CN107102093A - The detection method of compound pellet mattress cream - Google Patents
The detection method of compound pellet mattress cream Download PDFInfo
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- CN107102093A CN107102093A CN201710366383.8A CN201710366383A CN107102093A CN 107102093 A CN107102093 A CN 107102093A CN 201710366383 A CN201710366383 A CN 201710366383A CN 107102093 A CN107102093 A CN 107102093A
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Abstract
The invention provides a kind of detection method of the red mattress cream of compound, this method includes character, composition differentiates, check, assay, it is for the former examination criteria imperfection of the red mattress cream of the compound, in standard in addition to the detection of general indicator is carried out according to rules of preparations, there is no other any quality testing measures, drug quality cannot be monitored well, so as to add the discriminating to oriental wormwood and sweet wormwood, the discriminating of the red sage root and bark of ash, the discriminating of radix bupleuri, the discriminating of radix glycyrrhizae, add the assay to tanshinone IIA, the assay of tanshin polyphenolic acid B, the present invention can effectively detect whether illegal manufacturer throws less or do not throw corresponding medicinal raw material, also it have detected confusing sweet wormwood, bark of ash is mixed into, prevent misuse and abuse, so as to the effective quality for ensureing the red mattress cream of compound, make the drug safety effective, ensure the clinical efficacy of the medicine, safeguard the interests of patient.
Description
Technical field
The present invention relates to a kind of detection method of Chinese medicine preparation, the detection method of more particularly to a kind of red mattress cream of compound.
Background technology：
Compound pellet mattress cream be《The Sanitation Ministry medicine standard Traditional Chinese medicine historical preparation》Second medicine for recording, standard number is WS3-B-
0339-90, prescription is oriental wormwood, the red sage root, radix bupleuri, rhizoma imperatae, Radix Isatidis, radix glycyrrhizae, jujube, is the soft extract of pure Chinese medicine, it has
Clearing heat and promoting diuresis, the effect for the removing jaundice that detoxifies, clinically for treating acute infectious hepatitis, is clearing heat and promoting diuresis, solution in the market
Malicious removing jaundice, the common drug for treating acute infectious hepatitis, but because this standard is to formulate the upper end of the eighties in a century
, it is limited to technical merit and detecting instrument condition at that time, primary standard is still far from perfect, removes and carried out according to rules of preparations in standard
Outside the detection of general indicator, without other any quality testing measures, it cannot ensure drug quality well.
On the one hand, one of maximum medicinal material of consumption is oriental wormwood in the prescription of this medicine, accounts for a quarter of whole recipe quantity,
But oriental wormwood medicinal material is easily obscured with sweet wormwood and is used as medicine, and is had undesirable effect to drug quality and curative effect, oriental wormwood is that composite family is perennial
The seedling of herbaceous plant Artemisia capillaris or artemisia scoparia, taste bitter and cold, returns spleen, stomach, liver, gallbladder channel, main effect are exactly eliminating dampness and heat, are disappeared
Removing jaundice subcutaneous ulcer, also available for diseases such as wet sore itch, leaching stream yellow waters.And sweet wormwood is composite family annual herb plant sweet wormwood and artemisia annua
Herb, and using artemisia annua to be most common, most universal, nature and flavor bitter, acrid, cold, Return liver, courage, kidney channel, main effect are void of clearing up and discharging
Heat, cool blood preventing malaria, relieving summer-heat is antipyretic etc., for malaria fever and chills, fever due to yin deficiency, hectic fever due to yin labor illness, afternoon tidal fever, feverishness in palms and soles, hot summer weather
Diseases caused by external factors, fever without perspiration or have sweat, giddy headache, thirsty full and rapid pulse, or the warm heat disease later stage, the heresy of warm enter it is cloudy divide, night fever abating at dawn,
Heat moves back continuous low fever etc. after lossless card or warm heat disease.Both effects are different, it is necessary to have corresponding detection method to carry out
Monitoring.
On the other hand, one of maximum medicinal material of another consumption is the red sage root in the prescription of this medicine, also accounts for whole recipe quantity
A quarter, but red rooted salvia is often palmed off by bark of ash or is obscured and be used as medicine, and bad shadow is caused to drug quality and curative effect
Ring, bark of ash is the dry root of gentianaceae plant, the red sage root is the drying root and rhizome of labiate, but is due to both outward appearances
Character is quite similar, in recent years, is used it is often found that people someone obscures two kinds of medicinal materials, but both effects are different, it is necessary to
There is corresponding detection method to be monitored.
So, above mentioned problem can cause dosage dispensing of many illegal manufacturers when producing medicine not in strict accordance with prescription,
The high raw material of price is even reduced wantonly, causes the curative effect of medicine to be decreased obviously, the safe and effective of medicine is influenceed, and serious infringement is suffered from
The interests of person.
The content of the invention：
The purpose of the present invention, is to provide a kind of detection method of the red mattress cream of compound, this method is by increasing main Chinese medicinal materials oriental wormwood, pellet
Ginseng, radix bupleuri, the indentification by TLC of radix glycyrrhizae, and the indentification by TLC for easily obscuring medicinal material sweet wormwood, bark of ash is added, add
The assay of main component tanshinone IIA and tanshin polyphenolic acid B in red rooted salvia, so as to effectively monitor whether illegal manufacturer throws less
Or corresponding medicinal raw material is not thrown, also monitor confusing sweet wormwood, bark of ash and be mixed into, it is therefore prevented that misapplied and abuse, so as to have
The quality of the red mattress cream of monitoring compound of effect, makes the drug safety effective, it is ensured that the clinical efficacy of the medicine, safeguards the profit of patient
Benefit.
What the present invention was realized in：
The detection method for the red mattress cream of compound that the present invention is provided, including character, composition differentiate, checked, assay project, its
In, it is the discriminating to oriental wormwood and sweet wormwood, the discriminating of the red sage root and bark of ash, the discriminating of radix bupleuri, the discriminating of radix glycyrrhizae that composition, which differentiates, containing measurement
Surely it is assay, the assay of tanshin polyphenolic acid B to tanshinone IIA.
（1）Oriental wormwood and the discriminating of sweet wormwood：This product 1g, plus 8~12ml of methanol are taken, ultrasonically treated 20~40 minutes, is filtered, filter
Liquid is evaporated, and residue adds methanol 2ml to make dissolving, is used as need testing solution；Take escoparone contrast product, plus methanol that every 1ml is made and contains
0.4mg solution, is used as escoparone contrast product solution；Take Qinghaosu, plus methanol that every 1ml is made containing the molten of 0.2mg
Liquid, is used as Qinghaosu solution；Draw above-mentioned each 5 μ l of three kinds of solution, put respectively on same silica gel g thin-layer plate, with by
60~90 DEG C of petroleum ethers: ethyl acetate: acetone=6~8 of volume basis: 3~5: 1~3 be solvent, deploys, takes out, dry,
Inspected under the ultraviolet lamp for putting 365nm, in test sample chromatogram, on position corresponding with escoparone contrast product chromatogram, show phase
With the fluorescence spot of color；On position corresponding with Qinghaosu chromatogram, do not show the spot of same color；
（2）The discriminating of the red sage root and bark of ash：This product 2g is taken, add water 8~12ml, is stirred to dissolve, add diethyl ether 15~25ml, shaken,
Place 0.5~2 hour, point take ether layer, standby, raffinate is added diethyl ether 8~12ml again, and shaking is placed 0.5~2 hour, point take second
Ether layer, mixes with ether above, volatilizes, and residue adds ethyl acetate 1ml to make dissolving, is used as need testing solution；Take tanshinone IIA
Solution of every 1ml containing 0.3mg is made in reference substance, plus ethyl acetate, is used as tanshinone IIA reference substance solution；Take gentiamarin pair
Solution of every 1ml containing 0.2mg is made according to product, plus ethyl acetate, gentiamarin reference substance solution is used as；According to thin-layered chromatography examination
Test, draw above-mentioned each 4 μ l of three kinds of solution, put respectively on same silica gel g thin-layer plate, the benzene: ethyl acetate in terms of by volume=
20~22: 0.5~2 be solvent, deploys, takes out, dry, in test sample chromatogram, corresponding to tanshinone IIA reference substance chromatogram
On position, show the spot of same color；With on gentiamarin reference substance chromatogram relevant position, not showing the spot of same color；
（3）The discriminating of radix bupleuri：This product 3g is taken, add water 20~40ml, is stirred to dissolve, centrifuge, take supernatant, be added in 100~200
Mesh, 8g, internal diameter is 2.5~3cm, on the polyamide column A of wet method dress post, respectively with water, 20% ethanol and 50% ethanol each 80~
120ml is eluted, and is collected 50% ethanol eluate, is evaporated, residue adds methanol 1ml to make dissolving, is used as need testing solution；Separately take radix bupleuri
Control medicinal material 1g, is added water to cook 1.5 hours, and filtration, filtrate is concentrated into 10ml, is added in 100~200 mesh, 4g, internal diameter is 2cm, wet
On the polyamide column B of method dress post, eluted respectively with water 100ml and 50% ethanol 150ml, collect 50% ethanol eluate, be evaporated, it is residual
Slag adds methanol 1ml to make dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, the μ l of need testing solution 2~10 are drawn and right
According to the μ l of medicinal material solution 2, put respectively on same silica gel g thin-layer plate, the ethyl acetate: ethanol: water=11~13 in terms of by volume
: 1~3: 0.5~2 be solvent, deploys, takes out, dry, sprays with 10% ethanol solution of sulfuric acid of 5% paradime thylaminobenzaldehyde,
Hot blast is blown to spot development clearly, puts and is inspected under 365nm ultraviolet lamps, in test sample chromatogram, corresponding to control medicinal material chromatogram
Position on, show same color fluorescence spot；
（4）The discriminating of radix glycyrrhizae：This product 10g, plus 20~40ml of ethanol are taken, is stirred to dissolve, ultrasonically treated 10~30 minutes, filter
Cross, filtrate is evaporated, residue adds water 10~30 ml dissolvings, adjusts pH value to 2~3 with hydrochloric acid, 1~3 is extracted with ethyl acetate shaking
Secondary, 10~30 ml, combined ethyl acetate extract solution, are evaporated every time, and residue adds the ml of methanol 1 to make dissolving, is used as need testing solution；
Another extracting liquorice control medicinal material 1g, adds water appropriate, decocts 30 minutes, let cool, filter, filtrate is concentrated into 20ml, with it is water saturated just
Butanol shaking is extracted 2 times, and each 20ml merges n-butanol extracting liquid, with the water washing 2 times of n-butanol saturation, each 10ml,
N-butanol liquid is evaporated, and residue adds methanol 1ml to make dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, test sample is drawn
Solution and each 5~10 μ l of control medicinal material solution, put on same silica gel g thin-layer plate, the chloroform in terms of by volume respectively:
Methanol: water=38~42: 9~11: 0.5~2 be solvent, deploys, takes out, dry, and is sprayed with 5% vanillin-sulfuric acid solution,
105 DEG C are heated to spot development clearly, in test sample chromatogram, on position corresponding with control medicinal material chromatogram, show same color
Spot；
（5）The assay of tanshinone IIA：It is filler with octadecylsilane chemically bonded silica；Methanol in terms of by volume:
Water=75: 25 be mobile phase；Detection wavelength is 270nm；Number of theoretical plate is calculated by tanshinone IIA peak should be not less than 2000；Precision claims
Tanshinone IIA reference substance 10mg is taken, is put in 50ml brown measuring bottles, plus methanol is to scale, shakes up, then again from the mixed liquor shaken up
Middle precision measures 2ml, puts in 25ml brown measuring bottles, plus methanol is to scale, shakes up, and produces the μ g containing tanshinone IIA 16 in every 1ml
Reference substance solution；This product 2g is taken, is put in conical flask with cover, precision adds methanol 50ml, weighed weight, in power 120W, frequency
Ultrasonically treated 10~30 minutes under conditions of rate 59KHz, let cool, close plug, then weighed weight, less loss weight is supplied with methanol, is shaken
Even, filtration takes subsequent filtrate, produces need testing solution；It is accurate respectively to draw reference substance solution and each 5 μ l of need testing solution, injection
Liquid chromatograph, determines, produces；
Every gram of this product contains tanshinone IIA（C19H18O3）0.4mg must not be less than；
（6）The assay of tanshin polyphenolic acid B：It is filler with octadecylsilane chemically bonded silica；Methanol: second in terms of by volume
Nitrile: formic acid: water=30: be mobile phase at 10: 1: 59；Detection wavelength is 286nm；Number of theoretical plate is calculated by tanshin polyphenolic acid B peak to be not less than
2000；Tanshin polyphenolic acid B reference substance is weighed, plus solution of every 1ml containing 0.14mg is made in 75% methanol, produces reference substance solution；Take this
Product 2g, it is accurately weighed, put in conical flask with cover, precision adds 75% methanol 50ml, weighed weight, in power 120W, frequency
Ultrasonically treated 10~30 minutes under conditions of 59KHz, take out, let cool, then weighed weight, the weight of less loss is supplied with 75% methanol
Amount, shakes up, and filters, takes subsequent filtrate, produce need testing solution；It is accurate respectively to draw reference substance solution and each 10 μ of need testing solution
L, injects liquid chromatograph, determines, produces.
Every gram of (C containing tanshin polyphenolic acid B of this product36H30O16 ) 8mg must not be less than；
More specifically detection method：
（1）The discriminating of oriental wormwood and sweet wormwood is：This product 1g, plus methanol 10ml are taken, ultrasonically treated 30 minutes, is filtered, filtrate is evaporated, residual
Slag adds methanol 2ml to make dissolving, is used as need testing solution；Take escoparone contrast product, plus methanol that every 1ml is made containing the molten of 0.4mg
Liquid, is used as escoparone contrast product solution；Take Qinghaosu, plus methanol that solution of every 1ml containing 0.2mg is made, be used as green grass or young crops
Artemisin reference substance solution；Above-mentioned each 5 μ l of three kinds of solution are drawn, are put respectively on same silica gel g thin-layer plate, in terms of by volume
60~90 DEG C of petroleum ethers: ethyl acetate: acetone=7: be solvent at 4: 2, deploy, take out, dry, put under 365nm ultraviolet lamp
Inspect, in test sample chromatogram, on position corresponding with escoparone contrast product chromatogram, show the fluorescence spot of same color；
On position corresponding with Qinghaosu chromatogram, do not show the spot of same color；
（2）The discriminating of the red sage root and bark of ash is：This product 2g is taken, add water 10ml, is stirred to dissolve, add diethyl ether 20ml, shaken, place 1
Hour, point ether layer is taken, standby, raffinate is added diethyl ether 10ml again, and shaking is placed 1 hour, point take ether layer, with ether above
Mixing, is volatilized, residue adds ethyl acetate 1ml to make dissolving, is used as need testing solution；Take tanshinone IIA reference substance, plus ethyl acetate
Solution of every 1ml containing 0.3mg is made, tanshinone IIA reference substance solution is used as；Take gentiamarin reference substance, plus ethyl acetate system
Into solution of every 1ml containing 0.2mg, gentiamarin reference substance solution is used as；According to thin-layered chromatography experiment, above-mentioned three kinds of solution is drawn
Each 4 μ l, put on same silica gel g thin-layer plate, the benzene: ethyl acetate=21 in terms of by volume: 1 is solvent deploys respectively,
Take out, dry, in test sample chromatogram, with tanshinone IIA reference substance chromatogram relevant position, showing the spot of same color；
With on gentiamarin reference substance chromatogram relevant position, not showing the spot of same color；
（3）The discriminating of radix bupleuri is：This product 3g is taken, add water 30ml, is stirred to dissolve, centrifuge, take supernatant, be added in 100~200
Mesh, 8g, internal diameter is on 2.5~3cm, the polyamide column A of wet method dress post, to be washed respectively with water, 20% ethanol and each 100ml of 50% ethanol
It is de-, 50% ethanol eluate is collected, is evaporated, residue adds methanol 1ml to make dissolving, is used as need testing solution；Separately take radix bupleuri control medicinal material
1g, is added water to cook 1.5 hours, and filtration, filtrate is concentrated into 10ml, is added in 100~200 mesh, 4g, and internal diameter is 2cm, and wet method dress post is gathered
On acid amides post B, eluted respectively with water 100ml and 50% ethanol 150ml, collect 50% ethanol eluate, be evaporated, residue adds methanol
1ml makes dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, draw the μ l of need testing solution 2~10 and control medicinal material is molten
The μ l of liquid 2, put on same silica gel g thin-layer plate, the ethyl acetate: ethanol: water=12 in terms of by volume respectively: be expansion at 2: 1
Agent, deploys, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid of 5% paradime thylaminobenzaldehyde, it is clear that hot blast is blown to spot development
It is clear, put and inspected under 365nm ultraviolet lamps, in test sample chromatogram, on position corresponding with control medicinal material chromatogram, show same color
Fluorescence spot；
（4）The discriminating of radix glycyrrhizae is：This product 10g, plus ethanol 30ml are taken, is stirred to dissolve, ultrasonically treated 20 minutes, filtration, filtrate
It is evaporated, residue adds water 20 ml dissolvings, adjusts pH value to 2~3 with hydrochloric acid, is extracted 2 times with ethyl acetate shaking, 20 ml, is closed every time
And acetic acid ethyl acetate extract, it is evaporated, residue adds the ml of methanol 1 to make dissolving, is used as need testing solution；Another extracting liquorice control medicinal material 1g,
Add water appropriate, decoct 30 minutes, let cool, filter, filtrate is concentrated into 20ml, shake extraction 2 times with water saturated n-butanol, often
Secondary 20ml, merges n-butanol extracting liquid, and with the water washing 2 times of n-butanol saturation, each 10ml, n-butanol liquid is evaporated, and residue adds
Methanol 1ml makes dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, need testing solution and control medicinal material solution are drawn
Each 5~10 μ l, put on same silica gel g thin-layer plate, the chloroform: methanol: water=40 in terms of by volume: be at 10: 1 respectively
Solvent, deploys, and takes out, dries, and sprays with 5% vanillin-sulfuric acid solution, spot development is heated at 105 DEG C clearly, test sample
In chromatogram, on position corresponding with control medicinal material chromatogram, show the spot of same color；
（5）The assay of tanshinone IIA is：It is filler with octadecylsilane chemically bonded silica；First in terms of by volume
Alcohol: water=75: 25 be mobile phase；Detection wavelength is 270nm；Number of theoretical plate is calculated by tanshinone IIA peak should be not less than 2000；Essence
It is close to weigh tanshinone IIA reference substance 10mg, put in 50ml brown measuring bottles, plus methanol is to scale, shakes up, it is then mixed from what is shaken up again
Close precision in liquid and measure 2ml, put in 25ml brown measuring bottles, plus methanol is to scale, shakes up, and produces and contains tanshinone IIA in every 1ml
16 μ g reference substance solution；This product 2g is taken, is put in conical flask with cover, precision adds methanol 50ml, weighed weight, in power
Ultrasonically treated 20 minutes under conditions of 120W, frequency 59KHz, let cool, close plug, then weighed weight, less loss weight is supplied with methanol,
Shake up, filter, take subsequent filtrate, produce need testing solution；It is accurate respectively to draw reference substance solution and each 5 μ l of need testing solution, note
Enter liquid chromatograph, determine, produce；
Every gram of this product contains tanshinone IIA（C19H18O3）0.4mg must not be less than；
（6）The assay of tanshin polyphenolic acid B is：It is filler with octadecylsilane chemically bonded silica；Methanol in terms of by volume:
Acetonitrile: formic acid: water=30: be mobile phase at 10: 1: 59；Detection wavelength is 286nm；Number of theoretical plate is calculated by tanshin polyphenolic acid B peak should not be low
In 2000；Tanshin polyphenolic acid B reference substance is weighed, plus solution of every 1ml containing 0.14mg is made in 75% methanol, produces reference substance solution；Take
This product 2g, it is accurately weighed, put in conical flask with cover, precision adds 75% methanol 50ml, weighed weight, in power 120W, frequency
Ultrasonically treated 20 minutes under conditions of 59KHz, take out, let cool, then weighed weight, the weight of less loss is supplied with 75% methanol, is shaken
Even, filtration takes subsequent filtrate, produces need testing solution；It is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, injection
Liquid chromatograph, determines, produces；
Every gram of (C containing tanshin polyphenolic acid B of this product36H30O16 ) 8mg must not be less than.
Beneficial effects of the present invention：
The detection method of the red mattress cream of compound that the present invention is provided, by increase main Chinese medicinal materials oriental wormwood, the red sage root, radix bupleuri, radix glycyrrhizae it is thin
Layer chromatography differentiates, and adds and easily obscure the indentification by TLC of medicinal material sweet wormwood, bark of ash, add in red rooted salvia it is main into
Divide the assay of tanshinone IIA and tanshin polyphenolic acid B, it is former so as to effectively avoid illegal manufacturer from throwing less or do not throw corresponding medicinal material
Material, it is to avoid not in strict accordance with the dosage dispensing of prescription during production medicine, while also accurately monitored confusing sweet wormwood, the Qin
Macrophylla is mixed into, it is therefore prevented that is misapplied and is abused, it is to avoid it influences drug quality and curative effect, meanwhile, Components in Salvia miltiorrhiza in prescription is also
Effective monitoring is arrived, it is ensured that the clinical efficacy of the safe and effective and medicine of medicine, has safeguarded the interests of patient, the present invention is provided
The red mattress cream of compound detection method it is scientific and reasonable, practical.
Embodiment
Embodiment 1：
Prescription：Oriental wormwood 240g, red sage root 240g, radix bupleuri 120g, rhizoma imperatae 120g, Radix Isatidis 120g, radix glycyrrhizae 60g, jujube 60g.
Preparation method：The taste of the above seven, is added water to cook three times, collecting decoction, is stood, and filtration, filtrate is concentrated into relative density and is
1.30（Heat is surveyed）More than, plus sucrose 240g, heating fusing, mix, it is 1.10~1.30 to be concentrated into relative density, is produced.
Character：This product is the semifluid of sepia stiff, taste sweet tea, slight bitter.
Check：Relative density is 1.10~1.30.
Composition differentiates and assay：
（1）Oriental wormwood and the discriminating of sweet wormwood：This product 1g, plus methanol 8ml are taken, ultrasonically treated 20 minutes, filtration, filtrate was evaporated, residue
Plus methanol 2ml makes dissolving, is used as need testing solution；Take escoparone contrast product, plus methanol that solution of every 1ml containing 0.4mg is made,
It is used as escoparone contrast product solution；Take Qinghaosu, plus methanol that solution of every 1ml containing 0.2mg is made, be used as qinghaosu
Reference substance solution；Above-mentioned each 5 μ l of three kinds of solution are drawn, are put respectively on same silica gel g thin-layer plate, 60 in terms of by volume~
90 DEG C of petroleum ethers: ethyl acetate: acetone=6: be solvent at 3: 1, deploy, take out, dry, put and examined under 365nm ultraviolet lamp
Depending in test sample chromatogram, on position corresponding with escoparone contrast product chromatogram, showing the fluorescence spot of same color；With
On the corresponding position of Qinghaosu chromatogram, do not show the spot of same color；
（2）The discriminating of the red sage root and bark of ash：This product 2g is taken, add water 8ml, is stirred to dissolve, add diethyl ether 15ml, shaken, place 0.5 small
When, point ether layer is taken, standby, raffinate is added diethyl ether 8ml again, and shaking is placed 0.5 hour, point take ether layer, mixed with ether above
Close, volatilize, residue adds ethyl acetate 1ml to make dissolving, is used as need testing solution；Take tanshinone IIA reference substance, plus ethyl acetate system
Into solution of every 1ml containing 0.3mg, tanshinone IIA reference substance solution is used as；Gentiamarin reference substance, plus ethyl acetate is taken to be made
Per solution of the 1ml containing 0.2mg, gentiamarin reference substance solution is used as；According to thin-layered chromatography experiment, above-mentioned three kinds of solution is drawn each
4 μ l, put on same silica gel g thin-layer plate, the benzene: ethyl acetate=20 in terms of by volume: 0.5 is solvent deploys respectively,
Take out, dry, in test sample chromatogram, with tanshinone IIA reference substance chromatogram relevant position, showing the spot of same color；
With on gentiamarin reference substance chromatogram relevant position, not showing the spot of same color；
（3）The discriminating of radix bupleuri：This product 3g is taken, add water 20ml, is stirred to dissolve, centrifuges, take supernatant, be added in 100~200 mesh,
8g, internal diameter is on 2.5~3cm, the polyamide column A of wet method dress post, to be eluted respectively with water, 20% ethanol and each 80ml of 50% ethanol,
50% ethanol eluate is collected, is evaporated, residue adds methanol 1ml to make dissolving, is used as need testing solution；Radix bupleuri control medicinal material 1g separately is taken,
Add water to cook 1.5 hours, filter, filtrate is concentrated into 10ml, be added in 100~200 mesh, 4g, internal diameter is 2cm, wet method dress post it is poly-
On acid amides post B, eluted respectively with water 100ml and 50% ethanol 150ml, collect 50% ethanol eluate, be evaporated, residue adds methanol
1ml makes dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, draw the μ l of need testing solution 2~10 and control medicinal material is molten
The μ l of liquid 2, put on same silica gel g thin-layer plate, the ethyl acetate: ethanol: water=11 in terms of by volume respectively: be expansion at 1: 0.5
Agent, deploys, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid of 5% paradime thylaminobenzaldehyde, it is clear that hot blast is blown to spot development
It is clear, put and inspected under 365nm ultraviolet lamps, in test sample chromatogram, on position corresponding with control medicinal material chromatogram, show same color
Fluorescence spot；
（4）The discriminating of radix glycyrrhizae：This product 10g, plus ethanol 20ml are taken, is stirred to dissolve, ultrasonically treated 10 minutes, filtration, filtrate was steamed
Dry, residue adds water 10 ml dissolvings, adjusts pH value to 2~3 with hydrochloric acid, is extracted 1 time with ethyl acetate shaking, 10 ml, merges every time
Acetic acid ethyl acetate extract, is evaporated, and residue adds methanol 1ml to make dissolving, is used as need testing solution；Another extracting liquorice control medicinal material 1g, plus
Appropriate amount of water, decocts 30 minutes, lets cool, and filters, and filtrate is concentrated into 20ml, shakes extraction 2 times with water saturated n-butanol, every time
20ml, merges n-butanol extracting liquid, and with the water washing 2 times of n-butanol saturation, each 10ml, n-butanol liquid is evaporated, and residue adds first
Alcohol 1ml makes dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, need testing solution and control medicinal material solution each 5 are drawn
~10 μ l, put on same silica gel g thin-layer plate, the chloroform: methanol: water=38 in terms of by volume respectively: be exhibition at 9: 0.5
Agent is opened, is deployed, takes out, dries, is sprayed with 5% vanillin-sulfuric acid solution, spot development is heated at 105 DEG C clearly, test sample color
In spectrum, on position corresponding with control medicinal material chromatogram, show the spot of same color；
（5）The assay of tanshinone IIA：It is filler with octadecylsilane chemically bonded silica；Methanol in terms of by volume:
Water=75: 25 be mobile phase；Detection wavelength is 270nm；Number of theoretical plate is calculated by tanshinone IIA peak should be not less than 2000；Precision claims
Tanshinone IIA reference substance 10mg is taken, is put in 50ml brown measuring bottles, plus methanol is to scale, shakes up, then again from the mixed liquor shaken up
Middle precision measures 2ml, puts in 25ml brown measuring bottles, plus methanol is to scale, shakes up, and produces the μ g containing tanshinone IIA 16 in every 1ml
Reference substance solution；This product 2g is taken, is put in conical flask with cover, precision adds methanol 50ml, weighed weight, in power 120W, frequency
Ultrasonically treated 10 minutes under conditions of rate 59KHz, let cool, close plug, then weighed weight, less loss weight is supplied with methanol, is shaken up, is filtered
Cross, take subsequent filtrate, produce need testing solution；It is accurate respectively to draw reference substance solution and each 5 μ l of need testing solution, inject liquid phase color
Spectrometer, determines, produces.
Every gram of this product contains tanshinone IIA（C19H18O3）For 0.6mg.
（6）The assay of tanshin polyphenolic acid B：It is filler with octadecylsilane chemically bonded silica；First in terms of by volume
Alcohol: acetonitrile: formic acid: water=3O: 10: 1: 59 are mobile phase；Detection wavelength is 286nm；Number of theoretical plate is calculated by tanshin polyphenolic acid B peak should
It is not less than 2000；Tanshin polyphenolic acid B reference substance is weighed, plus solution of every 1ml containing 0.14mg is made in 75% methanol, produces reference substance molten
Liquid；This product 2g is taken, it is accurately weighed, put in conical flask with cover, precision adds 75% methanol 50ml, weighed weight, in power 120W,
Ultrasonically treated 10 minutes under conditions of frequency 59KHz, take out, let cool, then weighed weight, the weight of less loss is supplied with 75% methanol
Amount, shakes up, and filters, takes subsequent filtrate, produce need testing solution；It is accurate respectively to draw reference substance solution and each 10 μ of need testing solution
L, injects liquid chromatograph, determines, produces.
Every gram of (C containing tanshin polyphenolic acid B of this product36H30O16 ) it is 9.1mg.
Embodiment 2：
In the present embodiment, except the step in composition discriminating and assay（1）To step（6）Outside the difference of embodiment 1, remaining
Method all same.
Composition differentiates and assay：
（1）Oriental wormwood and the discriminating of sweet wormwood：This product 1g, plus methanol 12ml are taken, ultrasonically treated 40 minutes, filtration, filtrate was evaporated, residue
Plus methanol 2ml makes dissolving, is used as need testing solution；Take escoparone contrast product, plus methanol that solution of every 1ml containing 0.4mg is made,
It is used as escoparone contrast product solution；Take Qinghaosu, plus methanol that solution of every 1ml containing 0.2mg is made, be used as qinghaosu
Reference substance solution；Above-mentioned each 5 μ l of three kinds of solution are drawn, are put respectively on same silica gel g thin-layer plate, 60 in terms of by volume~
90 DEG C of petroleum ethers: ethyl acetate: acetone=8: be solvent at 5: 3, deploy, take out, dry, put and examined under 365nm ultraviolet lamp
Depending in test sample chromatogram, on position corresponding with escoparone contrast product chromatogram, showing the fluorescence spot of same color；With
On the corresponding position of Qinghaosu chromatogram, do not show the spot of same color；
（2）The discriminating of the red sage root and bark of ash：This product 2g is taken, add water 12ml, is stirred to dissolve, add diethyl ether 25ml, shaken, place 2 small
When, point ether layer is taken, standby, raffinate is added diethyl ether 12ml again, and shaking is placed 2 hours, point take ether layer, mixed with ether above
Close, volatilize, residue adds ethyl acetate 1ml to make dissolving, is used as need testing solution；Take tanshinone IIA reference substance, plus ethyl acetate system
Into solution of every 1ml containing 0.3mg, tanshinone IIA reference substance solution is used as；Gentiamarin reference substance, plus ethyl acetate is taken to be made
Per solution of the 1ml containing 0.2mg, gentiamarin reference substance solution is used as；According to thin-layered chromatography experiment, above-mentioned three kinds of solution is drawn each
4 μ l, put on same silica gel g thin-layer plate, the benzene: ethyl acetate=22 in terms of by volume: 2 be solvent, expansion takes respectively
Go out, dry, in test sample chromatogram, with tanshinone IIA reference substance chromatogram relevant position, showing the spot of same color；With
On gentiamarin reference substance chromatogram relevant position, do not show the spot of same color；
（3）The discriminating of radix bupleuri：This product 3g is taken, add water 40ml, is stirred to dissolve, centrifuges, take supernatant, be added in 100~200 mesh,
8g, internal diameter is on 2.5~3cm, the polyamide column A of wet method dress post, to be eluted respectively with water, 20% ethanol and each 120ml of 50% ethanol,
50% ethanol eluate is collected, is evaporated, residue adds methanol 1ml to make dissolving, is used as need testing solution；Radix bupleuri control medicinal material 1g separately is taken,
Add water to cook 1.5 hours, filter, filtrate is concentrated into 10ml, be added in 100~200 mesh, 4g, internal diameter is 2cm, wet method dress post it is poly-
On acid amides post B, eluted respectively with water 100ml and 50% ethanol 150ml, collect 50% ethanol eluate, be evaporated, residue adds methanol
1ml makes dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, draw the μ l of need testing solution 2~10 and control medicinal material is molten
The μ l of liquid 2, put on same silica gel g thin-layer plate, the ethyl acetate: ethanol: water=13 in terms of by volume respectively: be expansion at 3: 2
Agent, deploys, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid of 5% paradime thylaminobenzaldehyde, it is clear that hot blast is blown to spot development
It is clear, put and inspected under 365nm ultraviolet lamps, in test sample chromatogram, on position corresponding with control medicinal material chromatogram, show same color
Fluorescence spot；
（4）The discriminating of radix glycyrrhizae：This product 10g, plus ethanol 40ml are taken, is stirred to dissolve, ultrasonically treated 30 minutes, filtration, filtrate was steamed
Dry, residue adds water 30 ml dissolvings, adjusts pH value to 2~3 with hydrochloric acid, is extracted 3 times with ethyl acetate shaking, 30 ml, merges every time
Acetic acid ethyl acetate extract, is evaporated, and residue adds the ml of methanol 1 to make dissolving, is used as need testing solution；Another extracting liquorice control medicinal material 1g, plus
Appropriate amount of water, decocts 30 minutes, lets cool, and filters, and filtrate is concentrated into 20ml, shakes extraction 2 times with water saturated n-butanol, every time
20ml, merges n-butanol extracting liquid, and with the water washing 2 times of n-butanol saturation, each 10ml, n-butanol liquid is evaporated, and residue adds first
Alcohol 1ml makes dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, need testing solution and control medicinal material solution each 5 are drawn
~10 μ l, put on same silica gel g thin-layer plate, the chloroform: methanol: water=42 in terms of by volume respectively: be expansion at 11: 2
Agent, deploys, and takes out, dries, and sprays with 5% vanillin-sulfuric acid solution, spot development is heated at 105 DEG C clearly, test sample chromatogram
In, on position corresponding with control medicinal material chromatogram, show the spot of same color；
（5）The assay of tanshinone IIA：It is filler with octadecylsilane chemically bonded silica；Methanol in terms of by volume:
Water=75: 25 be mobile phase；Detection wavelength is 270nm；Number of theoretical plate is calculated by tanshinone IIA peak should be not less than 2000；Precision claims
Tanshinone IIA reference substance 10mg is taken, is put in 50ml brown measuring bottles, plus methanol is to scale, shakes up, then again from the mixed liquor shaken up
Middle precision measures 2ml, puts in 25ml brown measuring bottles, plus methanol is to scale, shakes up, and produces the μ g containing tanshinone IIA 16 in every 1ml
Reference substance solution；This product 2g is taken, is put in conical flask with cover, precision adds methanol 50ml, weighed weight, in power 120W, frequency
Ultrasonically treated 30 minutes under conditions of rate 59KHz, let cool, close plug, then weighed weight, less loss weight is supplied with methanol, is shaken up, is filtered
Cross, take subsequent filtrate, produce need testing solution；It is accurate respectively to draw reference substance solution and each 5 μ l of need testing solution, inject liquid phase color
Spectrometer, determines, produces.
Every gram of this product contains tanshinone IIA（C19H18O3）For 0.7mg.
（6）The assay of tanshin polyphenolic acid B：It is filler with octadecylsilane chemically bonded silica；First in terms of by volume
Alcohol: acetonitrile: formic acid: water=3O: 10: 1: 59 are mobile phase；Detection wavelength is 286nm；Number of theoretical plate is calculated by tanshin polyphenolic acid B peak should
It is not less than 2000；Tanshin polyphenolic acid B reference substance is weighed, plus solution of every 1ml containing 0.14mg is made in 75% methanol, produces reference substance molten
Liquid；This product 2g is taken, it is accurately weighed, put in conical flask with cover, precision adds 75% methanol 50ml, weighed weight, in power 120W,
Ultrasonically treated 30 minutes under conditions of frequency 59KHz, take out, let cool, then weighed weight, the weight of less loss is supplied with 75% methanol
Amount, shakes up, and filters, takes subsequent filtrate, produce need testing solution；It is accurate respectively to draw reference substance solution and each 10 μ of need testing solution
L, injects liquid chromatograph, determines, produces.
Every gram of (C containing tanshin polyphenolic acid B of this product36H30O16 ) it is 9.7mg.
Embodiment 3：
In the present embodiment, except the step in composition discriminating and assay（1）To step（6）Outside the difference of embodiment 1, remaining
Method all same.
（1）The discriminating of oriental wormwood and sweet wormwood is：This product 1g, plus methanol 10ml are taken, ultrasonically treated 30 minutes, filtration, filtrate was steamed
Dry, residue adds methanol 2ml to make dissolving, is used as need testing solution；Take escoparone contrast product, plus methanol that every 1ml is made containing 0.4mg
Solution, be used as escoparone contrast product solution；Take Qinghaosu, plus methanol that solution of every 1ml containing 0.2mg is made, make
For Qinghaosu solution；Above-mentioned each 5 μ l of three kinds of solution are drawn, are put respectively on same silica gel g thin-layer plate, with by volume
60~90 DEG C of petroleum ethers: ethyl acetate: acetone=7: be solvent at 4: 2 of meter, deploy, and take out, dry, put 365nm ultraviolet light
Inspected under lamp, in test sample chromatogram, on position corresponding with escoparone contrast product chromatogram, show the fluorescent spot of same color
Point；On position corresponding with Qinghaosu chromatogram, do not show the spot of same color；
（2）The discriminating of the red sage root and bark of ash is：This product 2g is taken, add water 10ml, is stirred to dissolve, add diethyl ether 20ml, shaken, place 1
Hour, point ether layer is taken, standby, raffinate is added diethyl ether 10ml again, and shaking is placed 1 hour, point take ether layer, with ether above
Mixing, is volatilized, residue adds ethyl acetate 1ml to make dissolving, is used as need testing solution；Take tanshinone IIA reference substance, plus ethyl acetate
Solution of every 1ml containing 0.3mg is made, tanshinone IIA reference substance solution is used as；Take gentiamarin reference substance, plus ethyl acetate system
Into solution of every 1ml containing 0.2mg, gentiamarin reference substance solution is used as；According to thin-layered chromatography experiment, above-mentioned three kinds of solution is drawn
Each 4 μ l, put on same silica gel g thin-layer plate, the benzene: ethyl acetate=21 in terms of by volume: 1 is solvent deploys respectively,
Take out, dry, in test sample chromatogram, with tanshinone IIA reference substance chromatogram relevant position, showing the spot of same color；
With on gentiamarin reference substance chromatogram relevant position, not showing the spot of same color；
（3）The discriminating of radix bupleuri is：This product 3g is taken, add water 30ml, is stirred to dissolve, centrifuge, take supernatant, be added in 100~200
Mesh, 8g, internal diameter is on 2.5~3cm, the polyamide column A of wet method dress post, to be washed respectively with water, 20% ethanol and each 100ml of 50% ethanol
It is de-, 50% ethanol eluate is collected, is evaporated, residue adds methanol 1ml to make dissolving, is used as need testing solution；Separately take radix bupleuri control medicinal material
1g, is added water to cook 1.5 hours, and filtration, filtrate is concentrated into 10ml, is added in 100~200 mesh, 4g, and internal diameter is 2cm, and wet method dress post is gathered
On acid amides post B, eluted respectively with water 100ml and 50% ethanol 150ml, collect 50% ethanol eluate, be evaporated, residue adds methanol
1ml makes dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, draw the μ l of need testing solution 2~10 and control medicinal material is molten
The μ l of liquid 2, put on same silica gel g thin-layer plate, the ethyl acetate: ethanol: water=12 in terms of by volume respectively: be expansion at 2: 1
Agent, deploys, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid of 5% paradime thylaminobenzaldehyde, it is clear that hot blast is blown to spot development
It is clear, put and inspected under 365nm ultraviolet lamps, in test sample chromatogram, on position corresponding with control medicinal material chromatogram, show same color
Fluorescence spot；
（4）The discriminating of radix glycyrrhizae is：This product 10g, plus ethanol 30ml are taken, is stirred to dissolve, ultrasonically treated 20 minutes, filtration, filtrate
It is evaporated, residue adds water 20 ml dissolvings, adjusts pH value to 2~3 with hydrochloric acid, is extracted 2 times with ethyl acetate shaking, 20 ml, is closed every time
And acetic acid ethyl acetate extract, it is evaporated, residue adds the ml of methanol 1 to make dissolving, is used as need testing solution；Another extracting liquorice control medicinal material 1g,
Add water appropriate, decoct 30 minutes, let cool, filter, filtrate is concentrated into 20ml, shake extraction 2 times with water saturated n-butanol, often
Secondary 20ml, merges n-butanol extracting liquid, and with the water washing 2 times of n-butanol saturation, each 10ml, n-butanol liquid is evaporated, and residue adds
Methanol 1ml makes dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, need testing solution and control medicinal material solution are drawn
Each 5~10 μ l, put on same silica gel g thin-layer plate, the chloroform: methanol: water=40 in terms of by volume: be at 10: 1 respectively
Solvent, deploys, and takes out, dries, and sprays with 5% vanillin-sulfuric acid solution, spot development is heated at 105 DEG C clearly, test sample
In chromatogram, on position corresponding with control medicinal material chromatogram, show the spot of same color；
（5）The assay of tanshinone IIA is：It is filler with octadecylsilane chemically bonded silica；With first by weight
Alcohol: water=75: 25 be mobile phase；Detection wavelength is 270nm；Number of theoretical plate is calculated by tanshinone IIA peak should be not less than 2000；Essence
It is close to weigh tanshinone IIA reference substance 10mg, put in 50ml brown measuring bottles, plus methanol is to scale, shakes up, it is then mixed from what is shaken up again
Close precision in liquid and measure 2ml, put in 25ml brown measuring bottles, plus methanol is to scale, shakes up, and produces and contains tanshinone IIA in every 1ml
16 μ g reference substance solution；This product 2g is taken, is put in conical flask with cover, precision adds methanol 50ml, weighed weight, in power
Ultrasonically treated 20 minutes under conditions of 120W, frequency 59KHz, let cool, close plug, then weighed weight, less loss weight is supplied with methanol,
Shake up, filter, take subsequent filtrate, produce need testing solution；It is accurate respectively to draw reference substance solution and each 5 μ l of need testing solution, note
Enter liquid chromatograph, determine, produce；
Every gram of this product contains tanshinone IIA（C19H18O3）For 0.7mg.
（6）The assay of tanshin polyphenolic acid B is：It is filler with octadecylsilane chemically bonded silica；With by weight
Methanol: acetonitrile: formic acid: water=30: be mobile phase at 10: 1: 59；Detection wavelength is 286nm；Number of theoretical plate is calculated by tanshin polyphenolic acid B peak
2000 should be not less than；Tanshin polyphenolic acid B reference substance is weighed, plus solution of every 1ml containing 0.14mg is made in 75% methanol, produces reference substance
Solution；This product 2g is taken, it is accurately weighed, put in conical flask with cover, precision adds 75% methanol 50ml, weighed weight, in power
Ultrasonically treated 20 minutes under conditions of 120W, frequency 59KHz, take out, let cool, then weighed weight, supply less loss with 75% methanol
Weight, shake up, filter, take subsequent filtrate, produce need testing solution；Accurate absorption reference substance solution and need testing solution are each respectively
10 μ l, inject liquid chromatograph, determine, produce.
Every gram of (C containing tanshin polyphenolic acid B of this product36H30O16 ) it is 7.2mg.
Claims (2)
1. a kind of detection method of the red mattress cream of compound, including character, composition differentiate, checked, assay, it is characterised in that：Into
Divide and differentiate it is the discriminating to oriental wormwood and sweet wormwood, the discriminating of the red sage root and bark of ash, the discriminating of radix bupleuri, the discriminating of radix glycyrrhizae, assay is
The assay of assay, tanshin polyphenolic acid B to tanshinone IIA；
Wherein：
（1）Oriental wormwood and the discriminating of sweet wormwood：This product 1g, plus 8~12ml of methanol are taken, ultrasonically treated 20~40 minutes, filtration, filtrate was steamed
Dry, residue adds methanol 2ml to make dissolving, is used as need testing solution；Take escoparone contrast product, plus methanol that every 1ml is made containing 0.4mg
Solution, be used as escoparone contrast product solution；Take Qinghaosu, plus methanol that solution of every 1ml containing 0.2mg is made, make
For Qinghaosu solution；Above-mentioned each 5 μ l of three kinds of solution are drawn, are put respectively on same silica gel g thin-layer plate, with by volume
60~90 DEG C of petroleum ethers: ethyl acetate: acetone=6~8 of meter: 3~5: 1~3 be solvent, deploys, takes out, dry, put
Inspect, in test sample chromatogram, on position corresponding with escoparone contrast product chromatogram, show identical under 365nm ultraviolet lamp
The fluorescence spot of color；On position corresponding with Qinghaosu chromatogram, do not show the spot of same color；
（2）The discriminating of the red sage root and bark of ash：This product 2g is taken, add water 8~12ml, is stirred to dissolve, add diethyl ether 15~25ml, shaken,
Place 0.5~2 hour, point take ether layer, standby, raffinate is added diethyl ether 8~12ml again, and shaking is placed 0.5~2 hour, point take second
Ether layer, mixes with ether above, volatilizes, and residue adds ethyl acetate 1ml to make dissolving, is used as need testing solution；Take tanshinone IIA
Solution of every 1ml containing 0.3mg is made in reference substance, plus ethyl acetate, is used as tanshinone IIA reference substance solution；Take gentiamarin pair
Solution of every 1ml containing 0.2mg is made according to product, plus ethyl acetate, gentiamarin reference substance solution is used as；According to thin-layered chromatography examination
Test, draw above-mentioned each 4 μ l of three kinds of solution, put respectively on same silica gel g thin-layer plate, the benzene: ethyl acetate in terms of by volume=
20~22: 0.5~2 be solvent, deploys, takes out, dry, in test sample chromatogram, corresponding to tanshinone IIA reference substance chromatogram
On position, show the spot of same color；With on gentiamarin reference substance chromatogram relevant position, not showing the spot of same color；
（3）The discriminating of radix bupleuri：This product 3g is taken, add water 20~40ml, is stirred to dissolve, centrifuge, take supernatant, be added in 100~200
Mesh, 8g, internal diameter is 2.5~3cm, on the polyamide column A of wet method dress post, respectively with water, 20% ethanol and 50% ethanol each 80~
120ml is eluted, and is collected 50% ethanol eluate, is evaporated, residue adds methanol 1ml to make dissolving, is used as need testing solution；Separately take radix bupleuri
Control medicinal material 1g, is added water to cook 1.5 hours, and filtration, filtrate is concentrated into 10ml, is added in 100~200 mesh, 4g, internal diameter is 2cm, wet
On the polyamide column B of method dress post, eluted respectively with water 100ml and 50% ethanol 150ml, collect 50% ethanol eluate, be evaporated, it is residual
Slag adds methanol 1ml to make dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, the μ l of need testing solution 2~10 are drawn and right
According to the μ l of medicinal material solution 2, put respectively on same silica gel g thin-layer plate, the ethyl acetate: ethanol: water=11~13 in terms of by volume
: 1~3: 0.5~2 be solvent, deploys, takes out, dry, sprays with 10% ethanol solution of sulfuric acid of 5% paradime thylaminobenzaldehyde,
Hot blast is blown to spot development clearly, puts and is inspected under 365nm ultraviolet lamps, in test sample chromatogram, corresponding to control medicinal material chromatogram
Position on, show same color fluorescence spot；
（4）The discriminating of radix glycyrrhizae：This product 10g, plus 20~40ml of ethanol are taken, is stirred to dissolve, ultrasonically treated 10~30 minutes, filter
Cross, filtrate is evaporated, residue adds water 10~30 ml dissolvings, adjusts pH value to 2~3 with hydrochloric acid, 1~3 is extracted with ethyl acetate shaking
Secondary, 10~30 ml, combined ethyl acetate extract solution, are evaporated every time, and residue adds methanol 1ml to make dissolving, is used as need testing solution；
Another extracting liquorice control medicinal material 1g, adds water appropriate, decocts 30 minutes, let cool, filter, filtrate is concentrated into 20ml, with it is water saturated just
Butanol shaking is extracted 2 times, and each 20ml merges n-butanol extracting liquid, with the water washing 2 times of n-butanol saturation, each 10ml,
N-butanol liquid is evaporated, and residue adds methanol 1ml to make dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, test sample is drawn
Solution and each 5~10 μ l of control medicinal material solution, put on same silica gel g thin-layer plate, the chloroform in terms of by volume respectively:
Methanol: water=38~42: 9~11: 0.5~2 be solvent, deploys, takes out, dry, and is sprayed with 5% vanillin-sulfuric acid solution,
105 DEG C are heated to spot development clearly, in test sample chromatogram, on position corresponding with control medicinal material chromatogram, show same color
Spot；
（5）The assay of tanshinone IIA：It is filler with octadecylsilane chemically bonded silica；Methanol in terms of by volume:
Water=75: 25 be mobile phase；Detection wavelength is 270nm；Number of theoretical plate is calculated by tanshinone IIA peak should be not less than 2000；Precision claims
Tanshinone IIA reference substance 10mg is taken, is put in 50ml brown measuring bottles, plus methanol is to scale, shakes up, then again from the mixed liquor shaken up
Middle precision measures 2ml, puts in 25ml brown measuring bottles, plus methanol is to scale, shakes up, and produces the μ g containing tanshinone IIA 16 in every 1ml
Reference substance solution；This product 2g is taken, is put in conical flask with cover, precision adds methanol 50ml, weighed weight, in power 120W, frequency
Ultrasonically treated 10~30 minutes under conditions of rate 59KHz, let cool, close plug, then weighed weight, less loss weight is supplied with methanol, is shaken
Even, filtration takes subsequent filtrate, produces need testing solution；It is accurate respectively to draw reference substance solution and each 5 μ l of need testing solution, injection
Liquid chromatograph, determines, produces；
（6）The assay of tanshin polyphenolic acid B：It is filler with octadecylsilane chemically bonded silica；Methanol: second in terms of by volume
Nitrile: formic acid：Water=30: be mobile phase at 10: 1: 59；Detection wavelength is 286nm；Number of theoretical plate is calculated by tanshin polyphenolic acid B peak to be not less than
2000；Tanshin polyphenolic acid B reference substance is weighed, plus solution of every 1ml containing 0.14mg is made in 75% methanol, produces reference substance solution；Take this
Product 2g, it is accurately weighed, put in conical flask with cover, precision adds 75% methanol 50ml, weighed weight, in power 120W, frequency
Ultrasonically treated 10~30 minutes under conditions of 59KHz, take out, let cool, then weighed weight, the weight of less loss is supplied with 75% methanol
Amount, shakes up, and filters, takes subsequent filtrate, produce need testing solution；It is accurate respectively to draw reference substance solution and each 10 μ of need testing solution
L, injects liquid chromatograph, determines, produces.
2. the detection method of the red mattress cream of compound according to claim 1, it is characterised in that：
Step（1）Described oriental wormwood and the discriminating of sweet wormwood are：This product 1g, plus methanol 10ml are taken, ultrasonically treated 30 minutes, is filtered,
Filtrate is evaporated, and residue adds methanol 2ml to make dissolving, is used as need testing solution；Take escoparone contrast product, plus methanol that every 1ml is made
Solution containing 0.4mg, is used as escoparone contrast product solution；Take Qinghaosu, plus methanol that every 1ml is made containing 0.2mg's
Solution, is used as Qinghaosu solution；Above-mentioned each 5 μ l of three kinds of solution are drawn, are put respectively on same silica gel g thin-layer plate, with
60~90 DEG C of petroleum ethers: ethyl acetate: acetone=7: be solvent at 4: 2 counted by volume, deploy, and take out, dry, put 365nm
Ultraviolet lamp under inspect, in test sample chromatogram, on the position corresponding with escoparone contrast product chromatogram, show same color
Fluorescence spot；On position corresponding with Qinghaosu chromatogram, do not show the spot of same color；
Step（2）The discriminating of the described red sage root and bark of ash is：This product 2g is taken, add water 10ml, is stirred to dissolve, add diethyl ether 20ml,
Shaking, is placed 1 hour, point takes ether layer, standby, and raffinate adds diethyl ether 10ml again, and shaking is placed 1 hour, point takes ether layer, with
Ether mixing above, is volatilized, residue adds ethyl acetate 1ml to make dissolving, is used as need testing solution；Tanshinone IIA reference substance is taken,
Plus solution of every 1ml containing 0.3mg is made in ethyl acetate, is used as tanshinone IIA reference substance solution；Gentiamarin reference substance is taken, plus
Solution of every 1ml containing 0.2mg is made in ethyl acetate, is used as gentiamarin reference substance solution；Tested according to thin-layered chromatography, in absorption
Each 4 μ l of three kinds of solution are stated, are put respectively on same silica gel g thin-layer plate, the benzene in terms of by volume：Ethyl acetate=21: 1 is exhibition
Agent is opened, is deployed, takes out, dries, in test sample chromatogram, with tanshinone IIA reference substance chromatogram relevant position, showing same color
Spot；With on gentiamarin reference substance chromatogram relevant position, not showing the spot of same color；
Step（3）The discriminating of described radix bupleuri is：This product 3g is taken, add water 30ml, is stirred to dissolve, centrifuge, take supernatant, be added in
100~200 mesh, 8g, internal diameter is on 2.5~3cm, the polyamide column A of wet method dress post, respectively with water, 20% ethanol and 50% ethanol
Each 100ml elutions, collect 50% ethanol eluate, are evaporated, residue adds methanol 1ml to make dissolving, is used as need testing solution；Separately take bavin
Hu control medicinal material 1g, is added water to cook 1.5 hours, and filtration, filtrate is concentrated into 10ml, is added in 100~200 mesh, 4g, internal diameter is 2cm,
On wet method dress post polyamide column B, eluted respectively with water 100ml and 50% ethanol 150ml, collect 50% ethanol eluate, be evaporated, it is residual
Slag adds methanol 1ml to make dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, the μ l of need testing solution 2~10 are drawn and right
According to the μ l of medicinal material solution 2, put respectively on same silica gel g thin-layer plate, the ethyl acetate: ethanol: water=12: 2: 1 in terms of by volume
For solvent, deploy, take out, dry, spray with 10% ethanol solution of sulfuric acid of 5% paradime thylaminobenzaldehyde, hot blast is blown to spot
Colour developing is clear, puts and is inspected under 365nm ultraviolet lamps, in test sample chromatogram, on position corresponding with control medicinal material chromatogram, shows phase
With the fluorescence spot of color；
Step（4）The discriminating of described radix glycyrrhizae is：This product 10g, plus ethanol 30ml are taken, is stirred to dissolve, ultrasonically treated 20 minutes,
Filtration, filtrate is evaporated, and residue adds water 20 ml dissolvings, and pH value is adjusted to 2~3 with hydrochloric acid, extracts 2 times with ethyl acetate shaking, every time
20 ml, combined ethyl acetate extract solution, are evaporated, and residue adds the ml of methanol 1 to make dissolving, is used as need testing solution；Another extracting liquorice pair
According to medicinal material 1g, add water appropriate, decoct 30 minutes, let cool, filter, filtrate is concentrated into 20ml, carried with the shaking of water saturated n-butanol
Take 2 times, each 20ml, merge n-butanol extracting liquid, with the water washing 2 times of n-butanol saturation, each 10ml, n-butanol liquid steams
Dry, residue adds methanol 1ml to make dissolving, is used as control medicinal material solution；According to thin-layered chromatography experiment, need testing solution and control are drawn
Each 5~10 μ l of medicinal material solution, put on same silica gel g thin-layer plate, the chloroform: methanol: water=40 in terms of by volume respectively
: be solvent at 10: 1, is deployed, and takes out, dries, and sprays with 5% vanillin-sulfuric acid solution, is heated to spot development at 105 DEG C clear
It is clear, in test sample chromatogram, on position corresponding with control medicinal material chromatogram, show the spot of same color；
Step（5）The assay of described tanshinone IIA is：It is filler with octadecylsilane chemically bonded silica；With by weight
Measure the methanol: water=75 than meter: 25 be mobile phase；Detection wavelength is 270nm；Number of theoretical plate is calculated by tanshinone IIA peak should not be low
In 2000；Precision weighs tanshinone IIA reference substance 10mg, puts in 50ml brown measuring bottles, plus methanol is to scale, shakes up, Ran Houzai
Precision measures 2ml from the mixed liquor shaken up, puts in 25ml brown measuring bottles, plus methanol is to scale, shakes up, and produces and contains in every 1ml
The μ g of tanshinone IIA 16 reference substance solution；This product 2g is taken, is put in conical flask with cover, precision addition methanol 50ml, weighed weight,
Ultrasonically treated 20 minutes under conditions of power 120W, frequency 59KHz, let cool, close plug, then weighed weight, supplied and subtracted with methanol
Weight loss, shakes up, filtration, takes subsequent filtrate, produces need testing solution；Accurate absorption reference substance solution and need testing solution are each respectively
5 μ l, inject liquid chromatograph, determine, produce；
Step（6）The assay of described tanshin polyphenolic acid B is：It is filler with octadecylsilane chemically bonded silica；With by weight
Than the methanol: acetonitrile: formic acid: water=30 of meter: be mobile phase at 10: 1: 59；Detection wavelength is 286nm；Number of theoretical plate presses tanshin polyphenolic acid B
Peak, which is calculated, should be not less than 2000；Tanshin polyphenolic acid B reference substance is weighed, plus solution of every 1ml containing 0.14mg is made in 75% methanol, produces
Reference substance solution；This product 2g is taken, it is accurately weighed, put in conical flask with cover, precision adds 75% methanol 50ml, weighed weight,
Ultrasonically treated 20 minutes under conditions of power 120W, frequency 59KHz, take out, let cool, then weighed weight, supplied with 75% methanol
The weight of less loss, shakes up, filtration, takes subsequent filtrate, produces need testing solution；Accurate absorption reference substance solution and test sample are molten respectively
Each 10 μ l of liquid, inject liquid chromatograph, determine, produce.
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