CN115403649A - Method for extracting and separating steroid saponin compounds from allium macrostemon - Google Patents
Method for extracting and separating steroid saponin compounds from allium macrostemon Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J71/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton is condensed with a heterocyclic ring
- C07J71/0005—Oxygen-containing hetero ring
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- G—PHYSICS
- G01—MEASURING; TESTING
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- G01N30/02—Column chromatography
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/08—Preparation using an enricher
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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Abstract
The invention relates to a method for extracting and separating steroid saponin compounds from allium macrostemon, belonging to the technical field of separation and purification of compound monomers. The invention takes allium macrostemon as a raw material, and obtains the steroid saponin compound monomer through extraction, concentration, chromatography, filtration, separation by a preparative high performance liquid chromatograph and product recovery. The separation method has the advantages of high separation efficiency, short production period, stable process, simple and convenient operation and low cost, and can realize the high-purity (more than 99 percent of the area normalization method) separation of the steroid saponin compounds.
Description
Technical Field
The invention relates to a method for extracting and separating compounds, in particular to a method for extracting and separating steroid saponin compounds from allium macrostemon, belonging to the technical field of separation and purification of compound monomers.
Background
The Bulbus Allii Macrostemi is dry bulb of Bulbus Allii Macrostemi or Bulbus Allii Macrostemi of Liliaceae, and is named as Bulbus Allii Macrostemi root, rhizoma Acori Calami, bulbus Allii Macrostemonis, herba Allium Sativi, and Bulbus Allii Macrostemonis, mainly produced in northeast and North China, and is harvested in summer and autumn, cleaned, removed fibrous root, steamed or scalded in boiling water, and sun-dried. The Bulbus Allii Macrostemi has various pharmacological effects, and its original plant can be used as both food and medicine. Some students considered that the source of the allium macrostemon plant carried by the traditional herbal is not the root of garlic a. Macrostemon Bunge but the allium macrostemon GDon of liliaceae through the examination of the herbal and the field investigation. At present, the Chinese pharmacopoeia takes the allium macrostemon and the small-root garlic as the source of the traditional Chinese medicine allium macrostemon. The longstamen onion bulb is pungent and bitter in taste and warm in nature, has the effects of activating yang, resolving masses, regulating qi and widening chest, is roar in the famous recipe of the heng dynasty, and uses longstamen onion bulb liquor decoction, the snakegourd longstamen bulb pinellia ternate decoction, the immature bitter orange longstamen onion bulb cassia twig decoction and the like which are recorded in the golden kui kou L ü e, as main drugs, and is used for treating diseases such as heartache and back pain, and is mainly used for treating diseases such as coronary heart disease, angina pectoris and the like clinically.
One of the main active ingredients of the allium macrostemon is steroid saponin, and so far, the total amount of the saponin separated from the allium macrostemon is more than forty. Macrostem onion was systematically studied in Peng army-Peng, wu Yan, jiang Yong and Chen Hai Feng.
At present, the steroid saponin compound monomer is refined by mainly adopting a silica gel column chromatography method, but the production period is long, the reagent waste is serious, and the product yield is low.
In the prior art, 30kg of fresh longstamen onion is crushed into homogenate and is extracted by ethanol with the volume fraction of 98% for 4 times, the liquid-material ratio is respectively 10; diluting with 5 times volume of water, adsorbing with D-101 macroporous resin column, eluting with 40% ethanol, and collecting eluate; concentrating under reduced pressure, detecting by TLC to obtain 125g of furosteroid total saponins, separating the furosteroid total saponins by silica gel column chromatography, performing gradient elution by ethyl acetate-ethanol-water (volume ratio of 5.9 to 0.1-6 46 H 74 O 23 ) Compound 2 (molecular formula C) 38 H 58 O 17 ) Compound 3 (molecular formula C) 57 H 96 O 28 ) Compound 4 (molecular formula C) 44 H 72 O 19 ) Compound 5 (molecular formula C) 43 H 70 O 19 ) And compound 6 (formula C) 33 H 56 O 10 ）。
CN101244190 discloses a method for preparing allium macrostemon total steroid saponin extract and its use, which comprises cutting allium macrostemon bulb into blocks, adding ethanol for extraction or reflux extraction, combining the extracts, concentrating under reduced pressure, standing or centrifuging, taking the lower layer precipitate, diluting with water, extracting with water saturated n-butanol, concentrating under reduced pressure to dryness, dissolving the residue with ethanol, adding acetone until the precipitate is not generated, filtering, and drying to obtain the allium macrostemon total steroid saponin extract. Wherein, the content of total steroidal saponins in the extract is more than 50 percent, the content of 2 compounds in the extract is more than 10 percent, and the structural formula of the compound (1) is as follows:
the structural formula of the compound (2) is as follows:
CN113058001A discloses a preparation method for sensitizing chemotherapy drug extracted from Bulbus Allii Macrostemi, 1) pulverizing and concentrating; 2) Preparing an extract; 3) Preparing the extract. The active micromolecules for improving the chemotherapy sensitivity are extracted from natural products, the effective part for improving the chemotherapy drug sensitivity can be obtained from the whole allium macrostemon, and the pharmacodynamic substance basis is provided for the clinical use of the allium macrostemon and the development of new drugs.
Disclosure of Invention
The invention aims to solve the problems in the prior art and provides a method for extracting and separating steroid saponin compounds from allium macrostemon. In the technical scheme, the allium macrostemon is taken as a raw material, and the steroid saponin compound monomer is finally obtained through 70% methanol extraction, concentration, chromatography, preparative high performance liquid chromatography separation and product recovery. The separation efficiency is high, the production period is short, the process is stable, the operation is simple and convenient, the cost is low, and the high-purity (more than 99 percent of the area normalization method) separation preparation of a large amount of steroid saponin compound monomers can be realized.
In order to achieve the technical purpose, the following technical scheme is proposed:
the technical scheme provides that: a method for extracting and separating steroid saponin compounds from Bulbus Allii Macrostemi, wherein the steroid saponin compounds are (25R) -26-O-beta-D-Glucopyranosyl-5 beta-furostan-3 beta, 22, 26-triol-3-O-beta-D-Glucopyranosyl- (1 → 2) -beta-D-galactopyranoside, CAS number 897386-27-5, and molecular formula C 45 H 76 O 19 Molecular weight 921.07, structural formula as follows:
the method specifically comprises the following steps:
and Y1, extraction: pulverizing Bulbus Allii Macrostemi into 1-4mm coarse powder, adding 70% methanol solution into per kilogram of Bulbus Allii Macrostemi coarse powder, extracting for 1-2 hr for 2-3 times, collecting, mixing, and filtering to obtain extractive solution;
and Y2, concentrating: concentrating the obtained extractive solution at 60 deg.C to obtain concentrated solution;
y3. Chromatography: introducing the obtained concentrated solution into an activated AB-8 macroporous resin column for enrichment, washing with purified water to colorless, then resolving with 70% methanol solution, recovering methanol, and obtaining the residual water solution as chromatographic solution;
and Y4, filtering: filtering the obtained chromatographic solution with a 0.45 μm filter membrane to obtain a filtrate containing steroid saponin compound monomers;
and Y5, separation: injecting the obtained filtrate into a preparative high performance liquid chromatograph, preparing the steroid saponin compound monomer, and collecting the preparation solution of the steroid saponin compound monomer;
wherein, the chromatographic conditions in the preparative high performance liquid chromatograph comprise:
packing of a chromatographic column: a C18 filler;
mobile phase: acetonitrile-0.1% aqueous formic acid V/V =24:76;
sample introduction amount: 1.0-1.5L/needle;
and (3) detection: an online evaporative light detector;
and Y6, product recovery: recovering methanol from the obtained preparation solution of the steroid saponin compound monomer, enriching the residual water solution with C18 and resolving with pure methanol, respectively concentrating at 45-55 deg.C to dry, and drying to obtain the steroid saponin compound monomer with purity of above 99%.
Further, before separation of the preparative high performance liquid chromatograph, the peak shape of the steroid saponin compound monomer in the preparative high performance liquid chromatograph is determined by a liquid-mass combination method, namely, the filtrate obtained in the step Y4 is taken for sample injection, preparation and separation of the steroid saponin compound monomer are carried out, and the corresponding peak shape of the steroid saponin compound monomer in the liquid chromatograph is determined according to a mass spectrum detection result.
The technical scheme also provides that: a method for detecting purity of steroid saponin compounds extracted from Bulbus Allii Macrostemi comprises adopting analytical High Performance Liquid Chromatograph (HPLC);
wherein, the chromatographic conditions in the analytical high performance liquid chromatograph comprise:
packing of a chromatographic column: octadecylsilane chemically bonded silica;
mobile phase: acetonitrile-0.05% formic acid V/V =24:76;
flow rate: 0.7mL/min;
and (3) detection: an evaporative light scattering detector;
column temperature: 35 ℃ is carried out.
By adopting the technical scheme, the beneficial technical effects brought are as follows:
1) According to the invention, the steroid saponin compound monomer is separated and purified by adopting the preparative high performance liquid chromatography system, so that a good separation effect is achieved, the steroid saponin compound monomer is pertinently collected by detecting through the evaporative light scattering detector, the target is clear, the resource waste caused by conventional column chromatography and the like is avoided, the product quality is convenient to control, and the product purity can reach more than 99%;
2) Because high performance liquid chromatography is higher to the requirements such as color and luster, purity of sample, the extract that obtains through simple processing can not directly advance kind, consequently, if before carrying out high performance liquid chromatography separation, do not carry out effectual pretreatment to advancing kind of solution, this not only makes the separation effect not good, but also probably causes serious influence to the instrument (if: shortening the life, etc.), thereby increasing the production cost of the product;
3) The invention aims at the physicochemical properties of various components in the allium macrostemon, carries out proper sequence collocation of the steps Y1-Y3 and proper parameter combination, effectively extracts components such as steroid saponin compound monomers and the like, and removes a large amount of impurities, thereby obtaining a sample solution which can enter a preparative high performance liquid chromatography system without causing great influence on the high performance liquid chromatography system, prolonging the service cycle of the system as much as possible, and saving the production cost;
4) In the high performance liquid chromatography separation process, the selection and combination of various chromatographic conditions are extremely important, and the peak emergence time, the peak shape and the like of the substance components are directly influenced. The invention determines specific chromatographic conditions through a large number of experimental researches and comparative analyses, thereby optimizing the peak emergence time, peak shape, separation effect and the like of the substance components and realizing the high-efficiency separation of the steroid saponin compound monomers.
Drawings
FIG. 1 is an HPLC chromatogram of a steroid saponin compound monomer in the present invention;
FIG. 2 is a diagram showing the results of the specificity test in example 4.
Detailed Description
In the following, the technical solutions in the embodiments of the present invention are clearly and completely described, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
In the following examples, the purity of the steroid saponin compound monomer of the final product is rechecked by adopting an analytical High Performance Liquid Chromatography (HPLC) method;
wherein, the chromatographic conditions in the analytical high performance liquid chromatograph comprise:
packing of a chromatographic column: octadecylsilane chemically bonded silica;
mobile phase: acetonitrile-0.05% formic acid V/V =24:76;
flow rate: 0.7mL/min;
and (3) detection: an evaporative light scattering detector;
column temperature: 35 ℃ is carried out.
Example 1
A method for extracting and separating steroid saponin compounds from allium macrostemon comprises the following process steps:
1) Extraction of
Pulverizing 1Kg of allium macrostemon into 1-4mm coarse powder, adding 10L of 70% methanol solution, extracting for 3 times, each for 2 hours, mixing, and filtering to obtain 36L of extractive solution;
2) Concentrating
Concentrating the extracting solution obtained in the step 1) to a small volume at 60 ℃, and obtaining 7L of concentrated solution;
3) AB-8 macroporous resin column chromatography
Enriching the concentrated solution obtained in the step 2) by using activated AB-8 macroporous resin, and after the enrichment is finished, washing by using 20L of water, and resolving by using 30L of methanol with the volume percentage concentration of 70%; after the methanol is recovered from the analysis solution, the total volume of the obtained concentrated solution is 6.5L;
4) Filtration
Filtering 6.5L of the concentrated solution obtained in the step 3) by using a 0.45-micron filter membrane to obtain 7L of filtrate;
5) High performance preparative liquid chromatography separation
The chromatographic column packing is: a C18 filler;
the mobile phase is as follows: acetonitrile-0.1% formic acid solution V/V =24:76;
sampling the filtrate obtained in the step 4), wherein the sampling amount is 1L per needle, detecting by an online evaporation light detector, and collecting 5L of steroid saponin compound monomers;
6) Product recovery
Recovering methanol from 5L of the steroid saponin compound monomer obtained in the step 6), enriching the residual aqueous solution by using C18, resolving by using methanol, concentrating to dryness at the temperature of 45-55 ℃ to obtain a high-purity solid of the steroid saponin compound, and drying to obtain 0.5g of the steroid saponin compound monomer.
The purity of the product is rechecked by an analytical High Performance Liquid Chromatography (HPLC) method, and the detection result is as follows: 99.25 percent of steroid saponin compound monomer.
Example 2
A method for extracting and separating steroid saponin compounds from allium macrostemon comprises the following process steps:
1) Extraction of
Pulverizing 10Kg of allium macrostemon into 1-4mm coarse powder, adding 110L of 70% methanol solution, extracting for 3 times, each for 2 hours, mixing, filtering to obtain 93L of extractive solution;
2) Concentrating
Concentrating the extracting solution obtained in the step 1) to a small volume at the temperature of 60 ℃ to obtain 12L of concentrated solution;
3) AB-8 macroporous resin column chromatography
Enriching the concentrated solution obtained in the step 2) by using activated AB-8 macroporous resin, and after the enrichment is finished, washing by using 85L of water, and resolving by using 130L of methanol with the volume percentage concentration of 70%; after the methanol is recovered from the analysis solution, 20L of the obtained concentrated solution is obtained;
4) Filtration
Filtering 20L of the concentrate obtained in step 3) with 0.45 μm filter membrane to obtain 21L of filtrate
5) High performance preparative liquid chromatography separation
The chromatographic column packing is: a C18 filler;
the mobile phase is as follows: acetonitrile-0.1% formic acid water solution V/V =24:76;
injecting sample into the filtrate obtained in the step 4), wherein the sample injection amount is 1L per needle, detecting by an online evaporation photodetector, and collecting steroid saponin compounds to prepare 70L of solution;
6) Product recovery
Preparing 70L of solution of the steroid saponin compounds obtained in the step 6), recovering methanol, enriching the residual aqueous solution by using C18, resolving by using methanol, concentrating to dryness at the temperature of 45-55 ℃ to obtain high-purity solid of the steroid saponin compounds, and drying to obtain 5.8g of the steroid saponin compounds.
The purity of the product is rechecked by an analytical High Performance Liquid Chromatography (HPLC) method, and the detection result is as follows: 99.35 percent of steroid saponin compounds.
Example 3
A method for extracting and separating steroid saponin compounds from allium macrostemon comprises the following process steps:
1) Extraction of
Pulverizing 20Kg of allium macrostemon into 1-4mm coarse powder, adding 200L of 70% methanol solution by volume, extracting for 3 times, each for 2 hours, mixing, filtering, and collecting 190L of extractive solution;
2) Concentrating
Concentrating the extracting solution obtained in the step 1) to a small volume at 60 ℃ to obtain 32L of concentrated solution;
3) AB-8 macroporous resin column chromatography
Enriching the concentrated solution obtained in the step 2) by using activated AB-8 macroporous resin, and after the enrichment is finished, washing by using 140L of water, and resolving by using 220L of methanol with the volume percentage concentration of 70%; after the methanol is recovered from the analysis solution, 20L of the obtained concentrated solution is obtained;
4) Filtration
Filtering 20L of the concentrated solution obtained in the step 3) by using a 0.45-micron filter membrane to obtain 22L of filtrate;
5) High performance preparative liquid chromatography separation
The chromatographic column packing is as follows: c18 filler;
the mobile phase is as follows: acetonitrile-0.1% formic acid solution V/V =24:76;
injecting sample into the filtrate obtained in the step 4), wherein the sample injection amount is 1L per needle, detecting by an online evaporation photodetector, and collecting 90L of steroid saponin compound preparation solution;
6) Product recovery
Preparing 90L of steroid saponin compound preparation solution obtained in the step 5), recovering methanol, enriching residual water solution by using C18, resolving by using methanol, concentrating to dryness at the temperature of 45-55 ℃ to obtain high-purity solid of the steroid saponin compound (the related HPLC chromatogram is shown in figure 1), and drying to obtain 13g of the steroid saponin compound.
The purity of the product is rechecked by an analytical High Performance Liquid Chromatography (HPLC) method, and the detection result is as follows: 99.41 percent of steroid saponin compounds.
Example 4
Based on example 3, the content measurement of chemical components in the allium macrostemon is applied to the quality control of the allium macrostemon medicinal material, so as to provide a precondition guarantee for the method for extracting and separating the steroidal saponins from the allium macrostemon. Therefore, researches such as chromatographic condition optimization, method determination, sample extraction method, method verification (specificity investigation, linear relation investigation, precision investigation, sample loading recovery rate and durability investigation), multi-batch sample detection and the like are carried out, and then a preparation method of a corresponding reference substance solution, a preparation method of a sample solution, chromatographic conditions and the like are obtained, wherein a result diagram of the specificity investigation is shown in fig. 2. To further illustrate the technology in the present application.
In the content measurement of chemical components in the allium macrostemon, the related method comprises the following steps:
1) Preparation of control solutions
Precisely weighing appropriate amount of steroid saponin monomer (CAS number: 897386-27-5) as reference, and adding 20% acetonitrile to obtain solution containing 0.4mg per 1 mL;
2) Preparation of test solution
Taking 2.5g of powder of the product (passing through a second sieve), precisely weighing, placing the powder in a conical flask with a plug, precisely adding 100mL of 55% methanol, carrying out ultrasonic treatment (power of 360W and frequency of 40 kHz) for 40min, cooling, shaking up, filtering, concentrating the filtrate under reduced pressure to dryness, precisely adding 25mL of 20% acetonitrile into residues, sealing the plug, weighing, carrying out ultrasonic treatment (power of 360W and frequency of 40 kHz) for 15min for dissolving, cooling, weighing again, complementing the weight loss by 20% acetonitrile, shaking up, filtering, and taking the subsequent filtrate;
3) Chromatographic conditions and System suitability test
Octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile-0.1% formic acid (24: 76) as the mobile phase; detecting by an evaporative light scattering detector at a flow rate of 0.7ml/min and a column temperature of 35 ℃;
4) Assay method
And respectively and precisely sucking 10 muL and 20 muL of the control solution and 20 muL of the test solution, injecting the test solution into a liquid chromatograph, measuring, and calculating by using an external standard two-point method logarithmic equation to obtain the test solution.
Claims (4)
1. A method for extracting and separating steroid saponin compounds from allium macrostemon is characterized in that the name of the steroid saponin compounds is
(25R) -26-O-beta-D-Glucopyranosyl-5 beta-furostan-3 beta, 22, 26-triol-3-O-beta-D-Glucopyranosyl- (1 → 2) -beta-D-galactopyranoside, CAS number 897386-27-5, molecular formula C 45 H 76 O 19 Molecular weight 921.07, structural formula as follows:
the method comprises the following steps:
and Y1, extraction: pulverizing Bulbus Allii Macrostemi into 1-4mm coarse powder, adding 70% methanol solution into per kilogram of Bulbus Allii Macrostemi coarse powder, extracting for 1-2 hr for 2-3 times, collecting, mixing, and filtering to obtain extractive solution;
and Y2, concentrating: concentrating the obtained extractive solution at 60 deg.C to obtain concentrated solution;
y3. Chromatography: introducing the obtained concentrated solution into an activated AB-8 macroporous resin column for enrichment, washing with purified water to colorless, then resolving with 70% methanol solution, recovering methanol, and obtaining the residual water solution as chromatographic solution;
and Y4, filtering: filtering the obtained chromatographic solution with a filter membrane of 0.45 μm to obtain a filtrate containing steroid saponin compound monomers;
and Y5, separation: injecting the obtained filtrate into a preparative high performance liquid chromatograph, preparing the steroid saponin compound monomer, and collecting the preparation solution of the steroid saponin compound monomer;
wherein, the chromatographic conditions in the preparative high performance liquid chromatograph comprise:
packing of a chromatographic column: c18 filler;
mobile phase: acetonitrile-0.1% aqueous formic acid V/V =24:76;
sample introduction amount: 1.0-1.5L/needle;
and (3) detection: an online evaporative light detector;
and Y6, product recovery: recovering methanol from the obtained preparation solution of the steroid saponin compound monomer, enriching the residual water solution with C18 and resolving with pure methanol, concentrating at 45-55 deg.C respectively to dry, and drying to obtain the steroid saponin compound monomer.
2. The method for extracting and separating the steroid saponin compounds from the allium macrostemon according to claim 1, wherein before the separation by the preparative high performance liquid chromatograph, the peak shape of the steroid saponin compound monomer in the preparative high performance liquid chromatograph is determined by a liquid-mass combination method, which specifically comprises:
and (5) sampling the filtrate obtained in the step Y4, preparing and separating the steroid saponin compound monomer, and determining the corresponding peak shape of the steroid saponin compound monomer in the liquid chromatogram according to the mass spectrum detection result.
3. The method for extracting and separating the steroid saponin compounds from the allium macrostemon according to claim 1, wherein the purity of the steroid saponin compound monomer is more than 99%.
4. The method for detecting the purity of the macrostem onion extracted and separated from the steroid saponin compounds as claimed in claim 1, 2 or 3, wherein the method for detecting the purity of the steroid saponin compound monomer adopts an analytical high performance liquid chromatograph;
wherein, the chromatographic conditions in the analytical high performance liquid chromatograph comprise:
packing of a chromatographic column: octadecylsilane chemically bonded silica;
mobile phase: acetonitrile and 0.05% formic acid, the volume ratio of acetonitrile to 0.05% formic acid being 24:76;
flow rate: 0.7mL/min;
and (3) detection: an evaporative light scattering detector;
column temperature: at 35 deg.c.
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| CN116559334A (en) * | 2023-06-16 | 2023-08-08 | 吉林省东方制药有限公司 | Quantitative detection method of alliin in blood stagnation removing capsule |
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| CN101244190A (en) * | 2008-01-31 | 2008-08-20 | 浙江大学 | Method of preparing allium macrostemon total sterides saponin extract and uses thereof |
| CN112300242A (en) * | 2020-10-26 | 2021-02-02 | 吉林大学 | Preparation method of furostanol saponin compound monomer |
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| CN101244190A (en) * | 2008-01-31 | 2008-08-20 | 浙江大学 | Method of preparing allium macrostemon total sterides saponin extract and uses thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116559334A (en) * | 2023-06-16 | 2023-08-08 | 吉林省东方制药有限公司 | Quantitative detection method of alliin in blood stagnation removing capsule |
| CN116559334B (en) * | 2023-06-16 | 2023-10-24 | 吉林省东方制药有限公司 | Quantitative detection method of alliin in blood stagnation removing capsule |
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