CN107589203B - Method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC - Google Patents
Method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC Download PDFInfo
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Abstract
The invention belongs to the field of plant component detection methods, and particularly relates to a method for simultaneously detecting three cannabinoids in hemp by SPE-HPLC. The invention applies the solid phase extraction-high performance liquid chromatography to the detection of the cannabinol compounds in the hemp, can simultaneously detect three cannabinol compounds in the hemp and carry out qualitative and quantitative analysis on the three cannabinol compounds. Firstly, determining high performance liquid chromatography conditions, drawing standard curves of three cannabinol compounds by detecting a mixed standard solution, and determining a linear regression equation; and performing solid-phase extraction and impurity removal on the hemp sample, and finally performing high performance liquid detection on the sample and calculating to obtain the contents of the three hemp phenolic compounds in the hemp. The solid-phase extraction method disclosed by the invention separates the cannabinol compounds to be detected from impurities, reduces the influence of the impurities on the detection, and provides a good foundation for liquid-phase detection. The detection method has good linear relation, precision, repeatability and accuracy in corresponding application range.
Description
Technical Field
The invention belongs to the field of plant component detection methods, and particularly relates to a method for simultaneously detecting three cannabinoids in hemp by SPE-HPLC.
Background
China hemp, one of the oldest crops on earth, is utilized by human as early as ten thousand years ago. Hemp bast fiber is one of the best natural fibers, is longer than cotton fiber, has stronger toughness and moisture absorption and better heat insulation effect, and is a superior raw material for textile and papermaking. With the development of modern science and technology, the active ingredients in hemp gradually attract attention, and among them, the cannabinoids are paid more attention because of more quantity and bioactive components. The cannabinol compounds mainly comprise Tetrahydrocannabinol (THC), Cannabidiol (CBD), Cannabinol (CBN) and cannabigerol (CBC).
Because of the wide variety of cannabinoids and the similar polarity of the cannabinoids, the prior art is difficult to effectively separate different cannabinoids, and the impurities mixed in the detection sample have great influence on the accuracy of the detection result because the impurities cannot be completely separated.
In the prior art, most of detection on cannabis phenolic compounds is limited to detection on THC or CBD chemical components in hemp seed oil, and most of detection methods are gas chromatography, wherein the THC detection documents are relatively more and the methods are more comprehensive. However, most of these detection methods are directed to single cannabinoids, and when a plurality of cannabinoids in hemp are required to be detected, respective pretreatment techniques and analysis methods are required, so that the detection is time-consuming and labor-consuming, and the cost is high.
Disclosure of Invention
The invention overcomes the defects of the prior art, applies solid phase extraction-high performance liquid chromatography (SPE-HPLC) to the detection of cannabinol compounds in the hemp, and provides a method for simultaneously detecting three cannabinol compounds in the hemp by using SPE-HPLC.
The invention discloses a method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC, which comprises the following steps:
firstly, determining the conditions of high performance liquid chromatography: the high performance liquid chromatography conditions are as follows:
c18 reverse phase column: 4.6X 150mm, 5 μm; column temperature: 40 ℃; detection wavelength: 220 nm; and (3) an elution mode: isocratic elution; mobile phase: methanol 0.1% formic acid 80: 20; flow rate: 1 mL/min; sample introduction amount: 20 mu L of the solution;
secondly, preparing a mixed standard solution: preparing a mixed standard solution which simultaneously contains three cannabinol compounds CBD, CBN and THC with the same mass concentration and the mass concentrations of which are 0.05 mu g/mL, 0.5 mu g/mL, 2 mu g/mL, 10 mu g/mL, 20 mu g/mL, 40 mu g/mL, 60 mu g/mL and 80 mu g/mL in sequence;
thirdly, determining a linear regression equation: sequentially detecting the mixed standard substance solution with each concentration prepared in the step two under the high performance liquid chromatography condition of the step one, determining the nature by retention time, drawing a standard curve according to the corresponding relation between the peak area size measured by each concentration and the concentration of the peak area, and respectively determining the linear regression equation of the three cannabinoids;
fourthly, preparing a solid phase extraction pretreatment sample: drying and crushing a hemp sample, putting hemp sample powder into a centrifugal tube, adding methanol, and extracting hemp phenolic compounds in the hemp sample powder by ultrasonic assistance; centrifuging the extract liquor, taking supernatant, and fixing the volume with methanol to obtain a solid-phase extraction pretreatment sample;
fifthly, solid phase extraction: adding the solid-phase extraction pretreatment sample obtained in the step four into a C18/500mg solid-phase small column by adopting a solid-phase extraction mode for retaining interferents, wherein a mobile phase is methanol, the flow rate is 1mL/min, the pressure is 0.1MPa, isocratic elution is carried out, and an effluent liquid is collected; leaching the solid-phase small column by using a small amount of chromatographic grade methanol, and collecting effluent liquid;
sixthly, detecting by high performance liquid chromatography: mixing the effluent liquid collected in the fifth step, re-fixing the volume by using methanol, detecting by adopting a high performance liquid chromatography condition in the first step after membrane filtration, measuring peak areas of all components in the sample, determining the peak areas qualitatively by using retention time, quantifying according to a linear regression equation determined in the third step, and calculating the concentrations of the three cannabinol compounds CBD, CBN and THC in the sample, thereby calculating the contents of the three cannabinol compounds CBD, CBN and THC in the hemp sample.
Furthermore, the preparation method of the mixed standard solution with different mass concentrations in the second step is to sequentially take 50 μ L, 40 μ L, 100 μ L, 200 μ L, 400 μ L, 600 μ L and 800 μ L from the mixed standard mother liquor which simultaneously contains THC, CBN and CBD and has the mass concentration of 100 μ g/mL in 100mL, 10mL, 2mL, 1mL and 1mL volumetric flasks and fix the volume with methanol.
Further, the preparation method of the mixed standard mother liquor is that 0.5mL of CBD, CBN and THC standard solutions with mass concentration of 1mg/mL are respectively put into a 5mL volumetric flask and the volume is determined by methanol.
Further, the retention time in the third step is as follows: CBD: 8.3min, CBN: 14.9min, THC: and (3) 18.6 min.
Further, the linear regression equation of the three cannabinol compounds in the third step is as follows:
CBD: Y74725X +38049, X applies: 0.05-80 mug/mL;
and (3) CBN: 116969X +24626, X applies: 0.05-60 mu g/mL;
THC: Y72297X +21790, range of use of X: 0.05-60 mu g/mL;
wherein: y is the peak area corresponding to the cannabinol compounds, and X is the mass concentration of the cannabinol compounds.
Further, the hemp sample in the fourth step is hemp leaves, and the drying and crushing treatment is to place the hemp leaves in a forced air drying oven for drying for 6 hours at 40 ℃, and to take a proper amount of dried hemp leaves to crush the hemp leaves into powder in a crusher.
Further, in the step four, 0.1g of hemp leaf powder is finely extracted in a 10mL centrifuge tube, 6mL of methanol is added, and ultrasonic assisted extraction is carried out for 30 min.
Further, the centrifugation treatment in the fourth step is 3000r/min for 5 min; the volume of the supernatant liquid which is determined by methanol is 10 mL.
Further, the solid phase column in the fifth step is activated by passing 5mL of chromatographic grade methanol through the column before use.
Further, the effluent of step six was mixed and re-metered to a volume of 25 mL.
The invention has the advantages that:
1. the invention establishes a method for simultaneously detecting three cannabinoids in hemp by using solid phase extraction-high performance liquid chromatography, can carry out qualitative and quantitative analysis on the cannabinoids in the hemp, and provides scientific basis for accurate judgment and rapid detection of the components and the content of the cannabinoids;
2. the method adopts the solid-phase extraction technology as a pretreatment link of high-efficiency liquid-phase detection, and separates the cannabinol compounds to be detected from impurity components by utilizing the good adsorption effect of the solid-phase small column on the impurity components such as chlorophyll, thereby reducing the influence of the impurity components on the detection and providing a good basis for the liquid-phase detection.
3. The C18 solid-phase small column has low adsorption rate for three cannabinol compounds, and is a good solid-phase small column of decoloration material; the solid phase extraction method has the advantages of simple operation, less solvent consumption, low cost and good separation effect.
4. Compared with the traditional method, the method disclosed by the invention realizes the simultaneous detection of the three cannabinol compounds, reduces the relative retention time of each cannabinol compound, can complete the quantitative detection of the three cannabinol compounds within 19 minutes, and improves the detection efficiency.
5. The chromatographic conditions established by the method have better separation degree on three cannabinoids, and the analysis conditions are suitable: linear regression equation R of three cannabinol compounds2All are larger than 0.999, which shows that the linear relation of the compounds in the corresponding application range is good; the detection method disclosed by the invention has the advantages that the precision experiment and the repeatability experiment RSD are within 1%, and the precision and the repeatability are good; the experimental recovery rate range of the sample-adding recovery rate of the detection method is 99% -113%, which shows that the accuracy is good.
6. The pretreatment of the sample in the invention adopts ultrasonic assisted extraction, which is more beneficial to the full dissolution of the cannabinoids and ensures the accuracy of the detection result.
Drawings
FIG. 1 is a CBD standard curve measured in step three of the example of the present invention;
FIG. 2 is a CBN standard curve measured in step three of the present invention;
FIG. 3 is a THC standard curve measured in step three of the example of the invention;
FIG. 4 is a high performance liquid chromatogram of a mixed standard solution with a cannabinoids content of 40 μ g/mL, as determined in step three of the example;
FIG. 5 is a high performance liquid chromatogram of three cannabinol compounds in hemp, measured in step six of the example of the invention;
FIG. 6 is a high performance liquid chromatogram of three cannabinol compounds in hemp obtained in Experimental example 1.
Detailed Description
The technical solutions of the present invention are further described below with reference to the following examples, but the present invention is not limited thereto, and any modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
The first embodiment is as follows: the invention discloses a method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC, which comprises the following steps:
firstly, determining the conditions of high performance liquid chromatography: the high performance liquid chromatography conditions are as follows:
c18 reverse phase column: 4.6X 150mm, 5 μm; column temperature: 40 ℃; detection wavelength: 220 nm; and (3) an elution mode: isocratic elution; mobile phase: methanol 0.1% formic acid 80: 20; flow rate: 1 mL/min; sample introduction amount: 20 μ L.
Secondly, preparing a mixed standard solution:
(1) preparing a mixed standard substance mother solution: respectively taking 0.5mL of THC, CBN and CBD standard substance solution with the mass concentration of 1mg/mL into a 5mL volumetric flask, and fixing the volume with methanol to prepare mixed standard substance mother liquor simultaneously containing THC, CBN and CBD with the same mass concentration;
(2) preparing a mixed standard solution: a mixed standard solution having a mass concentration of 0.05. mu.g/mL, 0.5. mu.g/mL, 2. mu.g/mL, 10. mu.g/mL, 20. mu.g/mL, 40. mu.g/mL, 60. mu.g/mL, or 80. mu.g/mL was prepared from the mixed standard mother solution in a volumetric flask containing 50. mu.L, 40. mu.L, 100. mu.L, 200. mu.L, 400. mu.L, 600. mu.L, or 800. mu.L in 100mL, 10mL, 2mL, 1mL, or 1mL in this order, and the volume was determined with methanol.
Thirdly, determining a linear regression equation: and (3) detecting the mixed standard substance solution with each concentration prepared in the step (II) by adopting a high performance liquid chromatography condition in the step (I), wherein the detection result of the mixed standard substance solution with the concentration of 40 mu g/mL is shown in a figure 4, and the detection result is characterized by retention time, and the retention time of three cannabinol compounds: CBD: 8.3min, CBN: 14.9min, THC: 18.6 min; drawing a standard curve according to the corresponding relation between the peak area size and the concentration of the peak area, wherein the standard curves of CBD, CBN and THC are sequentially shown in figures 1, 2 and 3, and respectively determining the linear regression equation of three cannabinol compounds:
CBD：Y＝74725X+38049，n＝8，R20.9992, X applies: 0.05-80 mug/mL;
CBN：Y＝116969X+24626，n＝7，R20.9995, X applies: 0.05-60 mu g/mL;
THC：Y＝72297X+21790，n＝7，R20.9992, X applies: 0.05-60 mu g/mL;
wherein: y is the peak area corresponding to cannabinol compound, X is the mass concentration of cannabinol compound, n is the number of different mixed standard substance concentration values when drawing standard curve, R is2Is a linear correlation coefficient.
Compared with the traditional method, the high performance liquid chromatography detection of the invention not only realizes the simultaneous detection of a plurality of cannabinols, but also reduces the relative retention time of each cannabinol, can complete the quantitative detection of three cannabinols within 19 minutes, and improves the detection efficiency. Linear regression equation R of three cannabinols2All are larger than 0.999, which indicates that the linearity relationship is good in the corresponding application range.
Fourthly, preparing a solid phase extraction pretreatment sample: drying hemp leaves in an air-blast drying oven at 40 ℃ for 6 hours, taking a proper amount of dried hemp leaves, crushing the hemp leaves in a crusher to powder, finely taking 0.1g of hemp leaf powder, putting the powder in a 10ml centrifugal tube, adding 6ml of methanol, performing ultrasonic assisted extraction for 30min, and centrifuging the extract for 5min at 3000 r/min; and taking the supernatant in a 10mL volumetric flask and fixing the volume by methanol to obtain a solid-phase extraction pretreatment sample.
Fifthly, solid phase extraction: adopting a solid phase extraction mode for retaining interferents, enabling a solid phase column to pass through a 5mL chromatographic grade methanol column for activating column effect before using, adding the solid phase extraction pretreatment sample obtained in the step four into a C18/500mg solid phase column, enabling a mobile phase to be methanol, enabling the flow rate to be 1mL/min, enabling the pressure to be 0.1MPa, carrying out isocratic elution, adsorbing and retaining green pigment components in the sample, enabling most phenolic substances to exist in an effluent liquid, and collecting the effluent liquid; leaching the solid-phase small column by using a small amount of chromatographic grade methanol, and collecting effluent liquid;
sixthly, detecting by high performance liquid chromatography: mixing the effluent liquid collected in the fifth step, re-metering the volume to 25mL by using methanol, detecting by adopting a high performance liquid chromatography condition in the first step after membrane filtration, measuring peak areas of all components in the sample according to a detection result shown in figure 5, determining the qualitative retention time, quantifying according to a linear regression equation determined in the third step, calculating the concentrations of three cannabinol compounds in the sample, and calculating the contents of the three cannabinol compounds in the hemp sample as follows: CBD: 1550 mug/g; and (3) CBN: 869 mu g/g; THC: 3875. mu.g/g.
Experimental example 1: experiment of adsorption rate of solid phase column to three cannabinol compounds in sample:
taking the solid-phase extraction pretreatment sample obtained in the fourth step of the embodiment as a test sample, filtering by using a 20-microliter membrane, detecting by using the high performance liquid chromatography condition of the first step of the embodiment, wherein the detection result is shown in fig. 6, measuring peak areas of components in the sample, determining the qualitative retention time, quantifying according to a linear regression equation determined in the third step, and calculating the concentrations of three cannabinol compounds in the sample, so that the contents of the three cannabinol compounds in the hemp sample without solid-phase extraction are respectively calculated as follows: CBD: 1781 μ g/g; and (3) CBN: 984 mu g/g; THC: 4693 ug/g.
The adsorption rate of the solid phase column on three cannabinol compounds is omega ═ a ÷ a × 100%, where a is the cannabinol compound content measured in experimental example 1 and a is the cannabinol compound content measured in example one.
The adsorption rates of the solid phase column for three cannabinol compounds are shown in table 1:
table 1:
as can be seen from Table 1, the adsorption rates of the solid phase column for the three cannabinol compounds are 11-17%, which shows that the solid phase column has very low adsorption rates for the three cannabinol compounds and can be used as a solid phase column with good decolorization material. Therefore, the method adopts a solid phase extraction mode for retaining interferents, takes a solid phase extraction technology as a pretreatment link of high performance liquid detection, has simple operation, less solvent consumption and low cost, effectively removes the colors (chlorophyll a and chlorophyll b) in the hemp leaves by utilizing the good adsorption effect of the solid phase column on the impurity components such as chlorophyll and the like, separates the hemp phenolic compounds to be detected from the impurity components, reduces the influence of the impurity components on the detection, and provides a good basis for the liquid detection.
Experimental example 2: precision experiment of SPE-HPLC method of the present invention
1 part of hemp leaf sample is finely taken, pretreatment is carried out according to the method of the first embodiment of the invention, the obtained detection sample is continuously injected for three times (N-1, N-2 and N-3), and the content data of three hemp phenolic compounds are shown in the table 2:
table 2:
as can be seen from Table 2, the RSD values of the precision experiments are all within 1 percent, which shows that the method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC has good precision.
Experimental example 3: repeatability experiment of SPE-HPLC method of the invention
6 parts of different hemp leaf samples (N1-N6) are extracted, pretreatment is carried out according to the method of the first embodiment of the invention, the obtained detection samples are sequentially subjected to sample injection detection, and the content data of three hemp phenolic compounds are shown in Table 3:
table 3:
as can be seen from Table 3, the RSD values of the repeatability tests are all within 1.05%, which shows that the method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC has good repeatability.
Experimental example 4: sample adding recovery rate experiment of SPE-HPLC method of the invention
Finely taking 3 different hemp leaf samples (N1, N2 and N3), respectively pretreating according to the method of the first embodiment of the invention to respectively obtain 25ml of detection samples, and carrying out high-efficiency liquid phase detection on the detection samples serving as substrates to obtain substrate values; taking 5mL of substrate detection samples, adding a standard substance containing three cannabinol compounds, wherein the addition amount of the three cannabinol compounds is 20 μ g, performing high performance liquid chromatography detection on the obtained samples as sample loading actual measurement samples to obtain actual measurement values, and the measured content data of the three cannabinol compounds are shown in Table 4:
table 4:
as can be seen from Table 4, the experimental recovery rate range of the sample recovery rate is 99% -113%, which shows that the method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC has good accuracy.
Claims (9)
1. A method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC is characterized by comprising the following steps:
firstly, determining the conditions of high performance liquid chromatography: the high performance liquid chromatography conditions are as follows:
c18 reverse phase column: 4.6X 150mm, 5 μm; column temperature: 40 ℃; detection wavelength: 220 nm; and (3) an elution mode: isocratic elution; mobile phase: methanol 0.1% formic acid 80: 20; flow rate: 1 mL/min; sample introduction amount: 20 mu L of the solution;
secondly, preparing a mixed standard solution: preparing a mixed standard solution which simultaneously contains three cannabinol compounds CBD, CBN and THC with the same mass concentration and the mass concentrations of which are 0.05 mu g/mL, 0.5 mu g/mL, 2 mu g/mL, 10 mu g/mL, 20 mu g/mL, 40 mu g/mL, 60 mu g/mL and 80 mu g/mL in sequence;
thirdly, determining a linear regression equation: sequentially detecting the mixed standard substance solution with each concentration prepared in the step two under the high performance liquid chromatography condition of the step one, determining the nature by retention time, drawing a standard curve according to the corresponding relation between the peak area size measured by each concentration and the concentration of the peak area, and respectively determining the linear regression equations of three cannabinol compounds, wherein the linear regression equations of the three cannabinol compounds are as follows:
CBD：Y＝74725X+38049，n＝8，R20.9992, X applies: 0.05-80 mug/mL;
CBN：Y＝116969X+24626，n＝7，R20.9995, X appliesThe range is as follows: 0.05-60 mu g/mL;
THC：Y＝72297X+21790，n＝7，R20.9992, X applies: 0.05-60 mu g/mL;
wherein: y is the peak area corresponding to cannabinol compound, X is the mass concentration of cannabinol compound, n is the number of different mixed standard substance concentration values when drawing standard curve, R is2Is a linear correlation coefficient;
fourthly, preparing a solid phase extraction pretreatment sample: drying and crushing a hemp sample, putting hemp sample powder into a centrifugal tube, adding methanol, and extracting hemp phenolic compounds in the hemp sample powder by ultrasonic assistance; centrifuging the extract liquor, taking supernatant, and fixing the volume with methanol to obtain a solid-phase extraction pretreatment sample;
fifthly, solid phase extraction: adding the solid-phase extraction pretreatment sample obtained in the step four into a C18/500mg solid-phase small column by adopting a solid-phase extraction mode for retaining interferents, wherein a mobile phase is methanol, the flow rate is 1mL/min, the pressure is 0.1MPa, isocratic elution is carried out, and an effluent liquid is collected; leaching the solid-phase small column by using a small amount of chromatographic grade methanol, and collecting effluent liquid;
sixthly, detecting by high performance liquid chromatography: mixing the effluent liquid collected in the fifth step, re-fixing the volume by using methanol, detecting by adopting a high performance liquid chromatography condition in the first step after membrane filtration, measuring peak areas of all components in the sample, determining the peak areas qualitatively by using retention time, quantifying according to a linear regression equation determined in the third step, and calculating the concentrations of the three cannabinol compounds CBD, CBN and THC in the sample, thereby calculating the contents of the three cannabinol compounds CBD, CBN and THC in the hemp sample.
2. The method according to claim 1, wherein the preparation method of the mixed standard solutions with different mass concentrations in step two comprises sequentially taking 50 μ L, 40 μ L, 100 μ L, 200 μ L, 400 μ L, 600 μ L, and 800 μ L from the mixed standard mother solution containing CBD, CBN, and THC and having a mass concentration of 100 μ g/mL in 100mL, 10mL, 2mL, 1mL, and 1mL volumetric flask and adding methanol to the volumetric flask to obtain the final volume.
3. The method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC as claimed in claim 2, wherein the preparation method of the mixed standard mother liquor is to take 0.5mL CBD, CBN, THC standard solutions with mass concentration of 1mg/mL respectively in a 5mL volumetric flask and fix the volume with methanol.
4. The method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC as claimed in any one of claims 1-3, wherein the retention time in step three is: CBD: 8.3min, CBN: 14.9min, THC: and (3) 18.6 min.
5. The method for simultaneous detection of three cannabinol compounds in hemp by SPE-HPLC as claimed in claim 4, wherein the hemp sample in step four is hemp leaves, the drying and pulverizing process comprises drying hemp leaves in a forced air drying oven at 40 ℃ for 6 hours, and pulverizing a proper amount of dried hemp leaves into powder in a pulverizer.
6. The method for simultaneous detection of three cannabinol compounds in hemp by SPE-HPLC as claimed in claim 5, wherein the ultrasound assisted extraction of step four is that 0.1g of hemp leaf powder is extracted into 10ml centrifuge tube, 6ml of methanol is added, and ultrasound assisted extraction is performed for 30 min.
7. The method for simultaneously detecting three cannabinol compounds in hemp by SPE-HPLC as claimed in claim 6, wherein the centrifugation treatment in step four is 3000r/min for 5 min; the supernatant was taken in a 10mL volumetric flask and made to volume with methanol.
8. The method for simultaneous detection of three cannabinol compounds in hemp by SPE-HPLC as claimed in claim 7, wherein the solid phase column of step five is activated by passing 5mL of chromatographic grade methanol through the column before using.
9. The method for simultaneous detection of three cannabinol compounds in hemp by SPE-HPLC as claimed in claim 8, wherein the effluent from step six is mixed and re-diluted to 25mL with methanol.
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| 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 |
| 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 |
| CN110646534B (en) * | 2019-09-24 | 2022-06-28 | 南京市产品质量监督检验院 | Detection method of cannabidiol in textile |
| CN112730697A (en) * | 2020-09-15 | 2021-04-30 | 中国标准化研究院 | Method for simultaneously detecting 5 cannabinol compounds by using HPLC-MS/MS |
| 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) |
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