CN108414650A - Fructus amomi discrimination method and its application and system - Google Patents
Fructus amomi discrimination method and its application and system Download PDFInfo
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Abstract
The present invention relates to a kind of fructus amomi discrimination method and its application and systems, belong to Chinese traditional medicine identification authentication technique field.The discrimination method includes the following steps：(1) preparation of reference substance solution：Precision weighs Bronyl acetate reference substance, and with organic solvent dissolving and constant volume is to get reference substance solution；(2) preparation of test solution：Fructus amomi test sample is taken, volatile oil is extracted, obtains test solution；(3) it identifies：Above-mentioned reference substance solution and test solution are drawn respectively, is injected separately into gas chromatograph for determination, and base identification is carried out to the fructus amomi test sample according to preassigned.This method can roll into a ball class, sand rice class and coarse powder class sample to seed and carry out base identification, overcome and differentiate the problem of fructus amomi true and false has difficulties in routine techniques.Foundation is provided for the identification and quality evaluation of fructus amomi, reference is provided for science, the comprehensive quality standard for formulating fructus amomi.
Description
Technical field
The present invention relates to Chinese traditional medicine identification authentication technique fields, more particularly to a kind of fructus amomi discrimination method and its application and are
System.
Background technology
Fructus amomi is Zingiber herbaceos perennial amomum viosum (Amomum villosum Lour.), green shell sand (Amomum
) or the dry mature fruit of SEMEN AMOMI LONGILIGULA (Amomum longiligulare) xanthioides.Its capsule ellipse, when ripe
Aubergine, brown after doing, seed polygonal have strong fragrance, bitter cool.It is distributed in Fujian China, Guangdong, Guangxi and cloud
South；Cultivation is wild in the shady wet place in mountainous region.Fruit hyoscine cures mainly the taste stagnation of the circulation of vital energy, place with the performance optimal of Guangdong amomum viosum
Food does not disappear, abdominal pain swollen ruffian, dysphagic vomiting, cold to rush down cold dysentery, is a kind of conventional Chinese medicine of integration of drinking and medicinal herbs.
There is the phenomenon that being palmed off as with nearly edge species in fructus amomi, can be reflected by peel character with arillate fructus amomi medicinal material
Not, but in commercially available fructus amomi there are a large amount of seed group's class samples and part sand rice and coarse powder class sample, such sample there is no technology
Means determine the true and false, and there is an urgent need for a kind of technical methods differentiating such sample.
Invention content
Based on this, it is necessary in view of the above-mentioned problems, provide a kind of fructus amomi discrimination method, the seed of fructus amomi can be rolled into a ball class,
Sand rice class and coarse powder class sample carry out base identification.
A kind of fructus amomi discrimination method, includes the following steps：
(1) preparation of reference substance solution：
Precision weighs Bronyl acetate reference substance, and with organic solvent dissolving and constant volume is to get reference substance solution；
(2) preparation of test solution：
Fructus amomi test sample is taken, volatile oil is extracted, obtains test solution；
(3) it identifies：Above-mentioned reference substance solution and test solution are drawn respectively, is injected separately into gas chromatograph for determination, are pressed
Base identification is carried out to the fructus amomi test sample according to preassigned.
Above-mentioned fructus amomi discrimination method, it is right by the way that analysis measurement will be carried out with gas-chromatography after the volatile oil extracting in fructus amomi
Seed group's class, sand rice class and coarse powder class sample carry out base identification, overcome in routine techniques and differentiate the fructus amomi true and false by shape
The problem of having difficulties.
In one of the embodiments, in the preparation of step (1) reference substance solution, also precision weighs linalool, camphor, indigo plant
Reference substance solution is made at least one of eucalyptus alcohol, caryophyllene oxide and Trans-caryophyllene reference substance together with Bronyl acetate.
It is qualitative to the progress of each ingredient using reference substance, have the advantages that accuracy is high.
In one of the embodiments, in the preparation of step (1) reference substance solution, the organic solvent is n-hexane, institute
State in reference substance solution, linalool, camphor, globulol, caryophyllene oxide, Bronyl acetate and Trans-caryophyllene concentration be
0.35-0.45mg/ml.Reference substance concentration is set as above range, there is preferable response in gas-chromatography, is conducive to analysis
The accuracy of differentiation.
In one of the embodiments, in the preparation of step (2) test solution, volatile oil is extracted in accordance with the following methods：
It takes fructus amomi test sample to be placed in flask, volatile oil determination apparatus is connected after adding water, from analyzer upper end, Jia Shui makes to be full of scale part,
And until overflow enters flask, reflux condensing tube is connected, is heated to reflux 4-6 hours, it is cooling, it stands, volatile oil extracting liquid is taken, with just
Hexane be solvent dilute 50-150 times to get.Method described above extracts volatile oil, has preferable extraction effect.
In one of the embodiments, in the preparation of step (2) test solution, the fructus amomi test sample includes kind
The fructus amomi sample of son group class, sand rice class and coarse powder class form.Above-mentioned discrimination method is not limited by the form of fructus amomi, no matter right
Seed group's class, or husky rice class or rough sort, there is preferable identification result.
In one of the embodiments, in step (3) identification, the condition of the gas chromatograph for determination is：
Chromatographic column：Agilent DB-1 quartz capillary columns, specification are 30m × 0.25mm × 0.25 μm；
Sample size：1μl；
Injector temperature：220℃-240℃；
Temperature program：60 DEG C of initial temperature is kept for 2 minutes, 120 DEG C is risen to 5 DEG C/min, after rising to 130 with 1 DEG C/min
DEG C, keep 5min；2000 DEG C are risen to 15 DEG C/min again, keeps 6min；
Detector：Fid detector, temperature are 240 DEG C -260 DEG C.
Said determination condition can detach the Multiple components in fructus amomi one by one, reach satisfied separating degree and peak type, full
Foot is subsequent to analyze and identify condition.
In one of the embodiments, in the step (3) identification, by by the guarantor of each chromatographic peak in test sample chromatogram
It stays time and the retention time of each chromatographic peak in reference substance chromatogram to be compared, each chromatographic peak in test sample chromatogram is carried out
Ingredient belongs to；
Or
When by by the retention time of each chromatographic peak in test sample chromatogram and the opposite reservation of Bronyl acetate chromatographic peak
Between compared, each chromatographic peak in test sample chromatogram is belonged to, it is specific as follows：
When the relative retention time of chromatographic peak and Bronyl acetate is 0.65-0.66, the then chromatography in test sample chromatogram
Peak is linalool；
When the relative retention time of chromatographic peak and Bronyl acetate is 0.715-0.735, the then color in test sample chromatogram
Spectral peak is camphor；
When the relative retention time of chromatographic peak and Bronyl acetate is 1.353-1.373, the then color in test sample chromatogram
Spectral peak is Trans-caryophyllene；
When the relative retention time of chromatographic peak and Bronyl acetate is 2.005-2.0205, the then color in test sample chromatogram
Spectral peak is globulol；
When in test sample chromatogram the relative retention time of chromatographic peak and Bronyl acetate be 2.0789-2.0989, then should
Chromatographic peak is caryophyllene oxide.
In one of the embodiments, in step (3) identification, the criterion is：By each ingredient in test sample
The peak area of chromatographic peak and the peak area ratio of Bronyl acetate are set as the limit value of the ingredient, are sentenced according to following standard
It is fixed：
When linalool limit value be more than or equal to 0.05, then judge the test sample for adulterant；
When linalool limit value be less than 0.05, and the limit value of caryophyllene oxide be more than or equal to 0.01, then judge the confession
Test product is adulterant；
When the limit value of linalool is less than 0.05, the limit value of caryophyllene oxide is less than 0.01, and the limit value of globulol
More than or equal to 0.08, then judge the test sample for Fructus Amomi；
When the limit value of linalool is less than 0.05, the limit value of caryophyllene oxide is less than 0.01, and the limit value of globulol is big
In equal to 0.01 and less than 0.08, then judge the test sample for class Fructus Amomi；
When the limit value of linalool is less than 0.05, the limit value of caryophyllene oxide is less than 0.01, and the limit value of globulol is small
In 0.01, the limit value of camphor is more than or equal to 0.85, and the limit value of Trans-caryophyllene is more than or equal to 0.025, then judges the confession
Test product is adulterant；
When the limit value of linalool is less than 0.05, the limit value of caryophyllene oxide is less than 0.01, and the limit value of globulol is small
In 0.01, the limit value of camphor is less than 0.85, and the limit value of Trans-caryophyllene is less than 0.025, then judges the test sample for sun
Fructus amomi or green shell sand benevolence.
Above-mentioned criterion has accuracy high, judges simple advantage.
The invention also discloses above-mentioned fructus amomi discrimination methods in seed group, fructus amomi, coarse powder class fructus amomi sample differentiate
Using.
The invention also discloses a kind of fructus amomi identification systems, using above-mentioned discrimination method, including pre-processing module, extraction
Module, detection module and identification module；
The pre-processing module is for configuring the reference substance solution；
The extraction module is used to extract the volatile oil of fructus amomi test sample, obtains test solution；
The detection module is used to carry out gas chromatographic analysis detection to reference substance solution and test solution；
The identification module obtains qualification result for the information that detection module obtains to be analyzed and handled.
Compared with prior art, the invention has the advantages that：
A kind of fructus amomi discrimination method of the present invention, by will be analyzed with gas-chromatography after the volatile oil extracting in fructus amomi
It measures, base identification is carried out to seed group class, sand rice class and coarse powder class sample, overcomes in routine techniques and sand is differentiated by shape
The problem of benevolence true and false has difficulties.Foundation is provided for the identification and quality evaluation of fructus amomi, is science, comprehensive formulation fructus amomi
Quality standard provides reference.
Description of the drawings
Fig. 1 is that reference substance chromatogram figure is mixed in embodiment 1；
Fig. 2 is separate sources test sample typical case's chromatogram in embodiment 1；
Fig. 3 is the criterion schematic diagram of fructus amomi sample in embodiment 1.
Specific implementation mode
To facilitate the understanding of the present invention, below with reference to relevant drawings to invention is more fully described.In attached drawing
Give presently preferred embodiments of the present invention.But the present invention can realize in many different forms, however it is not limited to this paper institutes
The embodiment of description.Keep the understanding to the disclosure more thorough on the contrary, purpose of providing these embodiments is
Comprehensively.
Unless otherwise defined, all of technologies and scientific terms used here by the article and belong to the technical field of the present invention
The normally understood meaning of technical staff is identical.Used term is intended merely to description tool in the description of the invention herein
The purpose of the embodiment of body, it is not intended that in the limitation present invention.
Agents useful for same in following embodiment is unless otherwise indicated commercially available.
Embodiment 1
Fructus amomi gas-chromatography discrimination method is studied.
One, the preparation of reference substance solution：
Take australene, down alkene, nopinene, beta-myrcene, limonene, γ-terpinene, α-terpinolene, linalool, camphor tree
Brain, isoborneol, borneol, Bronyl acetate, α-copaene, beta-elemene and appropriate Trans-caryophyllene, add n-hexane that concentration is made
Respectively be 0.4mg/ml (the wherein a concentration of 0.2mg/ml of beta-myrcene and α-copaene) mixed reference substance solution to get.
Two, the preparation of test solution：
Fructus amomi test sample is taken, extracts volatile oil by the following method：
It takes Seeds of Amomum Villosum to roll into a ball 30g, crushes, set in 500ml round-bottomed flasks, add water 300ml and bead number, shaking mixing
Afterwards, volatile oil determination apparatus is connected, until from analyzer upper end, Jia Shui makes to be full of scale part, and overflow enters flask, connection reflux
Condenser pipe is heated to reflux 5 hours, cooling, is stood 1 hour, is taken volatile oil extracting liquid 0.1ml, set in 10ml tool plug test tubes, add just
Hexane to scale, shake up to get.Take volatile oil 0.1ml, set in 10ml tool plug test tubes, add n-hexane to scale, shake up to get
Test solution.
Three, it detects：
1, analysis condition.
It is detected using Shimadzu GC 2010plus gas chromatographs, condition is：
Chromatographic column：Agilent DB-1 quartz capillary columns, specification are 30m × 0.25mm × 0.25 μm；
Sample size：1μl；
Injector temperature：30℃；
Temperature program：60 DEG C of initial temperature is kept for 2 minutes, 120 DEG C is risen to 5 DEG C/min, after rising to 130 with 1 DEG C/min
DEG C, keep 5min；2000 DEG C are risen to 15 DEG C/min again, keeps 6min；
Detector：Fid detector, temperature are 250 DEG C.
2, result.
2.1 mixing reference substance chromatograms.
Reference substance chromatogram is mixed as shown in Figure 1, figure label 1-15 is respectively australene, down alkene, nopinene, β-bay
Alkene, limonene, γ-terpinene, α-terpinolene, linalool, camphor, isoborneol, borneol, Bronyl acetate, α-copaene, β-
Elemene and Trans-caryophyllene.
It can be seen from the figure that there is preferable separating degree between each ingredient, and each chromatographic peak has preferable peak type, energy
Enough meet subsequent analysis requirement.
2.2 test sample chromatograms.
It takes the accurate samples of different sources collection Ji Yuan and totally 119 batches of fructus amomis is used as examination precise Identification fruit class sample
Product, including certified products fructus amomi (amomum viosum, green shell sand, SEMEN AMOMI LONGILIGULA) and common adulterant (long sequence fructus amomi, Amomum maricarpum Elm, red bean
It is cool), the typical chromatogram of different samples is obtained, as shown in Figure 2.
By comparing fructus amomi (amomum viosum, green shell sand, SEMEN AMOMI LONGILIGULA) and common adulterant (long sequence fructus amomi, Amomum maricarpum Elm, red bean
It is cool) characteristic spectrum, find 6 specific chromatographic peak (linalool, camphor, Bronyl acetate, indigo plants of representative and otherness
Eucalyptus alcohol, caryophyllene oxide, Trans-caryophyllene) (referring to table 1), using Bronyl acetate as base peak, 5 specific chromatographic peaks of final choice
Peak area ratio with Bronyl acetate is limit (referring to table 2), drafts the inspection method of fructus amomi.
The representative specific chromatographic peak list with otherness of table 1
The Characteristic chromatographic peak of 2 typical sample of table and the peak area ratio (limit value) of Bronyl acetate
3, the criterion of discrimination method.
By the above results, the criterion of fructus amomi sample discrimination method is formulated as shown in figure 3, specific as follows：
(1) linalool of fructus amomi (amomum viosum, green shell sand, SEMEN AMOMI LONGILIGULA) and Bronyl acetate chromatographic peak area ratio should be less than
0.05, caryophyllene oxide and Bronyl acetate chromatographic peak area ratio all should be less than 0.01；
(2) globulol of SEMEN AMOMI LONGILIGULA should be greater than 0.08 with Bronyl acetate chromatographic peak area ratio；
(3) globulol of amomum viosum and green shell sand should be less than 0.01, camphor and second with Bronyl acetate chromatographic peak area ratio
Sour borneol acetate chromatographic peak area ratio should be less than 0.85, Trans-caryophyllene and Bronyl acetate chromatographic peak area ratio should be less than 0.25.
Embodiment 2
A kind of fructus amomi discrimination method, includes the following steps：
One, the preparation of reference substance solution：
Precision weighs Bronyl acetate reference substance, and with organic solvent dissolving and constant volume is to get reference substance solution；
Two, the preparation of test solution：
Known source fructus amomi sample is taken, 28 batches of seed groups, sand rice and coarse powder class sample is made as test sample, according to implementation
The method of example 1 extracts volatile oil, obtains test solution；
Three, it identifies：It is accurate respectively to draw above-mentioned reference substance solution and test solution, it is injected separately into gas chromatograph, is pressed
It is measured according to the method for embodiment 1, and base identification is carried out to the fructus amomi test sample according to the standard of embodiment 1, as a result
As shown in the table.
3 seed of table rolls into a ball class, the judgement result of sand rice class and coarse powder class sample
Differentiate that result is as follows：
Above-mentioned differentiation result is compared with sample preparation source, all judgement results are accurate, i.e. judgement is accurate
Rate is 100%.
Each technical characteristic of embodiment described above can be combined arbitrarily, to keep description succinct, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, it is all considered to be the range of this specification record.
Several embodiments of the invention above described embodiment only expresses, the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the protection of the present invention
Range.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (10)
1. a kind of fructus amomi discrimination method, which is characterized in that include the following steps：
(1) preparation of reference substance solution：
Precision weighs Bronyl acetate reference substance, and with organic solvent dissolving and constant volume is to get reference substance solution；
(2) preparation of test solution：
Fructus amomi test sample is taken, volatile oil is extracted, obtains test solution；
(3) it identifies：Above-mentioned reference substance solution and test solution are drawn respectively, is injected separately into gas chromatograph for determination, according to pre-
Calibration is accurate to carry out base identification to the fructus amomi test sample.
2. fructus amomi discrimination method according to claim 1, which is characterized in that in the preparation of step (1) reference substance solution, also
Precision weighs at least one of linalool, camphor, globulol, caryophyllene oxide and Trans-caryophyllene reference substance, with acetic acid dragon
Reference substance solution is made in brain ester together.
3. fructus amomi discrimination method according to claim 2, which is characterized in that in the preparation of step (1) reference substance solution, institute
It is n-hexane to state organic solvent, in the reference substance solution, linalool, camphor, globulol, caryophyllene oxide, Bronyl acetate
Concentration with Trans-caryophyllene is 0.35-0.45mg/ml.
4. fructus amomi discrimination method according to claim 1, which is characterized in that in the preparation of step (2) test solution, press
Volatile oil is extracted according to following methods：Take fructus amomi for examinationProduct are placed in flask, volatile oil determination apparatus is connected after adding water, from analyzer
Until end plus water make to be full of scale part, and overflow enters flask, reflux condensing tube is connected, is heated to reflux 4-6 hours, it is cooling, it is quiet
Set, take volatile oil extracting liquid, using n-hexane as solvent dilute 50-150 times to get.
5. fructus amomi discrimination method according to claim 1, which is characterized in that the preparation of step (2) test solution
In, the fructus amomi test sample includes the fructus amomi sample of seed group class, sand rice class and coarse powder class form.
6. fructus amomi discrimination method according to claim 1, which is characterized in that in step (3) identification, the gas chromatograph
The condition of measurement is：
Chromatographic column：Agilent DB-1 quartz capillary columns, specification are 30m × 0.25mm × 0.25 μm；
Sample size：1μl；
Injector temperature：220℃-240℃；
Temperature program：60 DEG C of initial temperature is kept for 2 minutes, 120 DEG C is risen to 5 DEG C/min, after rising to 130 DEG C with 1 DEG C/min,
Keep 5min；2000 DEG C are risen to 15 DEG C/min again, keeps 6min；
Detector：Fid detector, temperature are 240 DEG C -260 DEG C.
7. according to claim 1-6 any one of them fructus amomi discrimination methods, which is characterized in that in step (3) identification, lead to
It crosses and carries out the retention time of each chromatographic peak in the retention time of each chromatographic peak in test sample chromatogram and reference substance chromatogram pair
Than belonging to each chromatographic peak in test sample chromatogram；
Or
By by the relative retention time of the retention time of each chromatographic peak in test sample chromatogram and Bronyl acetate chromatographic peak into
Row comparison carries out ingredient ownership to each chromatographic peak in test sample chromatogram, specific as follows：
When the relative retention time of chromatographic peak and Bronyl acetate is 0.65-0.66 in test sample chromatogram, then the chromatographic peak is
Linalool；
When the relative retention time of chromatographic peak and Bronyl acetate is 0.715-0.735, the then chromatographic peak in test sample chromatogram
For camphor；
When the relative retention time of chromatographic peak and Bronyl acetate is 1.353-1.373, the then chromatographic peak in test sample chromatogram
For Trans-caryophyllene；
When the relative retention time of chromatographic peak and Bronyl acetate is 2.005-2.0205, the then chromatographic peak in test sample chromatogram
For globulol；
When the relative retention time of chromatographic peak and Bronyl acetate is 2.0789-2.0989, the then chromatography in test sample chromatogram
Peak is caryophyllene oxide.
8. fructus amomi discrimination method according to claim 7, which is characterized in that in step (3) identification, the judgement mark
Standard is that the peak area of each ingredient chromatographic peak in test sample and the peak area ratio of Bronyl acetate are set as to the limit of the ingredient
Value, is judged according to following standard：
When linalool limit value be more than or equal to 0.05, then judge the test sample for adulterant；
When linalool limit value be less than 0.05, and the limit value of caryophyllene oxide be more than or equal to 0.01, then judge the test sample
For adulterant；
When the limit value of linalool is less than 0.05, the limit value of caryophyllene oxide is less than 0.01, and the limit value of globulol is more than
Equal to 0.08, then judge the test sample for Fructus Amomi；
When the limit value of linalool is less than 0.05, the limit value of caryophyllene oxide is less than 0.01, and the limit value of globulol is more than etc.
In 0.01 and be less than 0.08, then judge the test sample for class Fructus Amomi；
When the limit value of linalool is less than 0.05, the limit value of caryophyllene oxide is less than 0.01, and the limit value of globulol is less than
0.01, the limit value of camphor is more than or equal to 0.85, and the limit value of Trans-caryophyllene is more than or equal to 0.025, then judges this for examination
Product are adulterant；
When the limit value of linalool is less than 0.05, the limit value of caryophyllene oxide is less than 0.01, and the limit value of globulol is less than
0.01, the limit value of camphor is less than 0.85, and the limit value of Trans-caryophyllene is less than 0.025, then judges the test sample for spring
Fructus amomi or green shell sand benevolence.
9. claim 1-8 any one of them fructus amomi discrimination methods are in seed group, fructus amomi, coarse powder class fructus amomi sample differentiate
Using.
10. a kind of fructus amomi identification system, which is characterized in that use claim 1-8 any one of them discrimination methods, including preceding
Processing module, extraction module, detection module and identification module；
The pre-processing module is for configuring the reference substance solution；
The extraction module is used to extract the volatile oil of fructus amomi test sample, obtains test solution；
The detection module is used to carry out gas chromatographic analysis detection to reference substance solution and test solution；
The identification module obtains qualification result for the information that detection module obtains to be analyzed and handled.
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| CN111983092A (en) * | 2020-08-26 | 2020-11-24 | 劲牌有限公司 | Method for detecting fructus amomi medicinal material fingerprint |
| CN114755315A (en) * | 2021-07-23 | 2022-07-15 | 山东省中医药研究院 | Method for identifying amomum villosum or green-shell sand by using GC characteristic fingerprint |
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| CN110093443A (en) * | 2019-05-10 | 2019-08-06 | 河北中医学院 | A kind of kit and discrimination method identifying amomum viosum |
| CN110093443B (en) * | 2019-05-10 | 2023-08-01 | 河北中医学院 | Kit and method for identifying spring sand |
| CN111983092A (en) * | 2020-08-26 | 2020-11-24 | 劲牌有限公司 | Method for detecting fructus amomi medicinal material fingerprint |
| CN114755315A (en) * | 2021-07-23 | 2022-07-15 | 山东省中医药研究院 | Method for identifying amomum villosum or green-shell sand by using GC characteristic fingerprint |
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