CN106153814A - The discrimination method of Radix Salviae Miltiorrhizae in ostealgia medicated wine - Google Patents
The discrimination method of Radix Salviae Miltiorrhizae in ostealgia medicated wine Download PDFInfo
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- CN106153814A CN106153814A CN201610752416.8A CN201610752416A CN106153814A CN 106153814 A CN106153814 A CN 106153814A CN 201610752416 A CN201610752416 A CN 201610752416A CN 106153814 A CN106153814 A CN 106153814A
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Abstract
The invention discloses the discrimination method of Radix Salviae Miltiorrhizae in a kind of ostealgia medicated wine, be with tanshinone IIA reference substance for comparison, with ethyl phenylacetate=18～25 1～2 thin layer chromatography as developing solvent.The discrimination method of Radix Salviae Miltiorrhizae in the ostealgia medicated wine that the present invention provides, simple to operate, controllability is strong, can differentiate Components in Salvia miltiorrhiza in ostealgia medicated wine efficiently and accurately, the inventive method separating degree is high, highly sensitive, precision, good stability, favorable reproducibility, and negative control is noiseless, and different manufacturers lamellae and temperature, damp condition do not affect.Discrimination method of the present invention can carry out effective discriminating accurate, comprehensive, scientifical to Components in Salvia miltiorrhiza in ostealgia medicated wine, improves quality inspection standard, further ensure that the stable, safely, effectively of product quality.
Description
Technical field
The present invention relates to the discrimination method of Radix Salviae Miltiorrhizae in ostealgia medicated wine, particularly belong to Chinese medicine detection technique field.
Background technology
Ostealgia medicated wine be by Radix Aconiti Kusnezoffii Preparata, Herba Taxilli, Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae), Radix Clematidis, Rhizoma Polygoni Cuspidati, Caulis Trachelospermi, Rhizoma Smilacis Chinensis, Rhizoma Atractylodis (parched with bran),
Pine Nodular Branch, Radix Polygoni Multiflori Preparata, Caulis Sargentodoxae, Radix Salviae Miltiorrhizae, Ramulus Sambuci Williamsii, Herba Lycopodii, Fructus Chaenomelis, Rhizoma Chuanxiong, Radix Achyranthis Bidentatae, Herba Ephedrae, Cortex Periplocae, Flos Carthami, Radix Dipsaci
Make with Rhizoma Zingiberis.Ostealgia medicated wine has dispelling wind and relieving pain, relaxing muscles and tendons and activating QI and blood in the collateral effect, efficacy enhanced by wine, and wine helps medicine prestige, can quickly get through more
Resistance, unimpeded blood vessel, meridians, dissolve the blood stasis on cartilage, split adhesion organization, just as dissolving ice cube, gradually decompose calcification
The hypertrophy bony spur of deformation.In the quality standard of existing ostealgia medicated wine, its content only has medicated wine routine examination project, it is difficult to effectively
Ground controls drug quality.In ostealgia medicated wine side, Radix Salviae Miltiorrhizae is main component, differentiates it, beneficially the quality control of ostealgia medicated wine
System.Therefore, study and a kind of differentiate the method for Radix Salviae Miltiorrhizae in ostealgia medicated wine, it appears be particularly necessary.
Summary of the invention
For solving the deficiencies in the prior art, it is an object of the invention to provide the discriminating side of Radix Salviae Miltiorrhizae in a kind of ostealgia medicated wine
Method, it is possible to accurately differentiate Radix Salviae Miltiorrhizae in ostealgia medicated wine, simple to operate, separating degree is high.
Ostealgia medicated wine in the present invention, be by Radix Aconiti Kusnezoffii Preparata 50g, Herba Taxilli 50g, Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) 50g, Caulis Trachelospermi 25g, Radix Salviae Miltiorrhizae 25g,
Pine Nodular Branch 37.5g, Fructus Chaenomelis 25g, Herba Ephedrae 12.5g, Radix Clematidis 25g, Rhizoma Polygoni Cuspidati 37.5g, Rhizoma Smilacis Chinensis 25g, Rhizoma Zingiberis 6.25g, Radix Polygoni Multiflori Preparata
25g, Caulis Sargentodoxae 37.5g, Ramulus Sambuci Williamsii 50g, Herba Lycopodii 12.5g, Rhizoma Chuanxiong 12.5g, Radix Achyranthis Bidentatae 50g, Cortex Periplocae 50g, Flos Carthami 12.5g,
Radix Dipsaci 50g and Rhizoma Atractylodis (parched with bran) 12.5g makes.Above 22 tastes, are ground into coarse powder, according under fluid extract and extractum item
Percolation (" Chinese Pharmacopoeia " version general rule 0189 in 2015), make solvent by wine sugar liquid and (take Saccharum Sinensis Roxb. 430g, be dissolved in Chinese liquor
In 4300g), after impregnating 48 hours, with the speed slowly percolation of per kilogram crude drug 1～3mL per minute, collect percolate, stand,
Filter, to obtain final product.
In order to realize above-mentioned target, the present invention adopts the following technical scheme that:
The discrimination method of Radix Salviae Miltiorrhizae in ostealgia medicated wine, is with tanshinone IIA reference substance for comparison, with benzene-ethyl acetate=18
～25 1～2 be the thin layer chromatography of developing solvent.
The discrimination method of Radix Salviae Miltiorrhizae in aforementioned ostealgia medicated wine, comprises the following steps:
(1) preparation of need testing solution: take this product and add chloroform extraction, obtains chloroform liquid, is evaporated, and residue adds acetic acid
Ethyl ester makes it dissolve, as need testing solution；
(2) preparation of reference substance solution: take tanshinone IIA reference substance, adds ethyl acetate and makes in every 1mL red containing 2mg
The solution of ginseng ketone II A reference substance, as reference substance solution；
(3) differentiate: according to four the general rule 0502 thin layer chromatography tests of " Chinese Pharmacopoeia " version in 2015, draw reference substance molten
Liquid, need testing solution, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate=18～25 1～2 as developing solvent, and exhibition
Open, take out, dry, in test sample chromatograph, with on reference substance chromatograph relevant position, show the speckle of same color.
Specifically, the discrimination method of Radix Salviae Miltiorrhizae in aforementioned ostealgia medicated wine, comprise the following steps:
(1) preparation of need testing solution: take this product 95～105mL, adds chloroform 15～30mL every time, extracts 2～4
Secondary, merge chloroform liquid, be evaporated, residue adds 0.5～2mL ethyl acetate makes it dissolve, as need testing solution；
(2) preparation of reference substance solution: take tanshinone IIA reference substance, adds ethyl acetate and makes in every 1mL red containing 2mg
The solution of ginseng ketone II A reference substance, as reference substance solution；
(3) differentiate: according to four the general rule 0502 thin layer chromatography tests of " Chinese Pharmacopoeia " version in 2015, draw reference substance solution
8～12 μ L, need testing solution 8～12 μ L, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate=18～25 1～
2 is developing solvent, launches, takes out, dry, and in test sample chromatograph, with on reference substance chromatograph relevant position, shows the speckle of same color
Point.
Preferably, the discrimination method of Radix Salviae Miltiorrhizae in aforementioned ostealgia medicated wine, comprise the following steps:
(1) preparation of need testing solution: take this product 100mL, adds chloroform 15mL every time, extracts 3 times, merges three chloromethanes
Alkane liquid, is evaporated, and residue adds 0.5mL ethyl acetate makes it dissolve, as need testing solution；
(2) preparation of reference substance solution: take tanshinone IIA reference substance, adds ethyl acetate and makes in every 1mL red containing 2mg
The solution of ginseng ketone II A reference substance, as reference substance solution；
(3) differentiate: according to four the general rule 0502 thin layer chromatography tests of " Chinese Pharmacopoeia " version in 2015, draw reference substance solution
10 μ L, need testing solution 10 μ L, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate=19 1 as developing solvent, and exhibition
Open, take out, dry, in test sample chromatograph, with on reference substance chromatograph relevant position, show the speckle of same color.
In order to ensure discrimination method science of the present invention, reasonable, feasible, the discrimination method of the present invention is studied and examined
Examine.
Radix Salviae Miltiorrhizae is dry root and the rhizome of labiate Radix Salviae Miltiorrhizae.Having blood circulation promoting and blood stasis dispelling, inducing menstruation to relieve menalgia, clear away heart-fire relieving restlessness, cool
Effect of blood eliminating carbuncle.For obstruction of qi in the chest and cardialgia, gastral cavity abdomen hypochondriac pain, the accumulation of abdominal mass abdominal mass, pyretic arthralgia pain, dysphoria and insomnia, menoxenia, dysmenorrhea warp
Closing, skin infection swells and ache.Tanshinone IIA is its specificity composition in side, and therefore, this experiment will choose tanshinone IIA reference substance
As comparison, by the tanshinone IIA composition of Radix Salviae Miltiorrhizae in chloroform extraction this product, red rooted salvia is carried out thin layer discriminating, and
Investigate different point sample amount, different manufacturers lamellae, different temperatures, different humidity to the shadow of red rooted salvia thin layer chromatography in this product
Ring.Result of the test shows: with tanshinone IIA reference substance for comparison, with benzene-ethyl acetate=18～25 1～2 as developing solvent
Thin layer chromatography feature is obvious, and specificity is strong, can be as the discrimination method of Radix Salviae Miltiorrhizae in this product.
1, instrument and reagent
MS304s/01 electronic balance (METTLER TOLEDO (prunus mume (sieb.) sieb.et zucc. Teller-torr benefit)), P-1 type thin layer chromatograph developing cylinder
(company limited of Xinyi Instrument Factory, Shanghai), silica gel g thin-layer plate (Qingdao Haiyang chemical industry), 5ul point sample capillary tube (Tianjin Si Lida
Science and Technology Ltd.).
Ostealgia medicated wine (20141201,20141202,20141203, Jiangsu Xin Bang pharmaceutical Co. Ltd), tanshinone IIA pair
According to product (Nat'l Pharmaceutical & Biological Products Control Institute), chloroform (Xilong Chemical Co., Ltd, analytical pure), ethyl acetate
(Xilong Chemical Co., Ltd, analytical pure), benzene (Xilong Chemical Co., Ltd, analytical pure).
2, the investigation of discrimination method
2.1, the preparation method of need testing solution
In prior art, when Radix Salviae Miltiorrhizae differentiates, need testing solution obtains by adding EtOH Sonicate process.With prior art
Comparing, use chloroform as Extraction solvent in the present invention, extraction time is short, and extraction effect is good.Use two kinds of methods respectively
Preparing need testing solution, under equal conditions differentiate, the one group of separating degree using prior art is poor, and uses the present invention
One group of separating degree of method is high, favorable reproducibility.
2.2, the investigation of optimal point sample amount
Need testing solution and reference substance solution 4,6,8,10,12,16 μ L in difference aspiration step (1) and step (2), respectively
Point, on different silica gel g thin-layer plates, is investigated, and with benzene-ethyl acetate=19 1 as developing solvent, launches, takes out, dry.Its
In, test sample 10 μ L point sample amount best results, when point sample amount is more than 12 μ L, need testing solution position speckle occurs that hangover is existing
As, when point sample amount is less than 8 μ L, need testing solution position speckle is inconspicuous；During reference substance solution point sample amount 8～12 μ L effective,
The especially best results when 10 μ L, therefore selecting need testing solution point sample amount is 8～12 μ L, optimal point sample amount 10 μ L；Reference substance
Solution point sample amount 8～12 μ L, optimal point sample amount 10 μ L.
2.3, the investigation of developing solvent
In prior art, when Radix Salviae Miltiorrhizae differentiates, use chloroform-toluene-ethyl acetate-methyl alcohol-formic acid=6 4814
As developing solvent, and using benzene-ethyl acetate=18～25 1～2 in the present invention is developing solvent, selects two kinds of developing solvents respectively
Under equal conditions, Radix Salviae Miltiorrhizae in ostealgia medicated wine is differentiated.Result shows, uses the developing solvent of the present invention, and separating degree is higher.
3, specificity is investigated
Reference substance solution 10 μ L, Radix Salviae Miltiorrhizae negative control solution in need testing solution 10 μ L, step (2) in aspiration step (1)
10 μ L, are put on same silica gel g thin-layer plate by need testing solution, reference substance solution, negative control solution, respectively with benzene-acetic acid
Ethyl ester=19 1 are developing solvent, launch, and take out, dry.As shown in Figure 1.In test sample chromatograph, with reference substance chromatograph corresponding positions
Put, the speckle of aobvious same color, immaculate on relevant position in negative control chromatograph, show that the specificity of the method is good.
4, ruggedness is investigated
4.1 different manufacturers lamellaes
Reference substance solution 10 μ L in need testing solution 10 μ L, step (2) in aspiration step (1), by need testing solution, comparison
Product solution point sample respectively, changes the lamellae of different manufacturers, with benzene-ethyl acetate=19 1 as developing solvent, launches, takes out, dry in the air
Dry.In test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color, and it is thin to change different manufacturers
After laminate, without significant difference between each lamellae, show that changing different manufacturers lamellae differentiates without impact the thin layer of Radix Salviae Miltiorrhizae.As
Shown in Fig. 2.
4.2 humidity
Reference substance solution 10 μ L in need testing solution 10 μ L, step (2) in aspiration step (1), by need testing solution, comparison
Product solution point sample respectively, converts different humidity condition, with benzene-ethyl acetate=19 1 as developing solvent, launches, takes out, dry.Supply
In test product chromatograph, on position corresponding with reference substance chromatograph, after the speckle of aobvious same color, and conversion different humidity condition,
Without significant difference between each lamellae, show that converting different humidity differentiates without impact the thin layer of Radix Salviae Miltiorrhizae.As shown in Figure 3.
4.3, temperature
Reference substance solution 10 μ L in need testing solution 10 μ L, step (2) in aspiration step (1), by need testing solution, comparison
Product solution point sample respectively, converts condition of different temperatures, with benzene-ethyl acetate=19 1 as developing solvent, launches, takes out, dry.Supply
In test product chromatograph, on position corresponding with reference substance chromatograph, after the speckle of aobvious same color, and conversion condition of different temperatures,
Without significant difference between each lamellae, show that converting different temperatures differentiates without impact the thin layer of Radix Salviae Miltiorrhizae.As shown in Figure 4.
The invention have benefit that: the discrimination method of Radix Salviae Miltiorrhizae in the ostealgia medicated wine that the present invention provides, simple to operate, can
Control property is strong, it is possible to differentiate Components in Salvia miltiorrhiza in ostealgia medicated wine efficiently and accurately, and the inventive method separating degree is high, highly sensitive, accurate
Degree, good stability, favorable reproducibility, and negative control is noiseless, different manufacturers lamellae and temperature, damp condition the most not shadow
Ring.Discrimination method of the present invention can carry out effective discriminating accurate, comprehensive, scientifical to Components in Salvia miltiorrhiza in ostealgia medicated wine, improves
Quality inspection standard, further ensure that the stable, safely, effectively of product quality.
Accompanying drawing explanation
Fig. 1 be the present invention ostealgia medicated wine in the specificity identification color spectrogram of Radix Salviae Miltiorrhizae；
Fig. 2 is the chromatogram that in ostealgia medicated wine, Radix Salviae Miltiorrhizae uses different lamellae when differentiating；
Fig. 3 be in ostealgia medicated wine Radix Salviae Miltiorrhizae differentiate time different humidity under the conditions of chromatogram；
Chromatogram under condition of different temperatures when Fig. 4 is that in ostealgia medicated wine, Radix Salviae Miltiorrhizae differentiates；
The implication of reference in figure: Fig. 1: 1-test sample lot number 20141201,2-test sample lot number 20141202,3-supplies
Test product lot number 20141203,4-negative control, 5-tanshinone IIA reference substance；Fig. 2-Fig. 4: 1-test sample lot number 20141201,2-
Test sample lot number 20141202,3-test sample lot number 20141203,4-tanshinone IIA reference substance；Fig. 2: A-Qingdao Haiyang silica gel G
Lamellae (specification: 100 × 200mm), B-Yantai Chemical Industry Research Inst. silica gel g thin-layer plate (specification: 100 × 200mm)；Figure
3:A-humidity 32%, B-humidity 72%；Fig. 4: A-temperature 25 DEG C, B-temperature 8 DEG C.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is further introduced.
Embodiment 1
The discrimination method of Radix Salviae Miltiorrhizae in ostealgia medicated wine, comprises the following steps:
(1) preparation of need testing solution: take this product 95mL, adds chloroform 20mL every time, extracts 4 times, merges three chloromethanes
Alkane liquid, is evaporated, and residue adds 1mL ethyl acetate makes it dissolve, as need testing solution；
(2) preparation of reference substance solution: take tanshinone IIA reference substance, adds ethyl acetate and makes in every 1mL red containing 2mg
The solution of ginseng ketone II A reference substance, as reference substance solution；
(3) differentiate: according to four the general rule 0502 thin layer chromatography tests of " Chinese Pharmacopoeia " version in 2015, draw reference substance solution
8 μ L, need testing solution 8 μ L, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate=18 1 as developing solvent, launches,
Take out, dry, in test sample chromatograph, with on reference substance chromatograph relevant position, show the speckle of same color.
Embodiment 2
The discrimination method of Radix Salviae Miltiorrhizae in ostealgia medicated wine, comprises the following steps:
(1) preparation of need testing solution: take this product 105mL, adds chloroform 30mL every time, extracts 2 times, merges three chloromethanes
Alkane liquid, is evaporated, and residue adds 2mL ethyl acetate makes it dissolve, as need testing solution；
(2) preparation of reference substance solution: take tanshinone IIA reference substance, adds ethyl acetate and makes in every 1mL red containing 2mg
The solution of ginseng ketone II A reference substance, as reference substance solution；
(3) differentiate: according to four the general rule 0502 thin layer chromatography tests of " Chinese Pharmacopoeia " version in 2015, draw reference substance solution
12 μ L, need testing solution 12 μ L, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate=25 2 as developing solvent, and exhibition
Open, take out, dry, in test sample chromatograph, with on reference substance chromatograph relevant position, show the speckle of same color.
Embodiment 3
The discrimination method of Radix Salviae Miltiorrhizae in ostealgia medicated wine, comprises the following steps:
(1) preparation of need testing solution: take this product 100mL, adds chloroform 15mL every time, extracts 3 times, merges three chloromethanes
Alkane liquid, is evaporated, and residue adds 0.5mL ethyl acetate makes it dissolve, as need testing solution；
(2) preparation of reference substance solution: take tanshinone IIA reference substance, adds ethyl acetate and makes in every 1mL red containing 2mg
The solution of ginseng ketone II A reference substance, as reference substance solution；
(3) differentiate: according to four the general rule 0502 thin layer chromatography tests of " Chinese Pharmacopoeia " version in 2015, draw reference substance solution
10 μ L, need testing solution 10 μ L, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate=19 1 as developing solvent, and exhibition
Open, take out, dry, in test sample chromatograph, with on reference substance chromatograph relevant position, show the speckle of same color.
Embodiment 4
The discrimination method of Radix Salviae Miltiorrhizae in ostealgia medicated wine, comprises the following steps:
(1) preparation of need testing solution: take this product 98mL, adds chloroform 25mL every time, extracts 3 times, merges three chloromethanes
Alkane liquid, is evaporated, and residue adds 1mL ethyl acetate makes it dissolve, as need testing solution；
(2) preparation of reference substance solution: take tanshinone IIA reference substance, adds ethyl acetate and makes in every 1mL red containing 2mg
The solution of ginseng ketone II A reference substance, as reference substance solution；
(3) differentiate: according to four the general rule 0502 thin layer chromatography tests of " Chinese Pharmacopoeia " version in 2015, draw reference substance solution
11 μ L, need testing solution 9 μ L, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate=20 2 as developing solvent, and exhibition
Open, take out, dry, in test sample chromatograph, with on reference substance chromatograph relevant position, show the speckle of same color.
Claims (4)
1. the discrimination method of Radix Salviae Miltiorrhizae in ostealgia medicated wine, it is characterised in that: described discrimination method is to be right with tanshinone IIA reference substance
According to, with benzene-ethyl acetate=18～25 1～2 thin layer chromatography as developing solvent.
The discrimination method of Radix Salviae Miltiorrhizae in ostealgia medicated wine the most according to claim 1, it is characterised in that: concrete discrimination method bag
Include following steps:
(1) preparation of need testing solution: take this product and add chloroform extraction, obtains chloroform liquid, is evaporated, and residue adds ethyl acetate
It is made to dissolve, as need testing solution；
(2) preparation of reference substance solution: take tanshinone IIA reference substance, adds ethyl acetate and makes in every 1mL containing 2mg TANSHINONES
The solution of II A reference substance, as reference substance solution；
(3) differentiate: according to four the general rule 0502 thin layer chromatography tests of " Chinese Pharmacopoeia " version in 2015, draw reference substance solution, confession
Test sample solution, puts respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate=18～25 1～2 as developing solvent, launches, takes
Go out, dry, in test sample chromatograph, with on reference substance chromatograph relevant position, show the speckle of same color.
The discrimination method of Radix Salviae Miltiorrhizae in ostealgia medicated wine the most according to claim 2, it is characterised in that: concrete discrimination method bag
Include following steps:
(1) preparation of need testing solution: take this product 95～105mL, adds chloroform 15～30mL every time, extracts 2～4 times, closes
And chloroform liquid, it being evaporated, residue adds 0.5～2mL ethyl acetate makes it dissolve, as need testing solution；
(2) preparation of reference substance solution: take tanshinone IIA reference substance, adds ethyl acetate and makes in every 1mL containing 2mg TANSHINONES
The solution of II A reference substance, as reference substance solution；
(3) differentiate: according to four the general rule 0502 thin layer chromatography tests of " Chinese Pharmacopoeia " version in 2015, draw reference substance solution 8～
12 μ L, need testing solution 8～12 μ L, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate=18～25 1～2 be
Developing solvent, launches, and takes out, dries, and in test sample chromatograph, with on reference substance chromatograph relevant position, shows the speckle of same color.
4. according to the discrimination method of Radix Salviae Miltiorrhizae in the ostealgia medicated wine described in Claims 2 or 3, it is characterised in that: concrete discriminating side
Method comprises the following steps:
(1) preparation of need testing solution: take this product 100mL, adds chloroform 15mL every time, extracts 3 times, merges chloroform
Liquid, is evaporated, and residue adds 0.5mL ethyl acetate makes it dissolve, as need testing solution；
(2) preparation of reference substance solution: take tanshinone IIA reference substance, adds ethyl acetate and makes in every 1mL containing 2mg TANSHINONES
The solution of II A reference substance, as reference substance solution；
(3) differentiate: according to four the general rule 0502 thin layer chromatography tests of " Chinese Pharmacopoeia " version in 2015, draw reference substance solution 10 μ
L, need testing solution 10 μ L, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate=19 1 as developing solvent, launches,
Take out, dry, in test sample chromatograph, with on reference substance chromatograph relevant position, show the speckle of same color.
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Application publication date: 20161123 |