CN109580830A - The detection method of trans-anethole content in a kind of fennel seeds - Google Patents
The detection method of trans-anethole content in a kind of fennel seeds Download PDFInfo
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- CN109580830A CN109580830A CN201811627738.5A CN201811627738A CN109580830A CN 109580830 A CN109580830 A CN 109580830A CN 201811627738 A CN201811627738 A CN 201811627738A CN 109580830 A CN109580830 A CN 109580830A
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- 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
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Abstract
The invention discloses a kind of detection methods of trans-anethole content in fennel seeds, including, contrast solution preparation steps: taking trans-anethole reference substance, and ethyl acetate is added, and preparation obtains contrast solution；Test solution preparation steps: taking fennel seeds sample, is added ethyl acetate, weighed weight, obtains weight A, after ultrasonic treatment, it lets cool, then weighed weight, obtains weight B, the weight of A to B less loss is supplied with ethyl acetate, it shakes up, filters, take subsequent filtrate, obtain test solution；Determination step: taking contrast solution and test solution respectively, injects gas chromatograph, measurement.Detection method separating degree of the invention is high, is resistant to matrix interference, high sensitivity, and stability is high, and accuracy is high.
Description
Technical field
The present invention relates to a kind of detection methods of trans-anethole content in field of traditional Chinese medicine detection more particularly to fennel seeds.
Background technique
Currently, China constantly promotes the supervision of Chinese medicine and its prepared slice quality, traditional Chinese medicine is also again by vast
The favor of the people, traditional Chinese medicine at home using more and more extensive, the quality of Chinese medicine also more and more attention has been paid to.And medicinal material
And the effective content of characteristic component is one of evaluation and the important indicator for influencing its quality in medicine materical crude slice.For fennel seeds, instead
The content height of formula anethole is exactly the important indicator that its quality good or not is evaluated in detection.Trans-anethole in fennel seeds at present
The detection method of content detection is gas chromatography, and sample pre-treatments extraction mainly has extraction by steam distillation volatile oil and second
Acetoacetic ester ultrasonic extraction, chromatographic column temperature have constant temperature and temperature programming both of which, and sample introduction has shunting and not shunt mode etc..Due to
Different chromatographic apparatus, different chromatographic columns, different chromatographic parameters can all cause sample detection to be disturbed, and resolution ratio is low.
Summary of the invention
For overcome the deficiencies in the prior art, the purpose of the present invention is to provide trans-anethole contents in a kind of fennel seeds
Detection method, detection method separating degree is high, is resistant to matrix interference, high sensitivity, and stability is high, and accuracy is high.
The purpose of the present invention adopts the following technical scheme that realization:
The detection method of trans-anethole content in a kind of fennel seeds, including,
Contrast solution preparation steps: taking trans-anethole reference substance, and ethyl acetate is added, and preparation obtains contrast solution；
Test solution preparation steps: taking fennel seeds sample, is added ethyl acetate, weighed weight, obtains weight A, at ultrasound
It after reason, lets cool, then weighed weight, obtains weight B, the weight of A to B less loss is supplied with ethyl acetate, is shaken up, filter, take continuous filter
Liquid obtains test solution；
Determination step: taking contrast solution and test solution respectively, injects gas chromatograph, measurement.
Further, in contrast solution preparation steps, the reference substance of trans-anethole containing 0.4mg in every 1mL contrast solution.
Further, in test solution preparation steps, fennel seeds sample was the fennel after No. three sieves.
Further, in test solution preparation steps, fennel seeds sample 0.4-0.6g is taken.
Further, in test solution preparation steps, after taking fennel seeds sample, the additional amount of ethyl acetate is 20-30mL.
Further, in test solution preparation steps, the parameter of ultrasonic treatment are as follows: power 250-350W, frequency 35-
45kHz。
Further, in test solution preparation steps, the time of ultrasonic treatment is 25-35 minutes.
Further, in determination step, 2 μ L of contrast solution and test solution is taken respectively.
Further, in determination step, the chromatographic column of gas chromatograph is polyethylene glycol capillary column, column length 30m, interior
Diameter is 0.32mm, and film thickness is 0.25 μm.
Further, in determination step, the column temperature of gas chromatograph is 145 DEG C；Injection port is split sampling, and temperature is
165-200℃；Detector is flame ionization ditector, and temperature is 200 DEG C.
Compared with prior art, the beneficial effects of the present invention are:
The detection method of trans-anethole content in fennel seeds of the invention, the detection method overcome existing measurement side
Method is since different chromatographic apparatus, different chromatographic columns, different chromatographic parameters cause sample detection to be disturbed, resolution ratio is low
Defect, provides that a kind of separating degree is high, is resistant to matrix interference, high sensitivity, and stability is high, the high gas chromatographic detection of accuracy
Method.
Detailed description of the invention
Fig. 1 is the chromatogram of the detection method of embodiment 1；
Fig. 2 is the chromatogram of the detection method of comparative example 1.
Specific embodiment
In the following, being described further in conjunction with attached drawing and specific embodiment to the present invention, it should be noted that not
Under the premise of conflicting, new implementation can be formed between various embodiments described below or between each technical characteristic in any combination
Example.
The detection method of trans-anethole content in a kind of fennel seeds, including,
Contrast solution preparation steps: taking trans-anethole reference substance, and ethyl acetate is added, and preparation obtains contrast solution；
Test solution preparation steps: taking fennel seeds sample, is added ethyl acetate, weighed weight, obtains weight A, at ultrasound
It after reason, lets cool, then weighed weight, obtains weight B, the weight of A to B less loss is supplied with ethyl acetate, is shaken up, filter, take continuous filter
Liquid obtains test solution；
Determination step: taking contrast solution and test solution respectively, injects gas chromatograph, measurement.
As the mode that further carries out, in contrast solution preparation steps, the trans- fennel containing 0.4mg in every 1mL contrast solution
Fragrant brain reference substance.
As the mode that further carries out, in test solution preparation steps, fennel seeds sample was the fennel fruit after No. three sieves
Face powder end.
As the mode that further carries out, in test solution preparation steps, fennel seeds sample 0.4-0.6g is taken.
As the mode that further carries out, in test solution preparation steps, after taking fennel seeds sample, the addition of ethyl acetate
Amount is 20-30mL.
As the mode that further carries out, in test solution preparation steps, the parameter of ultrasonic treatment are as follows: power 250-
350W, frequency 35-45kHz.
As the mode that further carries out, in test solution preparation steps, the time of ultrasonic treatment is 25-35 minutes.
As the mode that further carries out, in determination step, 2 μ L of contrast solution and test solution is taken respectively.
As the mode that further carries out, in determination step, the chromatographic column of gas chromatograph is polyethylene glycol capillary column,
Column length is 30m, and internal diameter 0.32mm, film thickness is 0.25 μm.
As the mode that further carries out, in determination step, the column temperature of gas chromatograph is 145 DEG C；Injection port is to shunt
Sample introduction, temperature are 165-200 DEG C；Detector is flame ionization ditector, and temperature is 200 DEG C.
It is specific embodiment of the present invention below, used raw material, equipment etc. remove special limit in the following embodiments
It can be obtained by buying pattern outside fixed.
Embodiment 1:
The detection method of trans-anethole content in a kind of fennel seeds, including,
Contrast solution preparation steps: taking trans-anethole reference substance, and ethyl acetate is added, and preparation obtains contrast solution, often
The reference substance of trans-anethole containing 0.4mg in 1mL contrast solution；
Test solution preparation steps: taking 0.4g to cross the fennel after No. three sieves, and 20mL ethyl acetate is added, weighed heavy
Amount obtains weight A, is ultrasonically treated 26 minutes, the parameter of ultrasonic treatment are as follows: power 240W, frequency 40kHz are let cool, then weighed heavy
Amount, obtains weight B, the weight of A to B less loss is supplied with ethyl acetate, is shaken up, and filters, takes subsequent filtrate, obtain test solution；
Determination step: taking 2 μ L contrast solutions and test solution respectively, injects gas chromatograph, measurement.
Wherein, the chromatographic column of gas chromatograph is polyethylene glycol capillary column, column length 30m, internal diameter 0.32mm, film thickness
Degree is 0.25 μm.The column temperature of gas chromatograph is 145 DEG C；Injection port is split sampling, and temperature is 200 DEG C；Detector is hydrogen fire
Flame ionization detector, temperature are 200 DEG C.
Embodiment 2:
The detection method of trans-anethole content in a kind of fennel seeds, including,
Contrast solution preparation steps: taking trans-anethole reference substance, and ethyl acetate is added, and preparation obtains contrast solution, often
The reference substance of trans-anethole containing 0.4mg in 1mL contrast solution；
Test solution preparation steps: taking 0.5g to cross the fennel after No. three sieves, and 25mL ethyl acetate is added, weighed heavy
Amount obtains weight A, is ultrasonically treated 29 minutes, the parameter of ultrasonic treatment are as follows: power 270W, frequency 40kHz are let cool, then weighed heavy
Amount, obtains weight B, the weight of A to B less loss is supplied with ethyl acetate, is shaken up, and filters, takes subsequent filtrate, obtain test solution；
Determination step: taking 2 μ L contrast solutions and test solution respectively, injects gas chromatograph, measurement.
Wherein, the chromatographic column of gas chromatograph is polyethylene glycol capillary column, column length 30m, internal diameter 0.32mm, film thickness
Degree is 0.25 μm.The column temperature of gas chromatograph is 145 DEG C；Injection port is split sampling, and temperature is 200 DEG C；Detector is hydrogen fire
Flame ionization detector, temperature are 200 DEG C.
Embodiment 3:
The detection method of trans-anethole content in a kind of fennel seeds, including,
Contrast solution preparation steps: taking trans-anethole reference substance, and ethyl acetate is added, and preparation obtains contrast solution, often
The reference substance of trans-anethole containing 0.4mg in 1mL contrast solution；
Test solution preparation steps: taking 0.6g to cross the fennel after No. three sieves, and 30mL ethyl acetate is added, weighed heavy
Amount obtains weight A, is ultrasonically treated 34 minutes, the parameter of ultrasonic treatment are as follows: power 300W, frequency 40kHz are let cool, then weighed heavy
Amount, obtains weight B, the weight of A to B less loss is supplied with ethyl acetate, is shaken up, and filters, takes subsequent filtrate, obtain test solution；
Determination step: taking 2 μ L contrast solutions and test solution respectively, injects gas chromatograph, measurement.
Wherein, the chromatographic column of gas chromatograph is polyethylene glycol capillary column, column length 30m, internal diameter 0.32mm, film thickness
Degree is 0.25 μm.The column temperature of gas chromatograph is 145 DEG C；Injection port is split sampling, and temperature is 200 DEG C；Detector is hydrogen fire
Flame ionization detector, temperature are 200 DEG C.
Comparative example 1:
1 difference from Example 1 of comparative example is that comparative example 1 is using not shunt mode and carrier gas rate.Comparative example
Carrier gas rate is mainly reflected on chromatogram has shortened retention time, does not generate crucial effect to overall separation.
Effect assessment and performance detection
Each parameter of the measuring method of trans-anethole content is specifically such as by taking embodiment 1 as an example, in the fennel seeds of embodiment 1
Under:
The post case parameter of embodiment 1 is as follows:
The advantages of (rear operation) 180 DEG C of settings in 1 minute is: object is able to elution separation under predetermined chromatographic condition,
And corresponding curved flow of chromatography is collected, according to subsequent detection needs, sample characteristics of for example and chromatographic system characteristic to chromatographic system
It is purified, aging.
The advantages of equilibration time setting in 1 minute is: a capture program is complete before sample sequence starts and in sequence operation
Cheng Hou before next capture program operation sample introduction, to make system preferably be in original state, obtains better accuracy and again
Renaturation, then balanced for a period of time to system.
The parameter of preceding sample injector is as follows:
Using autosampler sample introduction advantage: cleaning journey according to sample characteristics of for example and cleaning solution placement location setting sample introduction needle
Sequence, suction-discharge rate guarantee sample introduction degree of purity and rate, prevent sample pollution, cross contamination, sub-sampling (sample introduction is slow),
Reduce sample diffusion etc..
SS injection port N2 parameter is as follows:
200 DEG C of temperature instantaneously all vaporizes after can guaranteeing sample feeding and will not decompose, crack.
The setting of pressure 11.047psi and split ratio (10:1) can guarantee that injection port has suitable pressure and selects properly
Split ratio so that flow rate of carrier gas (or linear velocity) reach setting value and control enter chromatographic column sample size so that sample exists
Nonoverload in chromatographic column, in the linear range, sample obtains better separating degree to chromatography response, guarantee detection repeatability and
Accuracy.
The parameter of chromatographic column is as follows:
Mean Speed is set as 35cm/sec: selecting suitable carrier gas rate is to reduce chromatographic column plate height, improve tower
Plate number makes the better separation detection of sample to improve column efficiency.On the other hand it under the conditions of guaranteeing reasonable separating degree, closes
Reason, which adjusts carrier gas rate, can also accelerate to detect speed, shorten detection time, improve the working efficiency of detection.
The parameter of preceding detector is as follows:
The parameter of signal is as follows:
Chromatographic column is described as follows: DB-WAX
Catalogue #:2407.37055
Model#:123-7032 Manufacturer:J&W
Internal diameter: 320.00 μm of length: 30.0m
The Film thickness:0.25 μm time: 1.429min
Maximum temperature: 250.0 DEG C.
Fig. 1 is the chromatogram of embodiment 1, and Fig. 2 is the chromatogram of comparative example 1, it can be seen that sample is pure from Fig. 1, Fig. 2
Only, other impurity in fennel seeds sample substrate are effectively removed, while object and impurity separating degree reach 1.5 or more,
Effectively object is separated with impurity.
Due to the different comparison diagrams 1 of sample introduction mode, Fig. 2 as it can be seen that in Fig. 1 on curved flow of chromatography object peak shape mark
Standard does not influence to integrate accuracy, can ensure that detection detection accuracy and repeatability.Sample overloads, on curved flow of chromatography in Fig. 2
Object peak shape is asymmetric, not only influences integral accuracy, can not ensure that sample can be quantified in the reasonable range of linearity
Detection.It can be seen that the present invention measures trans-anethole content in fennel seeds using shunt mode sample introduction, compared with not shunting,
Accurate, the advantages such as reproducible with detection.
The above embodiment is only the preferred embodiment of the present invention, and the scope of protection of the present invention is not limited thereto,
The variation and replacement for any unsubstantiality that those skilled in the art is done on the basis of the present invention belong to institute of the present invention
Claimed range.
Claims (10)
1. the detection method of trans-anethole content in a kind of fennel seeds, it is characterised in that including,
Contrast solution preparation steps: taking trans-anethole reference substance, and ethyl acetate is added, and preparation obtains contrast solution；
Test solution preparation steps: taking fennel seeds sample, is added ethyl acetate, weighed weight, obtains weight A, after ultrasonic treatment,
It lets cool, then weighed weight, obtains weight B, the weight of A to B less loss is supplied with ethyl acetate, is shaken up, filter, take subsequent filtrate, obtain
To test solution；
Determination step: taking contrast solution and test solution respectively, injects gas chromatograph, measurement.
2. the detection method of trans-anethole content in fennel seeds as described in claim 1, which is characterized in that contrast solution is matched
In step processed, the reference substance of trans-anethole containing 0.4mg in every 1mL contrast solution.
3. the detection method of trans-anethole content in fennel seeds as described in claim 1, which is characterized in that test solution is matched
In step processed, fennel seeds sample was the fennel after No. three sieves.
4. the detection method of trans-anethole content in fennel seeds as described in claim 1, which is characterized in that test solution is matched
In step processed, fennel seeds sample 0.4-0.6g is taken.
5. the detection method of trans-anethole content in fennel seeds as described in claim 1, which is characterized in that test solution is matched
In step processed, after taking fennel seeds sample, the additional amount of ethyl acetate is 20-30mL.
6. the detection method of trans-anethole content in fennel seeds as described in claim 1, which is characterized in that test solution is matched
In step processed, the parameter of ultrasonic treatment are as follows: power 250-350W, frequency 35-45kHz.
7. the detection method of trans-anethole content in fennel seeds as described in claim 1, which is characterized in that test solution is matched
In step processed, the time of ultrasonic treatment is 25-35 minutes.
8. the detection method of trans-anethole content in fennel seeds as described in claim 1, which is characterized in that determination step
In, 2 μ L of contrast solution and test solution is taken respectively.
9. the detection method of trans-anethole content in fennel seeds as described in claim 1, which is characterized in that determination step
In, the chromatographic column of gas chromatograph is polyethylene glycol capillary column, and column length 30m, internal diameter 0.32mm, film thickness is 0.25 μ
m。
10. the detection method of trans-anethole content in fennel seeds as described in claim 1, which is characterized in that determination step
In, the column temperature of gas chromatograph is 145 DEG C；Injection port is split sampling, and temperature is 165-200 DEG C；Detector be hydrogen flame from
Sonization detector, temperature are 200 DEG C.
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