A kind of detection method of hemorrhoid medicine index components content
CN105842373B (en) A method of establishing the finger-print of the pharmaceutical preparation of honeysuckle
CN104398642A (en) Preparation and quality detection method of compound prescription cortex phellodendri chinensis fluid
CN104730171B (en) Radix Paeoniae Rubra, Flos Lonicerae multi-target ingredient content assaying method in a kind of compound Chinese medicinal preparation
CN103207255A (en) Content detection method for Naoxintong capsule
CN106093262A (en) A kind of method of the finger printing of the pharmaceutical preparation setting up Radix Scrophulariae
CN109406645A (en) A kind of Antisathmatic oral liquid for child epheday intermedia, the detection method for frying semen armeniacae amarae, Radix Glycyrrhizae, radix scutellariae
CN106802327B (en) A method of it establishing youngster and rushes down the finger-print for stopping pharmaceutical preparation
CN108760945A (en) A kind of detection method of Qijiaoshenbai capsule
CN104155383B (en) The detection method of blue or green Pu granule
CN109633003A (en) Radix Polygalae mouth mountain ketone III and 3,6 in a kind of measurement Radix Polygalae, the method for-two mustard acyl cane sugar contents
CN101721475A (en) Quality control method for Yaotongning capsules
CN108627581A (en) Rhynchophyllin and isorhynchophylline content assaying method in a kind of Xiao ' erqixingcha particle
CN102068598A (en) Quality control method of Yangrong Baicao Wan for treating irregular menses caused by hemophthisis
CN106706835A (en) Quality detection method of trollius chinensis bunge effervescent tablets
CN103163272B (en) Quality control method of infant spleen tonifying medicament
CN110274970A (en) The method for building up for melting poor finger-print and its application in Yixiesheng capsule Quality Control
CN103575823B (en) The detection method of 8 kinds of chemical compositions in a kind of Tangminling preparation
CN106950289B (en) A kind of coronary disease treatment capsule one is surveyed comments detection method of content more
CN105004833A (en) Detection method for traditional Chinese medicine preparation for treating acute gouty arthritis and gout
CN104873686A (en) Quality detection method for ginseng antler brain-boosting capsule
CN109781884B (en) Establishing method of Qianliexin capsule fingerprint and fingerprint thereof
CN106168608A (en) The detection method of EGCG content in a kind of tea polyphenols
CN115343377A (en) Fingerprint spectrum of stomach-clearing coptis tablet and construction method and application thereof
CN104345115B (en) A kind of method for qualitative and quantitative detection of Kanggu-Zengsheng tablet

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
CB03 Change of inventor or designer information

Inventor after: Peng Hongying

Inventor after: Jiang Tao

Inventor after: Chen Xiaojian

Inventor after: Zhu Shaoxuan

Inventor after: Cheng Junlu

Inventor after: Jia Jialiang

Inventor before: Peng Hongying

Inventor before: Zhu Shaoxuan

Inventor before: Chen Xiaojian

Inventor before: Jiang Tao

Inventor before: Cheng Junlu

Inventor before: Jia Jialiang

CB03 Change of inventor or designer information
GR01 Patent grant
GR01 Patent grant