CN107064334B - The method of quality control of anthracene shellfish element - Google Patents
The method of quality control of anthracene shellfish element Download PDFInfo
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Abstract
The invention discloses a kind of method of quality control of anthracene shellfish element, especially using the mature dry fruit of Embelia laeta platymiscium Embelia laeta (Embelia laeta (Linn.) Mez) as raw material, the method extracted high-purity anthracene shellfish element and it is detected, its process is: after taking Embelia laeta plant extract, using Embelia laeta dry fruit as raw material, with extracted by ether, it is separated through silica gel column chromatography, with TLC combining data detection, collect the flow point containing anthracene shellfish element, high-efficient liquid phase technique separation is prepared with reverse phase again, analytic type high performance liquid chromatography detection is utilized to collected every a eluent, merge and retains high-purity anthracene shellfish element component, quality control is carried out using thin-layer chromatography and liquid chromatogram, this method can not only improve anthracene shellfish element purity, and production cost can be reduced.
Description
Technical field
The present invention relates to a kind of method of quality control fields of medicinal material chemical reference substance, and in particular to a kind of anthracene shellfish element mentions
It takes and method of quality control.
Background technique
Anthracene shellfish element is a kind of Diterpene glucoside class chemical component, is one of active components of plants and many plants and drug
The index ingredient of standard quality control.Current not yet corresponding national drug standards substance, both at home and abroad to anthracene shellfish element reference substance
System research have not been reported, it is right to anthracene shellfish element chemistry referring to the technical requirements of traditional Chinese chemical contrast (for assay)
It is studied according to product, establishes batch extracting technique, purity and the content of anthracene shellfish element chemical reference substance and the analysis of determination of foreign matter
Measuring method, to establish the technical standard of anthracene shellfish element chemical reference substance, for it as traditional Chinese chemical contrast and medicinal material and
The quality standard research of preparation provides scientific basic and guarantee.
The anthracene shellfish element reference substance of high-purity is the key problem in technology for controlling plant Embelia laeta and the quality containing Embelia laeta, numerous
Enterprise, scientific research and testing department require the anthracene shellfish element reference substance of high-purity, and the market demand is very big, since anthracene shellfish element is in medicinal material
In content it is low, extraction and separation technology require it is very high, difficult.The present invention carry out anthracene shellfish element Chemistry for Chinese Traditional Medicine standard items preparation and
The research of its Quality Control Technology, solves the problems, such as high-purity anthracene shellfish element chemical reference substance, the effect to the modernization of Chinese medicine be show and
It is clear to, there is great practical significance and learning value.
Anthracene shellfish element is the active material isolated from Embelia laeta plant, has the extracting method of document report anthracene shellfish element,
Such as: 1.[autograph] using hydrotropic agent from extracting embelin in common embelia fruit: establish it is a kind of with hydrotropic agent from common embelia fruit
The method for extracting embelin.Common embelia fruit dry, pulverize, and cumene sodium sulfonate (Na CS) aqueous solution is added, and stir, filtration.Filter
Liquid acid adding water, which is diluted to minimum hydrotropy concentration (MHC), precipitates embelin hereinafter, standing 60 min, and suspension centrifuge separation, which is rich in, presses
Shellfish extract.As a result optimum extraction condition is as follows, Extracting temperature: 25 ~ 30 DEG C, medicinal material Powder Particle Size: 100 mesh, hydrotropic agent
Concentration: 2.0 mol/L stir extraction time: 60 min.This method recovery rate > 90%, anthracene shellfish element purity > 85%.
The above method discusses the extraction separation of anthracene shellfish element, and separation purity is higher, but purity is not up to Chemistry for Chinese Traditional Medicine pair
According to the requirement of product, i.e. purity is greater than 98%, and separation process is cumbersome, and does not have anthracene shellfish element purity and method of quality control, is unable to satisfy
The needs of high-purity anthracene shellfish element chemical reference substance.
Summary of the invention
It is an object of the present invention to overcome the shortcomings of that existing anthracene shellfish element DNA purity is low, yield is low or high production cost,
And without system anthracene shellfish element extraction purification and method of quality control, provide the preparation and its quality control of a kind of high-purity anthracene shellfish element
Method, the anthracene shellfish element that this method is prepared is with high purity, high-quality, can be used as chemical reference substance or standard items, quality control
Method is effective, stable, uniform.
Crude drug source of the present invention:
Embelia laeta: the leaf of Myrsinacea Embelia laeta platymiscium Embelia laeta Embelia laeta (L.) Mez.
Anthracene shellfish element: from Embelia laeta extracting and developing, purification, purifying and be made, English name: Embelin.
Molecular formula: C17H26O4；
Structural formula:
What the method for quality control of anthracene shellfish element of the present invention was realized by following scheme:
The method of quality control of anthracene shellfish element can use high effective liquid chromatography for measuring, and chromatographic condition is as follows:
Chromatographic column is octadecyl silane filler；
It is respectively 10-90:90-10 with acetonitrile-water volume ratio, methanol-water volume ratio is that 20-80:90-10 is mobile phase；
Detection wavelength 280-310nm；
Flow velocity is 0.5~5 ml/min.
When preferably, using high effective liquid chromatography for measuring, chromatographic condition is as follows:
It is respectively 10-90:90-10 with acetonitrile-water volume ratio, methanol-water volume ratio is that 20:80-90:10 is mobile phase；
Flow velocity is 0.8~1.2 ml/min.
The method of quality control of anthracene shellfish element further includes thin-layered chromatography detection method, can be when prepared by anthracene shellfish element coarse crystallization
It waits and carries out rapid previewing survey, improve the quality of anthracene shellfish element coarse crystallization, quality monitoring can also be carried out after purification is crystallized,
This method method of anthracene shellfish element reference substance reference, chromatographic condition is, using silica G as chromatoplate, chromatoplate is soaked with 3% oxalic acid
It steeped, using toluene-ethyl acetate weight ratio 10:1 as solvent, petroleum ether: ethyl acetate weight ratio 10:1.2 is expansion
Agent, petroleum ether-toluene-ethyl acetate weight ratio 5:5:1 are solvent；Then concentrated sulfuric acid ethyl alcohol develops the color, and is placed under white light and examines
Depending in thin-layer chromatography being the single fluorescence spot of yellow under three development systems, then representing purity and meet the requirements, otherwise
It is undesirable.
Preferably, thin-layered chromatography detection method in the method for quality control of anthracene shellfish element are as follows:
The anthracene shellfish element sample solution of methanol 1mg/ml is taken, on same silica G chromatoplate, chromatoplate is soaked with 3% oxalic acid
It steeped, by different point sample amount gradient point samples, point sample amount is respectively 20 μ g, 40 μ g, 60 μ g, 80 μ g, 100 μ g；Respectively with first
Benzene-ethyl acetate weight ratio 10:1 is solvent, and petroleum ether: ethyl acetate weight ratio 10:1.2 is solvent, petroleum ether-
Toluene-ethyl acetate weight ratio 5:5:1 is solvent；It sets expansion cylinder to be unfolded respectively, exhibition is away from for 15cm, then concentrated sulfuric acid ethyl alcohol
Colour developing, is placed under white light and inspects, be yellow in the gradient point sample and three development system thin-layer chromatographys of 5 various concentrations
Single fluorescence spot.
The method of quality control of anthracene shellfish element of the present invention, which is characterized in that the anthracene shellfish element sample is by with lower section
Method is prepared:
1) it extracts: taking Embelia laeta maturation dry fruit, add diethyl ether extraction, is concentrated, filtering, through silica gel column chromatography, with petroleum ether-second
Acetate solution system carries out gradient elution, collects the eluent containing anthracene shellfish element, merges, and is concentrated to get anthracene shellfish element coarse crystallization；
2) it refines: isolating and purifying anthracene shellfish element coarse crystallization with preparative high performance liquid chromatography, with C-18 chromatographic column, cooperate first
Alcohol-water system carries out gradient elution, with methanol-water solution by after the dissolution of anthracene shellfish element coarse crystallization, injects preparative high-efficient liquid phase color
Spectrometer collects anthracene shellfish element solution of the purity 90% or more, be concentrated under reduced pressure to get.
Preferably, in the step 1), the eluent of various concentration is collected in grouping when gradient elution, and different eluents use
Thin-layered chromatography detection, with the method for anthracene shellfish element reference substance reference, chromatographic condition is, using silica G as chromatoplate, chromatoplate is used
3% oxalic acid impregnated, using toluene-ethyl acetate weight ratio 10:1 as solvent or petroleum ether: ethyl acetate weight ratio
10:1.2 is solvent or petroleum ether-toluene-ethyl acetate weight ratio 5:5:1 is solvent exhibition away from for 15cm；Then dense
The colour developing of sulfuric acid ethyl alcohol, is placed under white light and inspects, and selects in thin-layer chromatography, the single fluorescence spot of yellow eluent of high brightness,
Merge, is concentrated to get anthracene shellfish element coarse crystallization.
Preferably, in the step 2) purification, purity is collected respectively between 90%-98% and purity is greater than 98% or more
Eluent, be concentrated under reduced pressure, obtain anthracene shellfish element of the purity between 90%-98% and greater than 98% or more.
Preferably, in the step 2) purification, purity is collected after 90% or more anthracene shellfish element solution, using efficient liquid
Phase chromatography measures its purity, chromatographic condition again are as follows:
Chromatographic column is octadecyl silane filler；
It is respectively 10-90:90-10 with acetonitrile-water volume ratio, methanol-water volume ratio is that 20-80:90-10 is mobile phase；
Detection wavelength 280-310nm；
Flow velocity is 0.5~5 ml/min.
Compared with prior art, present invention substantive distinguishing features outstanding and significant progress are:
1, the anthracene shellfish element purity that the present invention extracts is very high, can achieve 98% or more, and purity meets chemical reference substance requirement,
It solves the supply problem of anthracene shellfish element chemical reference substance, provides scientific basic and guarantor are provided for the quality control of Embelia laeta medicinal material
Card.
2, the present invention has rational design, and simple process is extracted using aqueous solvent, and purity can be obtained after silica gel column chromatography
Higher anthracene shellfish element, most prepares the anthracene shellfish element chemical reference substance that purity reaches 98% or more through preparative high performance liquid chromatography afterwards,
Method is simple.
3, separating rate of the present invention is fast, with short production cycle, is suitble to industrialized production, there is good application prospect.
4, the present invention carries out purity test, assay and quality control using thin-layer chromatography and high performance liquid chromatography, really
Protect the quality of product.
By studying anthracene shellfish element chemical reference substance, the batch extracting technique, pure of anthracene shellfish element chemical reference substance is established
The analysis determining method of degree and content and determination of foreign matter is made to establish the technical standard of anthracene shellfish element chemical reference substance for it
Quality standard research for traditional Chinese chemical contrast and medicinal material and preparation provides scientific basic and guarantee.Result of study can be anthracene
Shellfish element chemical reference substance provides more complete Essential Chemistry foundation, grasps its chemical information and analysis and testing technology, is conducive to phase
The further development and utilization of product are closed, and for developing the peculiar product in China, develop the production with high-tech, high added value
Product improve the market competitiveness, it will generate potential and immeasurable social benefit and economic benefit.
Detailed description of the invention
It is to detect under 85:15 system that the final sample of Example 1, which is acetonitrile-acetic acid water volume ratio in mobile phase, respectively.
Fig. 1: anthracene shellfish plain color spectral peak uv absorption spectra；
Fig. 2: anthracene shellfish element chromatographic peak purity test three-dimensional spectrogram is shown to be single pure material peak；
Fig. 3: anthracene shellfish element DAD Peak homogeneity HPLC chromatogram (> 98%), chromatogram are single peak, are shown to be pure object
Mass peak；
Fig. 4: the HPLC of anthracene shellfish quality amount control detects chromatogram, and the content of principal component anthracene shellfish element is greater than 98.0%.Wherein scheme
Result table is analyzed in 4
Quantitative approach: normalization method
Specific embodiment
Embodiment 1: the preparation of high-purity anthracene shellfish element
Embelia laeta maturation dry fruit is taken, add diethyl ether extraction 3 times, and the amount of adding diethyl ether is 8 times of medicinal material weight every time, each extraction time
It is 2 hours, combined extract filters, and filtrate concentration obtains medicinal extract, adsorbs through silicagel column, with petroleum ether-ethyl acetate (0:100
~90:10) gradient elution, the elution fraction containing anthracene shellfish element is collected, concentration obtains anthracene shellfish element coarse crystallization.
Coarse crystallization is prepared into (chromatographic column: C-18 column with preparative RP-HPLC；Mobile phase: methanol: water volume ratio is
87:13；Detection wavelength is 290nm；Flow velocity is 5ml/min), respectively collect purity between 90%-98% and purity be greater than 98% with
On eluent, be concentrated under reduced pressure, obtain anthracene shellfish element of the purity between 90%-98% and greater than 98% or more.
All preparation solutions are detected with HPLC analysis method: chromatographic column C-18 column；Mobile phase is acetonitrile: acetic acid water body
Product is than being 85:15；Detection wavelength is 290nm；Flow velocity 1ml/min；Merge that retention time is identical and purity weight 90%~
Between 98% and purity is greater than the anthracene shellfish element preparation solution of 98% or more weight, is concentrated under reduced pressure, obtains purity in weight 90%~98%
Between and purity be greater than 98% or more anthracene shellfish element Red-brown powder.
Embodiment 2: the preparation of high-purity anthracene shellfish element
Embelia laeta maturation fruit is taken, add diethyl ether extraction 2 times, and the amount of adding diethyl ether is 10 times of medicinal material weight every time, when extracting every time
Between be 3 hours, filtration, merging filtrate, concentration, obtain medicinal extract, extract is adsorbed through silicagel column, with petroleum ether-ethyl acetate (0:
100~85:15) gradient elution, the elution fraction containing anthracene shellfish element is collected, concentration obtains anthracene shellfish element coarse crystallization.
Coarse crystallization is prepared into (chromatographic column: C-18 column with preparative RP-HPLC；Mobile phase methanol: water volume ratio is
80:20；Detection wavelength is 290nm；Flow velocity is 5ml/min), respectively collect purity between 90%-98% and purity be greater than 98% with
On eluent, be concentrated under reduced pressure, obtain anthracene shellfish element of the purity between 90%-98% and greater than 98% or more.
All preparation solutions are detected with HPLC analysis method: chromatographic column: C-18 column；Mobile phase acetonitrile: acetic acid water body
Product is than being 80:20；Detection wavelength is 290nm；Flow velocity is 1ml/min)；Merging retention time is identical and purity is in weight 90%
Between~98% and purity is greater than the anthracene shellfish element of 98% or more weight, is concentrated under reduced pressure, obtains purity between weight 90%~98%
And purity is greater than 98% or more anthracene shellfish element Red-brown powder.
Embodiment 3: the preparation of high-purity anthracene shellfish element
Embelia laeta maturation dry fruit is taken, add diethyl ether extraction 8 times, and the amount of adding diethyl ether is 2 times of medicinal material weight every time, each extraction time
It is 1 hour, filtration, merging filtrate is concentrated, obtains medicinal extract, extract is adsorbed through silicagel column, with petroleum ether-ethyl acetate (0:100
~80:20) gradient elution, the elution fraction containing anthracene shellfish element is collected, concentration obtains anthracene shellfish element coarse crystallization.
It is detected with HPLC, collects the flow point containing anthracene shellfish element, merged, concentration obtains anthracene shellfish element coarse crystallization.Coarse crystallization is used
Preparative RP-HPLC is prepared (chromatographic column: C-18 column；Mobile phase methanol: water volume ratio 90:10；Detection wavelength is
290nm；Flow velocity is 5ml/min), purity is collected respectively between 90%-98% and purity is greater than 98% or more eluent, decompression
Concentration obtains anthracene shellfish element of the purity between 90%-98% and greater than 98% or more.
All preparation solutions are detected with HPLC analysis method: chromatographic column: C-18 column；Mobile phase: acetonitrile: acetic acid water body
Product is than being 90:10；Detection wavelength is 290nm；Flow velocity is 1ml/min)；Merging retention time is identical and purity is in weight 90%
Between~98% and purity is greater than the anthracene shellfish element of 98% or more weight, is concentrated under reduced pressure, obtains purity between weight 90%~98%
And purity is greater than 98% or more anthracene shellfish element Red-brown powder.
Embodiment 4: the detection of anthracene shellfish element
Content made from Example 1-3 is detected in 98% or more anthracene shellfish element by following method of quality control:
1, thin-layered chromatography detection method:
The final sample of Example 1-3 adds methanol that solution of every 1ml containing 1mg is made respectively, is chromatography in same silica G
On plate, chromatoplate was impregnated with 3% oxalic acid, respectively the concentration gradient point sample different by 20 μ g, 40 μ g, 60 μ g, 80 μ g, 100 μ g；
It following solvent system is respectively adopted sets expansion cylinder and be unfolded respectively, exhibition is away from for 15cm:
Solvent system 1: with toluene-ethyl acetate weight ratio 10:1；
Solvent system 2: with toluene-ethyl acetate weight ratio 9:1；
Solvent system 3: petroleum ether: ethyl acetate weight ratio 10:1.2；
Solvent system 4: petroleum ether: ethyl acetate weight ratio 9:1；
Solvent system 5: petroleum ether-toluene-ethyl acetate weight ratio 5:5:1；
Solvent system 6: petroleum ether-toluene-ethyl acetate weight ratio 7:7:1；
It takes out, dries after expansion, iodine colour developing is set and inspected under white light；As a result in thin-layer chromatography, it is seen that yellow it is single
Fluorescence spot, 6 kinds of solvent systems, the gradient point sample of 3 samples, 5 various concentrations are single spot, have no impurity spot
Point.
2, high performance liquid chromatography:
Chromatographic condition and system suitability are filler with octadecylsilane chemically bonded silica；
Following flow phase system is respectively adopted, flow velocity is 1.0 ml/mins:
Flow phase system 1: acetonitrile-aqueous acetic acid volume ratio is 85:15；
Flow phase system 2: acetonitrile-aqueous acetic acid volume ratio is 90:10；
Flow phase system 3: acetonitrile-aqueous acetic acid volume ratio is 80:20；
Flow phase system 4: methanol-acetic acid water volume ratio is 87:13；
Flow phase system 5: methanol-acetic acid water volume ratio is 90:10；
Flow phase system 6: methanol-acetic acid water volume ratio is 80:20；
Detection wavelength is respectively 290nm, 310nm, and number of theoretical plate is calculated by the peak anthracene Bei Su should be not less than 3000.
This product that measuring method is taken at 105 DEG C of dryings to constant weights is appropriate, accurately weighed, is subject to six mobile phase systems respectively
System solution solution of every 1ml containing 3mg is made, shake up, precision measure 10 μ g inject liquid chromatograph, record chromatogram to it is main at
2.5 times or more of the peak retention time divided calculate content with area normalization method, and results sample measures anthracene shellfish cellulose content equal
98% or more.Determination of foreign matter, respectively in the chromatogram of not homologous ray record, in addition to solvent peak, impurity peak area summation result is equal
Less than 2.0%.
Peak purity detection: taking reference substance appropriate, presses two above flow phase system respectively, on high performance liquid chromatograph,
Peak homogeneity, HPLC chromatogram peak > 98% of anthracene shellfish element, chromatographic peak UV absorption light are carried out with diode array DAD detector
Spectrogram, three-dimensional map and 5 spectrograms are completely coincident, and are shown to be single pure material peak.
[structural identification]
UV λCDCl 3Max nm:290nm.
IRν KBr max cm-1 : 3300(-OH), 1640(carbonyl), 1608(phenyl ring).
Orange red crystallization, mp 141-143 DEG C.EI-MS m/z: 294[M]+, 154[M-C11H20]+ (100)。1H-NMR
(600MHz,CDCl3) δ ppm:6.03 (1H, s, H-6), 2.40 (2H, t, J=7.5 Hz, H-1ʹ), 1.43
(2H, m, H-2) , 1.28-1.30 (16H, m, H-3ʹ~10ʹ), 0.89(3H, t, J=6.6Hz, H-11ʹ)；13C-
NMR (150 MHz, CDCl3) (C-1) ~ 31.9 (C-10 of δ ppm:117.0 (C-3), 102.2 (C-6), 22.52
ʹ), 14.1 (C-11ʹ)。
The results showed that the anthracene shellfish element chemical reference substance of present invention separation, purifying, through infrared spectroscopy, ultraviolet spectra, core
Magnetic resonance, mass spectrum and Physico-chemical tests confirm chemical structure.It is detected through 6 development system TLC disclosed by the invention, 6 flowings
The HPLC of phase system and 2 different wave lengths detection, while purity test is done with DAD to chromatographic peak, show to meet Chinese medicine containing measurement
Surely the requirement of chemical reference substance is used, content is greater than 98%.
Claims (5)
1. a kind of method of quality control of anthracene shellfish element, it is characterised in that: using Embelia laeta platymiscium Embelia laeta maturation dry fruit as raw material,
High-purity anthracene shellfish element is extracted, then it is detected；
High-purity anthracene shellfish element the preparation method is as follows:
1) it extracts: taking Embelia laeta maturation dry fruit, add diethyl ether extraction, is concentrated, filtering, through silica gel column chromatography, with petroleum ether-acetic acid second
Ester solution system carries out gradient elution, collects the eluent containing anthracene shellfish element, merges, and is concentrated to get anthracene shellfish element coarse crystallization；
2) it refines: isolating and purifying anthracene shellfish element coarse crystallization with preparative high performance liquid chromatography, with C-18 chromatographic column, cooperate methanol-
Aqueous systems carry out gradient elution, with methanol-water solution by after the dissolution of anthracene shellfish element coarse crystallization, inject preparative high performance liquid chromatography
Instrument collects anthracene shellfish element solution of the purity 90% or more, be concentrated under reduced pressure to get；
The detection of the anthracene shellfish element includes carrying out qualitative detection using thin-layered chromatography and being determined using high performance liquid chromatography
Amount detection, detection process are as follows:
(1) carry out qualitative detection using thin-layered chromatography: with the method for anthracene shellfish element reference substance reference, chromatographic condition is, with silicon
Glue G is chromatoplate, and chromatoplate was impregnated with 3% oxalic acid, is respectively expansion with toluene-ethyl acetate weight ratio 10-15:1-2
Agent, petroleum ether: ethyl acetate weight ratio 10-15:1.2-2 is solvent, petroleum ether-toluene-ethyl acetate weight ratio 5-
8:5-8:1-2 is solvent；Then concentrated sulfuric acid ethyl alcohol develops the color, and is placed under white light and inspects, three development systems in thin-layer chromatography
Under be yellow single fluorescence spot, then represent purity and meet the requirements, it is otherwise undesirable；
The anthracene shellfish element sample solution of methanol 1mg/ml is taken, on same silica G chromatoplate, chromatoplate is impregnated with 3% oxalic acid
It crosses, by different point sample amount gradient point samples, point sample amount is respectively 20 μ g, 40 μ g, 60 μ g, 80 μ g, 100 μ g；Respectively with toluene-second
Acetoacetic ester weight ratio 10:1 is solvent perhaps petroleum ether: ethyl acetate weight ratio 10:1.2 is solvent or petroleum
Ether-toluene-ethyl acetate weight ratio 5:5:1 is solvent；It sets expansion cylinder to be unfolded respectively, exhibition is away from for 15cm, the then concentrated sulfuric acid
Ethyl alcohol colour developing, is placed under white light and inspects, in the gradient point sample and three development system thin-layer chromatographys of 5 various concentrations, be
The single fluorescence spot of yellow；
(2) quantitative detection: its chromatographic condition is carried out using high performance liquid chromatography are as follows:
Chromatographic column is octadecyl silane filler；
It is respectively 10-90:90-10 with acetonitrile-acetic acid water volume ratio, methanol-acetic acid water volume ratio is that 20-80:90-10 is flowing
Phase；
Detection wavelength 280-310nm；
Flow velocity is 0.5~5 ml/min；
Content and purity testing: respectively with 2 mobile phase solvent systems and 2 Detection wavelengths, chromatogram is recorded to principal component
2.5 times or more of peak retention time, with area normalization method calculate content, as a result system measurement sample size equal 98% with
On；Determination of foreign matter, respectively in the chromatogram of not homologous ray record, in addition to solvent peak, impurity peak area summation result is respectively less than
2.0%。
2. the method for quality control of anthracene shellfish element according to claim 1, it is characterised in that: described with acetonitrile-acetic acid water
Volume ratio is 85:15, and methanol-acetic acid water volume ratio is that 87:13 is mobile phase；
Detection wavelength 290nm；
Flow velocity is 0.8~1.2 ml/min.
3. the method for quality control of anthracene shellfish element according to claim 1, it is characterised in that:
In the step 1), the eluent of various concentration is collected in grouping when gradient elution, and different eluents use thin-layered chromatography
Detection, with the method for anthracene shellfish element reference substance reference, chromatographic condition are as follows: using silica G as chromatoplate, chromatoplate is impregnated with 3% oxalic acid
It crosses, using toluene-ethyl acetate weight ratio 10-15:1-2 as solvent or petroleum ether: ethyl acetate weight ratio 10-15:
It is solvent that 1.2-2, which is solvent or petroleum ether-toluene-ethyl acetate weight ratio 5-8:5-8:1-2, and exhibition is away from for 15cm；
The colour developing of concentrated sulfuric acid ethyl alcohol, is placed under white light and inspects, and selects the single fluorescence spot of the yellow elution of high brightness in thin-layer chromatography
Liquid merges, and is concentrated to get anthracene shellfish element coarse crystallization.
4. the method for quality control of anthracene shellfish element according to claim 1, it is characterised in that:
In the step 2) purification, purity is collected respectively between 90%-98% and purity is greater than 98% or more eluent, is subtracted
Pressure concentration obtains anthracene shellfish element of the purity between 90%-98% and greater than 98% or more.
5. the method for quality control of anthracene shellfish element according to claim 1, it is characterised in that:
In the step 2) purification, purity is collected after 90% or more anthracene shellfish element solution, is surveyed again using high performance liquid chromatography
Its fixed purity, chromatographic condition are as follows:
Chromatographic column is octadecyl silane filler；
It is respectively 10-90:90-10 with acetonitrile-acetic acid water volume ratio, methanol-acetic acid water volume ratio is that 20-80:90-10 is flowing
Phase；
Detection wavelength 280-310nm；
Flow velocity is 0.5~5 ml/min.
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| CN111562337A (en) * | 2019-02-13 | 2020-08-21 | 中国石油天然气股份有限公司 | Method and system for identifying polymer product |
| CN116898885B (en) * | 2023-07-25 | 2024-04-16 | 中国科学院西北高原生物研究所 | Water leaching active matter for inhibiting COVID-19 XBB 1.5 variant infection and application thereof |
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| RP-HPLC-DAD method for the estimation of embelin as marker in Embelia ribes and its polyherbal formulations;Soumya N. Madhavan等;《Biomed. Chromatogr.》;20100804;第25卷;第600-605页 * |
| THE PHARMACOGNOSTIC SPECIFICATION OF ARDISIA ELLIPTICA FRUITS AND THEIR EMBELIN CONTENTS BY TLC IMAGE ANALYSIS COMPARED TO TLC DENSITOMETRY;Yukongphan et al.;《BHST.》;20131102;第11卷(第2期);全文 * |
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