CN106955502A - A kind of method of Geniposidic acid and acteoside in ASE methods extraction plantain seed - Google Patents
A kind of method of Geniposidic acid and acteoside in ASE methods extraction plantain seed Download PDFInfo
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- CN106955502A CN106955502A CN201710247868.5A CN201710247868A CN106955502A CN 106955502 A CN106955502 A CN 106955502A CN 201710247868 A CN201710247868 A CN 201710247868A CN 106955502 A CN106955502 A CN 106955502A
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- extraction
- acteoside
- plantain seed
- ase
- geniposidic acid
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0288—Applications, solvents
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses the method for Geniposidic acid and acteoside in a kind of ASE methods extraction plantain seed, this method extraction time is shorter, reproducible, belongs to the extraction field of Geniposidic acid and acteoside.Plantain seed sample comminution is crossed No. two sieves, takes 1g to be mixed with 1g diatomite by this method；Mixture obtained by step 1 is loaded on and is placed with the ASE abstraction pools of fibrous filter membrane, plus diatomite is extremely parallel with pond mouthful；With methanol extraction, and extract is settled to 50ml with methanol.The present invention can substitute official method and Geniposidic acid in plantain seed and acteoside are extracted, and carry out assay.
Description
Technical field
The invention belongs to the extraction field of Geniposidic acid and acteoside, especially a kind of ASE methods extraction plantain seed
The method of middle Geniposidic acid and acteoside.
Background technology
《Chinese Pharmacopoeia》Version one prepares test sample method within 2015：The preparation of need testing solution takes this product powder (to cross two
Number sieve) about 1g, it is accurately weighed, put in conical flask with cover, precision adds 60% methanol 50ml, and weighed weight is heated to reflux 2 small
When, let cool, then weighed weight, the weight of less loss is supplied with 60% methanol, is shaken up, is filtered, is taken subsequent filtrate, produce.
This method return time is long, and the professional standards to technical staff require higher.
The content of the invention
For above-mentioned deficiency, the present invention is intended to provide Geniposidic acid and verbascose in a kind of ASE methods extraction plantain seed
The method of glycosides, this method extraction time is shorter, reproducible.
In order to realize above-mentioned technique effect, the technical scheme that the present invention is provided is such：A kind of ASE methods extract plantain seed
The method of middle Geniposidic acid and acteoside, comprises the steps successively：
Step 1：By plantain seed sample comminution, No. two sieves are crossed, take 1g to be mixed with 1g diatomite；
Step 2：By the mixture obtained by step 1 loaded on being placed with the ASE abstraction pools of fibrous filter membrane, plus diatomite to and pond
Mouth is parallel；
Step 3：With methanol extraction, and extract is settled to 50ml with methanol.
Preferably, the extraction conditions described in step 3 is：Extraction temperature is 80 DEG C, and extraction time is 8min；Cycle-index is
3 times；Flush volume is 60%；Purge time is 60s.
Preferably, the volume of the ASE abstraction pools described in step 2 is 10ml.
The present invention is compared with conventional method, and tool has the advantage that：
The present invention is once investigated to the solvent that Accelerate solvent extraction is used, and is had paper to mention and is extracted shaddock using 60% methanol
Skin glycosides relative amount is higher, and impurity is less, therefore to having carried out solvent investigation using methanol and 60% methanol, as a result show methanol with
60% methanol extracts plantain seed content unanimously, and impurity peaks are essentially identical, therefore uses the Extraction solvent consistent with pharmacopeia.
The present invention is investigated to extraction temperature, and extraction time will also be investigated, on above-mentioned experiment basis, I
Use 3 factor 3 horizontal quadrature experimental program L9 (33) experiment arrangement, investigated 3 factors to accelerated solvent extraction plantain seed
In Geniposidic acid, the influence of acteoside process.This experiment determines extraction temperature, static extracting time, cycle-index
And flush volume is investigation factor, it is determined that extracting from the optimal fast solvent of the Geniposidic acid in plantain seed, acteoside
Taking technique, so that it is determined that optimal extraction parameters.
Brief description of the drawings
Fig. 1 is the chromatogram of Geniposidic acid and acteoside reference substance；
Fig. 2 is the chromatogram of the need testing solution obtained by ASE methods are extracted；
Fig. 3 is the chromatogram of the need testing solution obtained by official method is extracted.
Embodiment
The claim of the present invention is described in further detail with reference to embodiment, but not constituted to this
Any limitation of invention, still any limited number of time modification made in the claims in the present invention protection domain, the power in the present invention
In the claimed scope of profit.
Embodiment 1
1st, instrument and equipment and reagent
1.1 instruments：
Electronic analytical balance (XA205DU), ASE350 Accelerate solvent extractions instrument (DIONEX companies of the U.S.), Thermo
U3000UHPLC liquid chromatographs
1.2 reagents：
Water：Meet one-level water as defined in GB/T 6682；
Methanol (CH4O)：Chromatographically pure；
Acetic acid (CH3COOH)：Analysis is pure.
2nd, method
The preparation of 2.1 reference substance solutions：
The preparation of reference substance solution takes Geniposidic acid reference substance, acteoside reference substance appropriate, accurately weighed, puts
In brown measuring bottle, plus every lml respectively mixed solutions containing 0.1mg are made in 60% methanol, produce.
The preparation of 2.2 need testing solutions
《Chinese Pharmacopoeia》Version one prepares test sample method within 2015：The preparation of need testing solution takes this product powder (to cross two
Number sieve) about 1g, it is accurately weighed, put in conical flask with cover, precision adds 60% methanol 50ml, and weighed weight is heated to reflux 2 small
When, let cool, then weighed weight, the weight of less loss is supplied with 60% methanol, is shaken up, is filtered, is taken subsequent filtrate, produce.
Accelerate solvent extraction method (ASE) prepares test sample method：The size-reduced machine of sample is crushed, and is crossed No. two sieves, is taken 1g, accurate
It is weighed, it is well mixed with about 1g diatomite, it is stand-by, it is moved into the ASE 10ml abstraction pools for putting fibrous filter membrane well in advance and adds
Appropriate diatomite, gently shaking is allowed to Chi Kou in the same horizontal line, tighten and cover on abstraction pool, be put into rapid extracting device
Row extraction.After extraction terminates, extract is transferred in 50ml volumetric flasks, extraction flask is cleaned with a small amount of methanol and is incorporated into appearance 3 times
In measuring bottle, using methanol constant volume to scale, produce.
2.3 ASE extraction conditions
Extraction temperature is that 80 DEG C of extraction times are 8min；Cycle-index is 3 times；Flush volume is 60%；Purge time is
60s。
2.4 chromatographic conditions and system suitability
A) instrument：The double ternary liquid phases (U-3000) of Thermo；
B) chromatographic column：Thermo Syncronis Dim. (mm) AQ, 1.7 μm, 50 × 2.1mm；
C) column temperature：40℃；
D) flow velocity：0.3mL/min；
E) mobile phase：Methanol-acetic acid-water (35:4:65)；
F) Detection wavelength：283nm；
Number of theoretical plate is calculated by Geniposidic acid peak should be not less than 3000.
2.5 determination methods
Determined according to high performance liquid chromatography (general rule 0512)；
It is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, liquid chromatograph is injected, determines, produces.
The requirement of 2.6 standard limited values
This product is calculated by dry product, containing Geniposidic acid (C16H22O10) 0.50%, acteoside must not be less than
(C29H36O15) 0.40% must not be less than.
2.7 calculate (external standard method)
C in formulaR--- reference substance solution concentration, unit is micro- gram per liter (mg/L)；
AX--- the peak area of test sample；
AR--- reference substance peak area.
Note：
It should be dismantled and cleaned out using preceding extraction bottom of pond portion, otherwise easily cause pressure instability；
Otherwise the filter paper of abstraction pool bottom should cause seepage in sealing ring；
Abstraction pool fills elastic moderate during sample, and too loose to be easily caused extract solution excessive；
Check whether gas cylinder air pressure reaches 1Mpa before start；
Using being cleaned out after terminating, abstraction pool will in time dry and (get rusty easily).
3rd, result
3.1 reappearance tests
Take the sample (lot number of identical lot number：20151001) 0.5g, it is totally 4 parts, accurately weighed, by above-mentioned ASE extracting methods
Need testing solution is extracted, sample size is 1 μ L, with above-mentioned chromatographic condition parallel laboratory test, measures Geniposidic acid and hair in sample
It is 1.2% that the content of stamen flower glucosides, which is shown in Table 1, RSD, and experiment shows that ASE extracting methods repeatability is good.
The Geniposidic acid of the ASE methods of table 1 extraction and the content of acteoside
| Repeatability 1 | Repeatability 2 | Repeatability 3 | Repeatability 4 | RSD% | |
| As a result % | 2.082 | 2.064 | 2.074 | 2.027 | 1.2 |
3.2 precision and stability test
Geniposidic acid reference substance, acteoside reference substance mixed solution are taken, continuous sample introduction 6 times records peak area,
Peak area RSD is 0.5%, shows that instrument precision is good.
Same need testing solution is taken, is placed at room temperature, is determined in accordance with the law respectively at 0,2,4,8,10h.As a result Astragaloside IV peak
The RSD of area is 0.2%, shows that need testing solution is stable in 12h.
3.3 samples difference instrument extract result and with official method results contrast (3 batches)
The sample of table 2 difference instrument extract result and with official method results contrast
| ASE1 | ASE2 | Pharmacopeia | |
| 150523 | 1.574 | 1.54 | 1.971 |
| 151001 | 1.698 | 1.581 | 1.902 |
| 1511024 | 0.964 | 1.095 | 1.713 |
| 20151001 | 2.073 | 1.626 | 2.139 |
4th, discuss：
The selection of 4.1 ASE Extraction solvents
The present invention is once investigated to the solvent that Accelerate solvent extraction is used, and is had paper to mention and is extracted shaddock using 60% methanol
Skin glycosides relative amount is higher, and impurity is less, therefore to having carried out solvent investigation using methanol and 60% methanol, as a result show methanol with
60% methanol extracts plantain seed content unanimously, and impurity peaks are essentially identical, therefore uses the Extraction solvent consistent with pharmacopeia.
The optimization of 4.2 ASE extraction conditions
Extraction temperature is investigated, and extraction time will also be investigated, on above-mentioned experiment basis, we use 3
Factor 3 horizontal quadrature experimental program L9 (33) experiment arrangement, has investigated 3 factors to the capital Buddhist nun in accelerated solvent extraction plantain seed
The influence of flat thuja acid, acteoside process.This experiment determines extraction temperature, static extracting time, cycle-index and flushing
Volume is investigation factor, refers to table 3.It is determined that extracting from the optimal fast solvent of the Geniposidic acid in plantain seed, acteoside
Taking technique, the results are shown in Table 4-6.
The factor level of table 3
The orthogonal experiment range analysis result of the Geniposidic acid of table 4
| Factor | Temperature | Time | Number of times | Experimental result | |
| Experiment 1 | 1 | 1 | 1 | 1 | 1.054 |
| Experiment 2 | 1 | 2 | 2 | 2 | 1.145 |
| Experiment 3 | 1 | 3 | 3 | 3 | 1.13 |
| Experiment 4 | 2 | 1 | 2 | 3 | 1.025 |
| Experiment 5 | 2 | 2 | 3 | 1 | 1.073 |
| Experiment 6 | 2 | 3 | 1 | 2 | 1.014 |
| Experiment 7 | 3 | 1 | 3 | 2 | 1.022 |
| Experiment 8 | 3 | 2 | 1 | 3 | 1.057 |
| Experiment 9 | 3 | 3 | 2 | 1 | 1.099 |
| Average 1 | 1.110 | 1.034 | 1.042 | 1.075 | |
| Average 2 | 1.037 | 1.092 | 1.090 | 1.060 | |
| Average 3 | 1.059 | 1.081 | 1.075 | 1.071 | |
| Extreme difference | 0.073 | 0.058 | 0.048 | 0.015 |
The orthogonal experiment range analysis result of the acteoside of table 5
| Factor | Temperature | Time | Number of times | Experimental result | |
| Experiment 1 | 1 | 1 | 1 | 1 | 0.821 |
| Experiment 2 | 1 | 2 | 2 | 2 | 0.885 |
| Experiment 3 | 1 | 3 | 3 | 3 | 0.89 |
| Experiment 4 | 2 | 1 | 2 | 3 | 0.835 |
| Experiment 5 | 2 | 2 | 3 | 1 | 0.858 |
| Experiment 6 | 2 | 3 | 1 | 2 | 0.834 |
| Experiment 7 | 3 | 1 | 3 | 2 | 0.81 |
| Experiment 8 | 3 | 2 | 1 | 3 | 0.809 |
| Experiment 9 | 3 | 3 | 2 | 1 | 0.827 |
| Average 1 | 0.865 | 0.822 | 0.821 | 0.835 | |
| Average 2 | 0.842 | 0.851 | 0.849 | 0.843 | |
| Average 3 | 0.815 | 0.850 | 0.853 | 0.845 | |
| Extreme difference | 0.050 | 0.029 | 0.032 | 0.010 |
The orthogonal experiment range analysis result of the Geniposidic acid of table 6 and acteoside total amount
Extraction time influences maximum to the content of Geniposidic acid and acteoside it can be seen from range analysis, its
Secondary is Extracting temperature and extraction time.Final to determine that optimum extraction process is combined as 80 DEG C of Extracting temperature, extraction time 8min is carried
Take number of times 3 times.
Above-described is only presently preferred embodiments of the present invention, all timess done in the range of the spirit and principles in the present invention
What modification, equivalent and improvement etc., should be included within the scope of the present invention.
Claims (3)
1. a kind of method of Geniposidic acid and acteoside in ASE methods extraction plantain seed, it is characterised in that under including successively
State step：
Step 1：By plantain seed sample comminution, No. two sieves are crossed, take 1g to be mixed with 1g diatomite；
Step 2：Mixture obtained by step 1 is loaded on and is placed with the ASE abstraction pools of fibrous filter membrane, plus diatomite is put down to Chi Kou
OK；
Step 3：With methanol extraction, and extract is settled to 50ml with methanol.
2. the method for Geniposidic acid and acteoside in a kind of ASE methods extraction plantain seed according to claim 1, its
It is characterised by, the extraction conditions described in step 3 is：Extraction temperature is 80 DEG C, and extraction time is 8min；Cycle-index is 3 times；Punching
It is 60% to wash volume；Purge time is 60s.
3. the method for Geniposidic acid and acteoside in a kind of ASE methods extraction plantain seed according to claim 1, its
It is characterised by, the volume of the ASE abstraction pools described in step 2 is 10ml.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113461749A (en) * | 2021-08-13 | 2021-10-01 | 许昌学院 | Method for extracting and preparing geniposide from plantago seeds |
Citations (3)
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| CN105738536A (en) * | 2016-04-22 | 2016-07-06 | 广西壮族自治区梧州食品药品检验所 | Extraction method for indirubin in folium isatidis |
| CN105929090A (en) * | 2016-04-22 | 2016-09-07 | 广西壮族自治区梧州食品药品检验所 | Method for extracting curculigoside in Curculigo orchioides |
| CN105954377A (en) * | 2016-04-22 | 2016-09-21 | 广西壮族自治区梧州食品药品检验所 | Extraction method for amentoflavone in Selaginella tamariscina |
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- 2017-04-17 CN CN201710247868.5A patent/CN106955502A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105738536A (en) * | 2016-04-22 | 2016-07-06 | 广西壮族自治区梧州食品药品检验所 | Extraction method for indirubin in folium isatidis |
| CN105929090A (en) * | 2016-04-22 | 2016-09-07 | 广西壮族自治区梧州食品药品检验所 | Method for extracting curculigoside in Curculigo orchioides |
| CN105954377A (en) * | 2016-04-22 | 2016-09-21 | 广西壮族自治区梧州食品药品检验所 | Extraction method for amentoflavone in Selaginella tamariscina |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113461749A (en) * | 2021-08-13 | 2021-10-01 | 许昌学院 | Method for extracting and preparing geniposide from plantago seeds |
| CN113461749B (en) * | 2021-08-13 | 2023-03-14 | 许昌学院 | Method for extracting geniposide from plantain seeds |
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Application publication date: 20170718 |