CN107589203A - A kind of method for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE HPLC - Google Patents
A kind of method for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE HPLC Download PDFInfo
- Publication number
- CN107589203A CN107589203A CN201710863944.5A CN201710863944A CN107589203A CN 107589203 A CN107589203 A CN 107589203A CN 201710863944 A CN201710863944 A CN 201710863944A CN 107589203 A CN107589203 A CN 107589203A
- Authority
- CN
- China
- Prior art keywords
- kinds
- spe
- fiber crops
- cannabinol compounds
- chinese fiber
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
The invention belongs to plant component detection method field, and in particular to a kind of method for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE HPLC.SPE HPLC is applied to the detection of cannabinol compounds in Chinese fiber crops by the present invention, can detect three kinds of cannabinol compounds in Chinese fiber crops simultaneously, and carry out qualitative and quantitative analysis to it.The present invention determines high-efficient liquid phase chromatogram condition first, draws the standard curve of three kinds of cannabinol compounds by the detection to hybrid standard product solution, determines equation of linear regression；SPE removal of impurities is carried out to Chinese fiber crops sample again, efficient liquid phase detection and the content by the way that three kinds of cannabinol compounds in Chinese fiber crops are calculated finally are carried out to sample.SPE of the present invention separates cannabinol compounds to be measured with impurity, reduces influence of the impurity to detection, and good basis is provided for liquid phase detection.Detection method linear relationship in the corresponding scope of application is good, precision, repeatability, the degree of accuracy are good.
Description
Technical field
The invention belongs to plant component detection method field, and in particular to one kind is detected in Chinese fiber crops simultaneously using SPE-HPLC
The method of three kinds of cannabinol compounds.
Background technology
As soon as Chinese fiber crops, before 10000 years, are utilized as one of crop most ancient on the earth by the mankind.Chinese fiber crops bast
Fiber is one of best natural fiber, and it is longer than cotton fiber, and toughness and hygroscopicity are stronger, and partiting thermal insulation effect is more preferable,
It is first-class textile and paper raw material.With the development of modernization science and technology, the active ingredient in Chinese fiber crops gradually causes concern, its
Middle cannabinol compounds factor amount and bioactive ingredients are more and be taken seriously.Cannabinol compounds mainly include tetrahydrochysene
The compositions such as cannabinol (THC), cannabidiol (CBD), cannabinol (CBN) and cannabigerol (CBC).
Because cannabinol compounds species is various, and these material polarity are similar, and it will be wherein different that prior art, which is difficult,
Cannabinol compounds composition is efficiently separated, and the impurity in detection sample is mixed in because that can not be completely separated to detection
As a result accuracy generates very big influence.
In the prior art most of detection on cannabinol compounds composition be all confined in the Chinese edestan oil THC or
The detection of CBD chemical analysis, detection method are mostly gas chromatography, and the detection document for THC is relatively more among these,
Method is more comprehensive.But these detection methods are detected for single cannabinol compounds mostly, when need detect the Chinese
, it is necessary to using respective pretreatment technology and analysis method during a variety of cannabinol compounds in fiber crops, detection is wasted time and energy, and
And consumption fund is big.
The content of the invention
, should by Solid phase extraction-HPLC (SPE-HPLC) instant invention overcomes above-mentioned the deficiencies in the prior art
Use the detection of cannabinol compounds in Chinese fiber crops, there is provided one kind detects three kinds of cannabinols in Chinese fiber crops simultaneously using SPE-HPLC
The method of class compound.
The present invention it is a kind of using SPE-HPLC simultaneously detect three kinds of cannabinol compounds in Chinese fiber crops method and step it is as follows：
First, high-efficient liquid phase chromatogram condition is determined：The high-efficient liquid phase chromatogram condition is：
C18 reversed-phase columns：4.6 × 150mm, 5 μm；Column temperature：40℃；Detection wavelength：220nm；Type of elution：Isocratic elution；
Mobile phase：Methanol:0.1% formic acid=80:20；Flow velocity：1mL/min；Sample size：20μL；
2nd, hybrid standard product solution is prepared：Prepare three kinds of cannabinol compounds for containing identical mass concentration simultaneously
CBD, CBN, THC and mass concentration be followed successively by 0.05 μ g/mL, 0.5 μ g/mL, 2 μ g/mL, 10 μ g/mL, 20 μ g/mL, 40 μ g/mL,
60 μ g/mL, 80 μ g/mL hybrid standard product solution；
3rd, equation of linear regression is determined：Each concentration hybrid standard product solution that step 2 is prepared is used into step 1 successively
High-efficient liquid phase chromatogram condition is detected, qualitative with retention time, and the size and its concentration of peak area are surveyed according to each concentration
Corresponding relation draws standard curve, and determines the equation of linear regression of three kinds of cannabinol compounds respectively；
4th, SPE pretreatment sample is prepared：Pulverization process is dried in Chinese fiber crops sample, take Chinese fiber crops sample powder in
Centrifuge tube simultaneously adds methanol, assists to extract the cannabinol compounds in Chinese fiber crops sample powder using ultrasound；Centrifugal treating extracts
Liquid simultaneously takes supernatant to obtain SPE pretreatment sample with methanol constant volume；
5th, SPE：Using the SPE pattern for retaining chaff interference, the SPE that step 4 is obtained pre-processes
Sample adds C18/500mg solid phase pillars, and mobile phase is methanol, flow velocity 1mL/min, pressure 0.1MPa, isocratic elution, is collected
Efflux；The solid phase pillar is eluted with a small amount of hplc grade methanol, collects efflux；
6th, high performance liquid chromatography detection：The efflux that step 5 is collected mixes and with methanol again constant volume, after membrane filtration
Detected using step 1 high-efficient liquid phase chromatogram condition, measure the peak area of each component in sample, root qualitative with retention time
The equation of linear regression determined according to step 3 quantifies, and calculates the concentration of three kinds of cannabinol compounds CBD, CBN, THC in sample,
Thus the content of three kinds of cannabinol compounds CBD, CBN, THC in Chinese fiber crops sample are calculated.
Further, the compound method of different quality concentration hybrid standard product solution is contained from simultaneously described in step 2
THC, CBN, CBD and mass concentration are to take 50 μ L, 50 μ L, 40 μ L, 100 μ successively in 100 μ g/mL hybrid standard product mother liquor
L, 200 μ L, 400 μ L, 600 μ L, 800 μ L are determined in 100mL, 10mL, 2mL, 1mL, 1mL, 1mL, 1mL, 1mL volumetric flask and with methanol
Hold.
Further, the compound method of the hybrid standard product mother liquor is that to take 0.5mL mass concentrations respectively be 1mg/mL
CBD, CBN, THC standard solution in 5mL volumetric flasks and with methanol constant volume.
Further, retention time described in step 3 is：CBD：8.3min, CBN：14.9min THC：18.6min.
Further, the equation of linear regression of three kinds of cannabinol compounds is described in step 3：
CBD：Y=74725X+38049, the X scope of application：0.05-80μg/mL；
CBN：Y=116969X+24626, the X scope of application：0.05-60μg/mL；
THC：Y=72297X+21790, X use range：0.05-60μg/mL；
Wherein：Y is the peak area corresponding to cannabinol compounds, and X is the mass concentration of cannabinol compounds.
Further, Chinese fiber crops sample described in step 4 is Chinese sesame slices, and the drying and crushing processing is that Chinese sesame slices are placed in into drum
Dried 6 hours for 40 DEG C in wind drying box, take the Chinese sesame slices after appropriate drying to be crushed to end in pulverizer.
Further, it is that essence takes Chinese sesame slices powder 0.1g to add in 10mL centrifuge tubes that ultrasound described in step 4, which assists extraction,
Enter 6mL methanol, ultrasound assists extraction 30min.
Further, centrifugal treating described in step 4 is 3000r/min centrifugations 5min；It is described to take supernatant with methanol constant volume
Volume be 10mL.
Further, solid phase pillar described in step 5 using it is preceding with 5mL hplc grade methanols cross post carry out activate post imitate.
Further, efflux described in step 6 mixes and the volume of constant volume is 25mL again.
The advantage of the invention is that：
1st, the present invention establishes one kind using Solid phase extraction-HPLC while detects three kinds of cannabinols in Chinese fiber crops
The method of class compound, qualitative and quantitative analysis can be carried out to cannabinol compounds in Chinese fiber crops, be cannabinol compounds
Composition and the accurate judgement of content and quick detection provide scientific basis；
2nd, the pre-treatment link that the present invention is detected using solid phase extraction techniques as efficient liquid phase, using solid phase pillar to leaf
The impurity components such as green element good adsorption effect, cannabinol compounds to be measured are separated with impurity component, reduce impurity into
Divide the influence to detection, good basis is provided for liquid phase detection.
3rd, C18 solid phases pillar of the present invention is low for the adsorption rate of three kinds of cannabinol compounds, is good decolouring material
Solid phase pillar；Solid phase extraction manipulations of the present invention are simple, consumption solvent is few, cost is low, good separating effect.
4th, detection reduces every kind of big again while the present invention realizes three kinds of cannabinol compounds compared with conventional method
The relative retention time of numb phenolic compound, the quantitative detection to three kinds of cannabinol compounds can be completed in 19 minutes, is carried
High detection efficiency.
5th, the chromatographic condition that this law is established is respectively provided with preferable separating degree, and analysis condition to three kinds of cannabinol compounds
Suitably：The equation of linear regression R of three kinds of cannabinol compounds of the invention20.999 is all higher than, shows that it is applicable model corresponding
It is good to enclose interior linear relationship；Detection method Precision Experiment, repeated experiment RSD show that it is accurate within 1%
Degree, repeatability are good；Detection method average recovery Assay recovery scope shows it between 99% -113%
The degree of accuracy is good.
6th, the pre-treatment of sample assists to extract using ultrasound in the present invention, and more conducively cannabinol compounds is abundant molten
Solution, ensure the accuracy of testing result.
Brief description of the drawings
Fig. 1 is the CBD standard curves that the step 3 of the embodiment of the present invention one measures；
Fig. 2 is the CBN standard curves that the step 3 of the embodiment of the present invention one measures；
Fig. 3 is the THC standard curves that the step 3 of the embodiment of the present invention one measures；
Fig. 4 is the hybrid standard that the cannabinol compounds content that the step 3 of the embodiment of the present invention one measures is 40 μ g/mL
The high-efficient liquid phase chromatogram of product solution；
Fig. 5 is the high performance liquid chromatography of three kinds of cannabinol compounds in the Chinese fiber crops that the step 6 of the embodiment of the present invention one measures
Figure；
Fig. 6 is the high-efficient liquid phase chromatogram of three kinds of cannabinol compounds in the Chinese fiber crops that experimental example 1 measures.
Embodiment
Technical scheme is described further with reference to embodiment, but is not limited thereto, it is every right
Technical solution of the present invention is modified or equivalent substitution, without departing from the spirit and scope of technical solution of the present invention, all should contain
Cover in protection scope of the present invention.
Embodiment one：A kind of method for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE-HPLC of the present invention
Step is as follows：
First, high-efficient liquid phase chromatogram condition is determined：The high-efficient liquid phase chromatogram condition is：
C18 reversed-phase columns：4.6 × 150mm, 5 μm；Column temperature：40℃；Detection wavelength：220nm；Type of elution：Isocratic elution；
Mobile phase：Methanol:0.1% formic acid=80:20；Flow velocity：1mL/min；Sample size：20μL.
2nd, hybrid standard product solution is prepared：
(1) hybrid standard product mother liquor is prepared：THC, CBN, CBD standard that 0.5mL mass concentrations are 1mg/mL are taken respectively
The hybrid standard product for containing identical mass concentration THC, CBN, CBD simultaneously are made in 5mL volumetric flasks and with methanol constant volume in product solution
Mother liquor；
(2) hybrid standard product solution is prepared：Taken successively from hybrid standard product mother liquor 50 μ L, 50 μ L, 40 μ L, 100 μ L,
200 μ L, 400 μ L, 600 μ L, 800 μ L are determined in 100mL, 10mL, 2mL, 1mL, 1mL, 1mL, 1mL, 1mL volumetric flask and with methanol
Hold, obtained mass concentration is followed successively by 0.05 μ g/mL, 0.5 μ g/mL, 2 μ g/mL, 10 μ g/mL, 20 μ g/mL, 40 μ g/mL, 60 μ g/
ML, 80 μ g/mL hybrid standard product solution.
3rd, equation of linear regression is determined：Each concentration hybrid standard product solution that step 2 is prepared is used into step 1 successively
High-efficient liquid phase chromatogram condition is detected, wherein the testing result of 40 μ g/mL hybrid standard product solution is as shown in figure 4, to retain
Time is qualitative, the retention time of three kinds of cannabinol compounds：CBD：8.3min, CBN：14.9min THC：18.6min；According to
The corresponding relation of size and its concentration that each concentration surveys peak area draws standard curve, and wherein CBD, CBN, THC standard is bent
Line is successively as shown in Figure 1, Figure 2, Figure 3 shows, and determine the equation of linear regression of three kinds of cannabinol compounds respectively：
CBD：Y=74725X+38049, n=8, R2=0.9992, the X scope of application：0.05-80μg/mL；
CBN：Y=116969X+24626, n=7, R2=0.9995, the X scope of application：0.05-60μg/mL；
THC：Y=72297X+21790, n=7, R2=0.9992, the X scope of application：0.05-60μg/mL；
Wherein：Y is the peak area corresponding to cannabinol compounds, and X is the mass concentration of cannabinol compounds, and n is
Draw different hybrid standard product concentration values numbers during standard curve, R2For linearly related property coefficient.
Detection reduces again while high performance liquid chromatography detection of the present invention realizes a variety of cannabinols compared with conventional method
The relative retention time of every kind of cannabinol, the quantitative detection to three kinds of cannabinols can be completed in 19 minutes, improves inspection
Survey efficiency.The equation of linear regression R of three kinds of cannabinols20.999 is all higher than, shows its linear relationship in the corresponding scope of application
Well.
4th, SPE pretreatment sample is prepared：Chinese sesame slices are placed in air dry oven into 40 DEG C to dry 6 hours, taken suitable
Chinese sesame slices after amount drying are crushed to end in pulverizer, and essence takes Chinese sesame slices powder 0.1g in 10ml centrifuge tubes, add 6ml first
Alcohol, ultrasound assist extraction 30min, and extract is centrifuged into 5min with 3000r/min；Supernatant is taken in 10mL volumetric flasks and with methanol
Constant volume, obtain SPE pretreatment sample.
5th, SPE：Using the SPE pattern for retaining chaff interference, solid phase pillar uses 5mL chromatographic grade first before using
Alcohol crosses post and carries out activating post effect, and the SPE pretreatment sample that step 4 is obtained adds C18/500mg solid phase pillars, flowing
It is mutually methanol, flow velocity 1mL/min, pressure 0.1MPa, isocratic elution, sample Green pigment composition is adsorbed to be retained, big portion
Divide aldehydes matter to be present in efflux, collect efflux；The solid phase pillar is eluted with a small amount of hplc grade methanol, received
Collect efflux；
6th, high performance liquid chromatography detection：The efflux that step 5 is collected mixes and is settled to 25mL, film again with methanol
Detected after filtering using step 1 high-efficient liquid phase chromatogram condition, testing result is as shown in figure 5, measure each component in sample
Peak area, it is qualitative with retention time, quantified according to the equation of linear regression that step 3 determines, calculate three kinds of cannabinoids in sample
The concentration of compound, the content for thus calculating three kinds of cannabinol compounds in Chinese fiber crops sample are respectively：CBD：1550μg/g；
CBN：869μg/g；THC：3875μg/g.
Experimental example 1：Solid phase pillar is tested to the adsorption rate of three kinds of cannabinol compounds in sample：
The SPE pretreatment sample that the step 4 of Example one obtains is test sample, is used after taking 20 μ L membrane filtrations
The high-efficient liquid phase chromatogram condition of the step 1 of embodiment one is detected, and testing result is as shown in fig. 6, measure each component in sample
Peak area, it is qualitative with retention time, quantified according to the equation of linear regression that step 3 determines, calculate three kinds of cannabinoids in sample
The concentration of compound, thus calculate the content difference of three kinds of cannabinol compounds in the Chinese fiber crops sample without SPE
For：CBD：1781μg/g；CBN：984μg/g；THC：4693μg/g.
For solid phase pillar to adsorption rate ω=(A-a) ÷ A × 100% of three kinds of cannabinol compounds, wherein A is experiment
The cannabinol compounds content that example 1 measures, a are the cannabinol compounds content that embodiment one measures.
Solid phase pillar is as shown in table 1 to the adsorption rate of three kinds of cannabinol compounds：
Table 1：
As shown in Table 1, solid phase pillar for the adsorption rate of three kinds of cannabinol compounds in 11-17%, show its for
The adsorption rate of these three cannabinol compounds is very low, can be as the solid phase pillar of good decolouring material.Therefore, it is of the invention
Using the SPE pattern for retaining chaff interference, the pre-treatment link detected using solid phase extraction techniques as efficient liquid phase, operation
Simply, consumption solvent is few, cost is low, and the good adsorption of the impurity components such as chlorophyll is acted on using solid phase pillar, effectively gone
Except the color (chlorophyll a and chlorophyll b) in Chinese sesame slices, cannabinol compounds to be measured are separated with impurity component, reduced
Influence of the impurity component to detection, good basis is provided for liquid phase detection.
Experimental example 2：The Precision Experiment of SPE-HPLC methods of the present invention
Essence takes 1 part of Chinese sesame slices sample, is pre-processed according to the method for the embodiment of the present invention one and gained is detected into sample
Three times (N-1, N-2, N-3), the content data for measuring three kinds of cannabinol compounds is as shown in table 2 for continuous sample introduction：
Table 2：
As shown in Table 2, Precision Experiment RSD values illustrate the present invention using SPE-HPLC while detect the Chinese within 1%
The method precision of three kinds of cannabinol compounds is good in fiber crops.
Experimental example 3：The repeated experiment of SPE-HPLC methods of the present invention
Essence takes 6 parts of different Chinese sesame slices samples (N1-N6), is pre-processed according to the method for the embodiment of the present invention one and by institute
Sample sample detection successively must be detected, the content data for measuring three kinds of cannabinol compounds is as shown in table 3：
Table 3：
As shown in Table 3, repeated experiment RSD values illustrate the present invention using SPE-HPLC while examined within 1.05%
The method repeatability for surveying three kinds of cannabinol compounds in Chinese fiber crops is good.
Experimental example 4：The average recovery experiment of SPE-HPLC methods of the present invention
Essence takes 3 parts of different Chinese sesame slices samples (N1, N2, N3), is located in advance according to the method for the embodiment of the present invention one respectively
Reason respectively obtains 25ml detection samples, detects sample using this and detects to obtain substrate value as substrate detection sample progress efficient liquid phase；Separately
Respectively take 5mL substrates detection sample and add the standard items simultaneously containing three kinds of cannabinol compounds thereto, make three kinds of hemps
The addition of phenolic compound is 20 μ g, and gained sample is detected as sample-adding actual measurement sample progress efficient liquid phase and surveyed
Value, the content data for measuring three kinds of cannabinol compounds are as shown in table 4：
Table 4：
As shown in Table 4, average recovery Assay recovery scope illustrates present invention utilization between 99% -113%
The method degree of accuracy that SPE-HPLC detects three kinds of cannabinol compounds in Chinese fiber crops simultaneously is good.
Claims (10)
- A kind of 1. method for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE-HPLC, it is characterised in that this method Step is as follows：First, high-efficient liquid phase chromatogram condition is determined：The high-efficient liquid phase chromatogram condition is：C18 reversed-phase columns：4.6 × 150mm, 5 μm；Column temperature：40℃；Detection wavelength：220nm；Type of elution：Isocratic elution；Flowing Phase：Methanol:0.1% formic acid=80:20；Flow velocity：1mL/min；Sample size：20μL；2nd, hybrid standard product solution is prepared：Prepare simultaneously containing identical mass concentration three kinds of cannabinol compounds CBD, CBN, THC and mass concentration are followed successively by 0.05 μ g/mL, 0.5 μ g/mL, 2 μ g/mL, 10 μ g/mL, 20 μ g/mL, 40 μ g/mL, 60 μ G/mL, 80 μ g/mL hybrid standard product solution；3rd, equation of linear regression is determined：Each concentration hybrid standard product solution that step 2 is prepared is efficient using step 1 successively Liquid phase chromatogram condition is detected, qualitative with retention time, and the size that peak area is surveyed according to each concentration is corresponding with its concentration Relation draws standard curve, and determines the equation of linear regression of three kinds of cannabinol compounds respectively；4th, SPE pretreatment sample is prepared：Pulverization process is dried in Chinese fiber crops sample, takes Chinese fiber crops sample powder in centrifugation Manage and add methanol, assist to extract the cannabinol compounds in Chinese fiber crops sample powder using ultrasound；Centrifugal treating extract is simultaneously Supernatant is taken to obtain SPE pretreatment sample with methanol constant volume；5th, SPE：Using the SPE pattern for retaining chaff interference, the SPE pretreatment sample that step 4 is obtained C18/500mg solid phase pillars are added, mobile phase is methanol, flow velocity 1mL/min, pressure 0.1MPa, isocratic elution, collects outflow Liquid；The solid phase pillar is eluted with a small amount of hplc grade methanol, collects efflux；6th, high performance liquid chromatography detection：The efflux that step 5 is collected mixes and with methanol again constant volume, is used after membrane filtration Step 1 high-efficient liquid phase chromatogram condition is detected, and measures the peak area of each component in sample, qualitative with retention time, according to step Rapid three equations of linear regression determined quantify, and calculate the concentration of three kinds of cannabinol compounds CBD, CBN, THC in sample, thus Calculate the content of three kinds of cannabinol compounds CBD, CBN, THC in Chinese fiber crops sample.
- A kind of 2. side for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE-HPLC according to claim 1 Method, it is characterised in that the compound method of different quality concentration hybrid standard product solution described in step 2 be from simultaneously contain CBD, CBN, THC and mass concentration are to take 50 μ L, 50 μ L, 40 μ L, 100 μ L, 200 successively in 100 μ g/mL hybrid standard product mother liquor μ L, 400 μ L, 600 μ L, 800 μ L are in 100mL, 10mL, 2mL, 1mL, 1mL, 1mL, 1mL, 1mL volumetric flask and with methanol constant volume.
- A kind of 3. side for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE-HPLC according to claim 2 Method, it is characterised in that the compound method of the hybrid standard product mother liquor is that to take 0.5mL mass concentrations respectively be 1mg/mL CBD, CBN, THC standard solution are in 5mL volumetric flasks and with methanol constant volume.
- 4. three kinds of cannabinol compounds in Chinese fiber crops are detected according to any described one kind of claim 1-3 simultaneously using SPE-HPLC Method, it is characterised in that retention time is described in step 3：CBD：8.3min, CBN：14.9min THC：18.6min.
- A kind of 5. side for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE-HPLC according to claim 4 Method, it is characterised in that the equation of linear regression of three kinds of cannabinol compounds is described in step 3：CBD：Y=74725X+38049, the X scope of application：0.05-80μg/mL；CBN：Y=116969X+24626, the X scope of application：0.05-60μg/mL；THC：Y=72297X+21790, X use range：0.05-60μg/mL；Wherein：Y is the peak area corresponding to cannabinol compounds, and X is the mass concentration of cannabinol compounds.
- A kind of 6. side for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE-HPLC according to claim 5 Method, it is characterised in that Chinese fiber crops sample described in step 4 is Chinese sesame slices, and the drying and crushing processing is that Chinese sesame slices are placed in into air blast to do Dried 6 hours for 40 DEG C in dry case, take the Chinese sesame slices after appropriate drying to be crushed to end in pulverizer.
- A kind of 7. side for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE-HPLC according to claim 6 Method, it is characterised in that it is that essence takes Chinese sesame slices powder 0.1g in 10ml centrifuge tubes that ultrasound described in step 4, which assists extraction, adds 6ml Methanol, ultrasound assist extraction 30min.
- A kind of 8. side for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE-HPLC according to claim 7 Method, it is characterised in that centrifugal treating described in step 4 is 3000r/min centrifugations 5min；The body that supernatant is taken with methanol constant volume Product is 10mL.
- A kind of 9. side for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE-HPLC according to claim 8 Method, it is characterised in that solid phase pillar described in step 5 using it is preceding with 5mL hplc grade methanols cross post carry out activate post imitate.
- A kind of 10. side for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE-HPLC according to claim 9 Method, it is characterised in that efflux described in step 6 mixes and the volume of constant volume is 25mL again.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710863944.5A CN107589203B (en) | 2017-09-21 | 2017-09-21 | Method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710863944.5A CN107589203B (en) | 2017-09-21 | 2017-09-21 | Method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107589203A true CN107589203A (en) | 2018-01-16 |
| CN107589203B CN107589203B (en) | 2020-06-02 |
Family
ID=61047671
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710863944.5A Expired - Fee Related CN107589203B (en) | 2017-09-21 | 2017-09-21 | Method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107589203B (en) |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109725080A (en) * | 2019-01-02 | 2019-05-07 | 中国农业科学院麻类研究所 | Cannabidiol, cannabidiolic acid, in tetrahydrocannabinol one or more of substances method for qualitative and quantitative detection |
| CN109917031A (en) * | 2019-02-28 | 2019-06-21 | 西藏育宁生物科技有限责任公司 | A method of cannabinoids content in measurement cannabidiol crude product |
| CN110596229A (en) * | 2019-09-23 | 2019-12-20 | 凯纳比斯药业有限公司 | Detection method for element impurities in hemp extract and hemp oil product |
| CN110632224A (en) * | 2019-09-16 | 2019-12-31 | 福建省中科生物股份有限公司 | Establishment method of hemp plant fingerprint spectrum |
| CN110646534A (en) * | 2019-09-24 | 2020-01-03 | 南京市产品质量监督检验院 | Method for detecting cannabidiol in textile |
| CN112710768A (en) * | 2020-12-10 | 2021-04-27 | 安徽瑞达健康产业有限公司 | Method for measuring content of dehydroxycannabidiol (DH-CBD) |
| CN112730696A (en) * | 2020-09-15 | 2021-04-30 | 中国标准化研究院 | Method for detecting 5 cannabinol compounds in cannabis sativa oil by using HPLC (high performance liquid chromatography) method |
| US11040932B2 (en) | 2018-10-10 | 2021-06-22 | Treehouse Biotech, Inc. | Synthesis of cannabigerol |
| US11084770B2 (en) | 2016-12-07 | 2021-08-10 | Treehouse Biotech, Inc. | Cannabis extracts |
| CN113533237A (en) * | 2021-09-15 | 2021-10-22 | 广州智汇生物科技有限公司 | Method for detecting illegal addition of cannabidiol in pet food |
| US11202771B2 (en) | 2018-01-31 | 2021-12-21 | Treehouse Biotech, Inc. | Hemp powder |
| CN115407000A (en) * | 2022-10-17 | 2022-11-29 | 中国计量大学 | Detection method of cannabidiol and tetrahydrocannabinol in industrial cannabis sativa |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101303335A (en) * | 2007-05-10 | 2008-11-12 | 上海医药工业研究院 | Method for identifying main cannabinol compounds in cannabis |
-
2017
- 2017-09-21 CN CN201710863944.5A patent/CN107589203B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101303335A (en) * | 2007-05-10 | 2008-11-12 | 上海医药工业研究院 | Method for identifying main cannabinol compounds in cannabis |
Non-Patent Citations (4)
| Title |
|---|
| C. RUSTICHELLI ET AL.: "Analysis of Cannabinoids in Fiber Hemp Plant Varieties (Cannabis Sativa L.) by High-Performance Liquid Chromatography", 《CHROMATOGRAPHIA》 * |
| 夏林波 等: "硅胶柱层析-RP-HPLC法同时测定火麻仁中3种大麻酚类化合物的含量", 《中国药房》 * |
| 王全林 等: "超高效液相色谱-串联质谱法测定火麻食品中特征大麻酚", 《理化检验-化学分册》 * |
| 肖培云 等: "工业大麻研究.Ⅲ.不同生长期工业大麻中四氢大麻酚和大麻二酚含量的测定方法比较", 《中国医药工业杂志》 * |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11084770B2 (en) | 2016-12-07 | 2021-08-10 | Treehouse Biotech, Inc. | Cannabis extracts |
| US11202771B2 (en) | 2018-01-31 | 2021-12-21 | Treehouse Biotech, Inc. | Hemp powder |
| US11040932B2 (en) | 2018-10-10 | 2021-06-22 | Treehouse Biotech, Inc. | Synthesis of cannabigerol |
| CN109725080A (en) * | 2019-01-02 | 2019-05-07 | 中国农业科学院麻类研究所 | Cannabidiol, cannabidiolic acid, in tetrahydrocannabinol one or more of substances method for qualitative and quantitative detection |
| CN109917031A (en) * | 2019-02-28 | 2019-06-21 | 西藏育宁生物科技有限责任公司 | A method of cannabinoids content in measurement cannabidiol crude product |
| CN110632224A (en) * | 2019-09-16 | 2019-12-31 | 福建省中科生物股份有限公司 | Establishment method of hemp plant fingerprint spectrum |
| CN110632224B (en) * | 2019-09-16 | 2022-02-15 | 福建省中科生物股份有限公司 | Establishment method of hemp plant fingerprint spectrum |
| CN110596229A (en) * | 2019-09-23 | 2019-12-20 | 凯纳比斯药业有限公司 | Detection method for element impurities in hemp extract and hemp oil product |
| CN110646534A (en) * | 2019-09-24 | 2020-01-03 | 南京市产品质量监督检验院 | Method for detecting cannabidiol in textile |
| CN112730696A (en) * | 2020-09-15 | 2021-04-30 | 中国标准化研究院 | Method for detecting 5 cannabinol compounds in cannabis sativa oil by using HPLC (high performance liquid chromatography) method |
| CN112710768A (en) * | 2020-12-10 | 2021-04-27 | 安徽瑞达健康产业有限公司 | Method for measuring content of dehydroxycannabidiol (DH-CBD) |
| CN113533237B (en) * | 2021-09-15 | 2021-11-16 | 广州智汇生物科技有限公司 | Method for detecting illegal addition of cannabidiol in pet food |
| CN113533237A (en) * | 2021-09-15 | 2021-10-22 | 广州智汇生物科技有限公司 | Method for detecting illegal addition of cannabidiol in pet food |
| CN115407000A (en) * | 2022-10-17 | 2022-11-29 | 中国计量大学 | Detection method of cannabidiol and tetrahydrocannabinol in industrial cannabis sativa |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107589203B (en) | 2020-06-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107589203A (en) | A kind of method for detecting three kinds of cannabinol compounds in Chinese fiber crops simultaneously using SPE HPLC | |
| CN105203657B (en) | Semen Ziziphi Spinosae reference extract and its preparation method and application | |
| CN101598707B (en) | Method for detecting various pesticide residue in oil-containing food | |
| CN107505405B (en) | Method for efficiently and rapidly extracting and measuring flavonoid pigment in Chinese rose petals | |
| CN102520079B (en) | Method for rapidly measuring content of solanesol in tobaccos by using UPLC (Ultra Performance Liquid Chromatography) | |
| CN108693257A (en) | The method for directly detecting tealeaves glucosides bound state aroma precursor substance | |
| CN112730696A (en) | Method for detecting 5 cannabinol compounds in cannabis sativa oil by using HPLC (high performance liquid chromatography) method | |
| CN109884207A (en) | A method of quick and precisely analyzing polyphenol content in rapeseed oil | |
| CN105445407B (en) | Detection method for fatty acid and vitamin E in idesia | |
| CN107271584A (en) | The capture method of carbonyls and tobacco-specific nitrosamine, extracting method and assay method in a kind of cigarette mainstream flue gas | |
| CN106501382A (en) | The extraction of mercury compound and detection method in a kind of flesh of fish | |
| CN107677745A (en) | The quality determining method of Desmodium styracifolium extractive of general flavone | |
| CN102980956A (en) | Quick extraction and detection method of 9-oxo-10,11-dehydroagerophorone | |
| CN107315058A (en) | A kind of method of total ginkgoic acid in detection ginkgo biloba succi | |
| CN103018384B (en) | A kind of Quality analysis and evaluation method of black tea and application thereof | |
| CN106442788B (en) | The method of discrimination of fresh tobacco leaves sample quality in a kind of tobacco metabolism group based on pigment | |
| CN104931594B (en) | The detection method of content of 5 hydroxymethyl furfural in a kind of Schisandra chinensis | |
| CN103822888B (en) | Catch up with the quality determining method of yellow grass | |
| CN109765322A (en) | The characteristic spectrum construction method and quality determining method of schizonepeta | |
| Weaver et al. | Determination of aflatoxins in botanical roots by a modification of AOAC official method SM 991.31: Single-laboratory validation | |
| CN109884199B (en) | Method for measuring content of flavonoid components in honey | |
| CN106370736A (en) | Method for simultaneous SMART column on-line purification and HPLC/UVE fluorescent detection of four aflatoxins in peanut product | |
| CN106324121A (en) | Two micro-nucleosides testing method for medicine samples of insect and herbage class | |
| CN104569253B (en) | The assay method of benzo [a] pyrene in a kind of cigarette mainstream flue gas | |
| CN110398560A (en) | A kind of UPLC method detecting atractylenolide Ⅰ and atractylodes lactone III in Rhizoma Atractylodis Macrocephalae drug |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20200602 Termination date: 20210921 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |