CN107179380A - Naozhenning TLC Identification - Google Patents
Naozhenning TLC Identification Download PDFInfo
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- CN107179380A CN107179380A CN201710474448.0A CN201710474448A CN107179380A CN 107179380 A CN107179380 A CN 107179380A CN 201710474448 A CN201710474448 A CN 201710474448A CN 107179380 A CN107179380 A CN 107179380A
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention discloses a kind of Naozhenning TLC Identification, belong to the Quality Control Technology field of Chinese medical extract and preparation.The invention provides the discriminating of dried orange peel in Naozhenning, test sample preparation method is easy, quick, accurate, environmental protection, target component good separating effect, makes printing operation be easy to progress, and point sample amount is controllable, it ensure that contained crude drug can quantify in test sample, separating degree is high, and sampling amount is small, and point sample amount is controllable, sensitivity is high, disturbs less.The invention provides the discriminating of moutan bark in Naozhenning, using methanol ultrasonic extraction, water-saturated n-butanol extraction, neutral alumina mixes sample, you can obtain test sample, and point sample amount is few, saves the time, and environmental pollution is few, operates more fast and convenient.The method specificity that the present invention is provided is strong, and reappearance, durability are good, the discriminating available for corresponding flavour of a drug in Naozhenning extract and preparation.
Description
Technical field
The present invention relates to a kind of Naozhenning TLC Identification, belong to the quality control skill of Chinese medical extract and preparation
Art field.
Background technology
Naozhenning be according to Patent No. 931007410 patent disclosed in prescription, be made be used for prevent and treat brain
Concussion, post-traumatic brain syndrome, the extract and Chinese patent drug of neurosurgery postoperative convalescence, can be made into granule, capsule, piece
All suitable dosage forms such as agent, soft capsule.In order to control the quality of Naozhenning preparation, we have carried out thin layer to it and have differentiated research.
Naozhenning particle is the particle being made according to patent prescription.There is sugared particle to be embodied at present《The People's Republic of China (PRC)
The Sanitation Ministry medicine standard Traditional Chinese medicine historical preparation（17th）》Page 230（Standard No.：WS3-B-3312-98）.Carried in the standard
Go out：Its prescription is：Radix Angelicae Sinensis 200g, glutinous rehmannia 200g, moutan bark 150g, Ligusticum wallichii 150g, earthworm 150g, red sage root 375g, Poria cocos 250g,
Dried orange peel 250g, caulis bambusae in taenian 250g, Semen Ziziphi Spinosae (parched) 250g, seed of Oriental arborvitae 250g；Preparation method is：The above ten simply, Radix Angelicae Sinensis, moutan bark, Ligusticum wallichii,
Dried orange peel distillating extracting oil, the another device of the aqueous solution after distillation is collected, seven tastes such as the dregs of a decoction and remaining red sage root add water to cook it is secondary, often
Secondary 1.5 hours, collecting decoction, filtration, filtrate merged with the above-mentioned aqueous solution, is concentrated into relative density 1.35~1.38 (55 DEG C)
Clear cream, adds appropriate sucrose and dextrin, with alcohol granulation, dries, adds above-mentioned volatile oil, mixes, 1000g is made and produces；
Specification is：Per packed 10g.Its discrimination method has 2, is specially：
（1）This product 10g is taken, finely ground, plus methanol 30mL, refluxing extraction 1 hour, filtration, filtrate is evaporated, and residue adds 0.3% hydrochloric acid molten
Liquid 20mL makes dissolving, puts and is flowed back 1 hour on boiling water bath, let cool, is extracted with benzene 20mL, discards benzene liquid, and sour water is full with 5g sodium chloride
With, shaking, with ethyl acetate extraction 2 times, each 20mL divides and takes ethyl acetate liquid, is evaporated, it is molten that residue adds absolute ethyl alcohol 1mL to make
Solution, is used as need testing solution.Separately take protocatechualdehyde reference substance, plus ethanol that solution of every 1mL containing 1mg is made, it is molten as reference substance
Liquid.According to thin-layered chromatography（Annex VIB）Experiment, draws each 5 μ L of above two solution, puts respectively on same silica gel g thin-layer plate,
With chloroform-acetone-formic acid（8:1:1）For solvent, deploy, take out, dry, put after being smoked in ammonia, the aobvious Huang of protocatechualdehyde spot
Color, then spray with 2% ferric trichloride ethanol solution.In test sample chromatogram, on position corresponding with reference substance chromatogram, show identical face
The spot of color.
（2）This product 10g is taken, finely ground, plus methanol 30mL, refluxing extraction 1 hour, filtration, filtrate is evaporated, and residue adds water
50mL, heating makes dissolving, lets cool, with extracted by ether 3 times, and each 15mL merges ether solution, extracted 3 times with 2% sodium carbonate liquor,
Each 15mL, point takes alkali lye, with salt acid for adjusting pH value to 2~3, is extracted with benzene 15mL, discards benzene liquid, then with extracted by ether 3 times,
Each 15mL, merges ether solution, volatilizes, residue adds methanol 1mL to make dissolving, is used as need testing solution.Forulic acid reference substance separately is taken,
Plus solution of every 1mL containing 1mg is made in methanol, is used as reference substance solution.According to thin-layered chromatography（Annex VIB）Experiment, draws above-mentioned
Each 5 μ L of two kinds of solution, put on same silica gel g thin-layer plate, with benzene-glacial acetic acid-methanol respectively（30:1:1）For solvent, exhibition
Open, take out, dry, put ultraviolet lamp（365nm）Under inspect.In test sample chromatogram, on position corresponding with reference substance chromatogram,
The fluorescence spot of aobvious same color.
Current sugar-free particle, the standard that performs is what State Food and Drug Administration issued：WS3- B-3312-98-1,
Every bag containing crude drug amount with《Drug Standard of Ministry of Public Health of the Peoples Republic of China Traditional Chinese medicine historical preparation（17th）》Page 230 is included
" Naozhenning particle（Standard No.：WS3-B-3312-98）Identical, discrimination method is also identical.
Naozhenning extract is the refined thing that is made of prescription disclosed in the patent according to Patent No. 931007410.
Naozhenning preparation be according to Patent No. 931007410 patent disclosed in prescription all suitable agent are made
The preparation of type.
By the further investigation of Shanxi Zhendong Anter Biology Pharmaceutical Co., Ltd., existing discrimination method is improved simultaneously
Increase, i.e., carried out discriminatory analysis to the red sage root, Radix Angelicae Sinensis, Ligusticum wallichii, spina date seed, dried orange peel five kinds of Chinese medicine, and disclosure simultaneously authorizes patent（Patent
Number：ZL 201310046420.9）, but in patent dried orange peel discrimination method complex steps, environmental pollution is more.
In current disclosed other medicinal materials delivered, preparation in the discrimination method of dried orange peel, test sample preparation method is cumbersome,
Need to be heated to reflux（Chinese patentCN200710072856.X）, or ethyl acetate extraction（Chinese patentCN201410222870.3）, or water-saturated n-butanol extraction（Chinese patentCN200910020802.8）, run counter to quality inspection
The principle of quick detection in survey, and using the above-mentioned aurantiamarin discrimination method with other disclosed medicinal materials or preparation to differentiate brain shake
It is higher containing sugar in equal not up to ideal states, or prepared test sample, it is impossible to control point sample amount, Huo Zhetong during peaceful preparation
Cross after solvent expansion, disturb more.In heretofore described Naozhenning dried orange peel thin-layer identification method, in test sample preparation process
By adding ethanol, unexpected effect is produced, above mentioned problem is solved, makes point sample amount controllable, and target component separation effect
It is really good, clear spot after colour developing.
Naozhenning is to treat cerebral concussion, unique medicine of post-traumatic brain syndrome, unique, determines its prescription
Uniqueness, flavour of a drug and ratio have difference, discriminating of the existing discrimination method on Paeoniflorin to moutan bark in Naozhenning
Have no and benefit, the still discrimination method without the root bark of tree peony in Naozhenning discrimination method.According to other existing published Chinese medicines or
The discrimination method of Paeoniflorin in person's preparation, prepared by test sample need heating organic solvent backflow（Chinese patentCN200610201379.8、CN200810117041.3）, or water-saturated n-butanol and the water successively extraction of n-butanol saturation
（Chinese patentCN200810046038.7）；Solvent after extraction is eluted by neutral alumina column（Chinese patentCN200810018308.3）, step is various, and not environmentally.
The content of the invention
The present invention is intended to provide a kind of Naozhenning TLC Identification, this method specificity is strong, reappearance, durability
It is good, differentiate available for the corresponding flavour of a drug in the Chinese medical extract and preparation.
Naozhenning is by the dried rhizome of rehmannia, Radix Angelicae Sinensis, Ligusticum wallichii, earthworm, the root bark of tree peony, the red sage root, Poria cocos, dried orange peel, caulis bambusae in taenian, Semen Ziziphi Spinosae (parched), cedar seed
The a herb of benevolence ten is constituted.The prescription of Naozhenning preparation is：
7-12 parts of dried rhizome of rehmannia 1-12 parts of Radix Angelicae Sinensis 5-10 parts of earthworm of 5-10 parts of Ligusticum wallichii
5-10 parts of root bark of tree peony 15-20 parts of red sage root 10-15 parts of dried orange peel of 10-15 parts of Poria cocos
6-11 parts of spina date seeds of caulis bambusae in taenian（Fry）6-11 parts of 6-11 parts of the seed of Oriental arborvitaes.
According to the prescription, the Naozhenning extract or preparation of the 11 taste Chinese medicines containing more than is made.
The invention provides above-mentioned Naozhenning TLC Identification, including any of following discrimination method or
Two kinds：
（1）The discriminating of dried orange peel in Naozhenning：
0.2 ~ 20g of Naozhenning extract or preparation, plus 5 ~ 100mL of methanol are taken, ultrasonically treated 10~60 minutes, filtration, filtrate was steamed
Dry, residue adds 1 ~ 10mL of methanol to make dissolving, adds 3 ~ 30mL absolute ethyl alcohols, filters, filtrate is evaporated, plus the mL systems of methanol 0.1 ~ 20
Into need testing solution；
Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution；
Test sample and each 0.5 ~ 20 μ L of reference substance solution is drawn to put respectively on same silica gel g thin-layer plate, using volume ratio as 71 ~
130:10~25:The mixed solvent of 5 ~ 19 acetate-methanol-water is that solvent is expanded at 4 ~ 6cm, takes out, dries, then
Using volume ratio as 9 ~ 30:5~20: 5~20:The upper solution of 0.5 ~ 2 toluene-ethyl acetate-formic acid-water is solvent, expansion
To 8 ~ 12cm, take out, dry；Spray is dried using mass fraction as 0.1 ~ 5% alchlor ethanol solution, puts and inspected under uviol lamp；
In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color；
（2）The discriminating of moutan bark in Naozhenning：
0.2 ~ 20g of Naozhenning extract or preparation, plus 5 ~ 100mL of methanol are taken, ultrasonically treated 10~60 minutes, filtration, filtrate was steamed
Dry, residue adds water 5~40mL dissolvings, is extracted 1~5 time with water-saturated n-butanol, 5~40mL, merges n-butanol liquid every time, steams
Dry, residue adds 0.1~dissolving of 20mL methanol, adds 0.1~5g of neutral alumina, shakes up, stand, take supernatant, is made for examination
Product solution；
Separately take Paeoniflorin reference substance, plus methanol that the reference substance solution containing 0.2 ~ 2mg/mL is made；
Draw need testing solution and each 0.5 ~ 20 μ L of reference substance solution, put respectively on same silica gel g thin-layer plate, using volume ratio as
19~59:5~20:The mixed solution of 0.5~5 chloroform-methanol-water is solvent, is deployed, and takes out, dries, and spray is with quality point
The vanillin-sulfuric acid solution of number 1 ~ 10% develops the color, and 60 ~ 120 DEG C are heated to clear spot；
In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color.
The vanillin-sulfuric acid solution compound method is：Vanillic aldehyde is taken, the ethanol solution of sulfuric acid that volume fraction is 10% is added to
In, the vanillin-sulfuric acid solution that mass fraction containing vanillic aldehyde is 1 ~ 10% is made.
The Naozhenning includes one in extract or tablet, capsule, particle, soft capsule, dispersible tablet, oral liquid formulations
Kind.
Further, described Naozhenning TLC Identification, including any of following discrimination method or
Two kinds：
（1）The discrimination method of dried orange peel is in Naozhenning：
0.4 ~ 10g of Naozhenning extract or preparation, plus 10 ~ 60mL of methanol are taken, ultrasonically treated 15~50 minutes, filtration, filtrate was steamed
Dry, residue adds 1 ~ 5mL of methanol to make dissolving, adds 3 ~ 15mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus 0.2 ~ 10mL methanol is made
Need testing solution；
Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution；
Draw test sample and each 1 ~ 10 μ L of reference substance solution are put on same silica gel g thin-layer plate, using volume ratio as 80 ~ 120 respectively:
12~22:The mixed solvent of 7 ~ 17 acetate-methanol-water is that solvent is expanded at 4 ~ 6cm, takes out, dries, then with body
Product is than being 12 ~ 28:8~18: 8~18:The upper solution of 0.6 ~ 2 toluene-ethyl acetate-formic acid-water is solvent, is expanded to
8.5 ~ 11.5cm, takes out, dries；Spray is dried with the alchlor ethanol solution of mass fraction 0.5 ~ 4.5%, is put and is examined under uviol lamp
Depending on；
In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color；
（2）The discriminating of moutan bark in Naozhenning：
0.4 ~ 10g of Naozhenning extract or preparation, plus 10 ~ 60mL of methanol are taken, ultrasonically treated 15~50 minutes, filtration, filtrate was steamed
Dry, residue adds water 8~30mL dissolvings, is extracted 1~4 time with water-saturated n-butanol, 10~30mL, merges n-butanol liquid every time, steams
Dry, residue adds 0.2 ~ dissolving of 10mL methanol, adds 0.2~3g of neutral alumina, shakes up, stands, take supernatant, test sample is made
Solution；
Separately take Paeoniflorin reference substance, plus methanol that the reference substance solution containing 0.4 ~ 1.8mg/mL is made；
Need testing solution and each 1 ~ 10 μ L of reference substance solution are drawn, is put respectively on same silica gel g thin-layer plate, using volume ratio as 20
~55:8~18:The mixed solution of 0.6~4 chloroform-methanol-water is solvent, is deployed, and takes out, dries, and is sprayed with mass fraction
2 ~ 8% vanillin-sulfuric acid solution develop the color, and 65 ~ 115 DEG C are heated to clear spot；
In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color.
Further, described Naozhenning TLC Identification, including any of following discrimination method or
Two kinds of person：
（1）The discrimination method of dried orange peel is in Naozhenning：
0.8 ~ 8g of Naozhenning extract or preparation, plus 20 ~ 50mL of methanol are taken, ultrasonically treated 20 ~ 40 minutes, filtration, filtrate was evaporated,
Residue adds 1.5 ~ 3.5mL of methanol to make dissolving, adds 4.5 ~ 10.5mL absolute ethyl alcohols, filters, filtrate is evaporated, plus 0.4 ~ 4mL of methanol
Dissolving, is made need testing solution；
Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution；
Draw test sample and each 2 ~ 5 μ L of reference substance solution are put on same silica gel g thin-layer plate, using volume ratio as 90 ~ 110 respectively:
15~20:10 ~ 15 acetate-methanol-water mixed solvent is solvent, is expanded at 4.5 ~ 5.5cm, takes out, dries, then
Using volume ratio as 15 ~ 25:6~15:6~15:The upper solution of 0.7 ~ 1.5 toluene-ethyl acetate-formic acid-water is solvent, expansion
To 9 ~ 11cm, take out, dry；Spray is dried with 1 ~ 4% alchlor ethanol solution, is put and inspected under 365nm uviol lamps；Test sample
In chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color.
（2）The discrimination method of moutan bark is in Naozhenning：
0.8 ~ 8g of Naozhenning extract or preparation, plus 20 ~ 50mL of methanol are taken, ultrasonically treated 20 ~ 40 minutes, filtration, filtrate was evaporated,
Residue adds water 10 ~ 25mL dissolvings, is extracted 2 ~ 4 times with water-saturated n-butanol, 18 ~ 25mL every time, merges n-butanol liquid, be evaporated, residual
Slag adds 0.4 ~ dissolving of 8mL methanol, adds 0.2 ~ 2g of neutral alumina, shakes up, stands, take supernatant, need testing solution is made；
It is another to take Paeoniflorin reference substance, plus methanol that the solution that every 1mL contains 0.5 ~ 1.5mg is made, it is used as reference substance solution；
Need testing solution and each 2 ~ 8 μ L of reference substance solution are drawn, is put respectively on same silica gel g thin-layer plate, using volume ratio as 30 ~
50:7~15:0.8 ~ 3 chloroform-methanol-water mixed solvent is solvent, is deployed, and takes out, dries, and spray is fragrant with mass fraction 3 ~ 7%
Oxalaldehyde sulfuric acid solution develops the color, and 80 ~ 110 DEG C are heated to clear spot；In test sample chromatogram, in position corresponding with reference substance chromatogram
On, show the spot of same color.
Further, described Naozhenning TLC Identification, including any of following discrimination method or
Two kinds of person：
（1）The discrimination method of dried orange peel is in Naozhenning：
3 ~ 5g of Naozhenning extract or preparation, plus 30 ~ 40mL of methanol are taken, ultrasonically treated 25 ~ 30 minutes, is filtered, filtrate is evaporated, residual
Slag adds 2 ~ 3mL of methanol to make dissolving, adds 6 ~ 9mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus 1 ~ 2mL of methanol dissolvings, is made for examination
Product solution；
Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution；
Draw test sample and each 2 ~ 4 μ L of reference substance solution are put on same silica gel g thin-layer plate, using volume ratio as 95 ~ 105 respectively:
15~20:10 ~ 15 acetate-methanol-water mixed solvent is solvent, is expanded at 4.5 ~ 5.5cm, takes out, dries, then
Using volume ratio as 18 ~ 22:8~12:8~12:The upper solution of 0.8 ~ 1.2 toluene-ethyl acetate-formic acid-water is solvent, expansion
To 10cm, take out, dry；Spray is dried with 2.5 ~ 3.5% alchlor ethanol solutions, is put and inspected under 365nm uviol lamps；Test sample
In chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color.
（2）The discrimination method of moutan bark is in Naozhenning：
3 ~ 5g of Naozhenning extract or preparation is taken, plus 30 ~ 40mL of methanol, ultrasonically treated 25 ~ 30 minutes, filtration, filtrate was evaporated,
Residue adds water 15 ~ 20mL dissolvings, is extracted 2 ~ 3 times with water-saturated n-butanol, 15 ~ 20mL every time, merges n-butanol liquid, be evaporated, residual
Slag adds 1 ~ dissolving of 2mL methanol, adds 0.3 ~ 1.5g of neutral alumina, shakes up, stands, take supernatant, need testing solution is made；
It is another to take Paeoniflorin reference substance, plus methanol that the solution that every 1mL contains 1 ~ 1.2mg is made, it is used as reference substance solution；
Need testing solution and each 2 ~ 4 μ L of reference substance solution are drawn, is put respectively on same silica gel g thin-layer plate, using volume ratio as 35 ~
45:9~12:1 ~ 2 chloroform-methanol-water mixed solvent is solvent, is deployed, and takes out, dries, and is sprayed with the vanillic aldehyde of mass fraction 4 ~ 6%
Sulfuric acid solution develops the color, and 90 ~ 105 DEG C are heated to clear spot；In test sample chromatogram, on position corresponding with reference substance chromatogram,
The spot of aobvious same color.
Dried orange peel discrimination method in the Naozhenning that the present invention is provided, test sample preparation method is easy, quick, accurate, environmental protection, saves
Gone to be heated to reflux, the step such as organic solvent extraction.By adding ethanol in test sample preparation process, unexpected effect is produced
Really, printing operation is made to be easy to progress, and point sample amount is controllable, it is ensured that contained crude drug can quantify in test sample, and separating degree is high, sampling
Amount is small, and point sample amount is few, and sensitivity is high, clear spot after colour developing, disturbs less.
The moutan bark discrimination method that the present invention is provided, easy, quick, accurate, environmental protection, eliminate be heated to reflux, Duo Zhongyou
The steps such as machine solvent extraction, macroporous resin column elution, use methanol ultrasonic extraction, water-saturated n-butanol extraction, neutral alumina
Sample is mixed, test sample can be obtained by simple operations, and point sample amount is few, saves the time, environmental pollution is few, discriminating is more accelerated
It is fast accurate.
Brief description of the drawings
Fig. 1 is moutan bark thin layer discriminating chromatogram in embodiment 9；
Fig. 2 is dried orange peel thin layer discriminating chromatogram in embodiment 9.
Embodiment
The present invention is further illustrated below by embodiment, but is not limited to following examples.
Embodiment 1：The thin-layer identification method of Naozhenning particle
According to standard WS3- B-3312-98 prepares the Naozhenning particle containing sucrose, and the discrimination method provided with the present invention is reflected
Not.
Prescription：Radix Angelicae Sinensis 200g, glutinous rehmannia 200g, moutan bark 150g, Ligusticum wallichii 150g, earthworm 150g, the red sage root 375g, Fu
Siberian cocklebur 250g, dried orange peel 250g, caulis bambusae in taenian 250g, Semen Ziziphi Spinosae (parched) 250g, seed of Oriental arborvitae 250g
Preparation method：The above ten simply, receive by Radix Angelicae Sinensis, moutan bark, Ligusticum wallichii, dried orange peel distillating extracting oil, the another device of the aqueous solution after distillation
Collection；Seven tastes such as the dregs of a decoction and remaining red sage root add water to cook it is secondary, 1.5 hours every time, collecting decoction, filtration, filtrate with it is above-mentioned water-soluble
Liquid merges, and is concentrated into the clear cream that relative density is 1.35~1.38 (55 DEG C), adds sucrose and dextrin, pelletize, dry, in addition
Volatile oil is stated, mixes, 1000g is made；Producing Naozhenning has sugared particle.
Differentiate：（1）Naozhenning particle 5g is taken, it is finely ground, plus methanol 30mL, ultrasonically treated 30 minutes, filtration, filtrate was evaporated,
Residue adds methanol 2mL to make dissolving, adds 6mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus methanol 1mL dissolvings, molten as test sample
Liquid；Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution.Draw test sample and reference substance solution is each
2 μ L are put on same silica gel g thin-layer plate, with acetate-methanol-water respectively（Volume ratio is 100:17:13）Mixed solvent is
Solvent, is expanded at 5cm, takes out, dries, then with toluene-ethyl acetate-formic acid-water（Volume ratio is 20:10:10:1）'s
Upper solution is solvent, is expanded to 10cm, takes out, dries.Spray is dried, put with the alchlor ethanol solution of mass fraction 3%
Inspected under 365nm uviol lamps.In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color.
（2）Naozhenning particle 5g is taken, it is finely ground, plus methanol 30mL, ultrasonically treated 30 minutes, filtration, filtrate was evaporated, and residue adds
Water 20mL dissolves, and is extracted 2 times with water-saturated n-butanol, each 20mL, merges n-butanol liquid, is evaporated, residue adds 1mL methanol molten
Solution, adds neutral alumina 0.5g, shakes up, and stands, takes supernatant, be used as need testing solution.Separately take Paeoniflorin reference substance, plus first
Solution of every 1mL containing 1mg is made in alcohol, is used as reference substance solution.Need testing solution and each 2 μ L of reference substance solution are drawn, respectively point
In on same silica gel g thin-layer plate, with chloroform-methanol-water（Volume ratio is 40:10:1）Mixed solvent is solvent, expansion, is taken
Go out, dry, the colour developing of the vanillin-sulfuric acid solution of mass fraction 5%, 105 DEG C are heated to clear spot.In test sample chromatogram, with it is right
According on the corresponding position of product chromatogram, show the spot of same color.
Glutinous rehmannia described in embodiment, is same medicinal material with the foregoing dried rhizome of rehmannia.
Embodiment 2：Naozhenning particle（Without sucrose）Thin-layer identification method
According to standard WS3- B-3312-98-1 prepares sucrose free Naozhenning particle, and the discrimination method provided with the present invention enters
Row differentiates.
Prescription：Radix Angelicae Sinensis 200g, glutinous rehmannia 200g, moutan bark 150g, Ligusticum wallichii 150g, earthworm 150g, the red sage root 375g, Fu
Siberian cocklebur 250g, dried orange peel 250g, caulis bambusae in taenian 250g, Semen Ziziphi Spinosae (parched) 250g, seed of Oriental arborvitae 250g
The above ten simply, collect by Radix Angelicae Sinensis, moutan bark, Ligusticum wallichii, dried orange peel distillating extracting oil, the another device of the aqueous solution after distillation；Medicine
Seven tastes such as slag and remaining red sage root add water to cook secondary, and 1.5 hours every time, collecting decoction, filtration, filtrate was closed with the above-mentioned aqueous solution
And, the clear cream that relative density is 1.35~1.38 (55 DEG C) is concentrated into, dextrin is added, pelletized, is dried, above-mentioned volatile oil is added,
Mix, 600g is made, Naozhenning sugar-free particle is produced.
Differentiate：（1）Naozhenning sugar-free particle 0.6g is taken, it is finely ground, plus methanol 60mL, ultrasonically treated 60 minutes, filtration, filtrate
It is evaporated, residue adds methanol 5mL to make dissolving, adds 30mL absolute ethyl alcohols, filtering, filtrate is evaporated, plus methanol 0.5mL dissolvings, as
Need testing solution；Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution.Draw test sample and control
Each 20 μ L of product solution are put on same silica gel g thin-layer plate, with acetate-methanol-water respectively（Volume ratio is 130:25:19）It is mixed
Bonding solvent is solvent, is expanded at 6cm, takes out, dries, then with toluene-ethyl acetate-formic acid-water（Volume ratio is 30:5:
5:3）Upper solution be solvent, be expanded to 12cm, take out, dry.Spray is dried in the air with the alchlor ethanol solution of mass fraction 5%
It is dry, put and inspected under 365nm uviol lamps.In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color
Point.
（2）Naozhenning sugar-free particle 0.6g is taken, it is finely ground, plus methanol 60mL, ultrasonically treated 60 minutes, filtration, filtrate was evaporated,
Residue add water 50mL dissolving, with water-saturated n-butanol extract 5 times, each 40mL, merge n-butanol liquid, be evaporated, residue adds 0.5mL
Methanol dissolves, and adds neutral alumina 0.2g, shakes up, and stands, takes supernatant, be used as need testing solution.Separately Paeoniflorin is taken to compare
Solution of every 1mL containing 0.2mg is made in product, plus methanol, is used as reference substance solution.Draw need testing solution and reference substance solution each 20
μ L, put on same silica gel g thin-layer plate, with chloroform-methanol-water respectively（Volume ratio is 59:20:5）Mixed solvent is solvent,
Expansion, takes out, dries, the colour developing of the vanillin-sulfuric acid solution of mass fraction 10%, and 120 DEG C are heated to clear spot.Test sample chromatogram
In, on position corresponding with reference substance chromatogram, show the spot of same color.
Embodiment 3：The thin-layer identification method of Naozhenning capsule
Prescription：Radix Angelicae Sinensis 220g, glutinous rehmannia 210g, moutan bark 160g, Ligusticum wallichii 140g, earthworm 130g, red sage root 330g, Poria cocos
260g, dried orange peel 260g, caulis bambusae in taenian 260g, Semen Ziziphi Spinosae (parched) 160g, seed of Oriental arborvitae 160g
Preparation method：Simply, Radix Angelicae Sinensis, moutan bark, Ligusticum wallichii, dried orange peel distillating extracting oil add other medicinal material water extractions, water to the above ten
After extraction, with 80% ethanol alcohol precipitation 2 times, paste is condensed into, starch is added and mixes, granulation, dry whole grain, add gained and wave
Hair oil, stirs, and Naozhenning capsule 200 is made, and every capsule fills 0.4g.
Differentiate：（1）Naozhenning capsule 's content 0.77g is taken, it is finely ground, plus methanol 10mL, ultrasonically treated 10 minutes, filtration,
Filtrate is evaporated, and residue adds methanol 0.5mL to make dissolving, adds 3mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus methanol 5mL dissolvings, makees
For need testing solution；Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution.Draw test sample and right
Put respectively on same silica gel g thin-layer plate, with acetate-methanol-water according to each 0.5 μ L of product solution（Volume ratio is 71:10:5）
Mixed solvent is solvent, is expanded at 4cm, takes out, dries, then with toluene-ethyl acetate-formic acid-water（Volume ratio is 9:5:
5:0.5）Upper solution be solvent, be expanded to 8cm, take out, dry.Spray is molten with the alchlor ethanol of mass fraction 0.1%
Liquid, dries, and puts and is inspected under 365nm uviol lamps.In test sample chromatogram, on position corresponding with reference substance chromatogram, show identical face
The spot of color..
（2）Naozhenning capsule 's content 0.75g is taken, it is finely ground, plus methanol 10mL, ultrasonically treated 10 minutes, filtration, filtrate was steamed
Dry, residue adds water 10mL dissolvings, is extracted 1 time with water-saturated n-butanol, each 10mL, merges n-butanol liquid, be evaporated, residue adds
10mL methanol dissolves, and adds neutral alumina 5g, shakes up, and stands, takes supernatant, be used as need testing solution.Separately take Paeoniflorin pair
Solution of every 1mL containing 1mg is made according to product, plus methanol, reference substance solution is used as.Draw need testing solution and reference substance solution is each
0.5 μ L, put on same silica gel g thin-layer plate, with chloroform-methanol-water respectively（Volume ratio is 19:5:0.5）Mixed solvent is exhibition
Agent is opened, is deployed, takes out, dries, the colour developing of the vanillin-sulfuric acid solution of mass fraction 1%, 60 DEG C are heated to clear spot.Test sample color
In spectrum, on position corresponding with reference substance chromatogram, show the spot of same color.
Embodiment 4：The thin-layer identification method of Naozhenning tablet
Prescription：Radix Angelicae Sinensis 175g, glutinous rehmannia 175g, moutan bark 150g, Ligusticum wallichii 75g, earthworm 75g, red sage root 300g, Poria cocos
250g, dried orange peel 250g, caulis bambusae in taenian 150g, Semen Ziziphi Spinosae (parched) 150g, seed of Oriental arborvitae 150g
Preparation method：Simply, Radix Angelicae Sinensis, moutan bark, Ligusticum wallichii, dried orange peel distillating extracting oil add other medicinal material water extractions, water to the above ten
After extraction, with 70% ethanol alcohol precipitation 2 times, be condensed into paste, add starch, sodium carboxymethyl starch mix, granulation, drying it is whole
Grain, adds gained volatile oil, stirs, tabletting.Naozhenning piece 200, every weight 0.5g is made.
Differentiate：（1）Naozhenning tablet 0.8g is taken, it is finely ground, plus methanol 30mL, ultrasonically treated 30 minutes, filtration, filtrate was steamed
Dry, residue adds methanol 2mL to make dissolving, adds 5mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus methanol 2mL dissolvings, is used as test sample
Solution；Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution.Draw test sample and reference substance solution
Each 2 μ L are put on same silica gel g thin-layer plate, with acetate-methanol-water respectively（Volume ratio is 98:20:15）Mixed solvent is
Solvent, is expanded at 5cm, takes out, dries, then with toluene-ethyl acetate-formic acid-water（Volume ratio is 22:15:15:2）'s
Upper solution is solvent, is expanded to 10cm, takes out, dries.Spray is dried, put with the alchlor ethanol solution of mass fraction 3%
Inspected under 365nm uviol lamps.In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color.
（2）Naozhenning tablet 0.8g is taken, it is finely ground, plus methanol 30mL, ultrasonically treated 30 minutes, filtration, filtrate was evaporated, residue
The 25mL that adds water dissolves, and is extracted 3 times with water-saturated n-butanol, each 15mL, merges n-butanol liquid, is evaporated, residue adds 5mL methanol molten
Solution, adds neutral alumina 2g, shakes up, and stands, takes supernatant, be used as need testing solution.Separately take Paeoniflorin reference substance, plus methanol
Solution of every 1mL containing 1.5mg is made, reference substance solution is used as.Need testing solution and each 5 μ L of reference substance solution are drawn, respectively point
In on same silica gel g thin-layer plate, with chloroform-methanol-water（Volume ratio is 35:9:1）Mixed solvent is solvent, is deployed, and is taken out,
Dry, the colour developing of the vanillin-sulfuric acid solution of mass fraction 3%, 100 DEG C are heated to clear spot.In test sample chromatogram, with reference substance
On the corresponding position of chromatogram, show the spot of same color.
Embodiment 5：The thin-layer identification method of Naozhenning soft capsule
Prescription：Radix Angelicae Sinensis 300g, glutinous rehmannia 300g, moutan bark 275g, Ligusticum wallichii 250g, earthworm 250g, red sage root 500g, Poria cocos
275g, dried orange peel 375g, caulis bambusae in taenian 275g, Semen Ziziphi Spinosae (parched) 275g, seed of Oriental arborvitae 275g
Preparation method：Simply, Radix Angelicae Sinensis, moutan bark, Ligusticum wallichii, dried orange peel distillating extracting oil add other medicinal material water extractions, water to the above ten
After extraction, with 80% ethanol alcohol precipitation 2 times, paste is condensed into, Naozhenning soft capsule 300 is made with dropping preparation method, per tablet
0.5g。
Differentiate：（1）Naozhenning soft capsule content 1g, plus methanol 25mL are taken, ultrasonically treated 20 minutes, filtration, filtrate was steamed
Dry, residue adds methanol 3mL to make dissolving, adds 10mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus methanol 1mL dissolvings, as trying
Product solution；Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution.Draw test sample and reference substance is molten
Each 1 μ L of liquid are put on same silica gel g thin-layer plate, with acetate-methanol-water respectively（Volume ratio is 120:20:17）Mixing is molten
Agent is solvent, is expanded at 5cm, takes out, dries, then with toluene-ethyl acetate-formic acid-water（Volume ratio is 21:11:10:
2）Upper solution be solvent, be expanded to 10cm, take out, dry.Spray is dried in the air with the alchlor ethanol solution of mass fraction 3.5%
It is dry, put and inspected under 365nm uviol lamps.In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color
Point.
（2）Naozhenning soft capsule content 0.7g, plus methanol 35mL are taken, ultrasonically treated 40 minutes, filtration, filtrate was evaporated,
Residue add water 25mL dissolving, with water-saturated n-butanol extract 3 times, each 20mL, merge n-butanol liquid, be evaporated, residue adds 2mL first
Alcohol dissolves, and adds neutral alumina 1g, shakes up, and stands, takes supernatant, be used as need testing solution.Paeoniflorin reference substance separately is taken, plus
Solution of every 1mL containing 0.8mg is made in methanol, is used as reference substance solution.Need testing solution and each 5 μ L of reference substance solution are drawn, point
Other point is on same silica gel g thin-layer plate, with chloroform-methanol-water（Volume ratio is 37:9:1）Mixed solvent is solvent, expansion,
Take out, dry, the colour developing of the vanillin-sulfuric acid solution of mass fraction 6%, 90 DEG C are heated to clear spot.In test sample chromatogram, with it is right
According on the corresponding position of product chromatogram, show the spot of same color
Embodiment 6：The thin-layer identification method of Naozhenning dispersible tablet
Prescription：Radix Angelicae Sinensis 250g, glutinous rehmannia 220g, moutan bark 200g, Ligusticum wallichii 100g, earthworm 100g, red sage root 400g, Poria cocos
330g, dried orange peel 300g, caulis bambusae in taenian 200g, Semen Ziziphi Spinosae (parched) 190g, seed of Oriental arborvitae 220g
Preparation method：Simply, Radix Angelicae Sinensis, moutan bark, Ligusticum wallichii, dried orange peel distillating extracting oil add other medicinal material water extractions, water to the above ten
After extraction, with 70% ethanol alcohol precipitation 2 times, paste is condensed into, appropriate sodium carboxymethyl starch, microcrystalline cellulose, micro mist is added
Silica gel, with alcohol granulation, drying, whole grain, add gained volatile oil, after mixing, tabletting.Naozhenning dispersible tablet 200 is made,
Every weight 0.65g.
Differentiate：（1）Naozhenning dispersible tablet 0.2g is taken, it is finely ground, plus methanol 20mL, ultrasonically treated 30 minutes, filtration, filtrate was steamed
Dry, residue adds methanol 1mL to make dissolving, adds 3mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus methanol 1mL dissolvings, is used as test sample
Solution；Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution.Draw test sample and reference substance solution
Each 1 μ L are put on same silica gel g thin-layer plate, with acetate-methanol-water respectively（Volume ratio is 112:19:15）Mixed solvent
For solvent, it is expanded at 5cm, takes out, dry, then with toluene-ethyl acetate-formic acid-water（Volume ratio is 21:13:13:
0.5）Upper solution be solvent, be expanded to 10cm, take out, dry.Spray is dried in the air with the alchlor ethanol solution of mass fraction 4%
It is dry, put and inspected under 365nm uviol lamps.In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color
Point.
（2）Naozhenning dispersible tablet 0.3g is taken, it is finely ground, plus methanol 20mL, ultrasonically treated 25 minutes, filtration, filtrate is evaporated, residual
Slag add water 10mL dissolving, with water-saturated n-butanol extract 2 times, each 15mL, merge n-butanol liquid, be evaporated, residue adds 1mL methanol
Dissolving, adds neutral alumina 0.3g, shakes up, and stands, takes supernatant, be used as need testing solution.Paeoniflorin reference substance separately is taken, plus
Solution of every 1mL containing 0.5mg is made in methanol, is used as reference substance solution.Need testing solution and each 2 μ L of reference substance solution are drawn, point
Other point is on same silica gel g thin-layer plate, with chloroform-methanol-water（Volume ratio is 50:19:3）Mixed solvent is solvent, expansion,
Take out, dry, the colour developing of the vanillin-sulfuric acid solution of mass fraction 3%, 112 DEG C are heated to clear spot.In test sample chromatogram, with
On the corresponding position of reference substance chromatogram, show the spot of same color.
Embodiment 7：The thin-layer identification method of Naozhenning oral liquid
Prescription：Radix Angelicae Sinensis 210g, glutinous rehmannia 210g, moutan bark 140g, Ligusticum wallichii 140g, earthworm 140g, red sage root 350g, Poria cocos
260g, dried orange peel 280g, caulis bambusae in taenian 240g, Semen Ziziphi Spinosae (parched) 230g, seed of Oriental arborvitae 230g
Preparation method：Simply, Radix Angelicae Sinensis, moutan bark, Ligusticum wallichii, dried orange peel distillating extracting oil add other medicinal material water extractions, water to the above ten
After extraction, with 70% ethanol alcohol precipitation 2 times, honey, polyoxyethylene sorbitan monoleate, sodium benzoate are added, with citron acid for adjusting pH value to 4.5
~5.5, stir evenly, filter, obtain Naozhenning oral liquid 1000mL, 5mL/ branch.
Differentiate：（1）Naozhenning oral liquid 5mL, plus methanol 30mL are taken, ultrasonically treated 10 minutes, is filtered, filtrate is evaporated, residual
Slag adds methanol 2mL to make dissolving, adds 8mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus methanol 1mL dissolvings, is used as need testing solution；
Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution.Draw test sample and each 2 μ L of reference substance solution
Put respectively on same silica gel g thin-layer plate, with acetate-methanol-water（Volume ratio is 98:15:13）Mixed solvent is expansion
Agent, is expanded at 4cm, takes out, dries, then with toluene-ethyl acetate-formic acid-water（Volume ratio is 18:11:11:1.5）It is upper
Layer solution is solvent, is expanded to 9cm, takes out, dries.Spray is dried, put with the alchlor ethanol solution of mass fraction 3%
Inspected under 365nm uviol lamps.In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color.
（2）Naozhenning oral liquid 5mL, plus methanol 30mL are taken, ultrasonically treated 10 minutes, filtration, filtrate was evaporated, and residue adds water
10mL dissolves, and is extracted 2 times with water-saturated n-butanol, each 15mL, merges n-butanol liquid, is evaporated, and residue adds 1mL methanol to dissolve,
Neutral alumina 0.5g is added, is shaken up, is stood, is taken supernatant, be used as need testing solution.Separately take Paeoniflorin reference substance, plus methanol
Solution of every 1mL containing 1mg is made, reference substance solution is used as.Draw need testing solution and each 2 μ L of reference substance solution, put respectively in
On same silica gel g thin-layer plate, with chloroform-methanol-water（Volume ratio is 38:9:1）Mixed solvent is solvent, is deployed, and takes out, dries in the air
It is dry, the colour developing of the vanillin-sulfuric acid solution of mass fraction 5%, 85 DEG C are heated to clear spot.In test sample chromatogram, with reference substance color
Compose on corresponding position, show the spot of same color.
Embodiment 8：The thin-layer identification method of children's Naozhenning particle
Prescription：Radix Angelicae Sinensis 200g, glutinous rehmannia 200g, moutan bark 150g, Ligusticum wallichii 150g, earthworm 150g, red sage root 375g, Poria cocos
250g, dried orange peel 250g, caulis bambusae in taenian 250g, Semen Ziziphi Spinosae (parched) 250g, seed of Oriental arborvitae 250g
Preparation method：The above ten simply, receive by Radix Angelicae Sinensis, moutan bark, Ligusticum wallichii, dried orange peel distillating extracting oil, the another device of the aqueous solution after distillation
Collection；Seven tastes such as the dregs of a decoction and remaining red sage root add water to cook it is secondary, 1.5 hours every time, collecting decoction, filtration, filtrate with it is above-mentioned water-soluble
Liquid merges, and is concentrated into the clear cream of relative density 1.35~1.38 (55 DEG C), adds sucrose and dextrin, pelletizes, and dries, and adds above-mentioned
Volatile oil, mixes, 1000g is made, produces.
Differentiate：（1）Children's Naozhenning particle 5g is taken, it is finely ground, plus methanol 20mL, ultrasonically treated 30 minutes, filtration, filtrate was steamed
Dry, residue adds methanol 1mL to make dissolving, adds 10mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus methanol 1mL dissolvings, as trying
Product solution；Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution.Draw test sample and reference substance is molten
Each 2 μ L of liquid are put on same silica gel g thin-layer plate, with acetate-methanol-water respectively（Volume ratio is 100:17:13）Mixing is molten
Agent is solvent, is expanded at 5cm, takes out, dries, then with toluene-ethyl acetate-formic acid-water（Volume ratio is 20:10:10:
1）Upper solution be solvent, be expanded to 10cm, take out, dry.Spray is dried in the air with the alchlor ethanol solution of mass fraction 3%
It is dry, put and inspected under 365nm uviol lamps.In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color
Point.
（2）Children's Naozhenning particle 5g is taken, it is finely ground, plus methanol 20mL, ultrasonically treated 30 minutes, filtration, filtrate is evaporated, residual
Slag add water 20mL dissolving, with water-saturated n-butanol extract 2 times, each 20mL, merge n-butanol liquid, be evaporated, residue adds 1mL methanol
Dissolving, adds neutral alumina 0.5g, shakes up, and stands, takes supernatant, be used as need testing solution.Paeoniflorin reference substance separately is taken, plus
Solution of every 1mL containing 1mg is made in methanol, is used as reference substance solution.Need testing solution and each 2 μ L of reference substance solution are drawn, respectively
Point is on same silica gel g thin-layer plate, with chloroform-methanol-water（Volume ratio is 40:10:1）Mixed solvent is solvent, expansion, is taken
Go out, dry, the colour developing of the vanillin-sulfuric acid solution of mass fraction 5%, 105 DEG C are heated to clear spot.In test sample chromatogram, with it is right
According on the corresponding position of product chromatogram, show the spot of same color.
Embodiment 9：Naozhenning particle prepared by Example 1 carries out Method validation.
（1）The Method validation that moutan bark differentiates：
The preparation of need testing solution：Naozhenning particle 5g is taken, it is finely ground, plus methanol 30mL, ultrasonically treated 30 minutes, filtration, filtrate
Be evaporated, residue add water 20mL dissolving, with water-saturated n-butanol extract 2 times, each 20mL, merge n-butanol liquid, be evaporated, residue adds
1mL methanol dissolves, and adds neutral alumina 0.5g, shakes up, and stands, takes supernatant, be used as need testing solution.
The preparation of reference substance solution：Solution of every 1mL containing 1mg is made in Paeoniflorin reference substance, plus methanol, molten as reference substance
Liquid.
The preparation of negative control solution：The negative control sample for lacking moutan bark is prepared in Naozhenning particle prescription ratio and technique
Product, take negative control sample 5g, and by the preparation method of need testing solution, negative control solution is made.
Determination method：Need testing solution and each 2 μ L of reference substance solution are drawn, is put respectively on same silica gel g thin-layer plate, with chlorine
Imitation-carbinol-water（Volume ratio is 40:10:1）Mixed solvent is solvent, is deployed, and takes out, dries, and is sprayed with the vanilla of mass fraction 5%
Aldehyde sulfuric acid solution develops the color, and 105 DEG C are heated to clear spot.
Result of the test：Three batches of test samples（Three batches of need testing solutions are done referred to herein as according to the particle of embodiment 1, quite
In three groups of parallel laboratory tests）In chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color；And negative control
In chromatogram, on position corresponding with reference substance chromatogram, not display dot.
Conclusion：This method specificity is strong, reproducible, the discriminating available for moutan bark in Naozhenning particle.
As shown in figure 1, sample 1 is Paeoniflorin reference substance, sample 2 ~ 4 is three test samples, and sample 5 is that moutan bark feminine gender is molten
Liquid.
（2）The Method validation that dried orange peel differentiates：
The preparation of need testing solution：Naozhenning particle 5g, plus methanol 30mL are taken, ultrasonically treated 30 minutes, filtration, filtrate was evaporated,
Residue adds methanol 2mL to make dissolving, adds 6mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus methanol 1mL dissolvings, molten as test sample
Liquid.
The preparation of reference substance solution：Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution.
The preparation of negative control solution：The negative control sample for lacking dried orange peel is prepared in Naozhenning particle prescription ratio and technique
Product, take negative control sample 5g, and by the preparation method of need testing solution, negative control solution is made.
Determination method：Draw test sample and each 2 μ L of reference substance solution are put on same silica gel g thin-layer plate, with acetic acid second respectively
Ester-methanol-water（Volume ratio is 100:17:13）Mixed solvent is solvent, is expanded at 5cm, take out, dry, then with toluene-
Acetic ether-methanoic acid-water（20:10:10:1）Upper solution be solvent, be expanded to 10cm, take out, dry.Spray is with quality
The alchlor ethanol solution of fraction 3%, dries, and puts and is inspected under 365nm uviol lamps.
Result of the test：Three batches of test samples（Ibid）In chromatogram, on position corresponding with reference substance chromatogram, show same color
Spot；And in negative control chromatogram, on position corresponding with reference substance chromatogram, not display dot.
Conclusion：This method specificity is strong, reproducible, the discriminating available for dried orange peel in Naozhenning particle.
As shown in Fig. 2 sample 1 ~ 3 is three batches of test samples, sample 4 is aurantiamarin reference substance, and sample 5 is that dried orange peel feminine gender is molten
Liquid.
Claims (8)
1. Naozhenning TLC Identification, it is characterised in that including any of following discrimination method or two kinds：
（1）The discriminating of dried orange peel in Naozhenning：
0.2 ~ 20g of Naozhenning extract or preparation, plus 5 ~ 100mL of methanol are taken, ultrasonically treated 10~60 minutes, filtration, filtrate was steamed
Dry, residue adds 1 ~ 10mL of methanol to make dissolving, adds 3 ~ 30mL absolute ethyl alcohols, filters, filtrate is evaporated, plus the mL systems of methanol 0.1 ~ 20
Into need testing solution；
Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution；
Test sample and each 0.5 ~ 20 μ L of reference substance solution is drawn to put respectively on same silica gel g thin-layer plate, using volume ratio as 71 ~
130:10~25:The mixed solvent of 5 ~ 19 acetate-methanol-water is that solvent is expanded at 4 ~ 6cm, takes out, dries, then
Using volume ratio as 9 ~ 30:5~20: 5~20:The upper solution of 0.5 ~ 3 toluene-ethyl acetate-formic acid-water is solvent, expansion
To 8 ~ 12cm, take out, dry；Spray is dried using mass fraction as 0.1 ~ 5% alchlor ethanol solution, puts and inspected under uviol lamp；
In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color；
（2）The discriminating of moutan bark in Naozhenning：
0.2 ~ 20g of Naozhenning extract or preparation, plus 5 ~ 100mL of methanol are taken, ultrasonically treated 10~60 minutes, filtration, filtrate was steamed
Dry, residue adds water 5~40mL dissolvings, is extracted 1~5 time with water-saturated n-butanol, 5~40mL, merges n-butanol liquid every time, steams
Dry, residue adds 0.1~dissolving of 20mL methanol, adds 0.1~5g of neutral alumina, shakes up, stand, take supernatant, is made for examination
Product solution；
Separately take Paeoniflorin reference substance, plus methanol that the reference substance solution containing 0.2 ~ 2mg/mL is made；
Draw need testing solution and each 0.5 ~ 20 μ L of reference substance solution, put respectively on same silica gel g thin-layer plate, using volume ratio as
19~59:5~20:The mixed solution of 0.5~5 chloroform-methanol-water is solvent, is deployed, and takes out, dries, and spray is with quality point
The vanillin-sulfuric acid solution of number 1 ~ 10% develops the color, and 60 ~ 120 DEG C are heated to clear spot；
In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color.
2. Naozhenning TLC Identification according to claim 1, it is characterised in that：The vanillin-sulfuric acid solution
Compound method is：Vanillic aldehyde is taken, is added in the ethanol solution of sulfuric acid that volume fraction is 10%, it is 1 that mass fraction containing vanillic aldehyde, which is made,
~ 10% vanillin-sulfuric acid solution.
3. Naozhenning TLC Identification according to claim 1, it is characterised in that：The Naozhenning is by giving birth to
Ground, Radix Angelicae Sinensis, Ligusticum wallichii, earthworm, the root bark of tree peony, the red sage root, Poria cocos, dried orange peel, caulis bambusae in taenian, Semen Ziziphi Spinosae (parched), a herb of the seed of Oriental arborvitae ten composition.
4. Naozhenning TLC Identification according to claim 3, it is characterised in that：The ten a herbs composition
Weight proportion be：
7-12 parts of dried rhizome of rehmannia 1-12 parts of Radix Angelicae Sinensis 5-10 parts of earthworm of 5-10 parts of Ligusticum wallichii
5-10 parts of root bark of tree peony 15-20 parts of red sage root 10-15 parts of dried orange peel of 10-15 parts of Poria cocos
6-11 parts of caulis bambusae in taenian 6-11 parts of the seed of Oriental arborvitae of 6-11 parts of Semen Ziziphi Spinosae (parched).
5. Naozhenning TLC Identification according to claim 1, it is characterised in that：The Naozhenning includes extracting
One kind in thing or tablet, capsule, particle, soft capsule, dispersible tablet, oral liquid formulations.
6. the Naozhenning TLC Identification according to any one of claim 1 ~ 5, it is characterised in that：Including following mirror
Any of other method or two kinds：
（1）The discriminating of dried orange peel in Naozhenning：
0.4 ~ 10g of Naozhenning extract or preparation, plus 10 ~ 60mL of methanol are taken, ultrasonically treated 15~50 minutes, filtration, filtrate was steamed
Dry, residue adds 1 ~ 5mL of methanol to make dissolving, adds 3 ~ 15mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus 0.2 ~ 10mL methanol is made
Need testing solution；
Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution；
Draw test sample and each 1 ~ 10 μ L of reference substance solution are put on same silica gel g thin-layer plate, using volume ratio as 80 ~ 120 respectively:
12~22:The mixed solvent of 7 ~ 17 acetate-methanol-water is that solvent is expanded at 4 ~ 6cm, takes out, dries, then with body
Product is than being 12 ~ 28:8~18:8~18:The upper solution of 0.6 ~ 2 toluene-ethyl acetate-formic acid-water is solvent, is expanded to
8.5 ~ 11.5cm, takes out, dries；Spray is dried with the alchlor ethanol solution of mass fraction 0.5 ~ 4.5%, is put and is examined under uviol lamp
Depending on；
In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color；
（2）The discriminating of moutan bark in Naozhenning：
0.4 ~ 10g of Naozhenning extract or preparation, plus 10 ~ 60mL of methanol are taken, ultrasonically treated 15~50 minutes, filtration, filtrate was steamed
Dry, residue adds water 8~30mL dissolvings, is extracted 1~4 time with water-saturated n-butanol, 10~30mL, merges n-butanol liquid every time, steams
Dry, residue adds 0.2 ~ dissolving of 10mL methanol, adds 0.2~3g of neutral alumina, shakes up, stands, take supernatant, test sample is made
Solution；
Separately take Paeoniflorin reference substance, plus methanol that the reference substance solution containing 0.4 ~ 1.8mg/mL is made；
Need testing solution and each 1 ~ 10 μ L of reference substance solution are drawn, is put respectively on same silica gel g thin-layer plate, using volume ratio as 20
~55:8~18:The mixed solution of 0.6 ~ 4 chloroform-methanol-water be solvent, deploy, take out, dry, spray with mass fraction 2 ~
8% vanillin-sulfuric acid solution develops the color, and 65 ~ 115 DEG C are heated to clear spot；
In test sample chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color.
7. Naozhenning TLC Identification according to claim 6, it is characterised in that：Including in following discrimination method
It is any or two kinds：
（1）0.8 ~ 8g of Naozhenning extract or preparation, plus 20 ~ 50mL of methanol are taken, ultrasonically treated 20 ~ 40 minutes, filtration, filtrate was steamed
Dry, residue adds 1.5 ~ 3.5mL of methanol to make dissolving, adds 4.5 ~ 10.5mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus methanol 0.4 ~
4mL dissolves, and need testing solution is made；
Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution；
Draw test sample and each 2 ~ 5 μ L of reference substance solution are put on same silica gel g thin-layer plate, using volume ratio as 95 ~ 110 respectively:
15~20:10 ~ 15 acetate-methanol-water mixed solvent is solvent, is expanded at 4.5 ~ 5.5cm, takes out, dries, then
Using volume ratio as 15 ~ 25:6~15:6~15:The upper solution of 0.7 ~ 1.5 toluene-ethyl acetate-formic acid-water is solvent, expansion
To 9 ~ 11cm, take out, dry；Spray is dried with 1 ~ 4% alchlor ethanol solution, is put and inspected under 365nm uviol lamps；Test sample color
In spectrum, on position corresponding with reference substance chromatogram, show the spot of same color；
（2）0.8 ~ 8g of Naozhenning extract or preparation, plus 20 ~ 50mL of methanol are taken, ultrasonically treated 20 ~ 40 minutes, filtration, filtrate was steamed
Dry, residue adds water 10 ~ 25mL dissolvings, is extracted 2 ~ 4 times with water-saturated n-butanol, 18 ~ 25mL, merges n-butanol liquid, be evaporated every time,
Residue adds 0.4 ~ dissolving of 8mL methanol, adds 0.2 ~ 2g of neutral alumina, shakes up, stands, take supernatant, need testing solution is made；
It is another to take Paeoniflorin reference substance, plus methanol that the solution that every 1mL contains 0.5 ~ 1.5mg is made, it is used as reference substance solution；
Need testing solution and each 2 ~ 8 μ L of reference substance solution are drawn, is put respectively on same silica gel g thin-layer plate, using volume ratio as 30 ~
50:7~15:0.8 ~ 3 chloroform-methanol-water mixed solvent is solvent, is deployed, and takes out, dries, and spray is fragrant with mass fraction 3 ~ 7%
Oxalaldehyde sulfuric acid solution develops the color, and 80 ~ 110 DEG C are heated to clear spot；In test sample chromatogram, in position corresponding with reference substance chromatogram
On, show the spot of same color.
8. Naozhenning TLC Identification according to claim 7, it is characterised in that：Including in following discrimination method
It is any or two kinds：
（1）3 ~ 5g of Naozhenning extract or preparation, plus 30 ~ 40mL of methanol are taken, ultrasonically treated 25 ~ 30 minutes, filtration, filtrate was steamed
Dry, residue adds 2 ~ 3mL of methanol to make dissolving, adds 6 ~ 9mL absolute ethyl alcohols, and filtering, filtrate is evaporated, plus 1 ~ 2mL of methanol dissolvings, is made
Need testing solution；
Separately take aurantiamarin reference substance plus methanol that saturated solution is made, be used as reference substance solution；
Draw test sample and each 2 ~ 4 μ L of reference substance solution are put on same silica gel g thin-layer plate, using volume ratio as 95 ~ 105 respectively:
15~20:10 ~ 15 acetate-methanol-water mixed solvent is solvent, is expanded at 4.5 ~ 5.5cm, takes out, dries, then
Using volume ratio as 18 ~ 22:8~12:8~12:The upper solution of 0.8 ~ 1.2 toluene-ethyl acetate-formic acid-water is solvent, expansion
To 10cm, take out, dry；Spray is dried with 2.5 ~ 3.5% alchlor ethanol solutions, is put and inspected under 365nm uviol lamps；Test sample
In chromatogram, on position corresponding with reference substance chromatogram, show the spot of same color；
（2）3 ~ 5g of Naozhenning extract or preparation is taken, plus 30 ~ 40mL of methanol, ultrasonically treated 25 ~ 30 minutes, filtration, filtrate was steamed
Dry, residue adds water 15 ~ 20mL dissolvings, is extracted 2 ~ 3 times with water-saturated n-butanol, 15 ~ 20mL, merges n-butanol liquid, be evaporated every time,
Residue adds 1 ~ dissolving of 2mL methanol, adds 0.3 ~ 1.5g of neutral alumina, shakes up, stands, take supernatant, need testing solution is made；
It is another to take Paeoniflorin reference substance, plus methanol that the solution that every 1mL contains 1 ~ 1.2mg is made, it is used as reference substance solution；
Need testing solution and each 2 ~ 4 μ L of reference substance solution are drawn, is put respectively on same silica gel g thin-layer plate, using volume ratio as 35 ~
45:9~12:1 ~ 2 chloroform-methanol-water mixed solvent is solvent, is deployed, and takes out, dries, and is sprayed with the vanillic aldehyde of mass fraction 4 ~ 6%
Sulfuric acid solution develops the color, and 90 ~ 105 DEG C are heated to clear spot；In test sample chromatogram, on position corresponding with reference substance chromatogram,
The spot of aobvious same color.
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Application publication date: 20170919 |