CN115469044A - Liquid chromatography-tandem mass spectrometry detection method for gingerol in ginger rhizome, overground stem and leaf - Google Patents
Liquid chromatography-tandem mass spectrometry detection method for gingerol in ginger rhizome, overground stem and leaf Download PDFInfo
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- 238000001514 detection method Methods 0.000 title claims abstract description 52
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- NLDDIKRKFXEWBK-AWEZNQCLSA-N gingerol Chemical compound CCCCC[C@H](O)CC(=O)CCC1=CC=C(O)C(OC)=C1 NLDDIKRKFXEWBK-AWEZNQCLSA-N 0.000 title claims abstract description 35
- 235000002780 gingerol Nutrition 0.000 title claims abstract description 33
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- FADFGCOCHHNRHF-VAWYXSNFSA-N [10]-Shogaol Chemical compound CCCCCCCCC\C=C\C(=O)CCC1=CC=C(O)C(OC)=C1 FADFGCOCHHNRHF-VAWYXSNFSA-N 0.000 claims description 4
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- 238000004885 tandem mass spectrometry Methods 0.000 claims description 3
- AIULWNKTYPZYAN-SFHVURJKSA-N (10)-Gingerol Chemical compound CCCCCCCCC[C@H](O)CC(=O)CCC1=CC=C(O)C(OC)=C1 AIULWNKTYPZYAN-SFHVURJKSA-N 0.000 claims description 2
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- LGZSMXJRMTYABD-MDZDMXLPSA-N [8]-Shogaol Chemical compound CCCCCCC\C=C\C(=O)CCC1=CC=C(O)C(OC)=C1 LGZSMXJRMTYABD-MDZDMXLPSA-N 0.000 claims description 2
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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
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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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
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- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
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Abstract
The invention discloses a liquid chromatography-tandem mass spectrometry detection method for gingerol in ginger rhizome, overground stem and leaf. The detection method comprises the following steps: s1, pretreating ginger tissues, crushing and sieving to obtain ginger powder; s2, adding methanol into the ginger powder for extraction, and centrifuging to obtain a supernatant; and S3, detecting the supernatant by adopting a liquid chromatography-tandem mass spectrometry method, namely realizing qualitative and quantitative detection of gingerol in the ginger, wherein ginger tissues are rhizomes, overground stems and leaves. The development of the detection method is an important premise and basis for the formation rule and mechanism research of the gingerol characteristic quality components in the ginger, and a reliable technical means is provided for variety breeding, quality evaluation and quality improvement of the ginger.
Description
Technical Field
The invention relates to a liquid chromatography-tandem mass spectrometry detection method for gingerol in ginger rhizome, overground stem and leaf, belonging to the technical field of detection.
Background
Gingerol in gingerHas strong anti-inflammatory, antibacterial and anticancer effects, is an important component of rhizoma Zingiberis recens with medicinal and health promoting effects, and mainly comprises gingerol, shogaol, and zingerone. At present, extraction methods of gingerol compounds mainly comprise squeezing, solvent extraction, column chromatography, supercritical CO 2 Extraction method, microwave extraction method, ultrasonic extraction method, enzymolysis method, etc., wherein the solvent extraction method commonly uses solvents such as acetonitrile, diethyl ether, acetone, methanol, ethanol, ethyl acetate, etc. The detection method mainly comprises stripping voltammetry, thin-layer chromatography, fluorescence analysis, ultraviolet-visible spectrophotometry, liquid Chromatography (LC), liquid chromatography-tandem mass spectrometry (LC-MS/MS) and the like. The total content of gingerol can only be quantified by stripping voltammetry, thin-layer chromatography, fluorescence analysis and ultraviolet-visible spectrophotometry, and all components cannot be analyzed; the content of gingerol can be quantified by using a standard substance by the liquid chromatography, but the steps and the calculation are more complicated; and the liquid chromatography-tandem mass spectrometry method can realize the qualitative and quantitative detection of each gingerol compound in ginger.
Disclosure of Invention
The invention aims to provide a liquid chromatography-tandem mass spectrometry detection method for gingerol in ginger, which is suitable for detecting gingerol in three tissues (rhizomes, overground stems and leaves) in ginger and provides a reliable technical means for variety breeding, quality evaluation and quality improvement of ginger.
The invention provides a liquid chromatography-tandem mass spectrometry detection method of gingerol in ginger, which comprises the following steps:
s1, pretreating ginger tissues, crushing and sieving to obtain ginger powder;
s2, adding methanol into the ginger powder for extraction, and centrifuging to obtain a supernatant;
and S3, detecting the supernatant by adopting a liquid chromatography-tandem mass spectrometry method, namely realizing qualitative and quantitative detection of gingerol in the ginger.
In the detection method, the ginger tissues comprise rhizomes, overground stems and leaves.
The detected gingerol comprises 8 compounds such as zingerone, 6-gingerol, 6-shogaol, 8-gingerol, 1-dehydro-6-zingerone, 10-gingerol, 8-shogaol and 10-shogaol.
In the detection method of the invention, in step S1, the pretreatment steps are as follows:
freeze-drying the ginger tissue after quick freezing, and carrying out quick freezing in liquid nitrogen;
the temperature of the freeze drying is-60 to-70 ℃;
the granularity of the ginger powder is 40-80 meshes;
grinding can be adopted for crushing;
can be stored in ultralow temperature condition (such as ultralow temperature refrigerator at-80 deg.C).
The freeze drying is carried out under the low temperature condition, the loss of volatile components in the sample is very small, and the quality components of the ginger sample can be retained to the maximum extent; and the drying is carried out under the vacuum condition, so that the oxygen in the air and the like can be prevented from generating chemical reaction with components in the sample. The gingerol compounds in ginger are unstable and can be converted into shogaols under thermal conditions, so that the gingerol compounds in ginger can be retained to a large extent by crushing after freeze drying.
In the detection method of the invention, the dosage of the methanol is as follows: 20-40 mL/g of the ginger powder, preferably 20mL/g;
the extraction comprises ultrasonic treatment and oscillation which are carried out in sequence;
the ultrasonic treatment time is 30-90 min, preferably 60min, and the temperature is 10-25 ℃, preferably 25 ℃;
the oscillation time is 60-90 min, preferably 60min.
In the detection method of the present invention, in step S2, the centrifugation conditions are as follows:
centrifuging at 4-10 deg.c and rotation speed of 10000-14000 rpm, preferably at 4 deg.c and 10000rpm for 10min;
before step S3, the supernatant was passed through a 0.22 μm filter.
In the detection method of the present invention, in step S3, the detection conditions of the liquid chromatography in the liquid chromatography-tandem mass spectrometry are as follows:
a chromatographic column: a C18 column;
column oven temperature: 35 ℃;
mobile phase: phase A is an aqueous solution containing 0.1% formic acid and 2mM ammonium formate, and phase B is a methanol solution containing 0.1% formic acid and 2mM ammonium formate;
gradient elution: 0-1min, 85% of phase A, 15% of phase B; 1-5min, 50% of phase A, 50% of phase B; 5-8 min,20% of phase A, 80% of phase B; 8-10min, 0% of phase A, 100% of phase B; 10-15min, 0% of phase A, 100% of phase B;
flow rate: 0.4mL/min;
sample introduction amount: 5 μ L.
In the detection method of the present invention, in step S3, the detection conditions of the mass spectrum in the liquid chromatography-tandem mass spectrometry are as follows:
type of ion source: electrospray ion source (H-ESI);
the scanning mode comprises the following steps: multiple Reaction Monitoring (MRM);
spray Voltage (Ion Spray Voltage): 3600V in positive ion mode and 2500V in negative ion mode;
mass spectrometry ion source temperature: 325 ℃;
sheath Gas (Sheath Gas): 30Arb;
assist Gas (Aux Gas): 10Arb;
tail Gas (Sweep Gas): 0Arb.
When the detection method is used for quantitative detection, a standard curve method (external standard method) can be adopted;
according to different polarities of compounds, methanol and dimethyl sulfoxide (DMSO) are respectively used as solvents to prepare standard substance stock solutions. Firstly, accurately weighing a standard substance (accurate to 0.01 mg) in a standard amount on an analytical balance, transferring the weighed standard substance into a 10mL volumetric flask, dissolving the standard substance by using methanol or DMSO, fixing the volume into the volumetric flask, preparing a finished standard substance stock solution, and placing the finished standard substance stock solution in a refrigerator at the temperature of-20 ℃ for storage. Diluting the mixed standard working solution to 1-2000 mug/L by methanol on the basis of the standard substance stock solution, and storing the mixed standard working solution in a refrigerator at the temperature of-20 ℃ for later use; each standard solution is prepared in situ.
The detection method has the following beneficial technical effects:
at present, most of detection methods for gingerol in ginger are quantitative analysis on one or more compounds in gingerol, shogaol and zingerone, discrimination and accuracy are low, a method for detecting the 8 gingerol compounds in ginger by adopting an LC-MS/MS method is not reported, and multiple reaction monitoring MRM mode quantitative detection has high sensitivity and good specificity. Therefore, the development of the detection method is an important premise and basis for researching the formation rule and mechanism of the gingerol characteristic quality components in the ginger, and a reliable technical means is provided for variety breeding, quality evaluation and quality improvement of the ginger.
Drawings
FIG. 1 is a statistical histogram of recovery for different extraction solvents.
Fig. 2 is a statistical histogram of recovery for different extraction temperatures.
Fig. 3 is a statistical histogram of recovery rate for different ultrasound extraction times.
Fig. 4 is a statistical histogram of recovery rates corresponding to different oscillation extraction times.
FIG. 5 is an ion pair chromatogram of 8 gingerols.
Fig. 6 is an ion pair chromatogram of water-acetonitrile as a mobile phase.
FIG. 7 is an ion pair chromatogram of ammonium acetate as mobile phase addition salt.
FIG. 8 is an ion pair chromatogram at a flow rate of 0.3 mL/min.
FIG. 9 is an ion pair chromatogram at a flow rate of 0.45 mL/min.
FIG. 10 is an ion pair chromatogram of 10min elution time.
FIG. 11 is an ion pair chromatogram of an elution time of 15 min.
Detailed Description
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The instrument and reagent information in the examples below are shown in tables 1 and 2.
TABLE 1 Main test instruments
TABLE 2 Main test reagents
Preparation of a standard solution: according to different polarities of the compounds, methanol and DMSO are respectively adopted as solvents to prepare standard substance stock solutions. Firstly, accurately weighing a standard substance (accurate to 0.01 mg) in a standard amount on an analytical balance, transferring the weighed standard substance into a 10mL volumetric flask, dissolving the standard substance by using a methanol or DMSO solution, fixing the volume of the standard substance into the volumetric flask, preparing a finished standard substance stock solution, and storing the stock solution in a refrigerator at the temperature of-20 ℃. Diluting the mixed standard working solution to 1-2000 mug/L by methanol on the basis of the standard substance stock solution, and storing the mixed standard working solution in a refrigerator at the temperature of-20 ℃ for later use; each standard solution is prepared just before use.
Examples 1,
1. Sample pretreatment and pretreatment
Pretreatment: removing large pieces of soil on the surface layer of the ginger, gently rubbing and washing in clear water by using a soft brush to remove the soil and rotten parts on the surface of the ginger, quickly freezing at least 500g of a clean ginger sample in liquid nitrogen, freeze-drying the quick-frozen sample by using a freeze dryer at-70 ℃, grinding and mixing different tissue samples of the ginger into powder, sieving by using a 60-mesh fine sieve, and placing the powder in an ultra-low temperature refrigerator at-80 ℃ for storage to be tested.
Sample pretreatment: accurately weighing 1.000+0.001g ginger different tissue samples by an analytical electronic balance, placing the samples in a 50mL centrifuge tube, then accurately adding 20mL methanol, carrying out vortex for 5min by a parallel vortex instrument, then carrying out ultrasonic extraction for 60min at normal temperature, then carrying out shaking extraction for 24h by a shaking table, centrifuging the extracted mixed solution for 10min at 4 ℃ and 10000rpm, and filtering the centrifuged supernatant with a 0.22 mu m filter membrane in a sample injection vial. Because the content of the compounds in the samples is different, high-content compounds are required to be diluted based on the measurement requirement of instrument protection, and therefore the extracting solution is diluted by 10 times, 50 times, 200 times and 1000 times respectively; the extracts after different dilutions were stored in a refrigerator at 4 ℃ to be tested.
2. Conditions of the apparatus
A chromatographic column: a phenomenox C18 column (phenomenox Co., USA) with a length of 100mm, an internal diameter of 2.1mm and a particle size of 2.6 μm;
column oven temperature: 35 ℃;
mobile phase used in the method: (phase a) 0.1% formic acid and 2mM aqueous ammonium formate solution, (phase B) 0.1% formic acid and 2mM methanolic ammonium formate solution;
method flow rate: 0.4mL/min;
mobile phase gradient elution: 0 to 1min,85% A (15% B); 1 to 5min,50% A (50% B); 5 to 8min,20% A (80% B); 8 to 10min,0% A (100% B); 10 to 15min,0% (100% B).
Sample introduction amount: 5 μ L.
The TSQ triple quadrupole mass spectrometry conditions were as follows:
ion source type: electrospray ion source (H-ESI);
the scanning mode is as follows: multiple Reaction Monitoring (MRM);
spray Voltage (Ion Spray Voltage): 3600V in positive ion mode and 2500V in negative ion mode;
mass spectrometry ion source Temperature (Temperature): 325 ℃;
sheath Gas (Sheath Gas): 30Arb;
assist Gas (Aux Gas): 10Arb;
tail Gas (Sweep Gas): 0Arb.
The ion pair of gingerol compound and the corresponding ion characteristic mass spectrum conditions of Collision Energy (CE), injection voltage (RF Lens) and the like are shown in Table 3.
TABLE 3 names, retention times, MRM ion pairs and Mass Spectrometry detection parameters of the various gingerol Compounds
3. Results of the experiment
(1) Pretreatment and optimization of instrument conditions
1) Optimization of pretreatment conditions
In the pretreatment method for ginger sample detection, pretreatment conditions such as extraction solvents (methanol, acetonitrile, water/methanol (1/1, v/v)), extraction temperature (10 ℃, room temperature, 30 ℃, 40 ℃), ultrasonic extraction time (30 min, 60min, 90min, 120min, 6h, 12h, 24 h), shaking extraction time (30 min, 60min, 90min, 120min, 6h, 12h, 24 h) and the like are optimized.
Gingerol compounds belong to nonpolar compounds, so organic solvents are required for extraction, three extraction solvents of methanol, acetonitrile and water/methanol (1/1, v/v) are compared in the experiment, and the results are shown in table 4. With the recovery rates as ordinate and different extraction solvents as abscissa, statistical histograms of the recovery rates of the 8 gingerol compounds when different extraction solvents were used were prepared, as shown in fig. 1. When acetonitrile is used as an extraction solvent, the extraction effect is poor. Methanol and water/methanol (1/1, v/v) extract similarly, but the direct use of methanol as the extraction solvent simplifies the experimental work. Therefore, methanol is used as the extraction solvent.
TABLE 4 Effect of different extraction solvents on gingerol recovery
Gingerol compounds are dehydrated and converted into the shogaol compounds under the high temperature condition, so that certain requirements are imposed on the extraction temperature, and the four extraction temperatures of 10 ℃, room temperature, 30 ℃ and 40 ℃ are compared in the experiment, and the results are shown in table 5 and figure 2. Experiments show that the extraction effect of gingerol is lower at 40 ℃, the extraction effect is lower than that at room temperature and 10 ℃ at 30 ℃, the extraction effect is not greatly different at room temperature and 10 ℃, and the experimental steps can be simplified by directly adopting room temperature for extraction. Therefore, the experiments were performed at room temperature.
TABLE 5 Effect of different extraction temperatures on gingerol recovery
Meanwhile, seven ultrasonic extraction times of 30min, 60min, 90min, 120min, 6h, 12h and 24h and oscillation extraction time are compared in the experiment, and the results are shown in table 6, fig. 3, table 7 and fig. 4. And (5) finding out the result through experiments. When the extraction is carried out for 30min in a short time, the extraction effect is lower than that of 60min, and when the extraction time is further prolonged, the recovery rate is not obviously improved. Therefore, the experiment is carried out by ultrasonic extraction for 60min and shaking extraction for 60min.
TABLE 6 Effect of different ultrasound extraction times on recovery
TABLE 7 Effect of different shaking extraction times on recovery
The optimal pretreatment conditions determined by experiments are as follows:
placing the sample in a 50mL centrifuge tube, then accurately adding 20mL methanol, carrying out vortex for 5min by using a parallel vortex instrument, carrying out ultrasonic extraction for 60min at normal temperature, then carrying out shaking extraction for 24h by using a shaking table, centrifuging the extracted mixed solution for 10min at 4 ℃ and 10000rpm, and filtering the centrifuged supernatant through a 0.22 mu m filter membrane in a sample injection vial.
2) Optimization of instrumentation test conditions
Based on liquid chromatography-triple quadrupole tandem mass spectrometry, conditions such as mobile phase (water-methanol, water-acetonitrile) and mobile phase addition salt (ammonium formate, ammonium acetate), flow rate (0.3 mL/min, 0.4mL/min, 0.45 mL/min), elution time and gradient (10 min, 15min, 20 min) are selected and optimized, the optimized detection condition of step 1 is determined, and ion pair chromatograms of 8 gingerol compounds under the optimized condition are shown in fig. 5.
Wherein, fig. 6 is an ion pair chromatogram of water-acetonitrile as a mobile phase; FIG. 7 is an ion pair chromatogram of ammonium acetate as mobile phase addition salt; FIG. 8 is an ion pair chromatogram at a flow rate of 0.3 mL/min; FIG. 9 is an ion pair chromatogram at a flow rate of 0.45 mL/min; FIG. 10 is an ion pair chromatogram of 10min elution time; FIG. 11 is an ion pair chromatogram of 15min elution time.
(2) Linear relationship, detection limit and quantitation limit
The Limit of detection (LOD), limit of quantification (LOQ) and linearity of the method of the present invention are evaluated. LODs and LOQs are 3 times and 10 times the signal-to-noise ratio (S/N), respectively; the standard curve was plotted for at least 7 concentrations in series.
Linear equation, linear range and correlation coefficient (r) of detection method established based on liquid chromatogram tandem mass spectrum 2 ) LODs and LOQs are shown in Table 8. As can be seen from Table 8, the linear equation of the detection method can be used for accurately, qualitatively and quantitatively detecting the target compound, the linear range is wide, and the qualitative and quantitative detection of each gingerol in the ginger sample can be met; wherein the linear correlation coefficient r 2 >0.9990. LODs and LOQs of zingerone are respectively 1000 mug/kg and 2000 mug/kg, LODs and LOQs of other compound components are respectively lower than 200 mug/kg and 400 mug/kg, and the detection requirement can be met. Compared with the traditional detection method, the method adopts an MRM scanning mode, has relatively higher sensitivity and more accurate quantitative result, and can provide more complete data support for gingerol content data in ginger.
TABLE 8 Standard curves, LODs and LOQs for ginger Compounds
(3) Recovery, matrix Effect and precision
The accuracy of the method was evaluated using the recovery rate of the spiked sample. Wherein the recovery rate calculation formula is as follows:
recovery = (detected amount-original amount)/addition amount × 100%
During the recovery evaluation, the scaling levels for L (low), M (medium) and H (high) 3 recoveries were designed, with the added compound levels being about 1/2, 1/1 and 3/2 of the original compound content in ginger. In common matrix substances, a matrix blank sample of gingerol is difficult to find, so that a slope method is adopted to evaluate the matrix effect, and the calculation formula of the slope method is as follows:
slope = substrate standard curve slope/solvent standard curve slope
When the slope is 0.8-1.2, the influence of the matrix effect on the method is small, and solvent standard sample can be used for quantification; when the slope is less than 0.8 or greater than 1.2, there is an inhibitory or reinforcing effect of the matrix effect on the method, and quantification using a matrix standard is required.
The recovery results of each compound in 3 different tissues of rhizome, overground stem and leaf of ginger are shown in table 9, and the matrix effect results are shown in table 10.
The recovery rates of 8 gingerol compounds in 3 tissues of rhizome, overground stem and leaf are respectively 63.5-122.3%, 64.2-123.2% and 60.5-127.8%, and the RSD of each compound is less than 20%, which shows that the instrument method and the pretreatment method meet the methodological evaluation and can be used for detecting 8 gingerol in 3 tissues of ginger. The matrix effect of the compounds in 3 tissues is respectively between 0.8-1, 0.8-1.2 and 0.8-1.2, which shows that the matrix effect of the method is not significant, the interference of the sample extracting solution has little influence on the test result, and the condition of adopting a solvent standard to carry out qualitative and quantitative determination is met. Therefore, the method is suitable for qualitative and quantitative detection of 8 gingerol compounds in rhizomes, overground stems and leaves of ginger.
TABLE 9 recovery and RSD of 8 gingerol compounds in 3 different tissues of ginger at 3 spiking levels
TABLE 10 matrix Effect of 8 gingerol Compounds in 3 different tissues of ginger
The invention establishes a liquid phase-tandem mass spectrometry (LC-MS/MS) for rapidly and accurately determining 8 gingerols in ginger, wherein the applicability of the detection method in three tissues (rhizomes, overground stems and leaves) of the ginger is evaluated by a methodology after establishment. The method adopts solvent standard in qualitative and quantitative determination of 8 gingerol compounds, and r of standard curve of 8 compounds 2 >0.9990 LODs of the method<1000 mug/kg, recovery rate of 3 tissue samples in ginger under the condition of adding standard level of matrix is 60.5% -127.8%, and RSD<20 percent. The methodological evaluation shows that the method can be suitable for detecting 8 gingerol in different tissues of the ginger, and provides a reliable technical means for evaluating and identifying the quality of the ginger.
Claims (10)
1. A liquid chromatography-tandem mass spectrometry detection method for gingerol in ginger comprises the following steps:
s1, pretreating ginger tissues, crushing and sieving to obtain ginger powder;
s2, adding methanol into the ginger powder for extraction, and centrifuging to obtain a supernatant;
and S3, detecting the supernatant by adopting a liquid chromatography-tandem mass spectrometry method, namely realizing qualitative and quantitative detection of gingerol in the ginger.
2. The detection method according to claim 1, characterized in that: the ginger tissue is rhizome, overground stem and leaf.
3. The detection method according to claim 1 or 2, characterized in that: the gingerol comprises zingerone, 6-gingerol, 6-shogaol, 8-gingerol, 1-dehydro-6-zingerone, 10-gingerol, 8-shogaol and 10-shogaol.
4. The detection method according to any one of claims 1 to 3, characterized in that: in step S1, the preprocessing steps are as follows:
quickly freezing the ginger tissues and then freeze-drying;
the temperature of the freeze drying is-60 to-70 ℃;
the ginger powder has a granularity of 40-80 meshes.
5. The detection method according to any one of claims 1 to 4, characterized in that: in the step S2, the volume content of the methanol in the methanol water solution is 0.1-0.2%;
the dosage of the methanol water solution is as follows: 20-40 mL/g of the ginger powder.
6. The detection method according to any one of claims 1 to 5, characterized in that: in the step S2, the extraction comprises ultrasonic treatment and oscillation which are sequentially carried out;
the ultrasonic treatment time is 30-90 min, and the temperature is 10-25 ℃;
the oscillation time is 60-90 min.
7. The detection method according to any one of claims 1 to 6, characterized in that: in step S2, the centrifugation conditions are as follows:
the temperature is 4-10 ℃, and the rotating speed is 10000-14000 rpm;
prior to step S3, the supernatant was passed through a 0.22 μm filter.
8. The detection method according to any one of claims 1 to 7, characterized in that: in step S3, the detection conditions of the liquid chromatography in the liquid chromatography-tandem mass spectrometry are as follows:
and (3) chromatographic column: a C18 column;
mobile phase: the phase A is an aqueous solution containing 0.1 percent of formic acid and 2mM ammonium formate, and the phase B is a methanol solution containing 0.1 percent of formic acid and 2mM ammonium formate;
gradient elution: 0-1min, 85% phase A, 15% phase B; 1-5min, 50% phase A, 50% phase B; 5-8 min,20% of phase A, 80% of phase B; 8-10min, 0% of phase A, 100% of phase B; 10 to 15min,0% of phase A, 100% of phase B.
9. The detection method according to any one of claims 1 to 8, characterized in that: in step S3, the detection conditions of mass spectrometry in the hplc-tandem mass spectrometry are as follows:
type of ion source: an electrospray ion source;
the scanning mode is as follows: monitoring multiple reactions;
spraying voltage: 3600V in positive ion mode and 2500V in negative ion mode;
mass spectrometry ion source temperature: 325 ℃;
sheath gas: 30Arb;
auxiliary gas: 10Arb;
tail gas: 0Arb.
10. The detection method according to any one of claims 1 to 9, characterized in that: and in the step S3, quantitatively detecting the gingerol by adopting a standard curve method.
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| CN114778728A (en) * | 2022-04-20 | 2022-07-22 | 中国医学科学院药用植物研究所云南分所 | Determination method for curcumin compound content |
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| CN114778728A (en) * | 2022-04-20 | 2022-07-22 | 中国医学科学院药用植物研究所云南分所 | Determination method for curcumin compound content |
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