CN110146600A - The preparation method and its detection method of psoralea corylifolia standard decoction - Google Patents
The preparation method and its detection method of psoralea corylifolia standard decoction Download PDFInfo
- Publication number
- CN110146600A CN110146600A CN201810148957.9A CN201810148957A CN110146600A CN 110146600 A CN110146600 A CN 110146600A CN 201810148957 A CN201810148957 A CN 201810148957A CN 110146600 A CN110146600 A CN 110146600A
- Authority
- CN
- China
- Prior art keywords
- psoralea corylifolia
- preparation
- standard decoction
- methanol
- water
- 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
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/487—Psoralea
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- 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
- G01N2030/042—Standards
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pathology (AREA)
- Immunology (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Mycology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Alternative & Traditional Medicine (AREA)
- Engineering & Computer Science (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The present invention relates to modern Chinese traditional medicine fields, and in particular to the preparation method and detection method of psoralea corylifolia standard decoction.The preparation method of psoralea corylifolia standard decoction of the present invention includes following step: taking corylifolia L, add water to cook, filtrate is obtained by filtration；Filtrate is concentrated and dried, is then lyophilized, the freeze-drying is divided into three phases: a. pre-freeze: pre-freezing temperature is -50 DEG C~-45 DEG C；B. primary drying: drying temperature is -30 DEG C~0 DEG C；C. redrying: drying temperature is 5~25 DEG C, obtains psoralea corylifolia standard decoction, the paste-forming rate of the psoralea corylifolia standard decoction is high, and by 6 months accelerated stability tests, content was relatively stable, and moisture content is relatively stable, and the resistance to moisture absorption does not generate impurity component；The present invention provides the method for the total content and total rate of transform of measurement psoralen and isopsorapen, and specificity is good, and repeatability is good, and stability is good.Characteristic spectrum method of the invention, using superelevation phase liquid phase, method is simple, can greatly shorten detection time, improves production efficiency.
Description
Technical field
The present invention relates to modern Chinese traditional medicine fields, and in particular to a kind of preparation method and its detection of psoralea corylifolia standard decoction
Method.
Background technique
Effects of Bu Gu rouge is the dry mature fruit of legumes psoraleae Psoralea corylifolia L., alias
Fructus psoaleae, Hu fragrant-flowered garlic etc., warm-natured, acrid flavour cure mainly kidney deficiency diarrhea, impotence, waist and knee crymodynia, the cold of insufficiency type on tcm clinical practice and breath with cough
Illness.Modern medicine, which shows psoralea corylifolia also, has a variety of pharmacology such as antitumor, antibacterial, anti-osteoporosis, antiestrogenic sample effect
The diseases such as activity, and treatment leucoderma, psoriasis.
Standard decoction is also known as standard decocting liquid, is a kind of traditional widely used administration form of clinic, standard decoction system abides by
Traditional Chinese medical theory is followed, standardizes according to clinical decoction decocting method and decocts, is separated by solid-liquid separation, preparation is suitably concentrated or through suitable side
Method is dry to be made, as measure Chinese medicinal granule whether the marker almost the same with clinical decoction.
Since standard decoction is connection traditional Chinese medicine medicine materical crude slice and modern Chinese herbal medicine preparation " bridge ", produced for control Chinese medicine terminal
The quality of product provides object of reference, for markization Chinese medicine difference administration form, it is ensured that homogeneity, the consistency of curative effect of quality provide
Object of reference conformity provides object of reference for evaluation different manufacturers product quality, therefore, Chinese medicine standard decoction quality standard
Formulation will provide basis for the formulation of the quality standard of the finished product of all water decoctions derived from medicine materical crude slice, but currently, not yet
About the preparation method of psoralea corylifolia standard decoction and the report of detection method.
Summary of the invention
For this purpose, the present invention, aiming to the above problems, provides a kind of preparation method of psoralea corylifolia standard decoction and its detection sides
Method is standardized with research on standard with the quality control to Chinese medicinal granule, realizes the quality control of Chinese medicinal granule entirety
System and effectively supervision.
The present invention provides a kind of preparation methods of psoralea corylifolia standard decoction, and it includes following step:
(1) corylifolia L is taken, is added water to cook, filtrate is obtained by filtration；
(2) filtrate in step (1) is concentrated and dried, is then lyophilized, the freeze-drying is divided into three phases: a. pre-freeze:
Pre-freezing temperature is -50 DEG C~-45 DEG C；B. primary drying: drying temperature is -30 DEG C~0 DEG C；C. redrying is parsed: dry
Dry temperature is 5~25 DEG C, obtains psoralea corylifolia standard decoction.
The present invention provides a kind of above-mentioned psoralea corylifolia standard decoction gross mass content and the detection method of total rate of transform, packets
Containing following step:
(1) preparation of reference solution:
Psoralea corylifolia reference substance, Isopsoralen are weighed, adds methanol that solution is made；
(2) preparation of test solution:
Psoralea corylifolia standard decoction is taken, solvent is added and extracts；
(3) efficient liquid phase chromatographic analysis
Using octadecylsilane chemically bonded silica as filler, mobile phase A is water, and Mobile phase B is methanol, and it is molten to draw reference substance
Liquid and test solution injection high performance liquid chromatograph are analyzed.
The present invention provides a kind of detection methods of the characteristic spectrum of above-mentioned psoralea corylifolia standard decoction, and it includes following steps
It is rapid:
(1) preparation of reference solution:
Psoralea corylifolia reference substance, appropriate Isopsoralen are weighed, adds methanol that solution is made；
(2) preparation of test solution:
It takes psoralea corylifolia standard decoction appropriate, solvent is added and extracts；
(3) superelevation liquid chromatogram is analyzed
It draws reference substance solution and test solution injects Ultra Performance Liquid Chromatography instrument, with octadecylsilane chemically bonded silica
For filler；Using methanol as mobile phase A, gradient elution is carried out by Mobile phase B of water phase, obtains the feature of psoralea corylifolia standard decoction
Map.
Specifically, the invention proposes following technical solutions.
The present invention provides a kind of preparation methods of psoralea corylifolia standard decoction, and it includes following step:
(1) corylifolia L is taken, is added water to cook, filtrate is obtained by filtration；
(2) filtrate in step (1) is concentrated and dried, is then lyophilized, the freeze-drying is divided into three phases: a. pre-freeze:
Pre-freezing temperature is -50 DEG C~-45 DEG C；B. primary drying: drying temperature is -30 DEG C~0 DEG C；C. redrying is parsed: dry
Dry temperature is 5~25 DEG C, obtains psoralea corylifolia standard decoction.
Preferably, for the preparation method, wherein in step (1), the number that adds water to cook is 1~3 time.
Preferably, for the preparation method, wherein the decoction number is 2 times, decocts for the first time plus 7~9 times are measured
Water, the water of preferably 8 times amounts, second decocts plus the water of 4~6 times of amounts, the water of preferably 6 times amounts.
Preferably, for the preparation method, wherein first time decocting time be 30~60min, preferably 60min,
Second of decocting time is 20~45min, preferably 45min.
Preferably, for the preparation method, wherein in step (1), the filtering uses 100~300 mesh screens
It is filtered, preferably 100 mesh screens are filtered.
Preferably, for the preparation method, wherein in step (2), the pre-freeze time be 3-6 hours, preferably 4
Hour；The primary drying time is 10-14 hours, preferably 14 hours；The redrying time is 5~7 hours, preferably 7 hours.
Preferably, for the preparation method, wherein in step (2), the drying temperature of the filtrate concentrate drying
It is 60~65 DEG C.
Preferably, for the preparation method, wherein the paste-forming rate of the psoralea corylifolia standard decoction be 17.10~
31.02%.
The present invention provides psoralea corylifolia standard decoction gross mass content and the detection methods of total rate of transform, and it includes following steps
It is rapid:
(1) preparation of reference solution:
Psoralea corylifolia reference substance, Isopsoralen are weighed, adds methanol that solution is made；
(2) preparation of test solution:
Psoralea corylifolia standard decoction is taken, solvent is added and extracts；
(3) efficient liquid phase chromatographic analysis
Using octadecylsilane chemically bonded silica as filler, mobile phase A is water, and Mobile phase B is methanol, and it is molten to draw reference substance
Liquid and test solution injection high performance liquid chromatograph are analyzed.
Preferably, for the detection method, wherein in step (2), the solvent is selected from Diluted Alcohol, 75% second
One of alcohol, 95% ethyl alcohol, 50% methanol, 75% methanol and methanol, preferably 75% ethyl alcohol.
Preferably, for the detection method, wherein in step (2), the extraction is using reflux, dissolution or ultrasound
One of extract, preferably ultrasonic extraction.
Preferably, for the detection method, wherein in step (2), the extraction time 15-45min of the extraction,
Preferably 30min.
Preferably, for the detection method, wherein based on dry product, the psoralen and isopsorapen
Gross mass content is 0.89~1.65%.
Preferably, for the detection method, wherein total rate of transform of the psoralen and isopsorapen is
18.58~34.52%.
The present invention provides the detection methods of the characteristic spectrum of the psoralea corylifolia standard decoction, and it includes following step:
(1) preparation of reference solution:
Psoralea corylifolia reference substance, Isopsoralen are weighed, adds methanol that solution is made；
(2) preparation of test solution:
Psoralea corylifolia standard decoction is taken, solvent is added and extracts；
(3) superelevation liquid chromatogram is analyzed:
It draws reference substance solution and test solution injects Ultra Performance Liquid Chromatography instrument, with octadecylsilane chemically bonded silica
For filler；Using methanol as mobile phase A, gradient elution is carried out by Mobile phase B of water phase, obtains the feature of psoralea corylifolia standard decoction
Map.
Preferably, for the detection method, wherein in step (2), the solvent selected from water, 95% ethyl alcohol,
One of 75% ethyl alcohol, Diluted Alcohol, methanol, 75% methanol, 50% methanol, preferably 95% ethyl alcohol.
Preferably, for the detection method, wherein in step (2), described extract is mentioned using refluxing extraction, ultrasound
One of take or dissolve extraction, preferably ultrasonic extraction.
Preferably, for the detection method, wherein in step (2), extraction time of the extraction is 15~
45min, preferably 30min.
Preferably, for the detection method, wherein in step (3), phosphoric acid of the water phase selected from water, 0.1%
Aqueous solution, 0.1% aqueous formic acid or 0.2% aqueous acetic acid, preferably 0.1% phosphate aqueous solution.
It is obtained by the present invention the utility model has the advantages that
1, the present invention provides a standard for the quality of Chinese medicinal granule, realizes the matter of Chinese medicinal granule entirety
Amount control and effectively supervision.
2, the present invention prepares psoralea corylifolia standard decoction using corylifolia L, and the paste-forming rate of the psoralea corylifolia standard decoction is
17.10~31.02%, paste-forming rate is higher, and by 6 months accelerated stability tests, content was relatively stable, and moisture content is relatively steady
It is fixed, the resistance to moisture absorption, and impurity component is not generated, preparation-obtained psoralea corylifolia standard decoction is subjected to high performance liquid chromatography point
Analysis, the method specificity is good, and repeatability is good, and stability is good.
3, the characteristic spectrum method that the present invention establishes, using superelevation phase liquid chromatogram, method is simple, divides a collection of preparation
18min is only used in analysis, can be greatly shortened and be tested and analyzed the time, and production efficiency is improved, so that large sample sampling Detection, real time monitoring is produced
The quality of product is possibly realized；In addition, existing conventional method need to consume organic solvent about 2L in organic solvent use aspect, and
The present invention only need its 1/10th, greatly reduce the consumption of solvent, be more energy-saving and environmentally friendly.
Detailed description of the invention
Fig. 1-1 is spectrogram corresponding to the psoralea corylifolia standard decoction of accelerated stability test in embodiment one；
Fig. 1-2 is that the psoralea corylifolia standard decoction of accelerated stability test in embodiment one accelerates spectrogram corresponding to 1 month；
Fig. 1-3 is that the psoralea corylifolia standard decoction of accelerated stability test in embodiment one accelerates spectrogram corresponding to 2 months；
Fig. 1-4 is that the psoralea corylifolia standard decoction of accelerated stability test in embodiment one accelerates spectrogram corresponding to 3 months；
Fig. 1-5 is that the psoralea corylifolia standard decoction of accelerated stability test in embodiment one accelerates spectrogram corresponding to 6 months；
Fig. 2 is the full wavelength scanner schematic diagram of psoralen；
Fig. 3 is the full wavelength scanner schematic diagram of Isopsoralen；
Fig. 4 is the height that the different solvents of test solution of gross mass content and total rate of transform are measured in embodiment one
Effect liquid phase chromatogram comparison diagram；
Fig. 5 is the height that the different extracting modes of test solution of gross mass content and total rate of transform are measured in embodiment one
Effect liquid phase chromatogram comparison diagram；
Fig. 6 is the height that the different extraction times of test solution of gross mass content and total rate of transform are measured in embodiment one
Effect liquid phase chromatogram comparison diagram；
Fig. 7 is the high performance liquid chromatography spectrogram of the gross mass content of the psoralen and Isopsoralen in embodiment one；
Fig. 8 is investigated to the specificity of the method for the test solution for measuring total content and total rate of transform in embodiment one
High-efficient liquid phase chromatogram；
Fig. 9 is the high-efficient liquid phase chromatogram different in flow rate that gross mass content and total rate of transform are measured in embodiment one；
Figure 10 is the high-efficient liquid phase chromatogram that the different column temperatures of gross mass content and total rate of transform are measured in embodiment one；
Figure 11 is the high-efficient liquid phase chromatogram that the different chromatographic columns of gross mass content and total rate of transform are measured in embodiment one；
Figure 12 is the ultra performance liquid chromatography figure that embodiment one measures different Detection wavelengths in the detection method of characteristic spectrum；
Figure 13-1 is the ultra high efficiency liquid phase color that embodiment one measures that mobile phase in the detection method of characteristic spectrum is methanol-water
Spectrogram；
Figure 13-2 is that mobile phase is-0.1% phosphate aqueous solution of methanol in the detection method of the measurement characteristic spectrum of embodiment one
Ultra performance liquid chromatography figure；
Figure 13-3 is that mobile phase is-0.1% aqueous formic acid of methanol in the detection method of the measurement characteristic spectrum of embodiment one
Ultra performance liquid chromatography figure；
Figure 13-4 is that mobile phase is-0.2 aqueous acetic acid of methanol in the detection method of the measurement characteristic spectrum of embodiment one
Ultra performance liquid chromatography figure；
Figure 14-1 is that embodiment one measures the superelevation that chromatographic column in the detection method of characteristic spectrum is BEH Shield RP18
Effect liquid phase chromatogram figure；
Figure 14-2 is that embodiment one measures the ultra high efficiency liquid phase color that chromatographic column in the detection method of characteristic spectrum is BEH C18
Spectrogram；
Figure 14-3 is that embodiment one measures the ultra high efficiency liquid that chromatographic column in the detection method of characteristic spectrum is Cortecs T3
Phase chromatogram；
Figure 15-1 is the ultra performance liquid chromatography figure that embodiment one measures that column temperature is 25 DEG C in the detection method of characteristic spectrum；
Figure 15-2 is the ultra performance liquid chromatography figure that embodiment one measures that column temperature is 30 DEG C in the detection method of characteristic spectrum；
Figure 15-3 is the ultra performance liquid chromatography figure that embodiment one measures that column temperature is 35 DEG C in the detection method of characteristic spectrum；
Figure 16-1 is that embodiment one measures the ultra high efficiency liquid phase color that flow velocity in the detection method of characteristic spectrum is 0.2ml/min
Spectrogram；
Figure 16-2 is that embodiment one measures the ultra high efficiency liquid phase that flow velocity in the detection method of characteristic spectrum is 0.25ml/min
Chromatogram；
Figure 16-3 is that embodiment one measures the ultra high efficiency liquid phase color that flow velocity in the detection method of characteristic spectrum is 0.3ml/min
Spectrogram；
Figure 17-1 is that embodiment one measures in the detection method of characteristic spectrum using water as the ultra high efficiency liquid phase color of Extraction solvent
Spectrogram；
Figure 17-2 is that embodiment one measures in the detection method of characteristic spectrum using 95% ethyl alcohol as the superelevation of Extraction solvent
Effect liquid phase chromatogram figure；
Figure 17-3 is that embodiment one measures in the detection method of characteristic spectrum using 75% ethyl alcohol as the superelevation of Extraction solvent
Effect liquid phase chromatogram figure；
Figure 17-4 is that embodiment one measures in the detection method of characteristic spectrum using Diluted Alcohol as the ultra high efficiency of Extraction solvent
Liquid chromatogram；
Figure 17-5 is the ultra high efficiency liquid in the detection method of the measurement characteristic spectrum of embodiment one using methanol as Extraction solvent
Phase chromatogram；
Figure 17-6 is that embodiment one measures in the detection method of characteristic spectrum using 75% methanol as the superelevation of Extraction solvent
Effect liquid phase chromatogram figure；
Figure 17-7 is that embodiment one measures in the detection method of characteristic spectrum using 50% methanol as the superelevation of Extraction solvent
Effect liquid phase chromatogram figure；
Figure 18-1 is that embodiment one measures the ultra high efficiency liquid phase that ultrasound treatment patterns are used in the detection method of characteristic spectrum
Chromatogram；
Figure 18-2 is that embodiment one measures the ultra high efficiency liquid phase that reflow treatment mode is used in the detection method of characteristic spectrum
Chromatogram；
Figure 18-3 is that embodiment one measures in the detection method of characteristic spectrum using the ultra high efficiency of oscillation dissolution process mode
Liquid chromatogram；
Figure 19-1 is the ultra high efficiency liquid phase color that embodiment one measures that extraction time in the detection method of characteristic spectrum is 15min
Spectrogram；
Figure 19-2 is the ultra high efficiency liquid phase color that embodiment one measures that extraction time in the detection method of characteristic spectrum is 30min
Spectrogram；
Figure 19-3 is the ultra high efficiency liquid phase color that embodiment one measures that extraction time in the detection method of characteristic spectrum is 45min
Spectrogram；
Figure 20 is the characteristic spectrum schematic diagram of preparation-obtained psoralea corylifolia standard decoction in embodiment one；
Figure 21 is the high performance liquid chromatography spectrogram of the gross mass content of the psoralen and Isopsoralen in embodiment two；
Figure 22 is the characteristic spectrum schematic diagram of preparation-obtained psoralea corylifolia standard decoction in embodiment two；
Figure 23 is the high performance liquid chromatography spectrogram of the gross mass content of the psoralen and Isopsoralen in embodiment three；
Figure 24 is the characteristic spectrum schematic diagram of preparation-obtained psoralea corylifolia standard decoction in embodiment three；
Figure 25 is the high performance liquid chromatography spectrogram of the gross mass content of the psoralen and Isopsoralen in example IV；
Figure 26 is the characteristic spectrum schematic diagram of preparation-obtained psoralea corylifolia standard decoction in example IV.
Specific embodiment
Term " Diluted Alcohol " is prepared using annex XVB test solution in " Chinese Pharmacopoeia one ", that is, takes ethyl alcohol 529ml,
Be diluted with water to 1000ml to get.This liquid contains C at 20 DEG C2H5OH should be 49.5%~50.5% (ml/ml).
As described above, it includes following step the present invention provides a kind of preparation method of psoralea corylifolia standard decoction:
(1) corylifolia L is taken, is added water to cook, filtrate is obtained by filtration；
(2) filtrate in step (1) is concentrated and dried, is then lyophilized, the freeze-drying is divided into three phases: a. pre-freeze:
Pre-freezing temperature is -50 DEG C~-45 DEG C；B. primary drying: drying temperature is -30 DEG C~0 DEG C；C. redrying is parsed: dry
Dry temperature is 5~25 DEG C, obtains psoralea corylifolia standard decoction.
In a kind of currently preferred specific embodiment, wherein in step (1), corylifolia L is adding decocting
Before boiling, it is broken to carry out appropriateness, and impregnated, soaking time 30min.
In a kind of currently preferred specific embodiment, wherein in step (1), the filtering is to filter while hot,
The filtering is filtered using 100~300 mesh screens, and preferably 100 mesh are filtered.
Preferably, the sieve is Taylor's sieve (Tyler Corporations, the U.S.) or filter cloth (new chemical industry Limited Liability public affairs in Xinxiang
Department).
In a kind of currently preferred specific embodiment, wherein in step (2), filtrate is concentrated under reduced pressure,
It is concentrated at being 60-65 DEG C in temperature, being concentrated into density is 1.03~1.05g/ml, and adjusting solid content is 10-14%.
In a kind of currently preferred specific embodiment, wherein in step (2), the pre-freeze time is 3-6 hours,
Preferably 6 hours；The primary drying time is 10-14 hours, preferably 14 hours；The redrying time is 5~8 hours, preferably
It is 7 hours.
In a kind of currently preferred specific embodiment, the biodiversity content of the psoralea corylifolia standard decoction is 2-
4%, preferably 2.2-3.6%.
The present invention provides a kind of psoralea corylifolia standard decoction gross mass content and the detection method of total rate of transform, packets
Containing following step:
(1) preparation of reference solution:
Psoralea corylifolia reference substance, appropriate Isopsoralen are weighed, adds methanol that solution is made；
(2) preparation of test solution:
It takes psoralea corylifolia standard decoction appropriate, solvent is added and extracts；
(3) efficient liquid phase chromatographic analysis
Using octadecylsilane chemically bonded silica as filler, mobile phase A is water, and Mobile phase B is methanol, and it is molten to draw reference substance
Liquid and test solution injection high performance liquid chromatograph are analyzed.
In a kind of currently preferred specific embodiment, wherein the volume ratio of the mobile phase A and Mobile phase B is
45:55。
In a kind of currently preferred specific embodiment, used chromatographic column when efficient liquid phase chromatographic analysis is carried out
It can be BEH Shield RP18 (Waters, 2.1 × 100mm, 1.7 μm), BEH C18 (Waters, 2.1 × 100mm, 1.7 μ
Or Cortecs T3 (Waters, 2.1 × 100mm, 1.7 μm), preferably BEH C18 (Waters, 2.1 × 100mm, 1.7 μ m)
m)。
In a kind of currently preferred specific embodiment, when carrying out efficient liquid phase chromatographic analysis, used chromatography
The column temperature of column can be 25 DEG C -35 DEG C, preferably 30 DEG C.
In a kind of currently preferred specific embodiment, wherein the reference solution the preparation method comprises the following steps: taking benefit
Bone fat element reference substance, appropriate Isopsoralen, micrometric measurement add methanol that the solution that every 1ml contains 20 μ g is made；
The test solution the preparation method comprises the following steps: taking psoralea corylifolia standard decoction, accurately weighed 0.1g sets stuffed conical flask
In, solvent 25ml is added in precision, and weighed weight is extracted using reflux or concussion dissolution or ultrasonic power, preferably ultrasonic
It extracts, then weighed weight, supplies less loss weight with solvent, shake up, filtrate is obtained by filtration.
In a kind of currently preferred specific embodiment, wherein used when carrying out efficient liquid phase chromatographic analysis
Flow velocity can be 0.2-0.3ml/min, preferably 0.25ml/min.
The present invention provides a kind of detection methods of the characteristic spectrum of psoralea corylifolia standard decoction, and it includes following step:
(1) preparation of reference solution:
Psoralea corylifolia reference substance, appropriate Isopsoralen are weighed, adds methanol that solution is made；
(2) preparation of test solution:
It takes psoralea corylifolia standard decoction appropriate, solvent is added and extracts；
(3) superelevation liquid chromatogram is analyzed
It draws reference substance solution and test solution injects Ultra Performance Liquid Chromatography instrument, with octadecylsilane chemically bonded silica
For filler；Using methanol as mobile phase A, gradient elution is carried out by Mobile phase B of water phase, obtains the feature of psoralea corylifolia standard decoction
Map.
In a kind of currently preferred specific embodiment, the reference solution the preparation method comprises the following steps: taking benefit
Bone fat element reference substance, appropriate Isopsoralen, micrometric measurement add methanol that the solution that every 1ml contains 20 μ g is made；
The test solution the preparation method comprises the following steps: taking psoralea corylifolia standard decoction, accurately weighed 0.1g sets stuffed conical flask
In, solvent 25ml is added in precision, and weighed weight is extracted using reflux or concussion dissolution or ultrasonic power, preferably ultrasonic
It extracts, then weighed weight, supplies less loss weight with solvent, shake up, filtrate is obtained by filtration.
In a kind of currently preferred specific embodiment, the water phase selected from water, 0.1% phosphate aqueous solution,
0.1% aqueous formic acid or 0.2% aqueous acetic acid, preferably 0.1% phosphate aqueous solution, wherein in 0~4min
Interior, mobile phase A is 5 → 60, and Mobile phase B is 95 → 40；In 4~14min, mobile phase A is 60 → 100, and Mobile phase B is 40 → 0；
In 14~14.1min, mobile phase A is 100 → 5, and Mobile phase B is 0 → 95；In 14.1~18min, mobile phase A 5, Mobile phase B
It is 95.
Below by preparation method and its detection method of the embodiment to psoralea corylifolia standard decoction of the present invention make into
One step explanation.Reagent used in embodiment, non-specified otherwise, be conventional reagent, wherein raw material used in embodiment
Information and the information of experimental facilities difference are as shown in Table 1 and Table 2:
Raw material information used in 1 present invention of table
| Raw material | Purity/lot number | Manufacturer |
| Psoralea corylifolia medicine materical crude slice | 201701 | It leaves one's hometown at Yunnan Province, family, Dehong prefecture Ruili City |
| Psoralen reference substance | 110739-201617 | National Institute for Food and Drugs Control |
| Isopsoralen reference substance | 110738-201614 | National Institute for Food and Drugs Control |
The information of experimental facilities used in 2 present invention of table
| Equipment | Model | Manufacturer |
| Mechanical fission cooking pot for herbs | YMW | Chaozhou port coin technique makes factory |
| Rotary Evaporators | Bμchi-R100 | Bu Qi Co., Ltd, Switzerland |
| Rotary Evaporators | YRE2000-B type | Yuhua Instrument Co., Ltd., Gongyi City |
| Vacuum pump using circulatory water | SHZ-DⅢ | Yuhua Instrument Co., Ltd., Gongyi City |
| Cryogenic liquid water circulating pump | DLSB-5/20 | Zhengzhou Greatwall Scientific Industrial & Trading Co., Ltd. |
| Freeze dryer | TRL-0.5 | Dalian Shuan Rui Science and Technology Ltd. |
| Cnc ultrasonic cleaning instrument | KQ-500DA | Kunshan Ultrasonic Instruments Co., Ltd. |
| Ultra Performance Liquid Chromatography instrument | VANQUISH | Sai Mo flies |
| Electronic balance | AL104/ML204 | Mettler-Toledo Instrument (Shanghai) Co., Ltd. |
| High performance liquid chromatograph | WatersACQUITY/HClass | Water generation Science and Technology Ltd. |
| Electric drying oven with forced convection | 101-2A | Shanghai perseverance scientific instrument Co., Ltd |
| High performance liquid chromatograph | Agilent1100 | Agilent Technologies, the U.S. |
| Electronic balance | HLD-30002 | Hangzhou You Heng weighing instruments Co., Ltd |
| Electronic balance | S225D | Sai Duolisi scientific instrument (Beijing) Co., Ltd |
| Digital display thermostat water bath | HH-4/HH-6 | Changzhou Pu Tian instrument manufacturing Co., Ltd |
Embodiment one
1. the preparation method of psoralea corylifolia standard decoction
(1) psoralea corylifolia medicine materical crude slice 100g is taken, is added water to cook twice, first decocts the water for adding 8 times of amounts, starts to decoct after impregnating 30min
It boils, opens intense fire (500W), after boiling, be adjusted to mild fire (200W) and decoct 60 minutes, decocted while hot with 100 mesh filter-cloth filterings, second
The water for adding 6 times of amounts, opens intense fire (500W), after boiling, is adjusted to mild fire (200W) and decocts 45min, carried out while hot with 100 mesh filter clothes
Filtering merges filtrate twice；
(2) by filtrate be transferred in Rotary Evaporators concentration (bath temperature: 65 DEG C, vacuum degree is -0.08~-
0.09MPa), being concentrated into density is 1.03g/ml, and adjusting solid content to 10% is dispensed into 10ml cillin bottle, partly jumped a queue, every bottle
Packing volume is 2.0ml, is transferred in vacuum freeze drier after packing, and pre-freeze is first carried out, and pre-freezing temperature is -45 DEG C,
The pre-freeze time is 4h, then carries out primary drying, and drying temperature is followed successively by -30 DEG C, and -20 DEG C, -10 DEG C, 0 DEG C, drying time divides
Not Wei 7h, 2h, 2h, 3h, then carry out redrying, drying temperature is followed successively by 5 DEG C, and 15 DEG C, 25 DEG C, drying time is respectively
2h, 2h, 3h take out, roll aluminium lid, obtain psoralea corylifolia standard decoction, and the paste-forming rate of the psoralea corylifolia presses formula are as follows: and paste-forming rate=
5ml concentrate is evaporated rear solid masses * concentrate gross mass/(5ml concentrate quality * medicine materical crude slice inventory) * 100% and is counted
It calculates, obtaining paste-forming rate is 31.02%, and the biodiversity content of the psoralea corylifolia standard decoction is calculated according to the following equation: water
Divide mass content=moisture content weight/sample weight * 100%, the biodiversity content for obtaining psoralea corylifolia standard decoction is 2.2%.
(3) accelerated stability test
Above-mentioned obtained psoralea corylifolia standard decoction is subjected to acceleration for stabilization experiment.
Accelerated stability test condition: 40 DEG C ± 2 DEG C, relative humidity 75% ± 5%
The result of 3 accelerated stability test of table
| Time | Content/% | Moisture/% |
| 0 | 1.21 | 2.25 |
| January | 1.21 | 2.18 |
| 2 months | 1.20 | 2.13 |
| March | 1.22 | 2.29 |
| June | 1.23 | 2.18 |
It can be seen that content is relatively stable, and moisture content is relatively stable from table 3 and Fig. 1-1 to 1-5, the resistance to moisture absorption, and not
Generate impurity component.
2. the psoralen and isopsorapen gross mass content of psoralea corylifolia standard decoction and the measuring method of total rate of transform
Wherein, the gross mass content of psoralen and isopsorapen=sample peak area * reference substance concentration * sample constant volume
Volume 50ml/ (reference substance peak area * standard decoction sampling amount 0.1g* (1- marks soup point)) * 100%
Total rate of transform of psoralen and isopsorapen=drying standard decoction psoralen, Isopsoralen total amount *
Standard decoction quality * (1- standard decoction moisture)/(dry medicine materical crude slice psoralen, Isopsoralen total amount * medicine materical crude slice inventory * (1-
Medicine materical crude slice moisture)) * 100%
The optimization of the condition of 2.1 pairs of detection methods
(1) preparation of reference substance solution
Psoralea corylifolia reference substance, appropriate Isopsoralen are taken, it is accurately weighed, add methanol that the solution that every 1ml contains 20 μ g is made, i.e.,
?.
(2) preparation of test solution
Psoralea corylifolia standard decoction sample in Example one is finely ground, takes about 0.1g, accurately weighed, sets stuffed conical flask
In, organic solvent 50ml is added in precision, and weighed weight extracts processing a period of time, then weighed weight, mended with organic solvent
Sufficient less loss weight, shakes up, filtration, take filtrate to get.
(3) efficient liquid phase chromatographic analysis
Using octadecylsilane chemically bonded silica as filler, with water-methanol (45:55) for mobile phase, in certain wavelength
Under detected, it is accurate respectively to draw reference substance solution and each 10 μ l of test solution, inject liquid chromatograph, be measured,
Theoretical cam curve is calculated by psoralen peak should be not less than 3000.
The experiment condition in detection method is investigated below:
A. the determination of Detection wavelength
Psoralea corylifolia reference substance, appropriate Isopsoralen are taken, it is accurately weighed, add methanol that the solution that every 1ml contains 20 μ g is made, i.e.,
?.The reference substance solution is taken, 10 μ l of sample introduction is analyzed, and is carried out full wavelength scanner and is recorded its ultraviolet figure, as a result such as Fig. 2 and Fig. 3
It is shown, figure it is seen that psoralen nearby has maximal ultraviolet absorption for 204nm, 244nm and 294nm in wavelength, and
From figure 3, it can be seen that Isopsoralen nearby has maximal ultraviolet absorption in 199nm, 246nm and 300nm, therefore, in conjunction with Fig. 2
With Fig. 3 as can be seen that being that psoralen and isopsorapen all has biggish UV absorption at 246nm in wavelength, therefore, choosing
Taking Detection wavelength is 246nm.
B. the preparation of test solution
The investigation of I different solvents
Psoralea corylifolia standard decoction 1.0g, finely ground, accurately weighed 0.1g prepared by Example one, sets tool plug cone by parallel 2 parts
In shape bottle, Diluted Alcohol, 75% ethyl alcohol, 95% ethyl alcohol, 50% methanol, 75% methanol, each 50ml of methanol is added in precision, and close plug claims
Determine weight, ultrasonic (power 250W, frequency 50kHz) is handled 30 minutes respectively, is let cool, then weighed weight, is mended with corresponding solvent
The weight of sufficient less loss shakes up, filters, take subsequent filtrate to get.It is accurate respectively to draw each 10 μ l of test liquid, liquid chromatograph is injected,
It is measured by above-mentioned chromatographic condition, obtains psoralen, Isopsoralen total content, the results are shown in Table 4 and Fig. 4.
The investigation of 4 different solvents of table
It can be seen that ethyl alcohol work of the psoralen and isopsorapen 75% in psoralea corylifolia standard decoction from table 4 and Fig. 4
Total content is higher when for solvent, and the total content of psoralen and isopsorapen reaches 1.352% as can be seen from Table 4, finally
The ethyl alcohol of selection 75% is as Extraction solvent.
The investigation of II different extracting modes
The psoralea corylifolia standard decoction 1.0g that Example one is prepared, finely ground, accurately weighed 0.1g, are set by parallel three parts
In stuffed conical flask, 75% ethyl alcohol 50ml, close plug, weighed weight, difference ultrasound (power 250W, frequency 50kHz) is added in precision
It handles 30 minutes, be heated to reflux 30 minutes, directly shaking dissolution, let cool, then weighed weight, the weight of less loss is supplied with 75% ethyl alcohol
Amount, shake up, filter, take subsequent filtrate to get.It is accurate respectively to draw each 10 μ l of test liquid, liquid chromatograph is injected, by above-mentioned chromatography
Condition measurement, obtains psoralen, Isopsoralen total content, as a result as shown in table 5 and Fig. 5.
The investigation of the different extracting modes of table 5
It can be seen that when using ultrasonic extraction mode from table 5 and Fig. 5, the total content of psoralen and isopsorapen
Highest, total content are up to 1.313%, therefore, ultrasound are taken to extract.
The investigation of III different extraction times
The psoralea corylifolia standard decoction about 0.1g that Example one is prepared, it is parallel 2 parts, accurately weighed, set tool plug taper
In bottle, 75% ethyl alcohol 50ml is added in precision, and close plug, weighed weight, ultrasonic (power 250W, frequency 50kHz) handles 15 points respectively
Clock, 30 minutes, 45 minutes, let cool, then weighed weight, the weight of less loss are supplied with 75% ethyl alcohol, shake up, filter, take subsequent filtrate,
To obtain the final product.It is accurate respectively to draw each 10 μ l of test liquid, liquid chromatograph is injected, by the measurement of above-mentioned chromatographic condition, obtains psoralen, different
Psoralen total content, the results are shown in Table 6 and Fig. 6.
The investigation of the different extraction times of table 6
From table 6 and Fig. 6 can be seen that at the extraction between in 30min, the total content of psoralen and isopsorapen compared with
Greatly, total content reaches 1.313%, therefore, uses extraction time for 30min.
It is detected using the above-mentioned condition optimized, result is as shown in fig. 7, according to spectrogram, gross mass content
The calculation formula of calculation formula and total rate of transform show that the gross mass content of psoralen and isopsorapen is 1.65%, mends
Total rate of transform of bone fat element and Isopsoralen is 34.52%.
The investigation of the methodology of 2.2 pairs of detection methods
A. the investigation of specificity
Psoralea corylifolia standard decoction test solution, the psoralen reference substance that accurate extraction embodiment one is prepared are molten
Liquid, Isopsoralen reference substance solution and each 10 μ l of methanol (negative control) inject liquid chromatograph, measure by chromatographic condition,
As a result see Fig. 8.
From figure 8, it is seen that the analysis method can correctly detect the gross mass content of psoralen and isopsorapen,
Not by the interference of Extraction solvent.
B. repetitive test
The psoralea corylifolia standard decoction that Example one is prepared, parallel 6 parts, sample confession is respectively prepared in every part of 2 needle of sample introduction
Test solution, 10 μ l of sample introduction, measures psoralen, Isopsoralen total amount, calculates its mass content and RSD, the results are shown in Table 7 respectively.
7 repetitive test of table
As can be seen from Table 7, the stable content of psoralen and isopsorapen, repeatability is preferable, RSD 0.86%.
C. stability test
Be prepared 1 part of sample of Example one, prepares test solution, temporally puts 10 μ l of sample introduction, measures peak face
Product value calculates its RSD, investigates the stability in 12 hours, the results are shown in Table 8 and table 9.
The stability test measurement result (psoralen) of 8 sample of table
The stability test measurement result (Isopsoralen) of 9 sample of table
The test solution of psoralea corylifolia standard decoction is good in 12 hours internal stabilities it can be seen from table 8 and table 9.
D. the investigation of flow velocity
1 part of sample of psoralea corylifolia standard decoction prepared by Example one is prepared into test solution, investigates 0.7ml/
Tri- flow velocitys of min, 0.8ml/min, 0.9ml/min, experimental result is as shown in table 10 and Fig. 9.
The investigation different in flow rate of table 10
| Serial number | Flow velocity (ml/min) | Psoralen, Isopsoralen total amount/% | RSD/% |
| 1 | 0.7 | 1.20 | |
| 2 | 0.8 | 1.20 | 0 |
| 3 | 0.9 | 1.20 |
It is preferable to can be seen that chromatographic peak separating effect under three kinds of flow velocitys from table 10 and Fig. 9.When flow velocity is 0.8ml/min
It is moderate the time required to analysis, therefore select flow velocity for 0.8ml/min.
E. the investigation of different column temperatures
1 part of the psoralea corylifolia standard decoction sample that Example one is prepared prepares test solution, investigates 25 DEG C, 30
DEG C, 35 DEG C of three temperature, experimental result is as shown in table 11 and Figure 10.
The investigation of the different column temperatures of table 11
| Serial number | Column temperature (DEG C) | Psoralen, Isopsoralen total amount/% | RSD/% |
| 1 | 25 | 1.20 | |
| 2 | 30 | 1.21 | 0.48 |
| 3 | 35 | 1.20 |
It is preferable to can be seen that chromatographic peak separating effect under three kinds of column temperatures from table 11 and Figure 10.Column temperature is analyzed when being 30 DEG C
Required time is moderate, it is contemplated that the tolerance of chromatographic column selects column temperature for 30 DEG C.
F. the investigation of different chromatographic columns
The psoralea corylifolia standard decoction sample that Example one is prepared, prepares test solution, is respectively adopted
Inertsil ODS-HL (Shimadzu, 250 × 4.6mm, 5 μm), ZORBAX SB-C18 (Agilent, 250 × 4.6mm, 5 μm),
Three kinds of chromatographic columns of Diamonsil C18 (2) (DiKMA, 250 × 4.6mm, 5 μm), divide psoralea corylifolia standard decoction sample
Analysis, experimental result is as shown in table 12 and Figure 11.
The comparison of the different chromatographic columns of table 12
| Serial number | Chromatographic column | Psoralen, Isopsoralen total amount/% | RSD/% |
| 1 | Inertsil | 1.21 | |
| 2 | ZORBAX | 1.20 | 0.45 |
| 3 | Diamonsil | 1.20 |
The separating effect that can be seen that three kinds of different model chromatographic columns from table 12 and Figure 11 is preferable, and retention time is moderate,
It is smaller to illustrate that chromatographic column influences the measurement result of sample, using Inertsil chromatographic column.
3. the detection method of the characteristic spectrum of psoralea corylifolia standard decoction
The optimization of the condition of 3.1 pairs of detection methods
(1) referring to the preparation of product solution
Psoralen reference substance, appropriate Isopsoralen are taken, it is accurately weighed, add methanol that the solution that every 1ml contains 20 μ g is made,
To obtain the final product.
(2) preparation of test solution
The psoralea corylifolia standard decoction that Example one is prepared, finely ground, accurately weighed 0.1g is set in stuffed conical flask,
Solvent 25ml is added in precision, and weighed weight extracts processing a period of time, and taking-up lets cool, is transferred in 50ml volumetric flask, adds
Solvent is settled to scale, shakes up, filtration, take filtrate to get.
(3) ultra performance liquid chromatography is analyzed
It is accurate respectively to draw reference substance solution and each 2 μ l of test solution, liquid chromatograph is injected, using octadecyl silicon
Alkane bonded silica gel is filler, using methanol as mobile phase A, carries out gradient elution according to the concentration of following table using water phase as Mobile phase B,
It is measured under certain Detection wavelength with certain column temperature and flow velocity, number of theoretical plate should be not less than by the calculating of psoralen peak
3000。
| Time (minute) | Mobile phase A (%) | Mobile phase B (%) |
| 0~4 | 5→60 | 95→40 |
| 4~14 | 60→100 | 40→0 |
| 14~14.1 | 100→5 | 0→95 |
| 14.1~18 | 5 | 95 |
The process conditions of detection method are optimized first, as follows:
1) determination of Detection wavelength
The psoralea corylifolia standard decoction that Example one is prepared, it is finely ground.About 0.1g is taken, it is accurately weighed, set tool plug taper
In bottle, 95% ethyl alcohol 25ml is added in precision, and weighed weight is ultrasonically treated 30min (250W, 40kHz), and taking-up is let cool, and is transferred to
In 50ml volumetric flask, add 95% ethyl alcohol to be settled to scale, shake up, takes subsequent filtrate as test liquid, record 190~400nm range
Interior absorption spectrum, as a result as shown in figure 12.
In figure 12 it can be seen that in 246nm or so, chromatographic peak that psoralea corylifolia standard decoction sample solution can detecte
At most, and the peak area of each chromatographic peak is larger, therefore selects 246nm for Detection wavelength.
2) investigation of flow visualizing
Using methanol as mobile phase A, analyzed by Mobile phase B of water phase, experimental result as shown in Figure 13-1 to 13-4,
In, the water phase is respectively water, 0.1% phosphate aqueous solution, 0.1% aqueous formic acid and 0.2% aqueous acetic acid.
Can be seen that from Figure 13-1 to 13-4 after the acid addition, peak type be improved significantly, by Figure 13-2, Figure 13-3 with
And Figure 13-4 is compared, it is known that it is preferable with the peak type of-0.1% phosphate aqueous solution of methanol, therefore, select-0.1% phosphoric acid of methanol
Aqueous solution is as flow visualizing.
3) investigation of different chromatographic columns
Using three kinds of difference filler chromatographic column BEH Shield RP18 (Waters, 2.1 × 100mm, 1.7 μm), BEH
C18 (Waters, 2.1 × 100mm, 1.7 μm), Cortecs T3 (Waters, 2.1 × 100mm, 1.6 μm) carry out characteristic spectrum
Analysis, separating effect is as shown in Figure 14-1 and 14-3.
It can be seen that replacement chromatographic column from Figure 14-1 and 14-3, do not influence the separation of characteristic peak.The present invention selects BEH
C18 (Waters, 2.1 × 100mm, 1.7 μm) is separated.
4) investigation of column temperature
With BEH C18 (Waters, 2.1 × 100mm, 1.7 μm) chromatographic column, having investigated column temperature respectively is 25 DEG C, 30 DEG C, 35
DEG C when sample separating effect, as a result as shown in Figure 15-1 to 15-3.
It can be seen that column temperature from Figure 15-1 to 15-3 not having much affect to the separation of sample.It is resistance in view of chromatographic column
By property, select column temperature for 30 DEG C.
5) investigation of flow velocity
Using BEH C18 (Waters, 2.1 × 100mm, 1.7 μm) chromatographic column, investigated respectively flow velocity be 0.2ml/min,
Sample separating effect when 0.25ml/min and 0.3ml/min, as a result as shown in Figure 16-1 to 16-3.
It is can be seen that from Figure 16-1 to 16-3 when flow velocity is 0.25ml/min, good separating effect, therefore, using flow velocity
It is separated for 0.25ml/min.
6) investigation of test sample Extraction solvent
The psoralea corylifolia standard decoction sample about 0.1g that Example one is prepared, it is accurately weighed, set 100ml conical flask
In, it is accurate respectively that aqueous solvent, 95% ethyl alcohol, 75% ethyl alcohol, Diluted Alcohol, methanol, 75% methanol, 50% methanol 25ml is added, surpass
Sonication (power 250W, frequency 40kHz) 30min, taking-up let cool, are transferred in 50ml volumetric flask, coordinative solvent is added to be settled to
Scale shakes up, then filtering is analyzed, result is as shown in Figure 17-1 to 17-7.
Water be can be seen that from Figure 17-1 to 17-7 as solvent, the characteristic peak extracted is less, and ethanol system is compared with first
For alcohol system, a chromatographic peak is had more in 1.3min or so, and difference is little between ethanol system, the present invention chooses 95%
Ethyl alcohol is as Extraction solvent.
7) investigation of test solution extracting mode
The psoralea corylifolia standard decoction sample about 0.1g that Example one is prepared, it is accurately weighed, set 100mL conical flask
In, 95% ethyl alcohol 25ml is added in precision, and ultrasound, reflux, mechanical shaking extraction 30min is respectively adopted, and taking-up is let cool, and is transferred to 50ml appearance
In measuring bottle, add 95% ethyl alcohol to be settled to scale, shake up, filtration is analyzed.As a result as shown in Figure 18-1 to 18-3 and table 13.
The different extracting modes of table 13 investigate (* seconds/sample weighting amount of peak area microvolt g)
| Extracting mode | Peak 1 | Peak 2 | Peak 3 | Peak 4 | Peak 5 |
| Ultrasonic 30min | 24170275 | 19598224 | 6420844 | 4923523 | 5073876 |
| Flow back 30min | 28648409 | 23049941 | 6432540 | 4993706 | 4819990 |
| Mechanical shaking extraction 30min | 27928481 | 22453067 | 6262781 | 4856755 | 4708373 |
From Figure 18-1 to 18-3 and table 13 is as can be seen that using three kinds of ultrasound, reflux, shaking resulting chromatographies of extracting mode
Peak number mesh is consistent, and ultrasonic extraction is different for different characteristic peak recovery rates from refluxing extraction mode, but difference is little, however
Ultrasonic treatment is simple to operation compared with refluxing extraction, and therefore, the present invention is extracted using ultrasound treatment patterns.
8) investigation of test solution extraction time
The psoralea corylifolia standard decoction sample about 0.1g that Example one is prepared, it is accurately weighed, set 100mL conical flask
In, 95% methanol 25ml is added in precision, is ultrasonically treated (power 250W, frequency 40kHz) 15min, 30min, 45min respectively, takes
It lets cool out, is transferred in 50ml volumetric flask, is settled to scale, shake up, filter to obtain the final product.Measurement result be shown in Table 14 and Figure 19-1 to
19-3。
The different extraction times of table 14 investigate (* seconds/sample weighting amount of peak area microvolt g)
| Extraction time | Peak 1 | Peak 2 | Peak 3 | Peak 4 | Peak 5 |
| 15min | 23461557 | 20327685 | 6351084 | 4858020 | 4690729 |
| 30min | 24640336 | 21347243 | 6409012 | 4916818 | 4707174 |
| 45min | 22369626 | 19431389 | 6333488 | 4854227 | 4668749 |
It is i.e. extractable complete to can be seen that ultrasonic extraction 30 minutes from table 14 and Figure 19-1 to 19-3, therefore, the present invention adopts
It is to extract for 30 minutes with extraction time.
The measurement of finger-print is carried out to standard decoction using the process conditions of above-mentioned optimization, as a result as shown in figure 20.
As seen from Figure 20, there should be 5 characteristic peaks in the characteristic spectrum of corylifolia L, peak corresponding with object of reference is
The peak S calculates the relative retention time of each characteristic peak Yu the peak S, wherein the relative retention time of peak (1) is 0.773, the phase of peak (2)
It is 0.806 to retention time, the relative retention time of peak (S) is 1.000, and the relative retention time of peak (4) is 1.032, peak (5)
Relative retention time be 2.072.
The methodological study of 3.2 pairs of detection methods
I precision is investigated
The psoralea corylifolia standard decoction sample that Example one is prepared is prepared by sample solution preparation method for examination
Liquid, sample introduction 6 times, 2 μ l, investigates the consistency of characteristic peak relative retention time and relative peak area every time, as a result such as table 15 and 16
It is shown.
15 precision experiment result of table (relative retention time)
| Number | Peak 1 | Peak 2 | Peak 3 (S) | Peak 4 | Peak 5 |
| 1 | 0.7711 | 0.8040 | 1.0000 | 1.0319 | 2.1335 |
| 2 | 0.7713 | 0.8042 | 1.0000 | 1.0320 | 2.1298 |
| 3 | 0.7715 | 0.8045 | 1.0000 | 1.0318 | 2.1284 |
| 4 | 0.7714 | 0.8044 | 1.0000 | 1.0318 | 2.1278 |
| 5 | 0.7717 | 0.8047 | 1.0000 | 1.0318 | 2.1279 |
| 6 | 0.7718 | 0.8047 | 1.0000 | 1.0318 | 2.1283 |
| RSD (%) | 0.032 | 0.034 | 0.000 | 0.009 | 0.103 |
16 precision experiment result of table (relative peak area)
| Number | Peak 1 | Peak 2 | Peak 3 (S) | Peak 4 | Peak 5 |
| 1 | 2.8774 | 2.2874 | 1.0000 | 0.6937 | 0.7190 |
| 2 | 2.7307 | 2.1804 | 1.0000 | 0.6791 | 0.6817 |
| 3 | 2.7728 | 2.2259 | 1.0000 | 0.6830 | 0.7008 |
| 4 | 2.7587 | 2.2206 | 1.0000 | 0.6799 | 0.6958 |
| 5 | 2.7608 | 2.2239 | 1.0000 | 0.6809 | 0.6966 |
| 6 | 2.7627 | 2.2270 | 1.0000 | 0.6807 | 0.6974 |
| RSD (%) | 1.839 | 1.537 | 0.000 | 0.799 | 1.720 |
From table 15 and table 16 as can be seen that each characteristic peak relative retention time and relative peak area RSD are respectively less than 2%, instrument
Device precision is good.
II repeatability is investigated
Totally 6 parts of the psoralea corylifolia standard decoction that Example one is prepared, confession is prepared by sample solution preparation method
Test solution, 2 μ l of sample introduction analysis, investigates the consistency of characteristic peak relative retention time and relative peak area, as a result such as table 17 and table 18
It is shown.
17 repeated experiment result (relative retention time) of table
| Number | Peak 1 | Peak 2 | Peak 3 (S) | Peak 4 | Peak 5 |
| 1 | 0.7720 | 0.8050 | 1.0000 | 1.0316 | 2.1279 |
| 2 | 0.7721 | 0.8051 | 1.0000 | 1.0318 | 2.1276 |
| 3 | 0.7721 | 0.8051 | 1.0000 | 1.0316 | 2.1276 |
| 4 | 0.7720 | 0.8050 | 1.0000 | 1.0318 | 2.1271 |
| 5 | 0.7721 | 0.8051 | 1.0000 | 1.0316 | 2.1264 |
| 6 | 0.7722 | 0.8052 | 1.0000 | 1.0318 | 2.1279 |
| RSD (%) | 0.007 | 0.007 | 0.000 | 0.008 | 0.027 |
18 repeated experiment result (relative peak area) of table
| Number | Peak 1 | Peak 2 | Peak 3 (S) | Peak 4 | Peak 5 |
| 1 | 2.7502 | 2.2140 | 1.0000 | 0.6801 | 0.6916 |
| 2 | 2.6891 | 2.1711 | 1.0000 | 0.6810 | 0.6962 |
| 3 | 2.7418 | 2.2098 | 1.0000 | 0.6790 | 0.6933 |
| 4 | 2.7403 | 2.2115 | 1.0000 | 0.6825 | 0.6893 |
| 5 | 3.0204 | 2.4172 | 1.0000 | 0.6822 | 0.6852 |
| 6 | 2.7533 | 2.2214 | 1.0000 | 0.6789 | 0.6970 |
| RSD (%) | 4.272 | 3.935 | 0.000 | 0.229 | 0.642 |
From table 17 and table 18 as can be seen that the relative retention time RSD of each characteristic peak is respectively less than 0.1%, but with respect to peak
The RSD of area illustrates that the retention time repeatability of this method is good between 0.0%-4.3%, relative peak area repeatability compared with
Difference.Due to relative peak area not being included in standard, the repeatability of this method is good.
III study on the stability
The psoralea corylifolia standard decoction that Example one is prepared, is prepared into test liquid by sample solution preparation method,
It is primary every 2 hours sample introductions, it measures 10 hours altogether, respectively 2 μ l of sample introduction, investigates characteristic peak relative retention time and relative peak area
Consistency, as a result as shown in table 19 and table 20.
19 stability experiment result (relative retention time) of table
| Number | Peak 1 | Peak 2 | Peak 3 (S) | Peak 4 | Peak 5 |
| 1 | 0.7721 | 0.8052 | 1.0000 | 1.0317 | 2.1250 |
| 2 | 0.7719 | 0.8051 | 1.0000 | 1.0317 | 2.1237 |
| 3 | 0.7725 | 0.8055 | 1.0000 | 1.0318 | 2.1233 |
| 4 | 0.7724 | 0.8054 | 1.0000 | 1.0317 | 2.1187 |
| 5 | 0.7726 | 0.8056 | 1.0000 | 1.0317 | 2.1111 |
| 6 | 0.7731 | 0.8059 | 1.0000 | 1.0318 | 2.1027 |
| RSD (%) | 0.053 | 0.035 | 0.000 | 0.005 | 0.417 |
20 stability experiment result (relative peak area) of table
| Number | Peak 1 | Peak 2 | Peak 3 (S) | Peak 4 | Peak 5 |
| 1 | 2.7440 | 2.2094 | 1.0000 | 0.6783 | 0.6906 |
| 2 | 2.7196 | 2.1900 | 1.0000 | 0.6793 | 0.6771 |
| 3 | 2.7530 | 2.2251 | 1.0000 | 0.6814 | 0.6911 |
| 4 | 2.7530 | 2.2350 | 1.0000 | 0.6801 | 0.6919 |
| 5 | 2.8160 | 2.3072 | 1.0000 | 0.6954 | 0.6952 |
| 6 | 2.7497 | 2.3495 | 1.0000 | 0.6985 | 0.6874 |
| RSD (%) | 1.612 | 2.752 | 0.000 | 1.312 | 0.916 |
From table 19 and table 20 as can be seen that the relative retention time of each characteristic peak and the RSD of relative peak area are respectively less than
3.0%, illustrate that test sample is good in 10h internal stability.
Embodiment two
1. the preparation method of psoralea corylifolia standard decoction
(1) psoralea corylifolia medicine materical crude slice 100g is taken, is added water to cook twice, first decocts the water for adding 7 times of amounts, opens intense fire, after boiling, adjusts
To be cooked by slow fire 60 minutes, the water for adding 4 times of amounts are decocted with 100 mesh filter-cloth filterings, second while hot, intense fire is opened, after boiling, is adjusted to text
Fire decocts 45min, is filtered while hot with 100 mesh filter clothes, merges filtrate twice；
(2) by filtrate be transferred in Rotary Evaporators concentration (bath temperature: 60 DEG C, vacuum degree is -0.08~-
0.09MPa), being concentrated into density is 1.05g/ml, and adjusting solid content to 14% is dispensed into 10ml cillin bottle, partly jumped a queue, every bottle
Packing volume is 2.0ml, is transferred in vacuum freeze drier after packing, and pre-freeze is first carried out, and pre-freezing temperature is -50 DEG C,
The pre-freeze time is 3h, then carries out primary drying, and drying temperature is followed successively by -30 DEG C, and -20 DEG C, -10 DEG C, 0 DEG C, drying time divides
Not Wei 7h, 2h, 2h, 3h, then carry out redrying, drying temperature is followed successively by 5 DEG C, and 15 DEG C, 25 DEG C, drying time is respectively
2h, 2h, 3h take out, roll aluminium lid, obtain psoralea corylifolia standard decoction, the paste-forming rate of the psoralea corylifolia is by the formula in embodiment one
It is calculated, obtaining paste-forming rate is 17.10%, and the biodiversity content of the psoralea corylifolia standard decoction is according in embodiment one
Formula is calculated, and the biodiversity content for obtaining psoralea corylifolia standard decoction is 3%.
2. the psoralen and isopsorapen gross mass content of psoralea corylifolia standard decoction and the detection method of total rate of transform
According to detection method described in embodiment one to the gross mass content of psoralen and isopsorapen and total transfer
Rate is measured, and content spectrogram is as shown in figure 21, public according to the calculating of gross mass content and total rate of transform in embodiment one
Formula show that the gross mass content of psoralen and isopsorapen is 0.89%, and total rate of transform is 18.58%.
3. the measuring method of the characteristic spectrum of psoralea corylifolia standard decoction
According to the characteristic spectrum of the measurement psoralea corylifolia standard decoction of detection method described in embodiment one, spectrogram such as Figure 22 institute
Show, it includes five peaks, respectively 0.774 (peak 1), 0.805 (peak 2), 1.000 (peak S), 1.033 (peaks 4), 2.081 (peaks 5).
Embodiment three
1. the preparation method of psoralea corylifolia standard decoction
(1) psoralea corylifolia medicine materical crude slice 100g is taken, is added water to cook twice, first decocts the water for adding 9 times of amounts, intense fire (500W) is opened, wait boil
After boiling, it is adjusted to mild fire (200W) and decocts 30 minutes, decoct the water for adding 4 times of amounts with 300 mesh filter-cloth filterings, second while hot, open intense fire
(500W) is adjusted to mild fire (200W) and decocts 20min, be filtered while hot with 300 mesh filter clothes, merge filtrate twice after boiling；
(2) by filtrate be transferred in Rotary Evaporators concentration (bath temperature: 65 DEG C, vacuum degree is -0.08~-
0.09MPa), being concentrated into density is 1.05g/ml, and adjusting solid content to 14% is dispensed into 10ml cillin bottle, partly jumped a queue, every bottle
Packing volume is 2.0ml, is transferred in vacuum freeze drier after packing, and pre-freeze is first carried out, and pre-freezing temperature is -45 DEG C,
The pre-freeze time is 6h, then carries out primary drying, and drying temperature is followed successively by -30 DEG C, and -20 DEG C, -10 DEG C, 0 DEG C, drying time divides
Not Wei 5h, 2h, 2h, 1h, then carry out redrying, drying temperature is followed successively by 5 DEG C, and 15 DEG C, 25 DEG C, drying time is respectively
2h, 2h, 1h take out, roll aluminium lid, obtain psoralea corylifolia standard decoction, the paste-forming rate of the psoralea corylifolia is by the formula in embodiment one
It is calculated, obtaining paste-forming rate is 18.35%, and the biodiversity content of the psoralea corylifolia is carried out according to the formula in embodiment one
It calculates, show that the biodiversity content of psoralea corylifolia standard decoction is 2.8%.
2. the psoralen and isopsorapen gross mass content of psoralea corylifolia standard decoction and the detection method of total rate of transform
According to detection method described in embodiment one to the gross mass content of psoralen and isopsorapen and total transfer
Rate is measured, and content spectrogram is as shown in figure 23, public according to the calculating of gross mass content and total rate of transform in embodiment one
Formula, as a result: the gross mass content of psoralen and isopsorapen is 1.22%, and total rate of transform is 24.76%.
3. the measuring method of the characteristic spectrum of psoralea corylifolia standard decoction
According to the characteristic spectrum of the measurement psoralea corylifolia standard decoction of detection method described in embodiment one, spectrogram such as Figure 24 institute
Show, it includes five peaks, respectively 0.773 (peak 1), 0.805 (peak 2), 1.000 (peak S), 1.032 (peaks 4), 2.087 (peaks 5).
Example IV
1. the preparation method of psoralea corylifolia standard decoction
(1) psoralea corylifolia medicine materical crude slice 100g is taken, is added water to cook twice, first decocts the water for adding 8 times of amounts, starts to decoct after impregnating 30min
It boils, opens intense fire (500W), after boiling, be adjusted to mild fire (200W) and decoct 45 minutes, decocted while hot with 200 mesh filter-cloth filterings, second
The water for adding 6 times of amounts, opens intense fire (500W), after boiling, is adjusted to mild fire (200W) and decocts 30min, carried out while hot with 200 mesh filter clothes
Filtering merges filtrate twice；
(2) by filtrate be transferred in Rotary Evaporators concentration (bath temperature: 65 DEG C, vacuum degree is -0.08~-
0.09MPa), being concentrated into density is 1.03g/ml, and adjusting solid content to 10% is dispensed into 10ml cillin bottle, partly jumped a queue, every bottle
Packing volume is 2.0ml, is transferred in vacuum freeze drier after packing, and pre-freeze is first carried out, and pre-freezing temperature is -45 DEG C,
The pre-freeze time is 4h, then carries out primary drying, and drying temperature is followed successively by -30 DEG C, and -20 DEG C, -10 DEG C, 0 DEG C, drying time divides
Not Wei 7h, 2h, 2h, 3h, then carry out redrying, drying temperature is followed successively by 5 DEG C, and 15 DEG C, 25 DEG C, drying time is respectively
2h, 2h, 3h take out, roll aluminium lid, obtain psoralea corylifolia standard decoction, the paste-forming rate of the psoralea corylifolia by embodiment one formula into
Row calculates, and obtaining paste-forming rate is 20.16%, the biodiversity content of the psoralea corylifolia standard decoction according to embodiment one formula into
Row calculates, and show that the biodiversity content of psoralea corylifolia standard decoction is 3.6%.
2. the psoralen and isopsorapen gross mass content of psoralea corylifolia standard decoction and the detection method of total rate of transform
According to detection method described in embodiment one to the gross mass content of psoralen and isopsorapen and total transfer
Rate is measured, and content spectrogram is as shown in figure 25, public according to the calculating of gross mass content and total rate of transform in embodiment one
Formula, as a result: the gross mass content of psoralen and isopsorapen is 1.43%, and total rate of transform is 28.91%.
3. the measuring method of the characteristic spectrum of psoralea corylifolia standard decoction
According to the characteristic spectrum of the measurement psoralea corylifolia standard decoction of detection method described in embodiment one, spectrogram such as Figure 26 institute
Show, it includes five peaks, respectively 0.774 (peak 1), 0.805 (peak 2), 1.000 (peak S), 1.033 (peaks 4), 1.978 (peaks 5).
Comparative example one
1. the preparation method of psoralea corylifolia standard decoction
Psoralea corylifolia standard decoction is prepared using the method being the same as example 1, difference is only that comparative example one using normal
The freeze-drying of rule is handled, and psoralea corylifolia standard decoction is obtained, and the paste-forming rate of psoralea corylifolia standard decoction is 22.19%, moisture
Mass content is calculated according to the formula of embodiment one, show that the biodiversity content of psoralea corylifolia standard decoction is up to 75-
90%, illustrate that the freeze drying process of this method cannot be removed effectively moisture.
2. the psoralen and isopsorapen total content of psoralea corylifolia standard decoction and the detection method of total rate of transform
According to detection method described in embodiment one to the gross mass content of psoralen and isopsorapen and total transfer
Rate is measured.As a result: the gross mass content of psoralen and isopsorapen is 1.32%, and total rate of transform is
26.07%.
3. the measuring method of the characteristic spectrum of psoralea corylifolia standard decoction
According to the characteristic spectrum of the measurement psoralea corylifolia standard decoction of detection method described in embodiment one, it includes five peaks,
Respectively 0.773 (peak 1), 0.805 (peak 2), 1.000 peak S, 1.032 (peaks 4), 2.086 (peaks 5).
4, the stability of psoralea corylifolia standard decoction
Psoralea corylifolia standard decoction saves 10 days at 5 DEG C, mouldy, not can be carried out and tests in next step.
Embodiment two to the obtained psoralea corylifolia standard decoction of example IV when carrying out acceleration for stabilization test, at 6 months
Interior content is relatively stable, and moisture content is relatively stable, the resistance to moisture absorption, and does not generate impurity component, is carrying out psoralen and different benefit
When the detection of the gross mass content of bone fat element and total rate of transform, method precision is high, and repeatability, stabilizability exists, and is mended
When the detection of the characteristic spectrum of bone fat standard decoction, the method precision is high, reproducible, being capable of stability presence.
The embodiment of the present invention one and comparative example one are compared as can be seen that using method conventional in the prior art
It is freezed, the moisture of standard decoction cannot be effectively removed, and resulting psoralea corylifolia standard decoction is saved 10 days at 5 DEG C and just sent out
It is mould, it not can be carried out the experiment of next step.
In conclusion the present invention is filtered to obtain filtrate using psoralea corylifolia is added water to cook, it is dry that filtrate is carried out concentration
Dry, be then lyophilized, freeze-drying is divided into three phases: a. pre-freeze: pre-freezing temperature is -50 DEG C~-45 DEG C；B. primary drying: dry
Dry temperature is -30 DEG C~0 DEG C；C. redrying is parsed: drying temperature is 5~25 DEG C, obtains psoralea corylifolia standard decoction, institute
The paste-forming rate of obtained psoralea corylifolia standard decoction is 17.10-31.02%, and paste-forming rate is higher, and carries out acceleration for stabilization experiment
When, content is relatively stable in 6 months, and moisture content is relatively stable, the resistance to moisture absorption, and does not generate impurity component, and stability is good,
Preparation-obtained psoralea corylifolia standard decoction is carried out psoralen and isopsorapen using efficient liquid phase chromatographic analysis always to contain
The detection of amount and total rate of transform, as a result: the gross mass content of psoralen and isopsorapen is 0.89-1.65%, is mended
Total rate of transform of bone fat element and Isopsoralen is 18.58-34.52%, and high using the detection method precision, weight
Renaturation is good, and sample test solution has good stability in 12 hours.
The present invention uses the characteristic spectrum of high effective liquid chromatography for measuring psoralea corylifolia standard decoction, the psoralea corylifolia standard soup
It include 5 characteristic peaks in agent, the relative retention time of each characteristic peak is within ± the 5% of specified value, and specified value 0.773
(peak 1), 0.806 (peak 2), 1.000 (peak S), 1.032 (peaks 4), 2.072 (peaks 5).It is high using the detection method precision,
Repeated preferable, sample test solution has good stability in 10 hours.
The above is only the preferred embodiment that the present invention is implemented, and not does limitation in any form to the present invention, all
The modifications, equivalent substitutions and improvements etc. done within the spirit and principles in the present invention are required to be included in protection of the invention
Within the scope of.
Claims (19)
1. a kind of preparation method of psoralea corylifolia standard decoction, it includes following step:
(1) corylifolia L is taken, is added water to cook, filtrate is obtained by filtration；
(2) filtrate in step (1) is concentrated and dried, is then lyophilized, the freeze-drying is divided into three phases: a. pre-freeze: pre-freeze
Temperature is -50 DEG C~-45 DEG C；B. primary drying: drying temperature is -30 DEG C~0 DEG C；C. redrying is parsed: dry temperature
Degree is 5~25 DEG C, obtains psoralea corylifolia standard decoction.
2. preparation method according to claim 1, wherein in step (1), the number that adds water to cook is 1~3 time.
3. preparation method according to claim 2, wherein the decoction number is 2 times, decocts for the first time plus 7~9 times are measured
Water, the water of preferably 8 times amounts, second decocts plus the water of 4~6 times of amounts, the water of preferably 6 times amounts.
4. preparation method according to claim 3, wherein first time decocting time be 30~60min, preferably 60min,
Second of decocting time is 20~45min, preferably 45min.
5. preparation method according to claim 1-4, wherein in step (1), the filtering using 100~
300 mesh screens are filtered, and preferably 100 mesh screens are filtered.
6. preparation method according to claim 1-5, wherein in step (2), the pre-freeze time is 3-6 hours,
Preferably 4 hours；The primary drying time is 10-14 hours, preferably 14 hours；The redrying time is 5~7 hours, preferably
It is 7 hours.
7. preparation method according to claim 1-6, wherein in step (2), what the filtrate was concentrated and dried
Drying temperature is 60~65 DEG C.
8. preparation method according to claim 1-7, wherein the paste-forming rate of the psoralea corylifolia standard decoction is
17.10~31.02%.
9. the detection method of any one of the claim 1-8 psoralea corylifolia standard decoction gross mass content and total rate of transform, packet
Containing following step:
(1) preparation of reference solution:
Psoralea corylifolia reference substance, Isopsoralen are weighed, adds methanol that solution is made；
(2) preparation of test solution:
Psoralea corylifolia standard decoction is taken, solvent is added and extracts；
(3) efficient liquid phase chromatographic analysis
Using octadecylsilane chemically bonded silica as filler, mobile phase A is water, and Mobile phase B is methanol, draw reference substance solution and
Test solution injection high performance liquid chromatograph is analyzed.
10. detection method according to claim 9, wherein in step (2), the solvent is selected from Diluted Alcohol, 75% second
One of alcohol, 95% ethyl alcohol, 50% methanol, 75% methanol and methanol, preferably 75% ethyl alcohol.
11. detection method according to claim 9 or 10, wherein in step (2), the extraction using reflux, dissolution or
One of ultrasound extracts, preferably ultrasonic extraction.
12. detection method according to claim 11, wherein in step (2), the extraction time 15- of the extraction
45min, preferably 30min.
13. according to the described in any item detection methods of claim 9-12, wherein based on dry product, the psoralen and different
The gross mass content of psoralen is 0.89~1.65%.
14. according to the described in any item detection methods of claim 9-13, wherein the psoralen and isopsorapen it is total
The rate of transform is 18.58~34.52%.
15. the detection method of the characteristic spectrum for the psoralea corylifolia standard decoction that any one of claim 1-8 the method is prepared,
It includes following step:
(1) preparation of reference solution:
Psoralea corylifolia reference substance, Isopsoralen are weighed, adds methanol that solution is made；
(2) preparation of test solution:
Psoralea corylifolia standard decoction is taken, solvent is added and extracts；
(3) superelevation liquid chromatogram is analyzed
It draws reference substance solution and test solution injects Ultra Performance Liquid Chromatography instrument, be to fill out with octadecylsilane chemically bonded silica
Fill agent；Using methanol as mobile phase A, gradient elution is carried out by Mobile phase B of water phase, obtains the characteristic pattern of psoralea corylifolia standard decoction
Spectrum.
16. detection method according to claim 15, wherein in step (2), the solvent selected from water, 95% ethyl alcohol,
One of 75% ethyl alcohol, Diluted Alcohol, methanol, 75% methanol, 50% methanol, preferably 95% ethyl alcohol.
17. detection method according to claim 15 or 16, wherein in step (2), it is described extract using refluxing extraction,
One of ultrasonic extraction or dissolution extraction, preferably ultrasonic extraction.
18. the described in any item detection methods of 5-17 according to claim 1, wherein in step (2), when the extraction of the extraction
Between be 15~45min, preferably 30min.
19. the described in any item detection methods of 5-18 according to claim 1, wherein in step (3), the water phase selected from water,
0.1% phosphate aqueous solution, 0.1% aqueous formic acid or 0.2% aqueous acetic acid, preferably 0.1% phosphoric acid water
Solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810148957.9A CN110146600B (en) | 2018-02-13 | 2018-02-13 | Preparation method and detection method of fructus psoraleae standard decoction |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810148957.9A CN110146600B (en) | 2018-02-13 | 2018-02-13 | Preparation method and detection method of fructus psoraleae standard decoction |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110146600A true CN110146600A (en) | 2019-08-20 |
| CN110146600B CN110146600B (en) | 2022-04-26 |
Family
ID=67588713
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810148957.9A Active CN110146600B (en) | 2018-02-13 | 2018-02-13 | Preparation method and detection method of fructus psoraleae standard decoction |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110146600B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113181683A (en) * | 2021-05-12 | 2021-07-30 | 山东宏济堂制药集团股份有限公司 | Radix pseudostellariae standard control substance and separation and purification method and detection method thereof |
| CN114113348A (en) * | 2020-08-26 | 2022-03-01 | 国药集团同济堂（贵州）制药有限公司 | Preparation method, characteristic spectrum and content determination method of rose standard decoction |
| CN114166954A (en) * | 2021-04-07 | 2022-03-11 | 国药集团同济堂（贵州）制药有限公司 | Preparation method and quality detection method of caulis et folium Periplocae Forrestii standard decoction |
| CN114515301A (en) * | 2020-11-19 | 2022-05-20 | 国药集团同济堂（贵州）制药有限公司 | Flatstem milkvetch seed standard decoction, preparation method and detection method |
| CN115737696A (en) * | 2022-11-22 | 2023-03-07 | 国药集团同济堂（贵州）制药有限公司 | Radix cynanchi wilfordii extract as well as preparation method, detection method and application thereof |
Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1115654A (en) * | 1994-07-19 | 1996-01-31 | 于延春 | Kidney-reinforcing oral medicine and its prepn |
| CN1425425A (en) * | 2002-12-27 | 2003-06-25 | 姚志平 | Medicine for preventing and curing osteoporosis and its preparing method |
| CN1839922A (en) * | 2006-01-24 | 2006-10-04 | 河南辅仁药业集团有限公司 | Traditional Chinese medicine preparation for treating leukoderma, psoriasis and its preparation method |
| CN101458237A (en) * | 2008-12-29 | 2009-06-17 | 北京同仁堂股份有限公司 | Method for detecting malaytea scurfpea fruit in malaytea scurfpea fruit containing preparations |
| CN101549010A (en) * | 2009-05-14 | 2009-10-07 | 浙江大学 | A preparing method and application of malaytea scurfpea fruit total glycosides extract |
| CN103083521A (en) * | 2013-02-20 | 2013-05-08 | 贵州同济堂制药有限公司 | Extraction method, separated extract and preparation of Xianlinggubao |
| CN105535199A (en) * | 2016-02-22 | 2016-05-04 | 林立朋 | Pharmaceutical preparation for treating polycystic ovarian syndromes accompanied with mammary gland hyperplasia |
| CN105687539A (en) * | 2016-03-14 | 2016-06-22 | 张艳娟 | Pharmaceutical preparation for treating low blocking antibody type recurrent spontaneous abortions |
| EP3061452A4 (en) * | 2013-10-21 | 2017-05-03 | Lunan Pharmaceutical Group Corporation | Use of icaritin in preparing medicament for preventing or treating hematocytopenia |
| CN106806599A (en) * | 2015-11-27 | 2017-06-09 | 宋淑芹 | A kind of Xianlinggubao side's prepared slices of Chinese crude drugs combination preparation and preparation method |
| CN107064347A (en) * | 2017-03-29 | 2017-08-18 | 广西壮族自治区梧州食品药品检验所 | A kind of ASE HPLC methods determine Psoralen, the method for Isopsoralen content in Psoralen ester |
| CN107456479A (en) * | 2017-08-02 | 2017-12-12 | 浙江大学 | A kind of preparation of psoralea corylifolia granule and detection method |
| US20170368083A1 (en) * | 2016-06-24 | 2017-12-28 | Infinitus (China) Company Ltd. | Ginsenoside composition, and preparation method and use thereof |
-
2018
- 2018-02-13 CN CN201810148957.9A patent/CN110146600B/en active Active
Patent Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1115654A (en) * | 1994-07-19 | 1996-01-31 | 于延春 | Kidney-reinforcing oral medicine and its prepn |
| CN1425425A (en) * | 2002-12-27 | 2003-06-25 | 姚志平 | Medicine for preventing and curing osteoporosis and its preparing method |
| CN1839922A (en) * | 2006-01-24 | 2006-10-04 | 河南辅仁药业集团有限公司 | Traditional Chinese medicine preparation for treating leukoderma, psoriasis and its preparation method |
| CN101458237A (en) * | 2008-12-29 | 2009-06-17 | 北京同仁堂股份有限公司 | Method for detecting malaytea scurfpea fruit in malaytea scurfpea fruit containing preparations |
| CN101549010A (en) * | 2009-05-14 | 2009-10-07 | 浙江大学 | A preparing method and application of malaytea scurfpea fruit total glycosides extract |
| CN103083521A (en) * | 2013-02-20 | 2013-05-08 | 贵州同济堂制药有限公司 | Extraction method, separated extract and preparation of Xianlinggubao |
| EP3061452A4 (en) * | 2013-10-21 | 2017-05-03 | Lunan Pharmaceutical Group Corporation | Use of icaritin in preparing medicament for preventing or treating hematocytopenia |
| CN106806599A (en) * | 2015-11-27 | 2017-06-09 | 宋淑芹 | A kind of Xianlinggubao side's prepared slices of Chinese crude drugs combination preparation and preparation method |
| CN105535199A (en) * | 2016-02-22 | 2016-05-04 | 林立朋 | Pharmaceutical preparation for treating polycystic ovarian syndromes accompanied with mammary gland hyperplasia |
| CN105687539A (en) * | 2016-03-14 | 2016-06-22 | 张艳娟 | Pharmaceutical preparation for treating low blocking antibody type recurrent spontaneous abortions |
| US20170368083A1 (en) * | 2016-06-24 | 2017-12-28 | Infinitus (China) Company Ltd. | Ginsenoside composition, and preparation method and use thereof |
| CN107064347A (en) * | 2017-03-29 | 2017-08-18 | 广西壮族自治区梧州食品药品检验所 | A kind of ASE HPLC methods determine Psoralen, the method for Isopsoralen content in Psoralen ester |
| CN107456479A (en) * | 2017-08-02 | 2017-12-12 | 浙江大学 | A kind of preparation of psoralea corylifolia granule and detection method |
Non-Patent Citations (3)
| Title |
|---|
| LUHUA ZHAO等: "Fingerprint analysis of Psoralea corylifolia L. by HPLC and LC–MS", 《JOURNAL OF CHROMATOGRAPHY B》 * |
| 张辉等: "健脾止泻合剂水提工艺优选", 《中医研究》 * |
| 王国振等: "癃闭舒片提取工艺优化和含量测定方法研究", 《河北中医药学报》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114113348A (en) * | 2020-08-26 | 2022-03-01 | 国药集团同济堂（贵州）制药有限公司 | Preparation method, characteristic spectrum and content determination method of rose standard decoction |
| CN114113348B (en) * | 2020-08-26 | 2023-10-17 | 国药集团同济堂（贵州）制药有限公司 | Preparation method of standard rose decoction, characteristic spectrum and content determination method |
| CN114515301A (en) * | 2020-11-19 | 2022-05-20 | 国药集团同济堂（贵州）制药有限公司 | Flatstem milkvetch seed standard decoction, preparation method and detection method |
| CN114515301B (en) * | 2020-11-19 | 2024-03-29 | 国药集团同济堂（贵州）制药有限公司 | Semen astragali Complanati standard decoction, and preparation method and detection method thereof |
| CN114166954A (en) * | 2021-04-07 | 2022-03-11 | 国药集团同济堂（贵州）制药有限公司 | Preparation method and quality detection method of caulis et folium Periplocae Forrestii standard decoction |
| CN114166954B (en) * | 2021-04-07 | 2023-10-17 | 国药集团同济堂（贵州）制药有限公司 | Preparation method and quality detection method of black bone vine standard decoction |
| CN113181683A (en) * | 2021-05-12 | 2021-07-30 | 山东宏济堂制药集团股份有限公司 | Radix pseudostellariae standard control substance and separation and purification method and detection method thereof |
| CN115737696A (en) * | 2022-11-22 | 2023-03-07 | 国药集团同济堂（贵州）制药有限公司 | Radix cynanchi wilfordii extract as well as preparation method, detection method and application thereof |
| CN115737696B (en) * | 2022-11-22 | 2024-04-23 | 国药集团同济堂（贵州）制药有限公司 | Radix cynanchi wilfordii extract and preparation method, detection method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110146600B (en) | 2022-04-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110146600A (en) | The preparation method and its detection method of psoralea corylifolia standard decoction | |
| CN106501434A (en) | A kind of HPLC finger print measuring methods of Double Harmonizing Decoction standard soup | |
| CN110455965A (en) | The preparation method and its HPLC fingerprint of pharmaceutical composition | |
| CN107271589A (en) | A kind of semen armeniacae amarae granule and its preparation and method of quality control | |
| CN103330758A (en) | Peony and liquorice soup formula granule, preparation method and detection method of peony and liquorice soup formula granule | |
| CN107456479B (en) | Preparation and detection method of fructus psoraleae formula granules | |
| CN103896890B (en) | A kind of extraction process of rographolide | |
| CN110161135A (en) | The preparation method and its detection method of teasel root standard decoction | |
| CN105777724A (en) | Method for extracting and purifying dehydrated safflower yellow B in safflower | |
| CN102702283B (en) | Method for quickly separating and preparing high-purity deoxyrhapontin and rhapontin | |
| CN102872177A (en) | Method for processing red ginseng | |
| CN109030641B (en) | Method for constructing fingerprint of Yangweishu granules | |
| CN111289648A (en) | Method for establishing traditional Chinese medicine compound preparation fingerprint and fingerprint thereof | |
| CN110196295A (en) | A kind of measuring method of psoralen in Shengjing tablet for invigoration, Isopsoralen content | |
| CN112098569B (en) | Preparation method and quality determination method of radix clematidis extract | |
| CN105055497A (en) | Sijunzi decoction tea and preparation process thereof | |
| CN107998187A (en) | A kind of concocting method of prepared fleece flower root | |
| CN101537056B (en) | Method for preparing Chinese medicinal preparation for treating external respiration infection of children | |
| CN111481647B (en) | Compound coptis composition and preparation process thereof | |
| CN108245561B (en) | Charred fructus crataegi, its preparation method and quality detection method | |
| CN114646721B (en) | Quality detection method of traditional Chinese medicine composition for benefiting brain and dredging collaterals | |
| CN105675741A (en) | Method for determining content of effective components in hair growing pill | |
| CN103006875B (en) | Processing method of honeyed schisandra chinensis | |
| CN104147140A (en) | Preparation method and quality control method of cortex fraxini formula granules | |
| CN103768494B (en) | One treats migrainous pharmaceutical composition and its preparation method and application |
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 |