CN105699539A - Method for extracting oleanolic acid in clematis root - Google Patents
Method for extracting oleanolic acid in clematis root Download PDFInfo
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- CN105699539A CN105699539A CN201610234712.9A CN201610234712A CN105699539A CN 105699539 A CN105699539 A CN 105699539A CN 201610234712 A CN201610234712 A CN 201610234712A CN 105699539 A CN105699539 A CN 105699539A
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- radix clematidis
- oleanolic acid
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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Abstract
The invention discloses a method for extracting oleanolic acid in clematis root, comprising the following steps: 1, pulverizing clematis root and screening; 2, weighing 1 part by weight of clematis root powder, and mixing with suitable quartz and for use; 3, adding 1 part by weight of diatomaceous earth into an extracting tank with a pre-placed filter membrane, mixing well, and adding suitable quartz sand to a position 2 mm below the horizon of an opening of the extracting tank; closing a cover of the extracting tank; 4, removing impurities from the mixture in the extracting tank; 5, after step 4, extracting the mixture in the extracting tank; acquiring an extract after extracting. The invention provides a method for extracting oleanolic acid in clematis root, and this method is simple to perform and high in extracting efficiency.
Description
Technical field
The present invention relates to test in laboratory field, particularly the extracting method of oleanolic acid in a kind of Radix Clematidis。
Background technology
Within 2015, pharmacopeia have recorded the method for oleanolic acid in Radix Clematidis of extracting: takes this product powder (crossing No. four sieves) about 4g, accurately weighed, put in apparatus,Soxhlet's, add ethyl acetate appropriate, it is heated to reflux 3 hours, discard acetic acid ethyl fluid, medicinal residues volatilize solvent, it is transferred in conical flask together with filtration paper cylinder, accurate addition Diluted Alcohol 50ml, weighed weight, it is heated to reflux 1 hour, let cool, weighed weight again, the weight of less loss is supplied with Diluted Alcohol, shake up, filter, precision measures subsequent filtrate 25ml, put and water-bath is evaporated, residue adds 2mol/L hydrochloric acid solution 30ml makes dissolving, it is heated to reflux 2 hours。Cooling down immediately, move in separatory funnel, with water 10ml gradation washing container, washing liquid is incorporated in separatory funnel。Adding ethyl acetate jolting to extract 3 times, each 15ml, combined ethyl acetate liquid, less than 70 DEG C are concentrated near dry, add methanol and dissolve, be transferred in 10ml measuring bottle, add methanol to scale, shake up, to obtain final product。
The method operating process of pharmacopeia is complicated, and the operant skill of operator is required height。
Summary of the invention
It is an object of the invention to provide the extracting method of oleanolic acid in a kind of Radix Clematidis, the method is simple to operate, and extraction efficiency is high。
Technical scheme provided by the invention is: the extracting method of oleanolic acid in a kind of Radix Clematidis, comprises the following steps:
Step 1: after Radix Clematidis is pulverized, sieve；
Step 2: weigh 1 weight portion Radix Clematidis powder, mix homogeneously with appropriate amount of quartz sand stand-by；
Step 3: be initially charged 1 weight portion kieselguhr in the abstraction pool putting filter membrane in advance well, adds appropriate quartz sand to reaching 2mm under abstraction pool saliva horizontal line after mix homogeneously；Cover the lid of abstraction pool；
Step 4: the mixture in abstraction pool is carried out remove impurity；
Remove impurity condition is: extractant is ethyl acetate；Extraction temperature 120 DEG C；Static extracting time 3min；Flush volume 100%；Quiet cycle number of times 2 times；
Step 5: the mixture in abstraction pool is extracted after terminating by step 4；Extraction is extracted liquid after terminating；
Extraction conditions is: extractant is Diluted Alcohol；Extraction temperature 100 DEG C；Static extracting time 5min；Flush volume 100%；Quiet cycle number of times 3 times；Described Diluted Alcohol is the percentage by volume of C2H5OH is 49.5%～50.5% when 20 DEG C。
In above-mentioned Radix Clematidis in the extracting method of oleanolic acid, the volume of described abstraction pool is 10ml。
In above-mentioned Radix Clematidis in the extracting method of oleanolic acid, in step 4 and step 5, the pressure of static extracting is 1500psi。
In above-mentioned Radix Clematidis in the extracting method of oleanolic acid, in step 4 and step 5, the purge time of static extracting is 60s。
In above-mentioned Radix Clematidis in the extracting method of oleanolic acid, also include step 6, described step 6 is particularly as follows: put extract and be evaporated in water-bath, residue adds 2mol/L hydrochloric acid solution makes it dissolve, cool down immediately after being heated to reflux 2 hours, moving in separatory funnel, with moisture time washing container, washing liquid is incorporated in separatory funnel；Adding ethyl acetate jolting to extract 3 times, combined ethyl acetate liquid, less than 70 DEG C are concentrated near dry, add methanol and dissolve, be transferred in 10ml measuring bottle, add methanol to scale, shake up, and under 15000r/min, centrifugal 3min, takes supernatant。
In above-mentioned Radix Clematidis in the extracting method of oleanolic acid, the instrument that described ASE extraction adopts is ASE350 Accelerate solvent extraction instrument。
The present invention is after adopting technique scheme, and it has the beneficial effect that
The extracting method of the present invention is simple to operate, and precision is consistent with the precision of official method, reproducible。
Accompanying drawing explanation
Fig. 1 is the rectangular plots of the chromatography validation verification of the present invention；
Detailed description of the invention
Below in conjunction with detailed description of the invention, technical scheme is described in further detail, but does not constitute any limitation of the invention。
Embodiment 1:
Size-reduced for Radix Clematidis machine is pulverized, excessively No. three sieves, about 1g, accurately weighed, mix homogeneously with appropriate amount of quartz sand, stand-by, filter membrane is put well in advance10ml abstraction pool is initially charged 1g kieselguhr, the rear sample adding mix homogeneously, add appropriate amount of quartz sand, gently jolting so as to Chi Kou 2mm place under same level line, tighten abstraction pool upper cover。Extract (include remove impurity and extract 2 steps, see ASE conditions method) terminate after, put and water-bath be evaporated, residue adds 2mol/L hydrochloric acid solution 30ml makes dissolving, is heated to reflux 2 hours。Cooling down immediately, move in separatory funnel, with water 10ml gradation washing container, washing liquid is incorporated in separatory funnel。Adding ethyl acetate jolting to extract 3 times, each 15ml, combined ethyl acetate liquid, less than 70 DEG C are concentrated near dry, add methanol and dissolve, be transferred in 10ml measuring bottle, add methanol to scale, shake up, and under 15000r/min, centrifugal 3min, takes supernatant, enters LC and extracts and get final product。
Extraction conditions (see table 1) is:
Table 1
Analysis method is LC liquid chromatography, liquid-phase chromatographic analysis condition:
A) the double; two ternary liquid phase chromatograph of instrument: ThermoU-3000
B) chromatographic column: ThermoAcclaimC302.1*150mm3 μm, i.e. carbon 30 chromatographic column, diameter 2.1mm, length 150mm, particle diameter 3 microns；
C) column temperature: 20 DEG C
D) flow velocity: 0.5mL/min
E) mobile phase: acetonitrile-water (volume ratio 62:38)
F) detection wavelength: 205nm
Chromatography validation verification:
Take oleanolic acid reference substance appropriate, accurately weighed, add mobile phase and make every 1ml solution containing 0.2mg, obtain pier fruit acid reference substance。
Take oleanolic acid reference substance appropriate, accurately weighed, put in brown measuring bottle, add methanol and make every 1ml solution containing oleanolic acid 1mg, then precision draws this solution 0.1 μ l, 0.2 μ l, 0.5 μ l, 0.8 μ l, 1.0 μ l entrance LC extractions respectively, and extracts according to said method, the results detailed in Table 2。Accompanying drawing 1 is corresponding with table 2, and accompanying drawing 1 abscissa indicated concentration, vertical coordinate represents peak area。
Table 2
The repeatability checking of the extracting process of 1.1 embodiments 1:
Take sample (lot number: 150410 (the Z)) 1.0g of identical lot number, totally 6 parts, accurately weighed, extracting need testing solution by ASE extracting method, sample size is 1 μ L, with above-mentioned chromatographic condition parallel test, record in sample the content of oleanolic acid in Table 2, RSD is 2.0%, and test shows that ASE extracting method repeatability is good, the results detailed in Table 3
Table 3
The degree of accuracy test of the extracting process of 1.2 embodiments 1
Adopting the ASE extracting process of embodiment 1 and the extracting process of pharmacopeia that Radix Clematidis is extracted, Radix Clematidis is 3 batches, and lot number is 150410 (Y) respectively, 1510124,150410 (Z)；
The ASE extracting process of the embodiment 1 Radix Clematidis parallel testing twice to same batch, respectively ASE1 and ASE2 represents。
Concrete test result is shown in table 4 below；
Table 4
The above-described presently preferred embodiments of the present invention that is only, all any amendment, equivalent replacement and improvement etc. made within the scope of the spirit and principles in the present invention, should be included within protection scope of the present invention。
Claims (6)
1. the extracting method of oleanolic acid in a Radix Clematidis, it is characterised in that comprise the following steps:
Step 1: after Radix Clematidis is pulverized, sieve；
Step 2: weigh 1 weight portion Radix Clematidis powder, mix homogeneously with appropriate amount of quartz sand stand-by；
Step 3: be initially charged 1 weight portion kieselguhr in the abstraction pool putting filter membrane in advance well, adds appropriate quartz sand to reaching 2mm under abstraction pool saliva horizontal line after mix homogeneously；Cover the lid of abstraction pool；
Step 4: the mixture in abstraction pool is carried out remove impurity；
Remove impurity condition is: extractant is ethyl acetate；Extraction temperature 120 DEG C；Static extracting time 3min；Flush volume 100%；Quiet cycle number of times 2 times；
Step 5: the mixture in abstraction pool is extracted after terminating by step 4；Extraction is extracted liquid after terminating；
Extraction conditions is: extractant is Diluted Alcohol；Extraction temperature 100 DEG C；Static extracting time 5min；Flush volume 100%；Quiet cycle number of times 3 times；Described Diluted Alcohol is the percentage by volume of C2H5OH is 49.5%～50.5% when 20 DEG C。
2. the extracting method of oleanolic acid in Radix Clematidis according to claim 1, it is characterised in that the volume of described abstraction pool is 10ml。
3. the extracting method of oleanolic acid in Radix Clematidis according to claim 1, it is characterised in that in step 4 and step 5, the pressure of static extracting is 1500psi。
4. the extracting method of oleanolic acid in Radix Clematidis according to claim 1, it is characterised in that in step 4 and step 5, the purge time of static extracting is 60s。
5. the extracting method of oleanolic acid in Radix Clematidis according to claim 1, it is characterized in that, also include step 6, described step 6 is particularly as follows: put extract and be evaporated in water-bath, residue adds 2mol/L hydrochloric acid solution makes it dissolve, and cools down immediately after being heated to reflux 2 hours, moves in separatory funnel, with moisture time washing container, washing liquid is incorporated in separatory funnel；Adding ethyl acetate jolting to extract 3 times, combined ethyl acetate liquid, less than 70 DEG C are concentrated near dry, add methanol and dissolve, be transferred in 10ml measuring bottle, add methanol to scale, shake up, and under 15000r/min, centrifugal 3min, takes supernatant。
6. the extracting method of oleanolic acid in Radix Clematidis according to claim 1, it is characterised in that the instrument that described ASE extraction adopts is ASE350 Accelerate solvent extraction instrument。
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107014923A (en) * | 2017-04-20 | 2017-08-04 | 广西壮族自治区梧州食品药品检验所 | A kind of method that ASE HPLC methods determine content of oleanolic acid in the root of Chinese clematis |
| CN107064384A (en) * | 2017-04-11 | 2017-08-18 | 广西壮族自治区梧州食品药品检验所 | A kind of method that ASE methods extract spinosin in spina date seed |
| CN107219315A (en) * | 2017-04-01 | 2017-09-29 | 广西壮族自治区梧州食品药品检验所 | A kind of method that ASE HPLC methods determine oleanolic acid and ursolic acid total content in Verbena officinalis |
| CN112098569A (en) * | 2019-06-17 | 2020-12-18 | 国药集团同济堂（贵州）制药有限公司 | Preparation method and quality determination method of radix clematidis extract |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103558303A (en) * | 2013-10-25 | 2014-02-05 | 贵州信邦制药股份有限公司 | Method for measuring content of oleanolic acid in glossy privet fruit and astragalus membranaceus healthy energy-strengthening preparation |
-
2016
- 2016-04-15 CN CN201610234712.9A patent/CN105699539A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103558303A (en) * | 2013-10-25 | 2014-02-05 | 贵州信邦制药股份有限公司 | Method for measuring content of oleanolic acid in glossy privet fruit and astragalus membranaceus healthy energy-strengthening preparation |
Non-Patent Citations (2)
| Title |
|---|
| 国家药典委员会: "《中华人民共和国药典 2015年版 一部》", 30 June 2015, 中国医药科技出版社 * |
| 王文俊 等: "一种测定女贞子中齐墩果酸和熊果酸的新方法", 《西北植物学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107219315A (en) * | 2017-04-01 | 2017-09-29 | 广西壮族自治区梧州食品药品检验所 | A kind of method that ASE HPLC methods determine oleanolic acid and ursolic acid total content in Verbena officinalis |
| CN107064384A (en) * | 2017-04-11 | 2017-08-18 | 广西壮族自治区梧州食品药品检验所 | A kind of method that ASE methods extract spinosin in spina date seed |
| CN107014923A (en) * | 2017-04-20 | 2017-08-04 | 广西壮族自治区梧州食品药品检验所 | A kind of method that ASE HPLC methods determine content of oleanolic acid in the root of Chinese clematis |
| CN112098569A (en) * | 2019-06-17 | 2020-12-18 | 国药集团同济堂（贵州）制药有限公司 | Preparation method and quality determination method of radix clematidis extract |
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