CN115326965B - Quality control method of Shenmu tranquilization capsule for improving deep sleep - Google Patents

Quality control method of Shenmu tranquilization capsule for improving deep sleep Download PDF

Info

Publication number
CN115326965B
CN115326965B CN202210981251.7A CN202210981251A CN115326965B CN 115326965 B CN115326965 B CN 115326965B CN 202210981251 A CN202210981251 A CN 202210981251A CN 115326965 B CN115326965 B CN 115326965B
Authority
CN
China
Prior art keywords
acid
content
solution
saponin
shenmu
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.)
Active
Application number
CN202210981251.7A
Other languages
Chinese (zh)
Other versions
CN115326965A (en
Inventor
李燕
雷晓林
张春礼
王海珍
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhouzhi Qingbao Pharmaceutical Co ltd
Original Assignee
Zhouzhi Qingbao Pharmaceutical Co ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Zhouzhi Qingbao Pharmaceutical Co ltd filed Critical Zhouzhi Qingbao Pharmaceutical Co ltd
Priority to CN202210981251.7A priority Critical patent/CN115326965B/en
Publication of CN115326965A publication Critical patent/CN115326965A/en
Application granted granted Critical
Publication of CN115326965B publication Critical patent/CN115326965B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • 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
    • 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
    • G01N2030/022—Column chromatography characterised by the kind of separation mechanism
    • G01N2030/027—Liquid chromatography
    • 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
    • G01N2030/062—Preparation extracting sample from raw material
    • 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
    • G01N2030/065—Preparation using different phases to separate parts of sample

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Steroid Compounds (AREA)

Abstract

The application relates to the technical field of quality control of Shenmu tranquilization capsules, and particularly discloses a quality control method of Shenmu tranquilization capsules for improving deep sleep. The quality control method comprises the following steps of detecting the content of the wild jujube seed saponin A in the content of the Shenmu Anshen capsule: s1, adding acid liquor into the content of the product to obtain acidolysis solution, wherein the acid liquor contains micromolecular organic acid and inorganic acid; s2, adding 50-70Vol.% ethanol solution into acidolysis solution, and fully vibrating and extracting; s3, extracting 50-70Vol.% ethanol solution in a split phase, evaporating the solvent, adding water into the residue for dissolution, and extracting with water saturated n-butanol; s4, removing the solvent, adding methanol into the residue for redissolution to obtain sample liquid to be detected, and detecting the content of the wild jujube seed saponin A by using a high performance liquid chromatography. The detection method has the advantage of accurately detecting the content of the wild jujube natural saponin A.

Description

Quality control method of Shenmu tranquilization capsule for improving deep sleep
Technical Field
The application relates to the technical field of quality control of Shenmu Anshen capsules, in particular to a quality control method of Shenmu Anshen capsules for improving deep sleep.
Background
The Shenmu Anshen capsule is a capsule product with the function of improving deep sleep, which is prepared from semen Platycladi, rhizoma Cyperi, rehmanniae radix, parched semen Ziziphi Spinosae, fructus Lycii, magnetitum preparata, os Draconis, concha Ostreae, fructus Schisandrae chinensis, cortex et radix Polygalae, borneolum Syntheticum, fructus crataegi and parched Massa Medicata Fermentata as main contents. The preparation method of the shenmu tranquilization capsule comprises the following steps: pulverizing calcined Magnetitum, os Draconis and Concha Ostreae together into fine powder, and sieving to obtain first mixed fine powder; pulverizing semen Platycladi, rhizoma Cyperi, rehmanniae radix, parched semen Ziziphi Spinosae, fructus Lycii, fructus Schisandrae chinensis, cortex et radix Polygalae, fructus crataegi and parched Massa Medicata Fermentata into fine powder, mixing with the first mixed fine powder, and sieving to obtain second mixed fine powder; grinding Borneolum Syntheticum, mixing with the second mixed fine powder, sieving, mixing, and making into capsule to obtain SHENMUSHEN Capsule. When the quality control is carried out on the product, since the wild jujube saponin A (from the stir-fried wild jujube seeds) is the main raw material for realizing the tranquillizing effect, the accurate detection of the content of the wild jujube saponin A is necessary.
The method for detecting the content of the wild jujube saponin A generally comprises the steps of degreasing, extracting and detecting; the organic solvent used in degreasing operation is diethyl ether, methanol, ethanol, etc., the organic solvent used in extraction can be water unsaturated solution or water saturated solution of n-butanol, and high performance liquid chromatography or spectrophotometry can be used for component detection.
However, the above-mentioned detection method has a problem of inaccurate detection results, and the main reason is that the separation and purification of the jujuboside a are insufficient in the early stage in the detection process, so that the detection results of the substances are inaccurate.
Disclosure of Invention
In order to improve the detection accuracy of the wild jujube seed saponin A, the application provides a quality control method of the Shenmu Anshen capsule for improving deep sleep.
The quality control method of the Shenmu tranquilization capsule for improving deep sleep provided by the application adopts the following technical scheme: the quality control method of the Shenmu tranquilization capsule for improving deep sleep comprises the following steps of detecting the content of the wild jujube seed saponin A in the content of the Shenmu tranquilization capsule, wherein the quality control method comprises the following steps of:
s1, acidolysis: adding acid liquor into the content of the product, and reacting to obtain acidolysis solution, wherein the weight-volume ratio of the content to the acid liquor is 0.21-0.46g/mL, and the acid liquor contains micromolecular organic acid and inorganic acid;
s2, phase separation extraction: adding 50-70Vol.% ethanol solution into acidolysis solution, and fully vibrating and extracting, wherein the weight-volume ratio of the content to the saturated 50-70Vol.% ethanol solution is 0.53-0.82g/mL;
s3, secondary extraction: taking 50-70Vol.% ethanol solution after split-phase extraction, evaporating the solvent, adding water into the residue for dissolution, and then extracting with water saturated n-butanol;
s4, preparing sample liquid and detecting: removing solvent in water saturated n-butanol extractive solution, adding methanol into residue, and redissolving to obtain sample solution to be detected, and detecting the content of semen Ziziphi Spinosae saponin A by high performance liquid chromatography.
In the scheme, the meaning of the weight-volume ratio of the content to the acid liquor in the step S1 is 0.21-0.46 g/mL: the weight of the corresponding content of each milliliter of acid liquor is 0.21-0.46 g.
In the quality control method of the Shenmu tranquilization capsule for improving the deep sleep, the effect of improving the deep sleep is only that of the Shenmu tranquilization capsule; the quality control method aims at the quality control method of the product of the shenmu tranquilization capsule.
The chemical components of the platycladi seed in the nerve-soothing capsule mainly comprise fatty oil, volatile oil, saponin and other components, and also comprise sitosterol, cypress alcohol, pinacolide and other compounds; the nutgrass galingale rhizome contains volatile essential oil, and the ingredients mainly comprise nutgrass galingale rhizome essence, fatty acid and the like; the main ingredients of rehmanniae radix are iridoid and its glycosides, saccharides and glycosides, amino acids, and various microelements etc.; the main components of the wild jujube seed saponin comprise fatty oil, flavonoid, triterpenes, alkaloids and organic acid, the main compounds of the wild jujube seed saponin comprise wild jujube seed saponin A, B, the flavonoid components are apigenin Huang Tongtan glycoside compounds, and the spinosin is the main flavonoid component of the wild jujube seed; the main components in the medlar are medlar polysaccharide, betaine and pigment; the main component of the magnet is ferroferric oxide, and contains impurities such as silicon, lead, titanium, phosphorus, manganese, calcium, chromium, barium, saw, magnesium and the like; the main components of Os Draconis are calcium carbonate and calcium phosphate, and also contain iron, potassium, sodium, chlorine, sulfate radical, etc.; the main components of the schisandra chinensis are schisandrin, vitamin C, resin, tannin, sugar and the like; the main chemical components of the polygala tenuifolia are organic matters and inorganic matters such as saponins, xanthone, oligosaccharide ester, alkaloid and the like; the borneol is mainly composed of D-borneol, and contains sesquiterpenes such as rhynchophylla, beta-elemene, caryophyllene and the like, and triterpene compounds such as oleanolic acid, maizhu acid, asiatic acid, borneol aromatic alcohol, lithocarpol and the like; the main components of the hawthorn are vitamin C, minerals, flavonoids, tannins, saccharides, organic acids, trace elements and the like.
When the content is directly subjected to the content detection of the wild jujube saponin A, the following steps are found: because the content contains complex and various components and contains a large amount of hydrophilic and lipophilic substances, the content of the spine date seed saponin A in the Shenmu Anshen capsule is influenced by a large amount of impurities in the substances. The main reason is that: when the content of the wild jujube seed saponin A in the shen mu tranquillizing capsule is extracted, the impurity substances can influence the distribution of the wild jujube seed saponin A in an organic phase, so that the wild jujube seed saponin A is difficult to completely extract, and the content of the wild jujube seed saponin A obtained by detection is low, namely the final detection result is inaccurate.
By adopting the technical scheme, firstly, by adding acid liquor, inorganic matters (water-soluble and water-insoluble, calcium carbonate, ferric oxide and the like) from magnetite and fossil fragments in the content can generate soluble salt (calcium salt, ferric salt and the like) under the action of the acid liquor, so that the negative influence on the full extraction of the spine date seed saponin A caused by the interaction (electrostatic adsorption, ion interaction, van der Waals force and the like) of salt ions and spine date seed saponin A is reduced as much as possible; meanwhile, flavonoid compounds can be precipitated and separated out due to an acidic environment, so that impurity separation is realized. Secondly, after 50-70vol.% of the ethanol solution is added, the acid solution after the reaction is a salt solution because a large amount of ions (metal ions, inorganic acid ions, organic acid ions) are also present in the acid solution after the reaction. The applicant found that the salt solution, when mixed with 50-70vol.% ethanol solution, spontaneously phase separated (i.e. the phase separation is stratified), forming a biphasic extraction system. At this time, the spinosa saponin A tends to be distributed in 50-70vol.% of the ethanol phase, and the impurities such as water-soluble proteins and polysaccharides tend to be distributed in the phase where the salt solution is located. The step can realize the separation of saponin, protein and polysaccharide. It should be noted here that the solute in the acid solution is not only an inorganic acid, but also a small molecular organic acid, because: 1. sufficient acid radical ions can fully react with inorganic impurities (calcium carbonate, ferric oxide and the like) to generate salt, namely salt solution is formed, so that the salt solution and 50-70Vol.% ethanol solution are matched to form a biphasic separation system in the later stage; 2. the organic acid radical formed after the reaction of the organic acid and the inorganic impurities exists in the salt solution, and because the main solvent of the phase is the salt solution, a large amount of polysaccharide, water-soluble protein and the like are distributed in the phase, in addition, because of the existence of the organic acid radical, the polarity of the phase is further improved, so that part of the weakly polar impurities (organic matters such as part of the weakly polar proteins) can be distributed in the phase, and the more sufficient separation of the polysaccharide, the water-soluble protein and other impurities and the wild jujube saponin A is realized.
In the scheme, inorganic salt is not additionally added, but the particularity of the content of the Shenmu tranquilization capsule is utilized, inorganic impurities are removed, a salt solution is generated at the same time, and the salt solution and 50-70Vol.% ethanol solution are mixed in a proper proportion to form a specific aqueous two-phase extraction system, so that the effective separation of the wild jujube saponin A and protein and polysaccharide impurities is improved, and the influence of the impurities on a detection result is obviously reduced. Therefore, by adopting the scheme, the detection result of the wild jujube saponin A is more accurate by fully extracting the wild jujube saponin A.
In addition, in the scheme, the relative dosage of the content, the acid liquor and the 50-70Vol.% ethanol solution is required to be noted, when the relative dosage of the acid liquor and the 50-70Vol.% ethanol solution is not in the dosage range, a split-phase extraction system is difficult to form 1, and further, the sufficient extraction and more accurate detection of the spina date seed saponin A are difficult to realize; or 2, a split-phase extraction system is formed, but the distribution amount of the wild jujube seed saponin A in 50-70Vol.% ethanol phase is less, so that the purposes of more fully extracting the wild jujube seed saponin A and more accurately detecting cannot be realized.
Optionally, the mass fraction of the small molecular organic acid in the acid liquor is 2-5%, and the mass fraction of the inorganic acid in the acid liquor is 5-10%.
Optionally, the small molecule organic acid is selected from any one or more of citric acid, formic acid, acetic acid, propionic acid and malic acid; the inorganic acid is sulfuric acid.
By adopting the technical scheme, the cooperation of the organic acid and the inorganic acid provides an acidic environment together, so that inorganic matters (water-soluble and water-insoluble, calcium carbonate, ferric oxide and the like) from magnetite and fossil fragments in the content are generated into soluble salts (calcium sulfate, ferric sulfate and the like) under the action of the acid as much as possible, and then a salt solution separated from 50-70Vol.% ethanol solution can be obtained during phase separation extraction. The small molecular organic acid with the dosage is selected and matched with the inorganic acid with the dosage, firstly, the acid environment provided by the acid liquor does not destroy the spinosa kernel saponin A in a large amount, but the purpose of providing a salt solution environment can be realized; in addition, the small molecular organic acid with the dosage can also achieve the aim of remarkably improving the polarity of acidolysis solution, and contributes to the distribution of partial weak-polarity impurities (organic matters such as partial weak-polarity proteins) in the acidolysis solution in the step S3, and further contributes to the full separation of polysaccharide, water-soluble proteins and other impurities and saponins.
Optionally, the small molecule organic acid is selected from any one or more of citric acid, acetic acid and propionic acid.
By adopting the technical scheme, when the three small molecular organic acids are adopted, the content of the finally detected spine date seed saponin A is higher, namely the detection result is more accurate.
Optionally, in the step S2, 50-70Vol.% of ethanol is added into acidolysis solution, and meanwhile, a distribution promoting agent with the weight percent of 6-15% of acid liquor is added; the distribution promoting agent is polyacrylic acid or polymethacrylic acid.
By adopting the technical scheme, polyacrylic acid or polymethacrylic acid is an amphiphilic polymer sensitive to pH, and is carboxyl containing a weak organic acid polymer substituent. Under acidic conditions, it appears to accept protons; in neutral and alkaline environments, it appears to give protons. Both polyacrylic acid and polymethacrylic acid are dissolved in water, and therefore, are present in acidolysis solution after addition. The applicant finds that the addition of the amphiphilic polymer can interact with small molecular organic acid to obviously change the distribution behavior of flavonoid impurities so as to further separate saponin from flavone, thereby further reducing the influence of the flavone on the detection accuracy of the wild jujube seed saponin A and further improving the detection accuracy of the wild jujube seed saponin A.
Optionally, the weight ratio of the small molecule organic acid to the partitioning agent is (0.2-0.5): 1.
Optionally, the degreasing solvent is selected from any one of 50-70% ethanol solution and diethyl ether.
Optionally, in step S3, the volume ratio of water to water saturated n-butanol solution is 1 (0.8-1.2).
Optionally, in step S4, the content of the wild jujube seed saponin a is detected by high performance liquid chromatography; the detection column is prepared by taking octadecylsilane chemically bonded silica as a filler; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and gradient elution is carried out.
Optionally, the content of the shenmu tranquilization capsule comprises the following raw materials in parts by weight:
10-25 parts of platycladi seed, 3-15 parts of nutgrass galingale rhizome, 10-25 parts of rehmannia root, 20-35 parts of stir-fried spina date seed, 10-25 parts of medlar, 10-25 parts of calcined magnetite, 10-25 parts of dragon bone, 10-25 parts of oyster, 10-25 parts of shizandra berry, 10-25 parts of polygala tenuifolia, 3-15 parts of borneol, 20-35 parts of hawthorn and 30-45 parts of stir-fried medicated leaven.
In summary, the application has the following beneficial effects:
1. the content of the wild jujube seed is more, except that the fried wild jujube seed also comprises twelve ingredients of platycladi seed, nutgrass galingale rhizome, rehmannia root, medlar, calcined magnetite, dragon bone, oyster, chinese magnoliavine fruit, thinleaf milkwort root-bark, borneol, hawthorn and fried medicated leaven, and the problem that the detection result is lower when the content of the wild jujube seed saponin A is detected by the existing method; therefore, the scheme is based on the problem that the content is firstly treated by acid liquor, and the split-phase extraction is realized by combining 50-70Vol.% ethanol solution, so that the influence of other impurities on the detection of the content of the wild jujube seed saponin A is eliminated as much as possible.
2. In the application, the acid liquor contains micromolecular organic acid and inorganic acid at the same time, and the inorganic acid is mainly used for promoting the formation of a phase-splitting system with ethanol solution; the small molecular organic acid is mainly used for changing the polarity of the salt solution so as to promote the separation of impurities and the spine date seed saponin.
3. The application further adds a distribution promoting agent to separate flavone from the wild jujube saponin A, thereby further removing impurities.
Detailed Description
The present application will be described in further detail with reference to examples.
High performance liquid chromatography detection method for wild jujube saponin A
When the method is used for detecting the content of the wild jujube seed saponin A in the content of the Shenmu Anshen capsule, the content is detected by a high performance liquid chromatography.
The detection column uses octadecylsilane chemically bonded silica gel as a filler; acetonitrile is taken as a mobile phase A, and water is taken as a mobile phase B; detecting by an evaporative light scattering detector; the theoretical plate number is not less than 2000 calculated according to the peak of the wild jujube seed saponin A.
In addition, gradient elution is adopted, and the gradient elution comprises the following procedures: within 0-10min, mobile phase A increased from 19% to 38% at constant speed, and mobile phase B decreased from 81% to 62% at constant speed; mobile phase a remained at 38% and mobile phase B remained at 62% over 10-25 min; the mobile phase A is increased from 38% to 65% at a constant speed and the mobile phase B is decreased from 62% to 35% at a constant speed within 25-30 min; in 30-35min, mobile phase A increased from 65% to 100% at constant speed, and mobile phase B decreased from 35% to 0% at constant speed.
Preparation example of Shenmu Anshen Capsule
The shenmu tranquilization capsule prescription comprises: 10-25 parts of platycladi seed, 3-15 parts of nutgrass galingale rhizome, 10-25 parts of rehmannia root, 20-35 parts of stir-fried spina date seed, 10-25 parts of medlar, 10-25 parts of calcined magnetite, 10-25 parts of dragon bone, 10-25 parts of oyster, 10-25 parts of shizandra berry, 10-25 parts of polygala tenuifolia, 3-15 parts of borneol, 20-35 parts of hawthorn and 30-45 parts of stir-fried medicated leaven.
The thirteen ingredients are crushed into fine powder together with calcined magnetite, dragon bone and oyster, and the fine powder is sieved to obtain first mixed fine powder; pulverizing semen Platycladi, rhizoma Cyperi, rehmanniae radix, parched semen Ziziphi Spinosae, fructus Lycii, fructus Schisandrae chinensis, cortex et radix Polygalae, fructus crataegi and parched Massa Medicata Fermentata into fine powder, mixing with the first mixed fine powder, and sieving to obtain second mixed fine powder; grinding Borneolum Syntheticum, mixing with the second mixed fine powder, sieving, mixing, and making into capsule to obtain SHENMUSHEN Capsule.
Preparation example 1
Shenmu tranquilization capsule prescription: 18g of platycladi seed, 9g of nutgrass galingale rhizome, 18g of rehmannia root, 27g of stir-fried spina date seed, 18g of medlar, 18g of calcined magnetite, 18g of dragon bone, 18g of oyster, 18g of Chinese magnoliavine fruit, 18g of polygala tenuifolia, 9g of borneol, 27g of hawthorn and 36g of stir-fried medicated leaven. In the prescription, the fried wild jujube accounts for 10.71 weight percent of the thirteen medicines.
The thirteen ingredients are crushed into fine powder together with calcined magnetite, dragon bone and oyster, and the fine powder is sieved by a 80-mesh sieve to obtain first mixed fine powder; pulverizing semen Platycladi, rhizoma Cyperi, rehmanniae radix, parched semen Ziziphi Spinosae, fructus Lycii, fructus Schisandrae chinensis, cortex et radix Polygalae, fructus crataegi and parched Massa Medicata Fermentata into fine powder, mixing with the first mixed fine powder, and sieving with 80 mesh sieve to obtain second mixed fine powder; grinding Borneolum Syntheticum, mixing with the second mixed fine powder, sieving with 80 mesh sieve, mixing, and making into capsule to obtain 1000 granule.
Examples
Example 1
A quality control method of SHENMUANSHEN Capsule for improving deep sleep comprises detecting the content of semen Ziziphi Spinosae saponin A in SHENMUANSHEN Capsule, and comprises the following steps:
s1, acidolysis: precisely weighing 10.05g of the content of the product, placing the product in a conical flask, then adding 48mL of acid liquor, and carrying out oscillation reaction for 5min to obtain acidolysis solution; the acid liquor is specifically as follows: the acid liquor contains 2wt% of acetic acid and 5wt% of sulfuric acid.
S2, phase separation extraction: and adding 19mL of 50Vol.% ethanol solution into the acidolysis solution, sufficiently shaking and extracting, and standing for 5min.
S3, secondary extraction: after the phase separation extraction, 50vol.% ethanol solution was taken, the solvent was evaporated to dryness, 30mL of water was added to the residue for dissolution, and then 24mL of water-saturated n-butanol was used for extraction.
S4, preparing sample liquid and detecting: removing the solvent in the water saturated n-butanol extracting solution, adding methanol into the residue for redissolution, and filtering the redissolution to obtain sample liquid to be detected; taking 10 mu L of sample liquid to be detected, adding the sample liquid into a high performance liquid chromatograph, and detecting the content of the wild jujube seed saponin A. The column, mobile phase and elution procedure for high performance liquid chromatography are as described above.
Example 2
A quality control method of SHENMUANSHEN Capsule for improving deep sleep comprises detecting the content of semen Ziziphi Spinosae saponin A in SHENMUANSHEN Capsule, and comprises the following steps:
s1, acidolysis: 9.95g of the content of the product is precisely weighed and placed in a conical flask, 22mL of acid liquor is added, and after shaking reaction is carried out for 5min, acidolysis solution is obtained; the acid liquor is specifically as follows: the acid solution contains 5wt% of formic acid and 10wt% of sulfuric acid.
S2, phase separation extraction: 70Vol.% ethanol solution 12mL is added into acidolysis solution, and the solution is fully vibrated and extracted and then is left stand for 5min.
S3, secondary extraction: after the phase separation extraction, 70vol.% ethanol solution was taken, the solvent was evaporated to dryness, 30mL of water was added to the residue for dissolution, and then 36mL of water-saturated n-butanol was used for extraction.
S4, preparing sample liquid and detecting: removing the solvent in the water saturated n-butanol extracting solution, adding methanol into the residue for redissolution, and filtering the redissolution to obtain sample liquid to be detected; taking 10 mu L of sample liquid to be detected, adding the sample liquid into a high performance liquid chromatograph, and detecting the content of the wild jujube seed saponin A. The column, mobile phase and elution procedure for high performance liquid chromatography are as described above.
Example 3
A quality control method of SHENMUANSHEN Capsule for improving deep sleep comprises detecting the content of semen Ziziphi Spinosae saponin A in SHENMUANSHEN Capsule, and comprises the following steps:
s1, acidolysis: precisely weighing 10.02g of the content of the product, placing the product in a conical flask, then adding 29mL of acid liquor, and carrying out shaking reaction for 5min to obtain acidolysis solution; the acid liquor is specifically as follows: the acid liquor contains 3.6wt% of citric acid and 8.4wt% of sulfuric acid.
S2, phase separation extraction: and adding 15mL of 65Vol.% ethanol solution into the acidolysis solution, sufficiently shaking and extracting, and standing for 5min.
S3, secondary extraction: after the phase separation extraction, 65vol.% ethanol solution was taken, the solvent was evaporated to dryness, 30mL of water was added to the residue for dissolution, and then 30mL of water-saturated n-butanol was used for extraction.
S4, preparing sample liquid and detecting: removing the solvent in the water saturated n-butanol extracting solution, adding methanol into the residue for redissolution, and filtering the redissolution to obtain sample liquid to be detected; taking 10 mu L of sample liquid to be detected, adding the sample liquid into a high performance liquid chromatograph, and detecting the content of the wild jujube seed saponin A. The column, mobile phase and elution procedure for high performance liquid chromatography are as described above.
Example 4
The difference between this example and example 3 is that when the content of the spinosad A in the content of the Shenmu Anshen capsule is detected, acetic acid is selected as the small molecular organic acid in the acid liquor in the step S1, the acetic acid content in the acid liquor is 3.6wt0.637%, and the rest is the same as in example 3.
Example 5
The difference between this example and example 3 is that propionic acid is selected as the small-molecule organic acid in the acid solution in step S1 when the content of the spinosad A in the content of the shen mu tranquilization capsule is detected, and the propionic acid content in the acid solution is 3.6wt% and the rest is the same as in example 3.
Example 6
The difference between this example and example 3 is that formic acid is selected as the small-molecule organic acid in the acid solution in step S1 when the content of the wild jujube seed saponin A in the content of the Shenmu Anshen capsule is detected, the content of the formic acid in the acid solution is 3.6wt%, and the rest is the same as example 3.
Example 7
The difference between this example and example 3 is that when the content of the spinosad A in the content of the Shenmu Anshen capsule is detected, the small molecular organic acid in the acid liquor in the step S1 is selected to be malic acid, the content of malic acid in the acid liquor is 3.6wt%, and the rest is the same as in example 3.
Example 8
The difference between this example and example 5 is that, when the content of the spinosad A in the content of Shenmu Anshen capsule is detected, in step S2, 65Vol.% ethanol solution is added to the acidolysis solution, and at the same time, polyacrylic acid with an acid solution of 10 wt.% is added, and the rest is the same as in example 5.
That is, in this scheme, step S2 is:
s2, phase separation extraction: 15mL of 65Vol.% ethanol solution is added into acidolysis solution, 4.1g of polyacrylic acid (the weight of acid liquor is about 41 g) is added, and the solution is fully vibrated and extracted and then is left stand for 5min.
Example 9
The difference between this example and example 8 is that, when the content of the spinosad A in the content of Shenmu Anshen capsule is detected, in step S2, 65Vol.% ethanol solution is added to the acidolysis solution, and at the same time, 10 wt.% polymethacrylic acid is added to the acidolysis solution, and the rest is the same as in example 8.
That is, in this scheme, step S2 is:
s2, phase separation extraction: to the acidolysis solution, 15mL of 65vol.% ethanol solution was added, followed by 4.1g of polymethacrylic acid (weight of the acid solution: about 41 g), followed by sufficient shaking extraction and standing for 5min.
Comparative example
Comparative example 1
A quality control method of SHENMUANSHEN Capsule for improving deep sleep comprises detecting the content of semen Ziziphi Spinosae saponin A in SHENMUANSHEN Capsule; the method for detecting the content of the wild jujube seed saponin A in the content of the shenmu tranquilization capsule comprises the following steps: s1, primary extraction: 10.05g of the content of the product is precisely weighed, placed in an conical flask, then 100mL of 50Vol.% ethanol solution is added, heated and refluxed in a water bath for 2 hours, cooled, then the volume loss is complemented by the 50Vol.% ethanol solution, and filtered.
S2, secondary extraction: 50mL of filtrate was taken, the solvent was evaporated, 30mL of water was added to the residue for dissolution, and then the mixture was extracted twice with 30mL of water-saturated n-butanol with shaking.
S3, preparing sample liquid and detecting: removing the solvent in the water saturated n-butanol extracting solution, adding methanol into the residue for redissolution, and filtering the redissolution to obtain sample liquid to be detected; taking 10 mu L of sample liquid to be detected, adding the sample liquid into a high performance liquid chromatograph, and detecting the content of the wild jujube seed saponin A. The column, mobile phase and elution procedure for high performance liquid chromatography are as described above.
Comparative example 2
The difference between this comparative example and comparative example 1 is that in step S1, 50vol.% of the ethanol solution is replaced with an equal volume of 70vol.% of the ethanol solution, otherwise identical to comparative example 1.
Specifically, step S1 is as follows:
s1, primary extraction: 10.04g of the content of the product is precisely weighed, placed in an conical flask, then 100mL of 70Vol.% ethanol solution is added, heated and refluxed in a water bath for 2 hours, cooled, then the volume loss is complemented by 70Vol.% ethanol solution, and filtered.
Comparative example 3
The difference between this comparative example and example 3 is that in the step S1, only 12wt% sulfuric acid was contained in the acid solution when the content of the zizyphus jujube saponins A in the content of the Shenmu Anshen capsule was detected, and the same as in example 3 was followed.
The step S1 in this comparative example is specifically:
s1, acidolysis: precisely weighing 10.02g of the content of the product, placing the product in a conical flask, then adding 29mL of acid liquor, and carrying out shaking reaction for 5min to obtain acidolysis solution; the acid liquor is sulfuric acid solution with the mass fraction of 12%.
Detection method
1. Content detection of wild jujube saponin A
1.1 Standard Curve drawing
Accurately weighing 10mg of semen Ziziphi Spinosae saponin A (C) 58 H 94 O 26 ) Dissolving the standard substance in a 50mL volumetric flask with methanol, fixing the volume, and shaking uniformly to obtain the standard solution. Then accurately sucking a proper amount of standard solution, diluting with methanol, preparing standard working solutions with mass concentrations of 10, 25, 50, 100 and 150 mug/mL respectively, measuring according to chromatographic conditions, taking the mass concentration (mug/mL) of the spina date seed saponin A as an abscissa, taking the corresponding peak area as an ordinate, and drawing a standard curve, wherein the obtained regression equation is as follows: y=0.0003026x+10.5321, r 2 ＝0.9983。
1.2 methodology investigation
1.2.1 precision test
Taking the sample liquid to be detected prepared in the embodiment 8 as a sample liquid, and continuously sampling for 6 times according to chromatographic conditions, wherein the result shows that: the relative peak area of each sample has an RSD of 1.13% and less than 3%, which proves that the experimental precision is good.
1.2.2 repeatability test
Taking sample liquid to be detected prepared in the embodiment 8 as a sample liquid, taking 6 parts of sample liquid respectively, and respectively sampling according to chromatographic conditions, wherein the result shows that: the RSD of each sample relative to the peak area was 2.06%, less than 3%, demonstrating good reproducibility of the experiment.
1.2.3 stability test
Taking the sample liquid to be detected prepared in the example 8 as a test liquid, and respectively measuring at 8 different time points of 0, 1, 2, 4, 8, 12, 18 and 24 hours according to chromatographic conditions, wherein the result shows that: the RSD of each sample relative to the peak area was 1.64% and less than 3%, demonstrating good stability of the experiment.
1.3 detection results of the content of Ziziphus Spinosae semen saponin A in different embodiments
In the examples and the comparative examples, the weight of the content of the Shenmu Anshen capsule is 10+/-0.05 g, and the weight of the content of the Shenmu Anshen capsule is 10.71wt percent of the fried spina date seed; namely, the weight of the stir-fried spine date seed in the content of the Shenmu tranquilization capsule is 1.07+/-0.005 g.
Preparation method of control detection sample one:
(1) Primary extraction: 1.07g of fried spina date seeds are precisely weighed, placed in an conical flask, then added with 100mL of 50Vol.% ethanol solution, heated in a water bath for refluxing for 2 hours, cooled, then complemented with volume loss by the 50Vol.% ethanol solution, and filtered.
(2) And (3) secondary extraction: 50mL of filtrate was taken, the solvent was evaporated, 30mL of water was added to the residue for dissolution, and then the mixture was extracted twice with 30mL of water-saturated n-butanol with shaking.
(3) Preparing sample liquid and detecting: removing the solvent in the water saturated n-butanol extracting solution, adding methanol into the residue for redissolution, and filtering the redissolution to obtain sample liquid to be detected; taking 10 mu L of sample liquid to be detected, adding the sample liquid into a high performance liquid chromatograph, and detecting the content of the wild jujube seed saponin A. The column, mobile phase and elution procedure for high performance liquid chromatography are as described above.
Preparation method of control detection sample II:
(1) Primary extraction: 1.07g of fried spina date seeds are precisely weighed, placed in an conical flask, then added with 100mL of 70Vol.% ethanol solution, heated in a water bath for refluxing for 2 hours, cooled, then complemented with volume loss by the 70Vol.% ethanol solution, and filtered.
(2) And (3) secondary extraction: 50mL of filtrate was taken, the solvent was evaporated, 30mL of water was added to the residue for dissolution, and then the mixture was extracted twice with 30mL of water-saturated n-butanol with shaking.
(3) Preparing sample liquid and detecting: removing the solvent in the water saturated n-butanol extracting solution, adding methanol into the residue for redissolution, and filtering the redissolution to obtain sample liquid to be detected; taking 10 mu L of sample liquid to be detected, adding the sample liquid into a high performance liquid chromatograph, and detecting the content of the wild jujube seed saponin A. The column, mobile phase and elution procedure for high performance liquid chromatography are as described above.
The high performance liquid chromatography detection results (peak areas) of the sample solutions to be detected obtained in the respective embodiments are substituted into the obtained standard curve to obtain the content of the spinosa saponin a in the respective embodiments, and the specific content is shown in table 1.
TABLE 1 detection of the content of the obtained Ziziphus jujuba saponins A in different embodiments
From the data of Table 1, it can be seen that the data of the content of zizyphus jujube saponin A in zizyphus jujube obtained in comparative example 1 (or comparative example 2) and comparative example 1 are compared: although the same method is used for detecting the content of the wild jujube seed and the wild jujube seed saponin A in the content of the product, the content of the wild jujube seed saponin A detected by the content of the product is obviously much lower. That is, other components in the content of the product can have influence on the content detection of the spine date seed saponin A, and further, more accurate spine date seed saponin A content is difficult to obtain by the existing method.
Therefore, the novel method is used for treating the content of the product, and the detection value of the wild jujube seed saponin A in the content of the product is obviously improved. In examples 8 and 9, a partitioning accelerator was added to the treatment of the content, and the addition of the partitioning accelerator further improved the detection value of the zizyphus jujube saponin a in the content: this is probably because even though only the semen Ziziphi Spinosae is treated separately and its content of the semen Ziziphi Spinosae saponin A is measured, since other components such as polysaccharide, protein and flavone in semen Ziziphi Spinosae are extracted simultaneously with the semen Ziziphi Spinosae saponin A, these components existing in semen Ziziphi Spinosae themselves will have an influence on the extraction of the semen Ziziphi Spinosae saponin A. Therefore, after the content is treated by adopting the scheme, proteins, polysaccharides and the like extracted from the content together with the wild jujube saponin A are separated, so that the detection amount of the wild jujube saponin A is further improved, and the detection value of the wild jujube saponin A with more accurate results is obtained.
2. Detection of total sugar and protein content in sample liquid to be detected obtained in different embodiments
2.1 method for detecting Total sugar
The sugar content detection method comprises the following steps: the detection of sugar content is carried out by using a phenol-sulfuric acid method, which utilizes polysaccharide to hydrolyze into monosaccharide under the action of sulfuric acid, rapidly dehydrates to generate a furfural derivative which can be condensed with phenol into a colored compound, and then obtains the sugar content in a sample according to the linear relation between absorbance (ultraviolet absorption wavelength at 490 nm) and sugar concentration.
Drawing a standard curve: developing and measuring absorbance of glucose standard substances with concentrations (g/L) of 0.02, 0.04, 0.06, 0.08 and 0.1 respectively according to the above method, linearly fitting with the concentration (g/L) of the glucose standard substance as abscissa X and absorbance value at 490nm as ordinate Y to obtain regressionThe equation is y=6.82x+0.04, r 2 ＝0.9996。
2.2 method for detecting protein
The method for detecting the protein content comprises the following steps: the detection of protein content was performed using coomassie brilliant blue method. The coomassie brilliant blue on-the-spot preparation method comprises the following steps: 100mg of Coomassie brilliant blue was added to 50mL of 95Vol.% ethanol solution and 100mL of 80 wt.% phosphoric acid solution, deionized water was set to 1000mL, and filtered for use. 5mL of coomassie brilliant blue solution is added into a sample to be detected, and after being mixed evenly, the sample is placed in a spectrophotometer to measure the absorbance at 595 nm.
Drawing a standard curve: preparing bovine serum albumin solutions with concentrations (g/L) of 0.02, 0.04, 0.06, 0.08 and 0.1 respectively, developing and measuring the absorbance according to the method, taking the concentration (g/L) of a bovine serum albumin standard substance as an abscissa X and the absorbance value at 595nm as an ordinate Y, and linearly fitting to obtain a regression equation of Y=2.96X+0.06, R 2 ＝0.997。
2.3 detection results
TABLE 2 polysaccharide and protein content in sample solutions to be tested obtained in different embodiments
The content of polysaccharide in the product is 14.36mg/g content, and the content of protein is 0.35g/g content. In the data results of Table 2, comparing examples 3-9 with comparative examples 1-2, the polysaccharide and protein content in the sample fluid to be tested can be significantly reduced after the content of the sample is treated by the present method.
3. Detection of total flavone content in sample liquid to be detected obtained in different embodiments
The method for detecting the flavone (the rutin standard substance is used for detecting the flavone content):
the detection method comprises the following steps: taking 3mL of sample liquid to be detected, adding 0.5mL of 5wt% NaNO 2 Shaking and standing for 6min, adding 0.5mL of 10wt% Al (NO) 3 ) 3 Shaking up and standing for 6min, adding 4mL 4wt% NaOH aqueous solution, waterConstant volume, shaking and standing for 15min. And finally, measuring the absorbance of the sample at the wavelength of 510nm by an ultraviolet-visible spectrophotometer.
Drawing a standard curve: the rutin standard substance is weighed and prepared into a standard solution with the concentration of 0.4mg/mL by absolute methanol. Precisely sucking 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0mL of the standard solution, respectively placing the standard solution and the standard solution into 10mL volumetric flasks, respectively adding absolute methanol until the total volume is 3mL, and measuring the absorbance of the standard solution with each concentration under the wavelength of 510nm by using an ultraviolet-visible spectrophotometer according to the method. Taking absorbance X as an abscissa and rutin standard solution concentration Y (mg/mL) as an ordinate to make a standard curve, and a regression equation is as follows: y=0.0932x+0.0031, r 2 ＝0.9991。
The flavone content of the sample liquid to be detected obtained in different embodiments is detected, and the specific results are shown in Table 3.
TABLE 3 content of flavonoids in sample solutions to be tested obtained in different embodiments
Description of the embodiments Example 3 Example 8 Example 9 Comparative example 1 Comparative example 2
Flavone content (mg/content) 3.23 2.13 2.32 7.74 7.88
The content of flavone in the product is 8.12 mg/g. In the data results in table 3, compared with examples 8 and 9, the addition of the partitioning agent can reduce the content of flavone in the sample liquid to be detected to a certain extent when the content is treated, so that the influence of flavone on the detection of the content of the spindly jujube saponin A is reduced.
The present embodiment is only for explanation of the present application and is not to be construed as limiting the present application, and modifications to the present embodiment, which may not creatively contribute to the present application as required by those skilled in the art after reading the present specification, are all protected by patent laws within the scope of claims of the present application.

Claims (6)

1. The quality control method of the Shenmu tranquilization capsule for improving deep sleep is characterized by comprising the following steps of detecting the content of the wild jujube seed saponin A in the content of the Shenmu tranquilization capsule:
s1, acidolysis: adding acid liquor into the content of the product, and reacting to obtain acidolysis solution, wherein the weight-volume ratio of the content to the acid liquor is 0.21-0.46g/mL, and the acid liquor contains micromolecular organic acid and inorganic acid;
s2, phase separation extraction: adding 50-70Vol.% ethanol solution into acidolysis solution, and fully vibrating and extracting, wherein the weight-volume ratio of the content to the saturated 50-70Vol.% ethanol solution is 0.53-0.82g/mL;
s3, secondary extraction: taking 50-70Vol.% ethanol solution after split-phase extraction, evaporating the solvent, adding water into the residue for dissolution, and then extracting with water saturated n-butanol;
s4, preparing sample liquid and detecting: removing solvent in water saturated n-butanol extractive solution, adding methanol into residue, and redissolving to obtain sample solution to be detected, and detecting the content of semen Ziziphi Spinosae saponin A by high performance liquid chromatography;
the mass fraction of small molecular organic acid in the acid liquor is 2-5%, and the mass fraction of inorganic acid in the acid liquor is 5-10%;
the small molecule organic acid is selected from any one or more of citric acid, formic acid, acetic acid, propionic acid and malic acid; the inorganic acid is sulfuric acid.
2. The method according to claim 1, wherein the small molecule organic acid is selected from any one or more of citric acid, acetic acid, and propionic acid.
3. The quality control method according to claim 1, wherein in step S2, 50 to 70vol.% ethanol is added to the acidolysis solution, and simultaneously, a partitioning accelerator is added in an amount of 6 to 15 wt.% of the acid solution; the distribution promoting agent is polyacrylic acid or polymethacrylic acid.
4. The method according to claim 3, wherein the weight ratio of the small molecule organic acid to the partitioning agent is (0.2-0.5): 1.
5. The method according to claim 1, wherein in step S3, the volume ratio of water to water-saturated n-butanol solution is 1 (0.8-1.2).
6. The quality control method according to claim 1, wherein in step S4, the content of the zizyphus jujube seed saponin a is detected by high performance liquid chromatography; the detection column is prepared by taking octadecylsilane chemically bonded silica as a filler; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and gradient elution is carried out.
CN202210981251.7A 2022-08-16 2022-08-16 Quality control method of Shenmu tranquilization capsule for improving deep sleep Active CN115326965B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202210981251.7A CN115326965B (en) 2022-08-16 2022-08-16 Quality control method of Shenmu tranquilization capsule for improving deep sleep

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202210981251.7A CN115326965B (en) 2022-08-16 2022-08-16 Quality control method of Shenmu tranquilization capsule for improving deep sleep

Publications (2)

Publication Number Publication Date
CN115326965A CN115326965A (en) 2022-11-11
CN115326965B true CN115326965B (en) 2023-09-15

Family

ID=83923750

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202210981251.7A Active CN115326965B (en) 2022-08-16 2022-08-16 Quality control method of Shenmu tranquilization capsule for improving deep sleep

Country Status (1)

Country Link
CN (1) CN115326965B (en)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103272018A (en) * 2013-05-10 2013-09-04 华南理工大学 Sedative and hypnotic active saponin components of spina date seed, as well as separation and purification method and application of sedative and hypnotic active saponin components

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103272018A (en) * 2013-05-10 2013-09-04 华南理工大学 Sedative and hypnotic active saponin components of spina date seed, as well as separation and purification method and application of sedative and hypnotic active saponin components

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
HPLC-ELSD测定枣仁安神胶囊中酸枣仁皂苷A的含量;何新荣;刘萍;孙艳;;中国药师(第04期);第497-499页 *
安神胶囊的薄层色谱鉴别;张晓红;王月茹;谢伟;;陕西中医(第09期);第1231-1232页 *
快速溶剂萃取联合高效液相色谱法测定不同产地酸枣仁中皂苷A、B的含量;丁轲;张彤楠;韩涛;;食品工业科技(第23期);第265-270页 *

Also Published As

Publication number Publication date
CN115326965A (en) 2022-11-11

Similar Documents

Publication Publication Date Title
DE69919981T2 (en) PROCESS FOR THE ISOLATION, RECOVERY AND CLEANING OF NON-POLAR EXTRACTS
CN109324126B (en) Method for simultaneously determining 9 chemical components in spina date seeds by using UPLC-MS/MS
CN103330758A (en) Peony and liquorice soup formula granule, preparation method and detection method of peony and liquorice soup formula granule
Yuniarti et al. Application of the natural deep eutectic solvent choline chloride-sorbitol to extract chlorogenic acid and caffeine from green coffee beans (Coffea canephora)
CN108008038A (en) Measure four kinds of remaining methods of Nitrofuran metabolites metabolite in gelatin type traditional Chinese medicine
Zhou et al. Application of mixed cloud point extraction for the analysis of six flavonoids in Apocynum venetum leaf samples by high performance liquid chromatography
CN104147054A (en) Ginkgo biloba leaf extract as well as preparation method and application thereof
CN115326965B (en) Quality control method of Shenmu tranquilization capsule for improving deep sleep
CN107884508B (en) Quality detection method of Yinhuang lung-clearing capsule
CN101869630B (en) Measurement method for content of atropine sulfate in traditional Chinese medicine suppository containing belladonna liquid extract
CN1981852A (en) Tall gastrodia tuber preparation with resuscitation-inducing function, its making and quality controlling method
CN104257768A (en) Application of total anthraquinone with various ingredients in stable and uniform proportion and composition of total anthraquinone to treatment of pancreatitis
CN108623642B (en) Deep eutectic solvent water mixture for synchronously extracting salidroside and tyrosol from rhodiola rosea, and preparation method and extraction method thereof
CN103083388B (en) Preparation method of fructus gleditsiae total saponins
CN111072747A (en) Ginsenoside and ultrasonic extraction method thereof
He et al. A new hollow fiber liquid/solid phase microextraction using Al 2 O 3 nanoparticles as the adsorbent for extraction of ginsenosides from an oral liquid
CN110243967B (en) Screening method of semen cuscutae estrogen-like action quality marker and construction and application of characteristic spectrum thereof
CN1335157A (en) Systemic separation process and prepn of ginkgetin, ginkgolide and bilobalide
CN107158113A (en) A kind of Chinese medicine composition, Its Preparation Method And Use
Yuan et al. Simultaneous determination of four active compounds in Centella asiatica by supramolecular solvent-based extraction coupled with high performance liquid chromatography-tandem mass spectrometry
Sawant et al. High-performance thin-layer chromatographic quantification of kaempferol and apigenin in the whole-plant powder of Sida spinosa Linn.
CN115508495B (en) Identification method of boiled peach kernels in Taohong Siwu decoction
CN113295816B (en) Thin layer chromatography detection method for Yihechun preparation
CN109270201B (en) HPLC (high performance liquid chromatography) characteristic spectrum of compound fructus momordicae lung-clearing preparation and construction method thereof
CN107389810A (en) A kind of content assaying method of strong drug composition

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant