CN106855549A - The pre-treating method of THC detection in a kind of fiery sesame oil - Google Patents
The pre-treating method of THC detection in a kind of fiery sesame oil Download PDFInfo
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- CN106855549A CN106855549A CN201710167665.5A CN201710167665A CN106855549A CN 106855549 A CN106855549 A CN 106855549A CN 201710167665 A CN201710167665 A CN 201710167665A CN 106855549 A CN106855549 A CN 106855549A
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- G01N30/02—Column chromatography
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- 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
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N30/02—Column chromatography
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Abstract
The invention discloses a kind of pre-treating method of THC detection in fiery sesame oil, belong to analysis determining technology field.The method of the present invention is extracted using acetonitrile, and the method for freezing processing and dispersive solid-phase extraction agent is purified to extract solution, and Instrumental Analysis is used for after filtering.Pre-treating method matrix is total to the too high deficiency of extractive content during the present invention compensate for existing safety standard, it is easy to operate, favorable reproducibility, the content of the common extract of extract solution mesostroma is down to 1.90mg/mL, up to 81.5%, the relative standard deviation of THC is 1.8%~4.7% to the common extract clearance of matrix, and the rate of recovery of THC has been effectively ensured, the impurity effectively eliminated in sample extracting solution is purified simultaneously, reduces the pollution to detecting instrument.
Description
Technical field
The present invention relates to a kind of pre-treating method of THC detection in fiery sesame oil, belong to analysis determining technology neck
Domain.
Background technology
Fiery fiber crops also known as hemp, Chinese fiber crops, are a kind of annual herb plants, be distributed widely in China Guangxi, Yunnan, Gansu,
The ground such as Heilungkiang, great edible and medical value.Fiery fiber crops seed containing 20~25% albumen, 20~30% carbohydrate, 25~
35% lipid, 10~15% fibers and abundant mineral matter.Content of polyunsaturated fatty acid is up to nearly 80% in fiery sesame oil, its
Linoleic acid and linolenic proportional balancing method, reach 3:1 or so, meet WHO recommendation less than or equal to 4:1 is optimal
The characteristics of ratio, it is beneficial to health.Fiery sesame oil has functions that to delay artery sclerosis, prevention cardiovascular and cerebrovascular disease, is value
The long-lived oil plant that must recommend.Abundant gamma-Linolenic acid also imparts fiery sesame oil industrial value very high, such as prepares skin care item
Deng.
Although fiery sesame oil is with high nutritive value, the material of health, wherein δ 9- are also unfavorable for containing some
THC is a kind of toxic component, acts on cardiovascular system and central nervous system, causes people to produce lax glad
Illusion or stupor of being sunk into sleep, long-term intake can cause spiritual degeneration, or even disablement.In existing detection technique, ginseng
The detection method in social public security standard GA/T 206-99 is examined, fiery sesame oil sample is by direct injected after n-hexane dissolution
In chromatographic apparatus, the process without extraction and removal of impurities forms the position such as serious matrix interference and pollution chromatographic column, detector.
And existing some other detection methods generally use solid phase extraction column as the method for decontamination substrate, when pre-treatment is operated
Between (needed for usual dedoping step for up to 1~2h) long, and there is high cost, wash-out solvent for use amount is big (produces 50
The discard solution of~150mL) the shortcomings of.Therefore, the pretreatment process that optimization is detected to THC in fiery sesame oil is for obtaining
More accurately testing result is obtained to be necessary.
The content of the invention
In order to make up the deficiencies in the prior art, the present invention provides a kind of pre-treatment side of THC detection in fiery sesame oil
Method, methods described includes that extraction, adsorption-edulcoration, nitrogen blow, redissolve and filters；It is described absorption be with aluminum oxide as cleaner, according to
The amount of 20~30mg aluminum oxide/g fire sesame oil shakes 50~70s to aluminum oxide is added in fiery sesame oil.
In one embodiment of the invention, the extraction is with acetonitrile as extractant, by 1g：The ratio of 8~12mL
Example is extracted to acetonitrile is added in fiery sesame oil at 20~25 DEG C, then at -10~30 DEG C of 60~240min of freezing.
In one embodiment of the invention, the extraction conditions is：It is accurate to weigh 1.00g fire sesame oil, add 8~
12mL acetonitrile solutions, 50~70s of vortex oscillation.
In one embodiment of the invention, the absorption is by 2.5~3.5mL by the liquid after extraction：20~
The ratio of 30mg adds aluminum oxide, and 2~4min is centrifuged after 50~70s of concussion.
In one embodiment of the invention, it is to use N that the nitrogen blows2Liquid is dried up.
In one embodiment of the invention, the redissolution is after adding tribenzylamine-hexane mixture to blow nitrogen
Material is redissolved.
In one embodiment of the invention, the tribenzylamine-hexane mixture concentration be 30 μ g tribenzylamines/mL just
Hexane.
In one embodiment of the invention, the filtering is filtered with 0.22 μm of organic filter membrane.
In one embodiment of the invention, the pre-treatment is comprised the following steps：
(1) 1.0g fire sesame oil is weighed in 50mL centrifuge tubes, adds 8~12mL acetonitrile solutions to be extracted, vortex oscillation
Mix, stand；
(2) the centrifuge tube level equipped with fiery sesame oil and acetonitrile solution in step (1) is stood 110 in -30~-10 DEG C~
130min；
(3) upper solution after stratification in step (2) is moved into the centrifuge tube equipped with 20~30mg neutral aluminas
In, 50~70s, centrifugation are shaken at 20~25 DEG C；
(4) supernatant after centrifugation in step (3) is transferred in centrifuge tube, in N2Lower drying is extremely residual without acetonitrile solution
Stay, add 0.5mL inner mark solutions (30 μ g tribenzylamines/mL n-hexanes) dissolvings；
(5) sample liquid obtained in step (4) is transferred in sample injection bottle through 0.22 μm of organic filter membrane.
Second object of the present invention is to provide a kind of detection method of THC in fiery sesame oil, and methods described includes
Following steps：(1) pre-treatment is carried out to fiery sesame oil using the pre-treating method；(2) dried up by every 2.5~3.5mL extracts
The ratio of 0.5mL is added to add inner mark solution afterwards；(3), gas chromatographic detection.
In one embodiment of the invention, the testing conditions are as follows：Using HP-5 (30m × 0.32mm × 0.25 μ
M) chromatographic column：Heating schedule：80 DEG C of holding 1min, rise to 230 DEG C, then rise to the speed of 5 DEG C/min with 30 DEG C/min speed
280 DEG C, keep 10min；Injector temperature：250 DEG C, detector temperature：300℃；Carrier gas is nitrogen, flow velocity 1.2mL/min；Enter
Sample amount：1μL；Split ratio：10：1.
The present invention also provides the pre-treating method answering in terms of the Product checking containing THC and its derivative
With.
Beneficial effect：Present invention pre-treating method of THC detection in fiery sesame oil simplifies pre-treatment step,
And the interference component such as triglycerides and pigment in fiery sesame oil can be effectively removed, the content of the common extract of extract solution mesostroma is down to
1.90mg/mL, up to 81.5%, the relative standard deviation of THC is 1.8%~4.7% to the common extract clearance of matrix,
There are simple to operate, saving solvent, energy-conserving and environment-protective simultaneously, the discard solution of generation is short less than 10mL, pre-treatment used time,
Sample after pre-treatment meets the requirement of detection.
Specific embodiment
2014 gas chromatographs, Japanese Shimadzu Corporation；Neutral alumina, 100~200 mesh, the rich star biology section of the big fine jade in Wuhan
Skill Co., Ltd.
Reagent：Tribenzylamine, lark prestige Science and Technology Ltd.；Acetonitrile, analyzes pure, Chemical Reagent Co., Ltd., Sinopharm Group；
Standard substance：The methyl alcohol standard liquid of THC, is 1mg/mL, Sigma Co., USA.
Embodiment 1
(1) pre-treatment：Accurate 1.0g (being accurate to 0.01g) the fire sesame oil that weighs adds 10mL acetonitriles in 50mL centrifuge tubes,
Vortex oscillation mixes 60s, stands.By centrifuge tube after horizontal positioned 120min in -20 DEG C of refrigerators, 3mL upper solutions are moved to
In 15mL centrifuge tubes containing 30mg neutral aluminas, 60s is shaken, 3.0min is centrifuged under conditions of 4000rpm, taken on 2mL
Clear liquid is in N2Lower drying, is dissolved with 0.5mL internal standards (30 μ g tribenzylamines/mL n-hexanes) solution.Resulting solution is analytical solution,
After through 0.22 μm of membrane filtration of polytetrafluoroethylene (PTFE), sample liquid is to be determined, and extract quality is altogether to measure sample liquid mesostroma
1.90mg/mL, repeated RSD are 1.25%.
(2) detection of sample：The standard liquid of different gradients is injected separately into gas-chromatography, is quantified with internal standard method
Analysis, i.e., carry out regression analysis with the THC concentration that compares of THC and the chromatographic peak area of internal standard substance, obtains
To standard curve.Sample extracting solution injection gas-chromatography is measured at identical conditions, tetrahydrochysene is big in measuring sample liquid
The ratio between chromatographic peak area of numb phenol and internal standard compound, brings standard curve into, obtains THC content in sample liquid, then basis
The Mass Calculation of sample representated by sample liquid obtains the content of THC in sample.
Wherein chromatographic condition is：Chromatographic column：HP-5(30m×0.32mm×0.25μm)；Heating schedule：80 DEG C of holdings
1min, 230 DEG C are risen to 30 DEG C/min speed, then rise to 280 DEG C with the speed of 5 DEG C/min, keep 10min；Injector temperature：
250 DEG C, detector temperature：300℃；Carrier gas is nitrogen, flow velocity 1.2mL/min；Sample size：1μL；Split ratio：10：1.
Regression analysis is carried out with the THC concentration that compares of THC and the chromatographic peak area of internal standard substance, is obtained
To standard curve：Y=(0.0225 ± 7.51e-5.0) x+ (0.0089 ± 0.0094), coefficient correlation is 0.9975.
Fiery sesame oil sample is chosen, the THC standard liquid of 30,60,90 μ g/g concentration levels is added respectively, and pressed
The determination of residual amount is carried out according to above-mentioned process step, concentration will be determined and be compared with the theoretical addition concentration of THC, obtained
THC TIANZHU XINGNAO Capsul, each pitch-based sphere parallel determination 6 times, obtains its relative standard deviation.THC is reclaimed
Rate 92.3%~94.9%, relative standard deviation is 1.8%~4.7%.
Target concentration is right with 10 times of signal to noise ratios (S/N) as detection limit using in the corresponding sample of 3 times of signal to noise ratios (S/N)
Used as quantitative limit, the detection of THC is limited to 0.72 μ g/mL to target concentration in the sample answered.
Embodiment 2
Accurate 1.0g (being accurate to 0.01g) the fire sesame oil that weighs adds 10mL acetonitriles in 50mL centrifuge tubes, and vortex oscillation is mixed
Even 60s, stands.By centrifuge tube after horizontal positioned 120min in -30 DEG C of refrigerators, 3mL upper solutions are moved to containing in 20mg
Property aluminum oxide 15mL centrifuge tubes in, shake 60s, 3.0min is centrifuged under conditions of 4000rpm, take 2mL supernatants in N2Under
Drying, is dissolved with 0.5mL internal standards (30 μ g tribenzylamines/mL n-hexanes) solution.Resulting solution is analytical solution, poly- through 0.22 μm
After the membrane filtration of tetrafluoroethene, sample liquid is to be determined, and extract quality is 2.03mg/mL, weight altogether to measure sample liquid mesostroma
Renaturation RSD is 1.92%.
Embodiment 3
Accurate 1.0g (being accurate to 0.01g) the fire sesame oil that weighs adds 10mL acetonitriles in 50mL centrifuge tubes, and vortex oscillation is mixed
Even 60s, stands.By centrifuge tube after horizontal positioned 120min in -20 DEG C of refrigerators, 3mL upper solutions are moved to containing in 25mg
Property aluminum oxide 15mL centrifuge tubes in, shake 50s, 3.0min is centrifuged under conditions of 4000rpm, take 2mL supernatants in N2Under
Drying, is dissolved with 0.5mL internal standards (30 μ g tribenzylamines/mL n-hexanes) solution.Resulting solution is analytical solution, poly- through 0.22 μm
After the membrane filtration of tetrafluoroethene, sample liquid is to be determined, and extract quality is 1.99mg/mL, weight altogether to measure sample liquid mesostroma
Renaturation RSD is 1.48%.
Embodiment 4
Accurate 1.0g (being accurate to 0.01g) the fire sesame oil that weighs adds 10mL acetonitriles in 50mL centrifuge tubes, and vortex oscillation is mixed
Even 60s, stands.By centrifuge tube after horizontal positioned 120min in -10 DEG C of refrigerators, 3mL upper solutions are moved to containing in 30mg
Property aluminum oxide 15mL centrifuge tubes in, shake 60s, 3.0min is centrifuged under conditions of 4000rpm, take 2mL supernatants in N2Under
Drying, is dissolved with 0.5mL internal standards (30 μ g tribenzylamines/mL n-hexanes) solution.Resulting solution is analytical solution, poly- through 0.22 μm
After the membrane filtration of tetrafluoroethene, sample liquid is to be determined, and extract quality is 2.06mg/mL, weight altogether to measure sample liquid mesostroma
Renaturation RSD is 2.15%.
Reference examples 1
Accurate 1.0g (being accurate to 0.01g) the fire sesame oil that weighs adds 10mL acetonitriles in 50mL centrifuge tubes, and vortex oscillation is mixed
Even 60s, stands.2mL supernatants are taken in N2Lower drying, is dissolved with 0.5mL internal standards (30 μ g tribenzylamines/mL n-hexanes) solution.Institute
Solution be analytical solution, through 0.22 μm of membrane filtration of polytetrafluoroethylene (PTFE) after, sample liquid is to be determined, measures base in sample liquid
Extract quality is 10.49mg/mL to matter altogether, and repeated RSD is 5.61%.
Reference examples 2
Accurate 1.0g (being accurate to 0.01g) the fire sesame oil that weighs adds 10mL acetonitriles in 50mL centrifuge tubes, and vortex oscillation is mixed
Even 60s, stands.By centrifuge tube after horizontal positioned 120min in -20 DEG C of refrigerators, 2mL supernatants are taken in N2Lower drying, uses
0.5mL internal standards (30 μ g tribenzylamines/mL n-hexanes) solution dissolves.Resulting solution is analytical solution, through 0.22 μm of polytetrafluoroethyl-ne
After the membrane filtration of alkene, sample liquid is to be determined, and extract quality is 5.17mg/mL, repeated RSD altogether to measure sample liquid mesostroma
It is 4.82%.
Reference examples 3
Pre-treatment：Accurate 1.0g (being accurate to 0.01g) the fire sesame oil that weighs adds 10mL acetonitriles, whirlpool in 50mL centrifuge tubes
Rotation vibration mixes 60s, stands.During 3mL upper solutions are moved into the 15mL centrifuge tubes containing 25mg neutral aluminas, 60s is shaken,
3.0min is centrifuged under conditions of 4000rpm, 2mL supernatants is taken in N2Lower drying, with 0.5mL internal standards, (30 μ g tribenzylamines/mL is just
Hexane) solution dissolving.Resulting solution is analytical solution, through 0.22 μm of membrane filtration of polytetrafluoroethylene (PTFE) after, sample liquid is to be measured
Fixed, extract quality is 5.98mg/mL altogether to measure sample liquid mesostroma, and repeated RSD is 4.29%.
Reference examples 4
Sample pre-treatments are carried out according to method same as Example 1, difference is, using florisil silica as adsorbent
Instead of aluminum oxide.Extract quality is 3.00mg/mL altogether to measure sample liquid mesostroma, and repeated RSD is 2.58%.
Reference examples 5
Sample pre-treatments are carried out according to method same as Example 1, difference is, using multi-walled carbon nano-tubes as absorption
Agent replaces aluminum oxide.Extract quality is 4.35mg/mL altogether to measure sample liquid mesostroma, and repeated RSD is 4.16%.
Although the present invention is disclosed as above with preferred embodiment, it is not limited to the present invention, any to be familiar with this skill
The people of art, without departing from the spirit and scope of the present invention, can do various changes with modification, therefore protection model of the invention
Enclose being defined of being defined by claims.
Claims (10)
1. the pre-treating method that THC is detected in a kind of fiery sesame oil, it is characterised in that in turn include the following steps：Extraction
Take, freezing processing, adsorption-edulcoration, nitrogen blow, redissolve and filters；The extraction is extracted with organic solvent；The freezing processing is
60~240min is freezed at -10~30 DEG C；The adsorption-edulcoration is with aluminum oxide as cleaner, according to 20~30mg aluminum oxide/g
The amount of fiery sesame oil carries out removal of impurities absorption to addition aluminum oxide in fiery sesame oil.
2. method according to claim 1, it is characterised in that the extraction is with acetonitrile as extractant, by 1g fire fiber crops
Oil：The ratio of 8~12mL acetonitriles adds acetonitrile, 50~70s of vortex oscillation under the conditions of 20~25 DEG C.
3. method according to claim 1 and 2, it is characterised in that to adding aluminum oxide, concussion 50 in the liquid after extraction
~70s.
4. method according to claim 3, it is characterised in that use N2By the liquid drying after adsorption-edulcoration.
5. the method according to claim 1 or 4, it is characterised in that after adding tribenzylamine-hexane mixture to blow nitrogen
Material redissolves.
6. method according to claim 5, it is characterised in that the tribenzylamine-hexane mixture concentration is 20~30 μ
G tribenzylamines/mL n-hexanes.
7. method according to claim 1, it is characterised in that filtered with 0.22 μm of organic filter membrane.
8. method according to claim 1, it is characterised in that the pre-treatment is successively as follows：
(1) 1.0g fire sesame oil is weighed, adds 8~12mL acetonitrile solutions to be extracted, vortex oscillation is mixed, and is stood；
(2) solution for finishing extraction in step (1) stands 110~130min in -30~-10 DEG C；
(3) to 20~30mg neutral aluminas are added in the upper solution after step (2) stratification, 50 are shaken at 20~25 DEG C
~70s, centrifugation；
(4) supernatant in collection step (3) after centrifugation, uses N2Drying is 30 μ to 0.5mL concentration without acetonitrile solution residual, is added
The solution dissolving of g tribenzylamines/mL n-hexanes；
(5) solution after step (4) is redissolved is through 0.22 μm of organic membrane filtration.
9. in a kind of fiery sesame oil THC detection method, methods described comprises the following steps：(1) claim 1 is applied
The pre-treating method carries out pre-treatment to fiery sesame oil；(2) gas chromatographic detection.
10. application of the method described in claim 1 in product of the detection containing THC and its derivative.
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Cited By (3)
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| CN109796311A (en) * | 2019-03-25 | 2019-05-24 | 黑龙江阳光工业大麻研究院 | A method of purifying cannabidiol from abandoned reagents |
| 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 |
| CN112285233A (en) * | 2020-10-22 | 2021-01-29 | 三明海关综合技术服务中心 | Pretreatment method for detecting cannabinoids new psychoactive substances in grease |
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Cited By (4)
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| 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 |
| CN109796311A (en) * | 2019-03-25 | 2019-05-24 | 黑龙江阳光工业大麻研究院 | A method of purifying cannabidiol from abandoned reagents |
| CN112285233A (en) * | 2020-10-22 | 2021-01-29 | 三明海关综合技术服务中心 | Pretreatment method for detecting cannabinoids new psychoactive substances in grease |
| CN112285233B (en) * | 2020-10-22 | 2022-05-13 | 三明海关综合技术服务中心 | Pretreatment method for detecting cannabinoids new psychoactive substances in grease |
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| CN106855549B (en) | 2019-11-26 |
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