CN105203657A - Spina date seed reference extract and preparation method and application thereof - Google Patents
Spina date seed reference extract and preparation method and application thereof Download PDFInfo
- Publication number
- CN105203657A CN105203657A CN201510585856.4A CN201510585856A CN105203657A CN 105203657 A CN105203657 A CN 105203657A CN 201510585856 A CN201510585856 A CN 201510585856A CN 105203657 A CN105203657 A CN 105203657A
- Authority
- CN
- China
- Prior art keywords
- date seed
- spina date
- reference extract
- preparation
- column
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to spina date seed reference extract and a preparation method and application thereof and belongs to the technical field of traditional Chinese medicine quality control. The preparation method of the spina date seed reference extract includes the following steps of smashing, degreasing, extracting, macroporous resin purifying, silica gel purifying and acquiring of the spina date seed reference extract finally. Spina date seed medicinal materials are degreased, extracted and purified through a specific method, finally the spina date seed reference extract with specific constituents is acquired, and the reference extract includes jujuboside A, jujuboside B and other saponin constituents and further includes spinosin, 6'''-ferula asafetida spinosin and other flavonoid constituents. The spina date seed reference extract serves as a reference to be used in quality control over a traditional Chinese medicine preparation containing spina date seed medicine flavor and has the advantage that constituent information is comprehensively fed back; when the spina date seed reference extract is used, the complex pretreatment step is not needed, inspection can be directly conducted after dissolution, and the advantages of being easy and convenient to operate and low in detection cost are achieved.
Description
Technical field
The present invention relates to quality control technologies for traditional Chinese medicine field, particularly relate to a kind of spina date seed reference extract and its preparation method and application.
Background technology
Spina date seed, another name jujube nucleus, mountain jujube kernel.It is the dry mature seed of Rhamnaceae plant wild jujube.To gather ripening fruits in the time of the year when autumn changes into winter, removing pulp and nucleocapsid, collect seed, dry and form.It is born in endroit, roadside, mainly originates in the ground such as Henan, Hebei, Shaanxi, Liaoning, Shanxi, Shandong, Yunnan.Gather during fruit maturation in autumn, remove pulp and stone, get seed, life is used or micro-stir-fry is used.
Spina date seed, as a kind of Chinese crude drug, records in Chinese Pharmacopoeia.Chinese Pharmacopoeia carries out indentification by TLC to the saponins of spina date seed medicinal material and flavones ingredient respectively: wherein, the thin-layer identification method of saponin component is: get this product powder 1g, add methyl alcohol 30ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, residue adds methyl alcohol 0.5ml makes dissolving, as need testing solution.Separately get Saponin A, B reference substance, add methyl alcohol respectively and make every 1ml respectively containing the mixed solution of 1mg, product solution in contrast.According to thin-layered chromatography (annex VI B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturated normal butyl alcohol for developping agent, launch, take out, dry, spray with 1% vanillin-sulfuric acid solution, inspect immediately.In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color.The thin-layer identification method of flavones ingredient is: get this product powder 1g, add sherwood oil (60 ~ 90 DEG C) 30ml, add hot reflux 2 hours, filter, discard sherwood oil liquid, the dregs of a decoction volatilize, and add methyl alcohol 30ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, residue adds methyl alcohol 2ml makes dissolving, as need testing solution.Separately get spina date seed control medicinal material 1g, be made in the same way of control medicinal material solution.Get spinosin reference substance again, add methyl alcohol and make the solution of every 1ml containing 0.5mg, product solution in contrast.Test according to thin-layered chromatography (annex VI B), draw each 2 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturated normal butyl alcohol for developping agent, launch, take out, dry, spray, with 1% vanillin-sulfuric acid solution, is inspected under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to reference substance chromatogram, aobvious identical blue-fluorescence spot.
But, in this discrimination method, adopt Saponin A, B reference substance and spinosin reference substance, spina date seed control medicinal material is contrast, respectively saponins and flavones ingredient in discriminating spina date seed, such method both improve testing cost, operated comparatively loaded down with trivial details again.
For the quality control of effective constituent in spina date seed medicinal material, Chinese Pharmacopoeia adopts high performance liquid chromatography to carry out assay to saponins and flavones ingredient respectively.But, in this content assaying method, the traditional Chinese medicine quality control model still followed conventional lines, namely with Saponin A, B or spinosin reference substance etc. in contrast, carry out Quality Evaluation Analysis to spina date seed, this mode evaluation index is relatively single, too relies on traditional Chinese chemical contrast, not environmentally, testing cost is high.
Summary of the invention
Based on this, be necessary for the problems referred to above, prepare a kind of spina date seed reference extract, and it can be used as tester to be applied in spina date seed and the quality control containing the Chinese medicine preparation of spina date seed, there is feedback component information comprehensive, and easy and simple to handle, that testing cost is low advantage.
A preparation method for spina date seed reference extract, comprises the following steps:
Pulverize: get spina date seed medicinal material, pulverize, obtain spina date seed meal;
Degreasing: get above-mentioned spina date seed meal, be placed in supercritical CO
2in the extraction kettle of extraction equipment, with supercritical CO
2the lipid component in spina date seed meal is removed in extraction, obtains the dregs of a decoction;
Extract: get the above-mentioned dregs of a decoction, adding concentration expressed in percentage by volume according to the amount of every gram of spina date seed medicinal material 6ml-10ml is 40%-80% ethanol water, adds hot reflux 1-4 hour, filter, obtain filtrate;
Macroporous resin purification: after above-mentioned filtrate is concentrated, loading adds on large pore resin absorption column, the ethanol water wash-out being 0-10% with the concentration expressed in percentage by volume of 7-11 column volume, discard, again with the ethanol water wash-out that the concentration expressed in percentage by volume of 3-5 column volume is 50%-70%, collect eluent, recycling design, obtains spina date seed reference extract crude product;
Silica gel purification: get above-mentioned spina date seed reference extract crude product, take concentration expressed in percentage by volume as the ethanol water dissolving of 50%-70%, add proper silica gel and mix sample, after solvent evaporated, loading is to silicagel column, first with the eluent ethyl acetate of 20-30 column volume, discards, again with the acetate-methanol wash-out that the volume ratio of 40-60 column volume is 70:30-50:50, collect eluent, recycling design, obtains spina date seed reference extract.
The preparation method of above-mentioned spina date seed reference extract, first by spina date seed with supercritical CO
2extraction, removes lipid component wherein, then extracts with 50%-70% ethanol water, then carry out purifying with macroreticular resin and silica gel successively, remove lengthy and jumbled composition, finally obtain the spina date seed reference extract with special component.Not only containing the saponin component such as Saponin A and Spine Date Seed jujubosideB in this extract, also containing spinosin and 6 " ' flavones ingredient such as-asafoetide acyl spinosin.Therefore, the spina date seed reference extract this preparation method obtained is used for spina date seed in contrast and contains in the quality control of spina date seed flavour of a drug Chinese medicine preparation, there is the comprehensive advantage of feedback component information, and, this spina date seed reference extract is used in contrast, without the need to carrying out loaded down with trivial details pre-treatment step, can directly dissolve rear detection, there is easy and simple to handle, that testing cost is low advantage.
Wherein in an embodiment, in described defatting step, following condition is adopted to carry out supercritical CO
2extraction: extraction kettle temperature is 40-60 DEG C, and pressure is 25-35Mpa, and the first separating still temperature is 40-50 DEG C, and pressure is 5.5-6.5Mpa, and the second separating still temperature is 30-40 DEG C, and pressure is 4-5Mpa, CO
2flow is 25-35L/ hour, extraction 2-4 hour.Adopt above-mentioned condition, the lipid impurities in spina date seed can be sloughed preferably.Further, supercritical CO is adopted
2the mode of extraction carries out degreasing, and compared with the degreasing of conventional oil ether, security is good, is beneficial to the application in large production, does not need operation between hoolivan, greatly improves the operability of its large production.
Wherein in an embodiment, the concrete steps of described macroporous resin purification are: after above-mentioned filtrate is concentrated, loading adds on large pore resin absorption column, with the water elution of 4-6 column volume, discard, then be the ethanol water wash-out of 10% with the concentration expressed in percentage by volume of 3-5 column volume, discard, again with the ethanol water wash-out that the concentration expressed in percentage by volume of 3-5 column volume is 50%-70%, collect eluent, recycling design, obtains spina date seed reference extract crude product.Adopt above-mentioned condition, there is good macroporous resin purification effect.
Wherein in an embodiment, the concrete steps of described macroporous resin purification are: after above-mentioned filtrate is concentrated, loading adds on D101 type large pore resin absorption column, with the water elution of 5 column volumes, discard, be the ethanol water wash-out of 10% again with the concentration expressed in percentage by volume of 4 column volumes, discard, be the ethanol water wash-out of 60% again with the concentration expressed in percentage by volume of 4 column volumes, collect eluent, recycling design is to dry, 60 DEG C of drying under reduced pressure 12 hours, are ground into fine powder, obtain spina date seed reference extract crude product.Adopt above-mentioned condition, there is best macroporous resin purification effect.
Wherein in an embodiment, in described silica gel purification step, fill in described silicagel column for particle diameter be 300 object purification on normal-phase silica gel, elution process is specially: first with the eluent ethyl acetate of 25 column volumes, discard, again with the acetate-methanol wash-out that the volume ratio of 17 column volumes is 70:30, collect eluent, the acetate-methanol wash-out being finally 50:50 with the volume ratio of 33 column volumes, collect eluent, merge the eluent that twice collection obtains, recycling design, obtain spina date seed reference extract.Described silicagel column can select commercially available silica gel post GraceRevelerissilicacartridges (columnsize12g).Adopt above-mentioned condition, there is best silica gel purification effect.
Wherein in an embodiment, in described silica gel purification step, by add proper silica gel mix sample and sample uniform filling after solvent evaporated in sebific duct, this sebific duct and low pressure are prepared liquid phase and are connected, prepare liquid phase with this low pressure and carry out wash-out.Prepare liquid phase with low pressure and carry out purifying, wash-out, have easy to operate, the advantage that elute effect is good.
Wherein in an embodiment, the concrete steps of described extraction are: adding concentration expressed in percentage by volume according to the amount of every gram of spina date seed medicinal material 8ml is 60% ethanol water, add hot reflux 2 times, each 1 hour, filter, merging filtrate.Adopt the method for gradation refluxing extraction, there is better extraction efficiency.
The invention also discloses a kind of spina date seed reference extract, prepared by the preparation method of above-mentioned spina date seed reference extract.
Above-mentioned spina date seed reference extract, wherein not only containing the saponin component such as Saponin A and Spine Date Seed jujubosideB, also containing spinosin and 6 " ' flavones ingredient such as-asafoetide acyl spinosin.Therefore, the spina date seed reference extract this preparation method obtained is used for spina date seed in contrast and contains in the quality control of spina date seed flavour of a drug Chinese medicine preparation, there is the comprehensive advantage of feedback component information, and, this spina date seed reference extract is used in contrast, without the need to carrying out loaded down with trivial details pre-treatment step, can directly dissolve rear detection, also there is easy and simple to handle, that testing cost is low advantage.
The invention also discloses a kind of application of spina date seed reference extract, above-mentioned spina date seed reference extract is used in spina date seed and the quality control containing the Chinese medicine preparation of spina date seed.
Above-mentioned spina date seed reference extract is used for spina date seed and contains in the quality control of spina date seed flavour of a drug Chinese medicine preparation, there is the comprehensive advantage of feedback component information, and, this spina date seed reference extract is used in contrast, without the need to carrying out loaded down with trivial details pre-treatment step, can directly dissolve rear detection, also there is easy and simple to handle, that testing cost is low advantage.
Wherein in an embodiment, described spina date seed and the method for quality control containing the Chinese medicine preparation of spina date seed are: described spina date seed reference extract, testing sample are detected with thin-layered chromatography and/or high performance liquid chromatography, and contrast differentiates;
The testing conditions of described thin-layered chromatography is:
Thin layer plate: silica gel thin-layer plate;
Developping agent: water-saturated n-butanol;
Developer and inspecting: with vanillin-sulfuric acid solution for developer is inspected, and/or inspect under being placed in daylight lamp, and/or inspect under the uviol lamp being placed in 365nm;
Described high performance liquid chromatography comprises flavonoids and detects and saponins detection;
Flavonoids testing conditions is:
Stationary liquid: C18 chromatographic column;
Column temperature: 20-30 DEG C;
Mobile phase: A phase is acetonitrile, B phase is water, carries out according to following gradient elution program:
Flow velocity: 1.0ml/min;
Detecting device: diode array detector;
Saponins testing conditions is:
Stationary liquid: C
18chromatographic column;
Column temperature: 30-40 DEG C;
Mobile phase: A phase is acetonitrile, B phase is water, carries out according to following gradient elution program:
Flow velocity: 1.0ml/min;
Detecting device: evaporative light-scattering detector.
Adopt above method can Multiple components well in thin-layer chromatography and liquid chromatography in separating acid jujube kernel, good degree of separation will be conducive to follow-up analysis and distinguishing.
Compared with prior art, the present invention has following beneficial effect:
The preparation method of a kind of spina date seed reference extract of the present invention, specific method is adopted to carry out degreasing to spina date seed medicinal material, Isolation and purification, finally obtain the spina date seed reference extract with special component, not only containing the saponin component such as Saponin A and Spine Date Seed jujubosideB in this extract, also containing spinosin and 6 " ' flavones ingredient such as-asafoetide acyl spinosin.Therefore, the spina date seed reference extract this preparation method obtained is used for spina date seed in contrast and contains in the quality control of spina date seed flavour of a drug Chinese medicine preparation, there is the comprehensive advantage of feedback component information, and, this Wild jujube seeds extract is used in contrast, without the need to carrying out loaded down with trivial details pre-treatment step, can directly dissolve rear detection, there is easy and simple to handle, that testing cost is low advantage.
The application of a kind of spina date seed reference extract of the present invention, above-mentioned spina date seed reference extract is used for spina date seed and contains in the quality control of spina date seed flavour of a drug Chinese medicine preparation, except there is above-mentioned advantage, also analytical approach is optimized, adopt this analytical approach, can Multiple components well in thin-layer chromatography and liquid chromatography in separating acid jujube kernel, good degree of separation will be conducive to follow-up analysis and distinguishing.
And, find out from the application of spina date seed reference extract, spina date seed reference extract can be applied to the qualitative and quantitative analysis of sedative jujube kernel capsule, ZAOREN ANSHEN YE and spina date seed granule, result accurately and reliably, its overall evaluation effect and reference substance, control medicinal material are suitable, are even better than alone reference substance evaluation method in contrast.In addition, using method and the reference substance of spina date seed reference extract are similar, directly dissolve after weighing, and especially when multicomponent assay, the preparation of reference extract solution is easier than the preparation of reference substance solution.Therefore, reference extract evaluation model has feasibility and certain advantage, is worthy of popularization.
Accompanying drawing explanation
Fig. 1 is spina date seed reference extract preparation method process chart in embodiment 1;
Fig. 2 is the HPLC collection of illustrative plates of the flavone compound of spina date seed reference extract in embodiment 1;
Fig. 3 is the HPLC collection of illustrative plates of the saponins compound of spina date seed reference extract in embodiment 1;
Fig. 4 inspects thin-layer chromatogram under daylight in embodiment 1;
Fig. 5 inspects thin-layer chromatogram under UV365nm in embodiment 1;
Fig. 6 is the thin-layer chromatogram inspected under sedative jujube kernel capsule and ZAOREN ANSHEN YE daylight in embodiment 2;
Fig. 7 is the thin-layer chromatogram inspected under sedative jujube kernel capsule and ZAOREN ANSHEN YE UV365nm in embodiment 2;
Fig. 8 is the thin-layer chromatogram inspected under spina date seed granule daylight in embodiment 3;
Fig. 9 is the thin-layer chromatogram inspected under spina date seed granule UV365nm in embodiment 3;
Figure 10-Figure 11 is the HPLC collection of illustrative plates of spina date seed granule and spina date seed reference extract in embodiment 3;
Figure 12 is the similarity schematic diagram in spina date seed reference extract chromatogram and minute interval, spina date seed granule chromatogram Figure 20 ~ 105 in embodiment 3;
Figure 13 is the thin-layer chromatogram inspected under different spina date seed reference extract daylight in comparative example;
Figure 14 is the thin-layer chromatogram inspected under different spina date seed reference extract UV365nm in comparative example.
Embodiment
For the ease of understanding the present invention, below with reference to relevant drawings, the present invention is described more fully.Preferred embodiment of the present invention is given in accompanying drawing.But the present invention can realize in many different forms, is not limited to embodiment described herein.On the contrary, provide the object of these embodiments be make the understanding of disclosure of the present invention more comprehensively thorough.
Unless otherwise defined, all technology used herein and scientific terminology are identical with belonging to the implication that those skilled in the art of the present invention understand usually.The object of term used in the description of the invention herein just in order to describe specific embodiment, is not intended to be restriction the present invention.Term as used herein "and/or" comprises arbitrary and all combinations of one or more relevant Listed Items.
The instrument used in following examples and reagent as follows:
One, instrument:
Supercritical CO
2extraction equipment (Huaan, Nantong supercritical extract company limited, model: HA221-50-06)
Two, reagent:
Spina date seed control medicinal material (Zhong Jian institute: 121517-201103);
Spinosin reference substance (Zhong Jian institute: 111869-201102);
6 " '-asafoetide acyl spinosin reference substance (Shanghai Tauto Biotechnology Co., Ltd.: 13111922);
Saponin A reference substance (Zhong Jian institute: 110734-200611);
Spine Date Seed jujubosideB reference substance (Zhong Jian institute: 110814-200607);
Sedative jujube kernel capsule (commercially available, lot number: 130202);
ZAOREN ANSHEN YE (commercially available, lot number: 3260034,3260036);
Spina date seed granule (commercially available, lot number: 503396T, 404236L, 402017L).
Embodiment 1
A kind of spina date seed reference extract, prepare by the following method, its technological process is as shown in Figure 1, specific as follows:
1, pulverize.
Get spina date seed medicinal material, pulverize, obtain spina date seed meal.
2, degreasing.
Get above-mentioned spina date seed meal 1kg, be placed in supercritical CO
2in the extraction kettle of extraction equipment, extract by following condition: extraction kettle temperature is 50 DEG C, pressure is 30Mpa, and the first separating still temperature is 45 DEG C, and pressure is 6.0Mpa, and the second separating still temperature is 35 DEG C, and pressure is 4.5Mpa, CO
2flow is 30L/ hour, extracts 3 hours.
Extraction obtains the dregs of a decoction after removing the lipid component in spina date seed meal.
3, extract.
Get the above-mentioned dregs of a decoction, adding concentration expressed in percentage by volume according to the amount of every gram of spina date seed medicinal material 8ml is 60% ethanol water, adds hot reflux 2 times, and each 1 hour, filter while hot, obtain filtrate, filtrate is orange red.
4, macroporous resin purification.
(1) concentrated: filtrate recycling ethanol is to about 2000ml (can produce sediment in concentration process, retain).
(2) adsorb: get the upper D101 type large pore resin absorption column (column internal diameter 4cm, high 20cm) of above concentrate supernatant 250ml (sediment with after appropriate water-soluble solution, with method loading).
(3) wash-out: first with the water elution of 1200ml (4.78 column volumes), discard, be the ethanol water wash-out of 10% again with the concentration expressed in percentage by volume of 1000ml (3.98 column volumes), discard, be the ethanol water wash-out of 60% again with the concentration expressed in percentage by volume of 1000ml (3.98 column volumes), collect eluent.
(4) concentrate drying: by above-mentioned eluent recycling design to dry, 60 DEG C of drying under reduced pressure 12 hours, are ground into fine powder, obtain spina date seed reference extract crude product.
5, silica gel purification.
(1) loading: get above-mentioned spina date seed crude product fine powder 2.0g, add appropriate 60% ethanol to dissolve, add silica gel (60 ~ 100 order) 10g and mix sample, solvent evaporated in water-bath, in uniform filling to blank sebific duct, set low in the standby liquid phase of compacting, connect prefabricated silicagel column (about 300 orders, 12g, internal diameter 2.5cm).
(2) setting elution process is: flow velocity is 30ml/min; First adopt eluent ethyl acetate 15min (about 25 column volumes), then ethyl acetate is adopted: methyl alcohol (70: 30) wash-out 10min (about 17 column volumes), finally adopt ethyl acetate: methyl alcohol (50: 50) wash-out 20min (about 33 column volumes), collect and combined ethyl acetate: methyl alcohol (70: 30) eluent and ethyl acetate: methyl alcohol (50: 50) eluent.
(3) concentrate drying: reclaim eluent solvent to dry, 60 DEG C of drying under reduced pressure 2 hours, obtain spina date seed reference extract.
According to the method described above, prepared the spina date seed reference extract of 10 batches, detected with high performance liquid chromatography.
One, test sample preparation.
1, the preparation of spina date seed reference extract solution.
The spina date seed reference extract 10mg of 1-10 batch in Example 1, dissolves with 10ml methyl alcohol, to obtain final product.
2, the preparation of reference substance solution.
The mixed solution of " '-asafoetide acyl spinosin reference substance appropriate, accurately weighed, add methyl alcohol and make every 1ml containing spinosin 0.2mg, 6 " '-asafoetide acyl spinosin 0.1mg that gets spinosin reference substance and 6, to obtain final product.
Get Saponin A reference substance and Spine Date Seed jujubosideB reference substance appropriate, accurately weighed, add methyl alcohol and make the mixed solution of every 1ml containing Saponin A 50 μ g, Spine Date Seed jujubosideB 30 μ g, to obtain final product.
Two, detect.
1, the mensuration of flavone compound.
Condition determination is as follows:
Stationary liquid: YMCHydrosphere-C
18chromatographic column (250 × 4.6mm, 5 μm);
Column temperature: 20 DEG C;
Mobile phase: A phase is acetonitrile, B phase is water, carries out according to following gradient elution program:
Table 1. gradient elution program
Flow velocity: 1.0ml/min;
Detecting device: diode array detector;
Determined wavelength: 335nm.
2, the mensuration of saponins compound.
Condition determination is as follows:
Stationary liquid: WatersX-bridgeC
18chromatographic column (250 × 4.6mm, 5 μm);
Column temperature: 35 DEG C;
Mobile phase: A phase is acetonitrile, B phase is water, carries out according to following gradient elution program:
Table 2. gradient elution program
Flow velocity: 1.0ml/min;
Detecting device: evaporative light-scattering detector.
Three, result.
Obtain spina date seed reference extract HPLC collection of illustrative plates as Figure 2-3, wherein, Fig. 2 is the HPLC collection of illustrative plates of flavone compound, and label is the chromatographic peak of A is spinosin (t
r=31.351min); Label is the chromatographic peak of B is 6 " '-asafoetide acyl spinosin (t
r=67.348min); Fig. 3 is the HPLC collection of illustrative plates of saponins compound, and label is the chromatographic peak of C is Saponin A (t
r=18.437min); Label is the chromatographic peak of D is Spine Date Seed jujubosideB (t
r=22.161min).
And with spinosin, 6 " '-asafoetide acyl spinosin, Saponin A and Spine Date Seed jujubosideB reference substance calculate this several composition content (with dry product calculate, content is in wt%), result is as shown in the table.
The content of each chemical composition and total amount (%) in table 3. different batches
As can be seen from the above results, adopt the spina date seed reference extract that the method for the present embodiment prepares, there is the spinosin of high level, 6 " '-asafoetide acyl spinosin, Saponin A and Spine Date Seed jujubosideB, and in spina date seed reference extract between different batches, each component content is relatively stable.
Again by the spina date seed reference extract of above-mentioned 10 batches, detect with thin-layered chromatography.
One, test sample preparation.
1, the preparation of spina date seed reference extract solution: the spina date seed reference extract 10mg of 1-10 batch in Example 1, dissolves with 1ml methyl alcohol, to obtain final product.
2, the preparation of reference substance solution: get spinosin reference substance, 6 again " '-asafoetide acyl spinosin reference substance, Saponin A reference substance and Spine Date Seed jujubosideB reference substance, add methyl alcohol respectively and make every 1ml respectively containing the solution of 1mg, product solution in contrast.
3, the preparation of spina date seed control medicinal material solution: get spina date seed control medicinal material 1g, add sherwood oil (60 ~ 90 DEG C) 30ml, add hot reflux 2 hours, filter, filter residue volatilizes, add methyl alcohol 30ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, medicinal material solution in contrast.
4, the preparation of spina date seed medicinal material solution is with spina date seed control medicinal material solution.
Two, detect.
Condition determination is as follows:
Thin layer plate: silica gel thin-layer plate (Merck silica gel 60 prefabricated board);
Developping agent: water-saturated n-butanol;
Developer and inspecting: dry rear spray with 1% vanillin-sulfuric acid solution, 80 DEG C of heating 2 minutes, inspect under putting daylight and ultraviolet lamp (365nm) respectively.
Three, result.
As shown in fig. 4-5, wherein: Fig. 4 inspects thin-layer chromatogram under daylight, Fig. 5 inspects thin-layer chromatogram (t:21 DEG C, RH:30%) under UV365nm to result.Label 1-9 is the spina date seed reference extract of above-mentioned lot number 1-9, label 10-11 is spina date seed medicinal material, label 12 is spina date seed control medicinal material, label 13 is spinosin reference substance, label 14 is 6 " ' asafoetide acyl spinosin reference substance; label 15 is Saponin A reference substance, and label 16 is Spine Date Seed jujubosideB reference substance.
As can be seen from spina date seed reference extract thin-layer chromatogram: the thin-layer chromatogram of spina date seed reference extract and spina date seed medicinal material collection of illustrative plates, spina date seed control medicinal material collection of illustrative plates spot correspondence are good, and profile information amount is enriched.
Embodiment 2
The spina date seed reference extract prepared by above-described embodiment 1 is used in sedative jujube kernel capsule, the discriminating of ZAOREN ANSHEN YE and assay.
One, TLC distinguish.
1, test sample preparation.
(1) preparation of sedative jujube kernel capsule, ZAOREN ANSHEN YE need testing solution.
Sedative jujube kernel capsule: get this product content 4g, add diethyl ether 50ml, adds hot reflux 30 minutes, filters, the dregs of a decoction volatilize solvent, add methyl alcohol 50ml, add hot reflux 30 minutes, filter, filtrate evaporate to dryness, the residue 20ml that adds water makes dissolving, extract 3 times with water saturated normal butyl alcohol jolting, each 25ml, merge normal butyl alcohol liquid, wash 2 times with ammonia solution, each 10ml, get normal butyl alcohol liquid, evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.
ZAOREN ANSHEN YE: get this product 20ml, by D101 type macroporous absorbent resin, (column internal diameter is 1.5cm, post height is 10cm), wash with water to colourless, discard eluent, use each 30ml of 70% methyl alcohol, methyl alcohol wash-out successively again, collect eluent, merge, evaporate to dryness, the residue 20ml that adds water makes dissolving, extract 3 times with water saturated normal butyl alcohol jolting, each 25ml, merge normal butyl alcohol liquid, 2 times are washed with ammonia solution, each 10ml, gets normal butyl alcohol liquid, evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.
(2) preparation of spina date seed reference extract solution is with embodiment 1.
(3) preparation of reference substance solution is with embodiment 1.
(4) preparation of spina date seed medicinal material and control medicinal material solution is with embodiment 1.
2, detect.
Condition determination is with embodiment 1.
3, result.
As Figure 6-Figure 7, wherein: Fig. 6 inspects thin-layer chromatogram under daylight, Fig. 7 inspects thin-layer chromatogram under UV365nm to result.Label 1 is sedative jujube kernel capsule (130202), label 2 is ZAOREN ANSHEN YE (3260034), label 3 is ZAOREN ANSHEN YE (3260036), label 4-9 is the spina date seed reference extract of above-mentioned lot number 1-6, label 10-11 is spina date seed medicinal material, label 12 is spina date seed control medicinal material, label 13 is spinosin reference substance, label 14 is 6 " '-asafoetide acyl spinosin reference substance; label 15 is Saponin A reference substance, and label 16 is Spine Date Seed jujubosideB reference substance.
As can be seen from above-mentioned thin-layer chromatogram: be applied to by spina date seed reference extract in the TLC distinguish of sedative jujube kernel capsule, ZAOREN ANSHEN YE, result display spina date seed reference extract collection of illustrative plates and sedative jujube kernel capsule, ZAOREN ANSHEN YE collection of illustrative plates show the fluorescence spot of same color.Spina date seed reference extract identification result and spina date seed control medicinal material similar, be better than alone reference substance differentiate effect.
Two, high performance liquid chromatography assay.
1, test sample preparation.
(1) preparation of sedative jujube kernel capsule, ZAOREN ANSHEN YE need testing solution.
Sedative jujube kernel capsule: get this product content under content uniformity item, mixing, porphyrize, get about 1g, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 50ml, close plug, weighed weight, ultrasonic process (power 380W, frequency 37kHz) 30 minutes (needing if desired to shake loose medicinal powder), lets cool, weighed weight again, supplies the weight of less loss, shakes up with 70% methyl alcohol, filter, get subsequent filtrate, to obtain final product.
ZAOREN ANSHEN YE: precision measures this product 3ml, puts in 10ml measuring bottle, adds 80% methyl alcohol to scale, violent jolting, filters, gets subsequent filtrate, to obtain final product.
(2) preparation of spina date seed reference extract solution.
Precision takes spina date seed reference extract and is about 4mg, puts in 10ml measuring bottle, adds methyl alcohol and dissolves and be diluted to scale, shake up, to obtain final product.
(3) preparation of reference substance solution.
The mixed solution of " '-asafoetide acyl spinosin reference substance appropriate, accurately weighed, add methyl alcohol and make every 1ml containing spinosin 0.2mg, 6 " '-asafoetide acyl spinosin 0.1mg that gets spinosin reference substance and 6, to obtain final product.
2, detect.
Detect with high performance liquid chromatography.
3, result.
Respectively with the spinosin and 6 in reference substance and spina date seed reference extract " content of '-asafoetide acyl spinosin calculated by peak area sedative jujube kernel capsule, spinosin and 6 in ZAOREN ANSHEN YE " '-asafoetide acyl spinosin, result is as shown in the table.
In table 4. sedative jujube kernel capsule, ZAOREN ANSHEN YE, the content results of each composition compares (n=2)
As can be seen from the above results, spina date seed reference extract is applied to the assay item of sedative jujube kernel capsule, ZAOREN ANSHEN YE, testing result shows: the result difference of reference extract computing method and reference substance computing method is little.Show that spina date seed reference extract can be applied to spinosin and 6 in sedative jujube kernel capsule and ZAOREN ANSHEN YE " ' assay of-asafoetide acyl spinosin, reliable results.
Embodiment 3
Spina date seed granule is spina date seed simple preparation, at present temporarily without quality standard.The method that the present invention has set up with reference to above-mentioned spina date seed reference extract, adopt the evaluation model of reference substance and reference extract to evaluate the quality of spina date seed granule in indentification by TLC, finger-print and assay three respectively, and compare the result difference of two kinds of evaluation models.
One, TLC distinguish
1, test sample preparation.
(1) preparation of spina date seed granule need testing solution.
Get this product porphyrize, get about 0.3g, add methyl alcohol 20ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.
(2) preparation of spina date seed reference extract solution is with embodiment 1.
(3) preparation of reference substance solution is with embodiment 1.
(4) preparation of spina date seed medicinal material and control medicinal material solution is with embodiment 1.
2, detect.
Condition determination is with embodiment 1.
3, result.
As illustrated in figs. 8-9, wherein: Fig. 8 inspects thin-layer chromatogram under daylight, Fig. 9 inspects thin-layer chromatogram (t:21 DEG C, RH:30%) under UV365nm to result.Label 1-3 is spina date seed granule (503396T, 404236L, 402017L), label 4-9 is the spina date seed reference extract of above-mentioned lot number 1-6, and label 10-11 is spina date seed medicinal material, and label 12 is spina date seed control medicinal material, label 13 is spinosin reference substance, label 14 is 6 " '-asafoetide acyl spinosin reference substance, label 15 is Saponin A reference substance, and label 16 is Spine Date Seed jujubosideB reference substance.
As can be seen from above-mentioned thin-layer chromatogram: spina date seed reference extract and each reference substance all can be used for the indentification by TLC of spina date seed granule, but the profile information amount that spina date seed reference extract is shown is abundanter, and identification result is similar to spina date seed control medicinal material.And the preparation method of spina date seed reference extract solution is consistent with the preparation method of reference substance solution, direct solubilizer dissolves, simple and convenient.
Two, finger-print.
1, test sample preparation.
(1) preparation of spina date seed granule need testing solution.
Get this product appropriate, porphyrize, gets about 0.3g, accurately weighed, puts in flat bottom flask, and precision adds methyl alcohol 20ml, weighed weight, adds hot reflux 1 hour, lets cool, more weighed weight, supplies the weight of less loss, shake up with methyl alcohol, filters, gets subsequent filtrate, to obtain final product.
(2) preparation of spina date seed reference extract solution is with embodiment 1.
2, detect.
Condition determination is as follows:
Stationary liquid: YMCHydrosphere-C
18chromatographic column (250 × 4.6mm, 5 μm);
Column temperature: 20 DEG C;
Mobile phase: A phase is acetonitrile, B phase is 0.1g/100ml phosphoric acid solution, carries out according to following gradient elution program:
Table 5. gradient elution program
Flow velocity: 1.0ml/min;
Detecting device: diode array detector;
Determined wavelength: 204nm.
3, result.
Number of theoretical plate calculates should be not less than 5000 by spinosin peak, obtain the HPLC collection of illustrative plates of Figure 10-11, wherein, in Figure 10, a figure is the HPLC collection of illustrative plates of spina date seed reference extract, in Figure 10, b figure and Figure 11 is respectively spina date seed granule (503396T, 404236L, 402017L) HPLC collection of illustrative plates.Label be 3 chromatographic peak be spinosin; Label be 4 chromatographic peak be 6 " '-asafoetide acyl spinosin, label be 9 chromatographic peak be Saponin A; Label be 10 chromatographic peak be Spine Date Seed jujubosideB.
As can be seen from Figure 10-11,10 Characteristic chromatographic peaks in spina date seed reference extract chromatogram all exist in spina date seed granule test sample chromatogram.In addition, because spina date seed reference extract is through purification process, this 10 chromatographic peaks corresponding with spina date seed granule mainly concentrate on the interval of 20 ~ 105 minutes, therefore gained chromatogram AIA formatted data is imported similarity evaluation software (2012.130723 version), calculate the similarity (the results are shown in Table 6 and Figure 12) of 20 ~ 105 minutes interval spina date seed reference extract chromatograms and spina date seed granule chromatogram, the similarity of both result displays is all more than 0.95, show that the correlativity of spina date seed reference extract and spina date seed granule finger-print is good.
Table 6 spina date seed reference extract and spina date seed granule fingerprint similarity result
Three, high performance liquid chromatography assay.
1, test sample preparation.
(1) the same fingerprint image spectral term of the preparation of spina date seed granule need testing solution.
(2) the same fingerprint image spectral term of the preparation of spina date seed reference extract solution.
(3) preparation of reference substance solution:
The mixed solution of " '-asafoetide acyl spinosin reference substance appropriate, accurately weighed, add methyl alcohol and make every 1ml containing spinosin 0.2mg, 6 " '-asafoetide acyl spinosin 0.1mg that gets spinosin reference substance and 6, to obtain final product.
Get Saponin A reference substance and Spine Date Seed jujubosideB reference substance appropriate, accurately weighed, add methyl alcohol and make the mixed solution of every 1ml containing Saponin A 50 μ g, Spine Date Seed jujubosideB 30 μ g, to obtain final product.
2, detect.
(1), spinosin and 6 " ' the mensuration of-asafoetide acyl spinosin.
Determined wavelength is 335nm, and all the other are with fingerprint image spectral term.The each 10 μ l of accurate absorption reference substance solution, reference extract solution and need testing solution respectively, injection liquid chromatography, measures, to obtain final product.
(2), the mensuration of Saponin A and Spine Date Seed jujubosideB.
According to the assay method of saponins compound in embodiment 1, accurate absorption reference substance solution 5 μ l and 25 μ l, reference extract solution 5 μ l and 25 μ l and need testing solution 20 μ l respectively, injection liquid chromatography, measures, calculate with external standard two-point method logarithmic equation, to obtain final product.
3, result.
(1), spinosin and 6 " ' the content of-asafoetide acyl spinosin.
In spina date seed granule, the content of each composition is as shown in the table.
In table 7 spina date seed granule, the content results of each composition compares (n=2)
As seen from the above table, Standard reference and reference extract method is adopted to calculate spinosin and 6 in spina date seed granule respectively " assay of ' content of-asafoetide acyl spinosin; the measurement result difference of two kinds of methods is little (relative average debiation is below 3.0%); show that spina date seed reference extract can be applied to spinosin and 6 in spina date seed granule " '-asafoetide acyl spinosin, reliable results.
(2), the content of Saponin A and Spine Date Seed jujubosideB.
In spina date seed granule, the content of each composition is as shown in the table.
In table 8 spina date seed granule, the content results of each composition compares (n=2)
As seen from the above table, Standard reference and reference extract method is adopted to calculate the content of Saponin A and Spine Date Seed jujubosideB in spina date seed granule respectively, the measurement result difference of two kinds of methods is little (relative average debiation is below 3.0%), show that spina date seed reference extract can be applied to the assay of Saponin A and Spine Date Seed jujubosideB in spina date seed granule, reliable results.
Comparative example
The spina date seed reference extract of this comparative example, adopts following different preparation method to prepare:
Spina date seed reference extract A, its preparation method is substantially identical with the preparation method of embodiment 1, and difference is not through defatting step in the preparation process of this Wild jujube seeds extract A, with macroreticular resin and silica gel purification after extracting directly.
Spina date seed reference extract B, its preparation method is substantially identical with the preparation method of embodiment 1, and difference is in the preparation process of this spina date seed reference extract B not through macroreticular resin and silica gel purification.
Spina date seed reference extract C, its preparation method is substantially identical with the preparation method of embodiment 1, and difference is do not have degreasing in the preparation process of this spina date seed reference extract C, also not through macroreticular resin and silica gel purification.
One, contrasting character's characteristic
The spina date seed reference extract prepared in the spina date seed reference extract this comparative example prepared and embodiment 1 carries out proterties Property comparison, and result is as shown in the table.
Table 9 character pair ratio
| Sample | Dissolubility in methyl alcohol | Draw moist |
| The reference extract that embodiment 1 obtains | Whole dissolving | Have draw moist |
| Spina date seed reference extract A | Whole dissolving | Have draw moist |
| Spina date seed reference extract B | There is insolubles | Have draw moist |
| Spina date seed reference extract C | There is insolubles | Have draw moist |
As can be seen from the above table, the dissolubility of spina date seed reference extract B and C in methyl alcohol is poor, has insolubles; Spina date seed reference extract B and C has and draws moist, not easily preserves.And the spina date seed reference extract that embodiment 1 obtains all dissolves in methyl alcohol, draw and moistly also to improve, easily preserve.
Two, thin-layer chromatography is contrasted.
The spina date seed reference extract prepared in the spina date seed reference extract this comparative example prepared and embodiment 1 detects with thin-layered chromatography, and detection method is the same, and testing result as illustrated by figs. 12-13.
Wherein: Figure 13 inspects thin-layer chromatogram under daylight, Figure 14 inspects thin-layer chromatogram (t:24 DEG C, RH:35%) under UV365nm.Label 1-4,10 is above-mentioned spina date seed reference extract C, label 5-9,11-12 is above-mentioned spina date seed reference extract B, and label 13 is spina date seed control medicinal material, and label 14 is Saponin A reference substance, label 15 is Spine Date Seed jujubosideB reference substance, and label 16 is spinosin reference substance.
As can be seen from the thin-layer chromatogram of Figure 13-14: compared with embodiment 1 collection of illustrative plates (Fig. 4-Fig. 5), in spina date seed reference extract B and C thin layer collection of illustrative plates, a lot of at the spot of the impurity components such as the region grease near solvent front, the clear spot degree of flavonoids and saponin active ingredient is inadequate, affects the identification result of thin-layer chromatography.
Three, component content is contrasted.
The spina date seed reference extract prepared in spina date seed reference extract A, B and the C this comparative example prepared and embodiment 1 detects with high performance liquid chromatography, and detection method is the same, and testing result is as shown in the table.
Each component content (%) in table 10 different spina date seed reference extract
As can be seen from the above table, the spina date seed reference extract obtained with embodiment 1 compares, Saponin A contained in spina date seed reference extract A and the total content of Spine Date Seed jujubosideB content and index composition all lower; And spina date seed reference extract B and C, content and the total content of each index composition are all very low; And in embodiment 1 content of each index composition and total content all higher.
Each technical characteristic of the above embodiment can combine arbitrarily, for making description succinct, the all possible combination of each technical characteristic in above-described embodiment is not all described, but, as long as the combination of these technical characteristics does not exist contradiction, be all considered to be the scope that this instructions is recorded.
The above embodiment only have expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but can not therefore be construed as limiting the scope of the patent.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (10)
1. a preparation method for spina date seed reference extract, comprises the following steps:
Pulverize: get spina date seed medicinal material, pulverize, obtain spina date seed meal;
Degreasing: get above-mentioned spina date seed meal, be placed in supercritical CO
2in the extraction kettle of extraction equipment, with supercritical CO
2the lipid component in spina date seed meal is removed in extraction, obtains the dregs of a decoction;
Extract: get the above-mentioned dregs of a decoction, adding concentration expressed in percentage by volume according to the amount of every gram of spina date seed medicinal material 6ml-10ml is 40%-80% ethanol water, adds hot reflux 1-4 hour, filter, obtain filtrate;
Macroporous resin purification: after above-mentioned filtrate is concentrated, loading adds on large pore resin absorption column, the ethanol water wash-out being 0-10% with the concentration expressed in percentage by volume of 7-11 column volume, discard, again with the ethanol water wash-out that the concentration expressed in percentage by volume of 3-5 column volume is 50%-70%, collect eluent, recycling design, obtains spina date seed reference extract crude product;
Silica gel purification: get above-mentioned spina date seed reference extract crude product, take concentration expressed in percentage by volume as the ethanol water dissolving of 50%-70%, add proper silica gel and mix sample, after solvent evaporated, loading is to silicagel column, first with the eluent ethyl acetate of 20-30 column volume, discards, again with the acetate-methanol wash-out that the volume ratio of 40-60 column volume is 70:30-50:50, collect eluent, recycling design, obtains spina date seed reference extract.
2. the preparation method of spina date seed reference extract according to claim 1, is characterized in that, in described defatting step, adopts following condition to carry out supercritical CO
2extraction: extraction kettle temperature is 40-60 DEG C, and pressure is 25-35Mpa, and the first separating still temperature is 40-50 DEG C, and pressure is 5.5-6.5Mpa, and the second separating still temperature is 30-40 DEG C, and pressure is 4-5Mpa, CO
2flow is 25-35L/ hour, extraction 2-4 hour.
3. the preparation method of spina date seed reference extract according to claim 1, it is characterized in that, the concrete steps of described macroporous resin purification are: after above-mentioned filtrate is concentrated, loading adds on large pore resin absorption column, with the water elution of 4-6 column volume, discard, be the ethanol water wash-out of 10% again with the concentration expressed in percentage by volume of 3-5 column volume, discard, again with the ethanol water wash-out that the concentration expressed in percentage by volume of 3-5 column volume is 50%-70%, collect eluent, recycling design, obtains spina date seed reference extract crude product.
4. the preparation method of spina date seed reference extract according to claim 3, it is characterized in that, the concrete steps of described macroporous resin purification are: after above-mentioned filtrate is concentrated, loading adds on D101 type large pore resin absorption column, with the water elution of 5 column volumes, discard, be the ethanol water wash-out of 10% again with the concentration expressed in percentage by volume of 4 column volumes, discard, be the ethanol water wash-out of 60% again with the concentration expressed in percentage by volume of 4 column volumes, collect eluent, recycling design is to dry, 60 DEG C of drying under reduced pressure 12 hours, be ground into fine powder, obtain spina date seed reference extract crude product.
5. the preparation method of spina date seed reference extract according to claim 1, it is characterized in that, in described silica gel purification step, fill in described silicagel column for particle diameter be 300 object purification on normal-phase silica gel, elution process is specially: first with the eluent ethyl acetate of 25 column volumes, discard, again with the acetate-methanol wash-out that the volume ratio of 17 column volumes is 70:30, collect eluent, the acetate-methanol wash-out being finally 50:50 with the volume ratio of 33 column volumes, collect eluent, merge the eluent that twice collection obtains, recycling design, obtain spina date seed reference extract.
6. the preparation method of spina date seed reference extract according to claim 5, it is characterized in that, in described silica gel purification step, by add proper silica gel mix sample and sample uniform filling after solvent evaporated in sebific duct, this sebific duct and low pressure are prepared liquid phase be connected, prepare liquid phase with this low pressure and carry out wash-out.
7. the preparation method of spina date seed reference extract according to claim 1, is characterized in that, the concrete steps of described extraction are: adding concentration expressed in percentage by volume according to the amount of every gram of spina date seed medicinal material 8ml is 60% ethanol water, add hot reflux 2 times, each 1 hour, filter, merging filtrate.
8. a spina date seed reference extract, is characterized in that, is prepared by the preparation method of the spina date seed reference extract described in any one of claim 1-7.
9. an application for spina date seed reference extract, is characterized in that, is used for by spina date seed reference extract according to claim 8 in spina date seed and the quality control containing the Chinese medicine preparation of spina date seed.
10. the application of spina date seed reference extract according to claim 9, it is characterized in that, described spina date seed and be containing the method for quality control of Chinese medicine preparation of spina date seed: described spina date seed reference extract, testing sample are detected with thin-layered chromatography and/or high performance liquid chromatography, contrast differentiates;
The testing conditions of described thin-layered chromatography is:
Thin layer plate: silica gel thin-layer plate;
Developping agent: water-saturated n-butanol;
Developer and inspecting: with vanillin-sulfuric acid solution for developer is inspected, and/or inspect under being placed in daylight lamp, and/or inspect under the uviol lamp being placed in 365nm;
Described high performance liquid chromatography comprises flavonoids and detects and saponins detection;
Flavonoids testing conditions is:
Stationary liquid: C
18chromatographic column;
Column temperature: 20-30 DEG C;
Mobile phase: A phase is acetonitrile, B phase is water, carries out according to following gradient elution program:
Flow velocity: 1.0ml/min;
Detecting device: diode array detector;
Saponins testing conditions is:
Stationary liquid: C
18chromatographic column;
Column temperature: 30-40 DEG C;
Mobile phase: A phase is acetonitrile, B phase is water, carries out according to following gradient elution program:
Flow velocity: 1.0ml/min;
Detecting device: evaporative light-scattering detector.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510585856.4A CN105203657B (en) | 2015-09-15 | 2015-09-15 | Semen Ziziphi Spinosae reference extract and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510585856.4A CN105203657B (en) | 2015-09-15 | 2015-09-15 | Semen Ziziphi Spinosae reference extract and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105203657A true CN105203657A (en) | 2015-12-30 |
| CN105203657B CN105203657B (en) | 2017-03-29 |
Family
ID=54951448
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510585856.4A Active CN105203657B (en) | 2015-09-15 | 2015-09-15 | Semen Ziziphi Spinosae reference extract and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105203657B (en) |
Cited By (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105670798A (en) * | 2016-03-02 | 2016-06-15 | 塔里木大学 | Extraction method of jujube kernel volatile oil |
| CN105670784A (en) * | 2016-03-02 | 2016-06-15 | 塔里木大学 | Method for extracting and purifying jujube kernel volatile oil |
| CN105687368A (en) * | 2016-03-02 | 2016-06-22 | 塔里木大学 | Application of jujube kernel volatile oil in mouse hair growth |
| CN105779116A (en) * | 2016-05-05 | 2016-07-20 | 广州合诚三先生物科技有限公司 | Comprehensive extraction method for effective ingredients of spina date seeds |
| CN106723021A (en) * | 2016-11-28 | 2017-05-31 | 武汉哈福科技有限公司 | A kind of functional food for improving the health care of sleep |
| CN106814157A (en) * | 2017-01-17 | 2017-06-09 | 广州浩意万医药科技有限公司 | The preparation method of rascal reference extract |
| CN106841433A (en) * | 2017-01-17 | 2017-06-13 | 广州浩意万医药科技有限公司 | A kind of rascal reference extract and its application |
| CN107573399A (en) * | 2017-09-21 | 2018-01-12 | 上海诗丹德标准技术服务有限公司 | A kind of preparation method of Saponin A reference substance |
| CN108181400A (en) * | 2018-01-25 | 2018-06-19 | 南京中医药大学 | The method that UHPLC-QQQ-MS/MS measures 6 kinds of chemical composition contents in spina date seed decocting liquid simultaneously |
| CN109223993A (en) * | 2018-10-10 | 2019-01-18 | 中华全国供销合作总社济南果品研究院 | The method of saponin(e is extracted in a kind of garden stuff processing waste |
| CN109521138A (en) * | 2018-12-07 | 2019-03-26 | 浙江工业大学 | Rapid identification method for spina date seeds and fried spina date seeds |
| CN109879919A (en) * | 2019-02-20 | 2019-06-14 | 滨州医学院 | A method of three kinds of flavonoid glycosides of separation preparation from semen ziziphi spinosae |
| CN110045039A (en) * | 2019-05-16 | 2019-07-23 | 辽宁仙草堂药业有限公司 | A kind of quality determining method of ZAOREN ANSHEN YE |
| CN110376318A (en) * | 2018-04-13 | 2019-10-25 | 广州万正药业有限公司 | A kind of semen ziziphi spinosae reference extract and its preparation method and application |
| CN110857939A (en) * | 2018-08-23 | 2020-03-03 | 完美（中国）有限公司 | Detection method of traditional Chinese medicine composition with sleep improvement function |
| CN113109495A (en) * | 2021-05-24 | 2021-07-13 | 北京大学 | Quality detection method of jujube kernel nerve-soothing capsules based on thin-layer chromatography |
| CN113214349A (en) * | 2021-05-14 | 2021-08-06 | 华南理工大学 | Spina date seed saponin, preparation thereof and application thereof in preparing hypoglycemic drugs |
| CN113341007A (en) * | 2021-05-24 | 2021-09-03 | 北京大学 | Method for measuring contents of multiple components in whole Chinese date seed nerve-soothing capsule based on HPLC (high performance liquid chromatography) characteristic spectrum |
| CN115575551A (en) * | 2022-09-20 | 2023-01-06 | 贵州中医药大学 | Detection method of rhizoma bletillae, preparation method of rhizoma bletillae control extract and application of rhizoma bletillae control extract |
-
2015
- 2015-09-15 CN CN201510585856.4A patent/CN105203657B/en active Active
Non-Patent Citations (7)
| Title |
|---|
| CHUNYAN NIU 等: "Quantitative analysis and chromatographic fingerprinting of the semen zizyphi spinosae by ultra-high-performance liquid chromatography coupled with diode-array detector", 《J.SEP.SCI.》 * |
| JINGJING LIU 等: "Simultaneous analysis and identification of main bioactive constituents in extract of Zizyphus jujuba var. sapinosa (Zizyphi spinosi semen)by high-performance liquid chromatography–photodiode array detection–electrospray mass spectrometry", 《TALANTA》 * |
| 杜志敏 等: "不同来源酸枣仁有效成分的对照分析", 《广东药学院学报》 * |
| 王筱亮 等: "多指标正交试验优选酸枣仁最佳提取工艺", 《中国中药杂志》 * |
| 陈学林 等: "超滤法纯化酸枣仁提取物的工艺研究", 《西北师范大学学报（自然科学版）》 * |
| 高剑峰 等 3: "酸枣仁中总黄酮的含量测定及提取工艺研究", 《现代中药研究与实践》 * |
| 鲍康德 等: "HPLC-ELSD 法同时测定酸枣仁中黄酮及皂苷类成分", 《中国天然药物》 * |
Cited By (31)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105670784A (en) * | 2016-03-02 | 2016-06-15 | 塔里木大学 | Method for extracting and purifying jujube kernel volatile oil |
| CN105687368A (en) * | 2016-03-02 | 2016-06-22 | 塔里木大学 | Application of jujube kernel volatile oil in mouse hair growth |
| CN105670798A (en) * | 2016-03-02 | 2016-06-15 | 塔里木大学 | Extraction method of jujube kernel volatile oil |
| CN105687368B (en) * | 2016-03-02 | 2020-02-14 | 塔里木大学 | Application of jujube kernel volatile oil in mouse hair growth |
| CN105670784B (en) * | 2016-03-02 | 2020-01-03 | 塔里木大学 | Extraction and purification method of jujube kernel volatile oil |
| CN105670798B (en) * | 2016-03-02 | 2020-01-03 | 塔里木大学 | Extraction method of jujube kernel volatile oil |
| CN105779116A (en) * | 2016-05-05 | 2016-07-20 | 广州合诚三先生物科技有限公司 | Comprehensive extraction method for effective ingredients of spina date seeds |
| CN106723021A (en) * | 2016-11-28 | 2017-05-31 | 武汉哈福科技有限公司 | A kind of functional food for improving the health care of sleep |
| CN106841433B (en) * | 2017-01-17 | 2019-06-11 | 广州浩意万医药科技有限公司 | A kind of green peel reference extract and its application |
| CN106814157A (en) * | 2017-01-17 | 2017-06-09 | 广州浩意万医药科技有限公司 | The preparation method of rascal reference extract |
| CN106841433A (en) * | 2017-01-17 | 2017-06-13 | 广州浩意万医药科技有限公司 | A kind of rascal reference extract and its application |
| CN106814157B (en) * | 2017-01-17 | 2018-11-20 | 广州浩意万医药科技有限公司 | The preparation method of green peel reference extract |
| CN107573399B (en) * | 2017-09-21 | 2020-08-11 | 上海诗丹德标准技术服务有限公司 | Preparation method of jujuboside A reference substance |
| CN107573399A (en) * | 2017-09-21 | 2018-01-12 | 上海诗丹德标准技术服务有限公司 | A kind of preparation method of Saponin A reference substance |
| CN108181400B (en) * | 2018-01-25 | 2020-08-04 | 南京中医药大学 | Method for simultaneously measuring contents of 6 chemical components in spina date seed water decoction by UHP L C-QQQ-MS/MS |
| CN108181400A (en) * | 2018-01-25 | 2018-06-19 | 南京中医药大学 | The method that UHPLC-QQQ-MS/MS measures 6 kinds of chemical composition contents in spina date seed decocting liquid simultaneously |
| CN110376318A (en) * | 2018-04-13 | 2019-10-25 | 广州万正药业有限公司 | A kind of semen ziziphi spinosae reference extract and its preparation method and application |
| CN110857939A (en) * | 2018-08-23 | 2020-03-03 | 完美（中国）有限公司 | Detection method of traditional Chinese medicine composition with sleep improvement function |
| CN110857939B (en) * | 2018-08-23 | 2022-03-18 | 完美（中国）有限公司 | Detection method of traditional Chinese medicine composition with sleep improvement function |
| CN109223993A (en) * | 2018-10-10 | 2019-01-18 | 中华全国供销合作总社济南果品研究院 | The method of saponin(e is extracted in a kind of garden stuff processing waste |
| CN109521138A (en) * | 2018-12-07 | 2019-03-26 | 浙江工业大学 | Rapid identification method for spina date seeds and fried spina date seeds |
| CN109521138B (en) * | 2018-12-07 | 2020-11-13 | 浙江工业大学 | Rapid identification method for spina date seeds and fried spina date seeds |
| CN109879919A (en) * | 2019-02-20 | 2019-06-14 | 滨州医学院 | A method of three kinds of flavonoid glycosides of separation preparation from semen ziziphi spinosae |
| CN109879919B (en) * | 2019-02-20 | 2022-03-15 | 滨州医学院 | Method for separating and preparing three flavonoid glycosides from spina date seeds |
| CN110045039A (en) * | 2019-05-16 | 2019-07-23 | 辽宁仙草堂药业有限公司 | A kind of quality determining method of ZAOREN ANSHEN YE |
| CN113214349A (en) * | 2021-05-14 | 2021-08-06 | 华南理工大学 | Spina date seed saponin, preparation thereof and application thereof in preparing hypoglycemic drugs |
| CN113109495A (en) * | 2021-05-24 | 2021-07-13 | 北京大学 | Quality detection method of jujube kernel nerve-soothing capsules based on thin-layer chromatography |
| CN113341007A (en) * | 2021-05-24 | 2021-09-03 | 北京大学 | Method for measuring contents of multiple components in whole Chinese date seed nerve-soothing capsule based on HPLC (high performance liquid chromatography) characteristic spectrum |
| CN113341007B (en) * | 2021-05-24 | 2023-10-10 | 北京大学 | HPLC (high Performance liquid chromatography) characteristic spectrum-based method for measuring content of all ingredients of jujube kernel nerve-soothing capsules |
| CN115575551A (en) * | 2022-09-20 | 2023-01-06 | 贵州中医药大学 | Detection method of rhizoma bletillae, preparation method of rhizoma bletillae control extract and application of rhizoma bletillae control extract |
| CN115575551B (en) * | 2022-09-20 | 2023-09-26 | 贵州中医药大学 | Bletilla striata detection method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105203657B (en) | 2017-03-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105203657A (en) | Spina date seed reference extract and preparation method and application thereof | |
| CN107843677B (en) | Radix paeoniae rubra control extract and preparation method and application thereof | |
| CN102749348B (en) | Method for identifying active components in medicinal plant | |
| CN102998411B (en) | A kind of detection method of ageratum dripping pill | |
| CN104306745A (en) | Quality control method for rhizoma gastrodiae capsule | |
| CN106198837A (en) | The quality determining method of old cough with asthma sheet | |
| CN103869041A (en) | Quality control method for cough-relieving oral pharmaceutical composition | |
| CN109164178B (en) | Quality detection and identification method for gardenia and processed products thereof | |
| CN103323558B (en) | Ginkgo biloba preparation analysis sample rapid preparation method based on solid phase extraction technology | |
| CN102980956A (en) | Quick extraction and detection method of 9-oxo-10,11-dehydroagerophorone | |
| CN102608248B (en) | Relinqing granules and polygonum capitatum thin-layer fingerprint chromatogram determination method | |
| CN108088715B (en) | Moutan bark reference extract and preparation method and application thereof | |
| CN103822888B (en) | Catch up with the quality determining method of yellow grass | |
| CN102818863B (en) | Method for identifying proanthocyanidins in ginkgo leaf preparation | |
| CN1879756B (en) | Quality control method of kidney-replenishing blood-nourishing soft capsule | |
| CN108037200B (en) | Quality detection method of kidney nourishing and tranquilizing pills | |
| CN101703610A (en) | Quality detection method of Qingnao antihypertensive tablet | |
| CN106596759B (en) | A kind of HPLC analysis method of Ramulus et folium taxi cuspidatae extract and its preparation | |
| CN103616344B (en) | A kind of detection method of Tibetan medicine material Corydalis impatiens (Pall.) Fisch | |
| CN103969356B (en) | A kind of discrimination method of the finger printing of red rooted salvia | |
| CN105616946A (en) | Preparation for treating cough, preparation method and quality control method thereof | |
| CN109884199A (en) | Method for measuring content of flavonoid components in honey | |
| CN101632804B (en) | Quality control method for wind-dispelling heat-dissipating capsules | |
| CN104133028B (en) | A kind of method for building up of madder granule efficient liquid-phase chromatograph finger print atlas | |
| Wang et al. | HPLC fingerprint combined with quantitation of main effective components and chemometrics as an efficient method for quality evaluation of Oviductus Ranae |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |