CN106198791B - Method that is a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood - Google Patents
Method that is a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood Download PDFInfo
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- CN106198791B CN106198791B CN201610506609.5A CN201610506609A CN106198791B CN 106198791 B CN106198791 B CN 106198791B CN 201610506609 A CN201610506609 A CN 201610506609A CN 106198791 B CN106198791 B CN 106198791B
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Abstract
The invention discloses methods that is a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood, it is ultrasonically treated sample to be tested with methanol and does blank solution simultaneously, the hybrid standard working solution with concentration gradient is prepared with methanol, wherein the concentration range of agalloch eaglewood tetrol is 20~200 μ g/mL, the concentration range of benzylacetone is 0.5~5 μ g/mL, using gas chromatography-mass spectrometry to standard working solution, blank solution and sample solution carry out analysis detection, obtain related chromatogram, qualitative analysis is carried out with the retention time and characteristic ion abundance ratio of object in sample solution and standard working solution, quantitative analysis is carried out with external standard method, finally draw standard curve and result of calculation.The advantages that present invention operation is quick, simple and practicable, and object can measure simultaneously, the measurement of agalloch eaglewood tetrol and benzylacetone suitable for agalloch eaglewood.The method of the present invention high sensitivity, reproducible, the rate of recovery is high.
Description
【Technical field】
The present invention relates to methods that is a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood, belong to detection method technology
Field.
【Background technology】
Agalloch eaglewood is the timber that Isolated From Thymelaeaceae Species agalloch eaglewood or suspension culture of Aquilaria sinensis contain resin, main product in China, Japan, India and its
His country in Southeast Asia, has effects that promoting qi circulation and relieving pain, warming middle energizer to arrest vomiting, gas of receiving are relievingd asthma, cures mainly the swollen bored pain of chest and abdomen, gastrofrigid vomiting oh
Inverse, the kidney deficiency circulation of vital energy in the wrong direction breathes heavily the diseases such as anxious, is the main prescription raw medicinal material that hundred kinds of Chinese patent drugs are cured in China.In addition to medicinal, agalloch eaglewood or name
Expensive natural perfume material and stacte raw material, the market demand is huge.But in nature, only when eaglewood is stimulated or damages
When hindering, agalloch eaglewood is just will produce, Edgeworthia chrysantha process generally requires the even years up to a hundred decades, with ecological environment destruction and agalloch eaglewood
The over-exploitation of resource, eaglewood resource is increasingly exhausted, is now listed in national second class protection wild plant, and be loaded into《Generation
Conservation of nature alliance of boundary endangered species Red List》, therefore far can not meet the needs of market, cause agalloch eaglewood price to be climbed year by year
It rises.Presently commercially available agalloch eaglewood medicinal material is very different, and there are a large amount of agalloch eaglewood inferior goods, and even adulterant or suspension culture of Aquilaria sinensis timber is impregnated fragrant
Counterfeit drug and dulcet tree material such as cypress etc. fill generation made of material, utter misery post-processing, serious impact on medical effect.
The ingredients such as chromone class, aromatic and sesquiterpenoids are mainly contained in agalloch eaglewood, have 125 agalloch eaglewood product of document pair, its
His 25 kinds of dulcet tree materials and the agallochum material of health are detected, and as a result prove only can just detect chromone class in agalloch eaglewood
Ingredient agalloch eaglewood tetrol and aromatic component benzylacetone.It follows that agalloch eaglewood tetrol and benzylacetone are the characteristic components of agalloch eaglewood,
Its content can be used as the important evidence of agalloch eaglewood authenticity and quality evaluation.Therefore, version in 2015《Chinese Pharmacopoeia》Provide agalloch eaglewood
Four alcohol content of middle agalloch eaglewood is not less than 0.10%.
Currently, it is respectively liquid chromatography and gas-chromatography to measure the method for agalloch eaglewood tetrol and benzylacetone content in agalloch eaglewood
Method, but the method that there is no while measuring two kinds of compounds in agalloch eaglewood.Although liquid chromatography and gas chromatography have stronger
Analysis ability, but each constituent content in the measurement product that they can only be quantitatively are detached, but for quickly and accurately Qualitive test
It has some limitations, and big using liquid-phase chromatographic analysis solvent consumption, testing cost is high.The present invention uses gas-chromatography-
Tandem mass spectrum method, can it is accurate simultaneously, quickly and easily measure agalloch eaglewood tetrol and benzylacetone content in agalloch eaglewood.
【Invention content】
The technical problem to be solved by the present invention is to overcome the deficiencies of the prior art and provide one kind can quickly, accurately determine
Property differentiate, and the method that agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood can be measured simultaneously.The method of the present invention uses gas chromatography-mass spectrum
The advantages of combination detection method has easy to operate, high sensitivity, reproducible, and target compound measures simultaneously.
The present invention to achieve the above object, using following technical scheme：
Measuring instrument and equipment used in the present invention are：Gas chromatograph-mass spectrometer (GC-MS), supersonic wave cleaning machine, electronics
Balance.
Method that is a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood, it is characterised in that include the following steps：
A, testing sample solution is prepared：Agalloch eaglewood sample ultra-pure water is cleaned and is dried, No. three sieves is smashed it through, accurately weighs
0.3g is accurate to 0.0001g in being placed in plug conical flask, and 12~15mL of methanol is added in precision, and weighed weight impregnates 1h, frequency
40kHz, power 300W are ultrasonically treated 50min, let cool, then weighed weight, the weight of loss is supplied with methanol, is shaken up, and centrifuge, quiet
It sets, takes supernatant liquor up to sample solution, it is to be measured, while doing blank solution；
B, standard working solution is prepared：With agalloch eaglewood tetrol of the methanol preparation with concentration gradient and with the benzyl of concentration gradient
Benzylacetone hybrid standard working solution, the wherein concentration range of agalloch eaglewood tetrol are 20~200 μ g/mL, the concentration model of benzylacetone
It encloses for 0.5~5 μ g/mL；
C, gas chromatography-mass spectrometry analysis：Using gas chromatograph-mass spectrometer (GC-MS) to standard working solution, blank solution and
Sample solution carries out analysis detection, obtains related chromatogram, in sample solution and standard working solution when the reservation of object
Between and characteristic ion abundance ratio carry out qualitative analysis, with external standard method carry out quantitative analysis；
Wherein the gas chromatography-mass spectrum condition is：
Chromatographic column：HP-5MS capillary columns, 30m × 0.25mm × 0.25 μm；Input mode：Splitless injecting samples, injection port
Temperature：250℃；Carrier gas：He, 99.999%, column flow 1.2mL/min；Column temperature uses temperature programming：40 DEG C of initial temperature is protected
1min is held, 210 DEG C are risen to 10 DEG C/min, 1min is kept, then 220 DEG C are risen to 2 DEG C/min, keeps 1min, then with 8 DEG C/min
300 DEG C are risen to, 4min is kept；Sample size：1μL；
Mass Spectrometry Conditions are：Ion source temperature：250℃；Ionization pattern：Electron impact ionization；Mass spectrum transmission line temperature 280
℃；Solvent delay time 3.0min；
Scan mode:Selected ion monitoring scans (SIM), and the monitoring period to each ion is 50ms；
D. it draws standard curve and result calculates：Standard working solution is introduced into gas chromatograph-mass spectrometer (GC-MS), with external standard
Method carries out quantitative analysis, and the concentration of the peak area of target compound quota ion and target compound is carried out linear regression point
Analysis, obtains standard curve equation of linear regression, and related coefficient need to be not less than 0.999；The sample solution of preparation is measured, is surveyed
The peak area of target compound in sample solution is obtained, unary linear regression equation is substituted into and obtains target compound in sample solution
Content acquires agalloch eaglewood tetrol and benzylacetone in sample respectively further according to following calculation formula (1) and calculation formula (2) conversion
Content：
Calculation formula (1) is：
Calculation formula (2) is：
In formula：
X1--- the content of agalloch eaglewood tetrol in sample, unit %；
X2--- the content of benzylacetone in sample, unit mg/kg；
C --- the concentration of target compound in sample solution, unit are μ g/mL；
C0--- the concentration of target compound in blank solution, unit are μ g/mL；
V --- sample solution constant volume, unit are milliliter (mL)；
The quality that M --- sample weighs, unit g.
When being calculated according to formula, with the average value of parallel determinations twice for final measurement result, as a result retaining 3 has
Three number of effect.
The targeted agalloch eaglewood of the present invention includes import agalloch eaglewood, Home-made occluder and the agalloch eaglewood using the production of artificial Edgeworthia chrysantha technology.
For the present invention when preparing standard working solution, the gradient concentration of agalloch eaglewood tetrol is 20 μ g/mL, 40 μ g/mL, 80 μ g/
The gradient concentration of mL, 120 μ g/mL, 160 μ g/mL, 200 μ g/mL, benzylacetone are 0.5 μ g/mL, 1 μ g/mL, 2 μ g/mL, 3 μ g/
mL、4μg/mL、5μg/mL.Specifically preparation method is：
0.1g benzylacetone standard items accurately are weighed, 0.1mg is accurate to, is placed in 100mL volumetric flasks, simultaneously with methanol dissolving
Constant volume is configured to the benzylacetone standard reserving solution I of a concentration of 1mg/mL；0.02g agalloch eaglewood tetrol standard items accurately are weighed, accurately
It to 0.1mg, is placed in 10mL volumetric flasks, is dissolved with methanol, while accurately pipetting 0.500mL benzylacetones standard reserving solution I and setting
In same 10mL volumetric flasks and constant volume, it is configured to hybrid standard storing solution II, the wherein concentration of agalloch eaglewood tetrol and benzylacetone
Respectively 2mg/mL and 50 μ g/mL；The accurate hybrid standard storage for pipetting 0.1mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL
Standby liquid II, is respectively placed in 10mL volumetric flasks, with methanol constant volume to scale to get standard working solution.
In the present invention, when selected ion monitoring scans, agalloch eaglewood tetrol and benzylacetone be quantitative, qualitative ion and its abundance
Than see the table below 1：
Table 1：Agalloch eaglewood tetrol and benzylacetone quantify and qualitative selection ion table
。
The present invention is in drawing standard curve and result calculating, the selection ion color of target compound and standard items in sample
Spectral peak at identical retention time (± 0.5%) occurs, and the mass-to-charge ratio of corresponding mass spectrometric fragment ion is consistent with standard items,
Abundance ratio should meet compared with standard items：
When relative abundance ＞ 50%, allow ± 10% deviation；
When relative abundance 20%~50%, allow ± 15% deviation；
When relative abundance 10%~20%, allow ± 20% deviation；
When relative abundance≤10%, allow ± 50% deviation；Target analytes can be confirmed at this time.
The present invention compares ultrasound and is heated to reflux two kinds of extracting modes, the results showed that using methanol as Extraction solvent, ultrasound
The object content of 50min extractions and the object content for being heated to reflux 5h extractions are almost the same.Ultrasonic extracting method is quickly high
Effect, and can be in normal-temperature operation, therefore select ultrasonic extraction mode.
The present invention selects methanol as Extraction solvent in the preparation process of testing sample solution, with ethyl alcohol, acetone, dichloro
Methane, ethyl acetate, acetonitrile, hexamethylene, anhydrous ether, petroleum ether are compared, and the extraction efficiency of methanol is optimal.Therefore it is preferred that methanol is made
For Extraction solvent.
The present invention has investigated influence of the sample broke degree to measurement result, the results showed that after grain size is less than No. three sieves, subtracts
Small particle, the measurement result of target compound is without significant changes in sample, and the relative standard deviation of measurement result is less than 5%,
Sample homogeneity disclosure satisfy that the measurement requirement of the present invention.Therefore, No. three sieves are crossed when the preferred sample pre-treatments of the present invention.
The present invention has investigated influence of the solvent dosage to measurement result in the preparation process of testing sample solution, as a result table
Bright in the case where solvent dosage is 12~15mL, extraction efficiency is preferable, is further added by solvent dosage, and extraction efficiency variation is little.
Therefore select solvent dosage for 12~15mL from the point of view of cost-effective.
The present invention has investigated influence of the extraction time to measurement result in the preparation process of testing sample solution, as a result table
Bright extraction time is more than 50min, and extraction efficiency is not much different.Therefore consider that the selective extraction time is from time-saving angle
50min。
The present invention has selected suitable standard working solution range, the survey of agalloch eaglewood tetrol and benzylacetone suitable for agalloch eaglewood
It is fixed.
The present invention is during gas chromatography-mass spectrometry analysis, in order to obtain more preferably separating effect, and avoids other groups
Divide and benzylacetone and agalloch eaglewood tetrol interfered, through repeatedly debugging optimization, heats up before having selected 210 DEG C and after 220 DEG C very fast,
The slower temperature program that heats up between 210 DEG C~220 DEG C is detected.
In the present invention, when obtaining the reservation of benzylacetone and agalloch eaglewood tetrol standard items by full scan mode (SCAN) first
Between and full scan spectrogram, then respectively extract benzylacetone and agalloch eaglewood tetrol mass spectrogram, and combine mass spectrogram fragment ion phase
Information is closed, determines the qualitative, quantitative ion of target compound.
Compared with prior art, the present invention has the following advantages：
The present invention provides a kind of gas chromatography-mass spectrography measure agalloch eaglewood in agalloch eaglewood tetrol and benzylacetone method,
The advantages that detection method has operation quick, simple and practicable, and object can measure simultaneously, suitable for agalloch eaglewood agalloch eaglewood tetrol with
The measurement of benzylacetone.
The present invention suitable Pretreatment in order to obtain uses extracting mode, Extraction solvent, sample broke degree, solvent
Amount and extraction time optimize and select, and use can provide qualitative and quantitative information gas chromatography-mass spectrography simultaneously
Instrument (GC/MS) is used as detecting instrument, in agalloch eaglewood sample agalloch eaglewood tetrol and benzylacetone be measured, the results showed that standard is bent
Line linear dependence is good, and the detection of agalloch eaglewood tetrol is limited to 80mg/kg, relative standard deviation 3.47%, average recovery of standard addition
Between 92.6%~97.5%, the detection of benzylacetone is limited to 2mg/kg, relative standard deviation 4.04%, and average mark-on returns
For yield 94.7%~103% between, this shows this method high sensitivity, reproducible, and the rate of recovery is high.
【Description of the drawings】
Fig. 1 is the chromatogram of standard working solution；
Fig. 2 is the chromatogram of blank solution；
Fig. 3 is the chromatogram of sample solution；
Fig. 4 is the full scan chromatogram of standard working solution；
Fig. 5 is four alcohol extracting ion massspectrum figure of agalloch eaglewood；
Fig. 6 is benzylacetone extraction ion massspectrum figure.
【Specific implementation mode】
Further more detailed description is done to the present invention below in conjunction with specific implementation mode.
Embodiment 1：
Key instrument and reagent：
TSQ Quantum XLS Series gas chromatographs-mass spectrometer and its work station (matching Mo Feishier companies in the U.S.)；
UA22MFD supersonic wave cleaning machines (Chinese WIGGENS companies)；SIGMA 2-16K desk centrifuges (German sartoriussigma
Company)；CP 225D type electronic balance (sensibility reciprocals：0.0001g, Beijing Sai Duolisi companies).
Agalloch eaglewood tetrol standard items (96.8%, National Institute for Food and Drugs Control), benzylacetone standard items (98.0%,
Aladdin reagent (Shanghai) Co., Ltd.), methanol, ethyl alcohol, acetone, dichloromethane, ethyl acetate, acetonitrile, hexamethylene, anhydrous second
Ether, petroleum ether (chromatography alcohol, German Merck companies).The useful water of institute is ultra-pure water (resistivity is less than 18.2 μ s/cm).
Prepare testing sample solution：
Agalloch eaglewood sample ultra-pure water is cleaned and is dried, No. three sieves is smashed it through, accurately weighs 0.3g and be placed in conical flask with cover
In, it is accurate to 0.0001g.Methanol 15mL is added in precision, and weighed weight impregnates 1h, is ultrasonically treated (frequency 40kHz, power
300W) 50min is let cool, then weighed weight, and the weight of loss is supplied with methanol, is shaken up, and is centrifuged, and is stood, is taken supernatant liquor to obtain the final product
Sample solution, it is to be measured, while doing blank test.
Prepare standard working solution：
0.1020g benzylacetone standard items accurately are weighed, are placed in 100mL volumetric flasks, with methanol dissolving and constant volume, are prepared
At the benzylacetone standard reserving solution I of a concentration of 1mg/mL；0.0207g agalloch eaglewood tetrol standard items accurately are weighed, are placed in 10mL appearances
It in measuring bottle, is dissolved with methanol, while accurately pipetting 0.500mL benzylacetones standard reserving solution I and being placed in same 10mL volumetric flasks
And constant volume, it is configured to hybrid standard storing solution II, the wherein concentration of agalloch eaglewood tetrol and benzylacetone is respectively 2mg/mL and 50 μ g/
mL；The accurate hybrid standard storing solution II for pipetting 0.1mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL, is placed in 10mL capacity
In bottle, with methanol constant volume to scale to get standard working solution, the wherein gradient concentration of agalloch eaglewood tetrol is 20 μ g/mL, 40 μ g/
The gradient concentration of mL, 80 μ g/mL, 120 μ g/mL, 160 μ g/mL, 200 μ g/mL, benzylacetone are 0.5 μ g/mL, 1 μ g/mL, 2 μ
g/mL、3μg/mL、4μg/mL、5μg/mL。
Gas chromatography-mass spectrometry analysis：
Analysis detection is carried out to standard working solution, blank solution and sample solution using gas chromatograph-mass spectrometer (GC-MS),
Related chromatogram is obtained, sees Fig. 1, Fig. 2 and Fig. 3, abscissa indicates that retention time, ordinate indicate response in figure.With sample
The retention time and characteristic ion abundance ratio of solution and object in standard working solution carry out qualitative analysis, are carried out with external standard method
Quantitative analysis, the gas chromatography-mass spectrum condition are：
Chromatographic column：HP-5MS capillary columns, 30m × 0.25mm × 0.25 μm；Input mode：Splitless injecting samples, injection port
Temperature：250℃；Carrier gas：He, 99.999%, column flow 1.2mL/min；Column temperature uses temperature programming：40 DEG C of initial temperature is protected
1min is held, 210 DEG C are risen to 10 DEG C/min, 1min is kept, then 220 DEG C are risen to 2 DEG C/min, keeps 1min, then with 8 DEG C/min
300 DEG C are risen to, 4min is kept；Sample size：1μL；
Mass Spectrometry Conditions are：Ion source temperature：250℃；Ionization pattern：Electron impact ionization；Mass spectrum transmission line temperature 280
℃；Solvent delay time 3.0min；
Scan mode is that Salbutamol Selected Ion Monitoring scans (SIM), wherein agalloch eaglewood tetrol and benzylacetone be quantitative, qualitative ion and
Its abundance ratio is shown in Table 1, and the monitoring period to each ion is 50ms.
Draw standard curve and result of calculation：
Standard working solution is introduced into gas chromatograph-mass spectrometer (GC-MS), quantitative analysis is carried out with external standard method, by target chemical combination
The peak area of object and the concentration of target compound carry out linear regression analysis, obtain standard curve equation of linear regression, phase relation
Number need to be not less than 0.999；The sample solution of preparation is measured, the peak area of target compound in sample solution, generation are measured
Enter unary linear regression equation and obtain the content of target compound in sample solution, according to following calculation formula (1) and calculates public
Formula (2) converts acquires the content of agalloch eaglewood tetrol and benzylacetone in sample respectively：
Calculation formula (1) is：
Calculation formula (2) is：
In formula：
X1--- the content of agalloch eaglewood tetrol in sample, unit %；
X2--- the content of benzylacetone in sample, unit mg/kg；
C --- the concentration of target compound in sample solution, unit are μ g/mL；
C0--- the concentration of target compound in blank solution, unit are μ g/mL；
V --- sample solution constant volume, unit mL；
The quality that M --- sample weighs, unit g；
With the average value of parallel determinations twice for final measurement result, as a result retain 3 effective three numbers.
The range of linearity of the method for the present invention, detection limit and quantitative limit：
Standard working solution is introduced into gas chromatograph-mass spectrometer (GC-MS), quantitative analysis is carried out with external standard method, by target chemical combination
The peak area of object and the concentration of target compound carry out linear regression analysis, obtain the standard curve range of linearity, regression equation, phase
Relationship number, further accordance with the spectrogram of each component quota ion after standard working solution sample introduction, in combination with sample solution spectrogram, then with
Three times signal-to-noise ratio (S/N=3) calculates detection limit, is computed the detection limit that assessment obtains benzylacetone and agalloch eaglewood tetrol.As a result see
Table 2.
Table 2：The range of linearity, regression equation, related coefficient and detection limit
The method of the present invention precision and recovery test：
It is parallel to weigh 7 parts of agalloch eaglewood sample, 7 independent experiments, the relative standard deviation of measurement result are carried out as stated above
(RSD) it is method precision, the RSD of agalloch eaglewood tetrol and benzylacetone is respectively 3.47% and 4.04%.Another parallel weigh can
9 parts of agalloch eaglewood sample carries out the mark-on reclaims experiment of high, medium and low 3 concentration levels, the average recovery rate of target compound respectively
Between 92.6%~103%, the results are shown in tables 3.
Table 3：Method precision and the rate of recovery
The optimization and selection of Pretreatment：
One, suitable Pretreatment in order to obtain, to extracting mode, Extraction solvent, sample broke degree, solvent dosage and
Extraction time optimizes and selects.
1. the optimization and selection of extracting mode：
The present invention compares ultrasound and is heated to reflux two kinds of extracting modes, the results showed that using methanol as Extraction solvent, ultrasound
The target compound content of 50min extractions and the object measured value for being heated to reflux 5h extractions are almost the same.Because of ultrasonic extraction side
Method rapidly and efficiently, and can in normal-temperature operation, therefore select ultrasonic extraction mode, refer to table 4.
Table 4：The optimization and selection of extracting mode
2. the optimization and selection of Extraction solvent：
The present invention uses methanol, ethyl alcohol, acetone, dichloromethane, ethyl acetate, acetonitrile, hexamethylene, anhydrous ether and stone respectively
Oily ether is detected sample as Extraction solvent.Measurement result shows methanol, ethyl alcohol, dichloromethane and ethyl acetate to heavy
The extraction efficiency of fragrant tetrol is apparently higher than acetone, acetonitrile, hexamethylene, anhydrous ether and petroleum ether, and when selection methanol, ethyl alcohol,
When dichloromethane and ethyl acetate are as Extraction solvent, the measurement result no significant difference of agalloch eaglewood tetrol；Methanol and acetone are to benzyl
The extraction efficiency of benzylacetone is apparently higher than ethyl alcohol, dichloromethane, ethyl acetate, acetonitrile, hexamethylene, anhydrous ether and petroleum ether,
And when selecting methanol and acetone as Extraction solvent, the measurement result no significant difference of benzylacetone.It is therefore preferable that methanol is made
For the Extraction solvent of the present invention.Refer to table 5.
Table 5：The optimization and selection of Extraction solvent
Note:：" ND " indicates that target compound is not detected.
3. the optimization and selection of sample broke degree：
By agalloch eaglewood sample comminution, after crossing No.1 sieve, No. two sieves, No. three sieves, No. four sieves respectively, examined as stated above
It surveys.As known from Table 6, when grain size is more than No. three sieves, the relative standard deviation (RSD) of measurement result is excessive, the weight of testing result
Renaturation cannot meet the measurement requirement of the present invention, and when grain size is less than No. three sieves, sample homogeneity increases, measurement result
RSD is less than 5%, disclosure satisfy that the measurement requirement of the present invention.When grain size is less than No. three sieves, after crossing No. three sieves, No. four sieves
Sample is measured, the measurement result no significant difference of target compound.Therefore, No. three are crossed when the preferred sample pre-treatments of the present invention
Sieve.
Table 6：The optimization and selection of sample comminution degree
4. the optimization and selection of solvent dosage：
Accurate addition methanol 6,9,12,15,18mL are detected the present invention respectively in agalloch eaglewood sample.Measurement result table
It is bright, after methanol usage is more than 9mL, increase methanol usage, the measurement result of agalloch eaglewood tetrol still has an increase, and the survey of benzylacetone
Result is determined without significant change；After methanol usage is more than 12mL, increase methanol usage, the measurement knot of agalloch eaglewood tetrol and benzylacetone
Fruit has no significant change, therefore from the point of view of cost-effective, select solvent dosage for 12~15mL.Refer to table 7.
Table 7：The optimization and selection of solvent dosage
5. the optimization and selection of extraction time：
The present invention is in the preparation process of testing sample solution, and ultrasound 30,40,50,60,70min are detected respectively.It surveys
It is fixed the result shows that, upon extracting between more than 40min after, increase extraction time, the measurement result of agalloch eaglewood tetrol still has an increase, and benzyl
The measurement result of benzylacetone is without significant change；Between upon extracting more than 50min after, increase extraction time, agalloch eaglewood tetrol and benzyl
The measurement result of acetone has no significant change, therefore considers from time-saving angle, and the selective extraction time is 50min.Refer to table
8。
Table 8：The optimization and selection of extraction time
Two, the optimization and selection of gas-chromatography -- mass spectrometer parameter：
Suitable gas-chromatography -- mass spectrometer parameter in order to obtain, to temperature program, qualitative ion and quota ion
It optimizes and selects.
1. temperature program optimizes：
When actual sample detects, when heating rate is very fast, 210 DEG C or so have many aromatic components and sesquialter note component
Appearance is concentrated, retention time is closer to, and in order to obtain more preferably separating effect, and avoids other components to agalloch eaglewood tetrol and benzyl
Benzylacetone interferes, and through repeatedly debugging optimization, heats up before the present invention has selected 210 DEG C and after 220 DEG C very fast, and 210 DEG C~220
The slower temperature program that heats up between DEG C is detected.
2. the optimization and selection of qualitative, quantitative ion：
In the present invention, the full scan spectrogram of agalloch eaglewood tetrol and benzylacetone is obtained by full scan mode (SCAN) first,
See Fig. 4, abscissa indicates that retention time, ordinate indicate response in figure；Extract benzylacetone and agalloch eaglewood tetrol respectively again
Mass spectrogram, is shown in Fig. 5 and Fig. 6 respectively, and abscissa indicates that mass number, ordinate indicate abundance in figure.According to the fragment in mass spectrogram
Ion selects the characteristic ion fragment that abundance is relatively high, mass number is larger and interference is few, final determining target compound
Qualitative, quantitative ion, is shown in Table 1.
Embodiment 2：
According to assay method described in embodiment 1,10 batches of commercially available agalloch eaglewood samples are detected, agalloch eaglewood tetrol and benzyl
The content of acetone is shown in Table 9.As shown in Table 9, the mark that the content of agalloch eaglewood tetrol and benzylacetone is selected in the present invention in sample solution
Within the scope of quasi- working curve, the measurement of standard working solution agalloch eaglewood tetrol and benzylacetone suitable for agalloch eaglewood.
Table 9：The content of agalloch eaglewood tetrol and benzylacetone in 10 batches of agalloch eaglewood samples
。
Claims (8)
1. method that is a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood, it is characterised in that include the following steps：
A, testing sample solution is prepared：Agalloch eaglewood sample ultra-pure water is cleaned and is dried, No. three sieves is smashed it through, accurately weighs 0.3g
It has been placed in plug conical flask, has been accurate to 0.0001g, 12~15mL of methanol is added in precision, and weighed weight impregnates 1h, is ultrasonically treated
50min is let cool, then weighed weight, and the weight of loss is supplied with methanol, is shaken up, and is centrifuged, and is stood, is taken supernatant liquor up to sample
Solution, it is to be measured, while doing blank solution；
B, standard working solution is prepared：The agalloch eaglewood tetrol with concentration gradient and the benzyl with concentration gradient third are prepared with methanol
Ketone hybrid standard working solution, the wherein concentration range of agalloch eaglewood tetrol are 20~200 μ g/mL, and the concentration range of benzylacetone is
0.5~5 μ g/mL；
C, gas chromatography-mass spectrometry analysis：Using gas chromatograph-mass spectrometer (GC-MS) to standard working solution, blank solution and sample
Solution carries out analysis detection, obtains related chromatogram, with the retention time of object in sample solution and standard working solution and
Characteristic ion abundance carries out quantitative analysis than carrying out qualitative analysis, with external standard method；
Wherein the gas chromatography-mass spectrum condition is：
Chromatographic column：HP-5MS capillary columns, 30m × 0.25mm × 0.25 μm；Input mode：Splitless injecting samples, injector temperature：
250℃；Carrier gas：He, 99.999%, column flow 1.2mL/min；Column temperature uses temperature programming：40 DEG C of initial temperature is kept
1min rises to 210 DEG C with 10 DEG C/min, keeps 1min, then rise to 220 DEG C with 2 DEG C/min, keeps 1min, then with 8 DEG C/min liters
To 300 DEG C, 4min is kept；Sample size：1μL；
Mass Spectrometry Conditions are：Ion source temperature：250℃；Ionization pattern：Electron impact ionization；280 DEG C of mass spectrum transmission line temperature；It is molten
Agent delay time 3.0min；
Scan mode：Selected ion monitoring scans, and the monitoring period to each ion is 50ms；
D. it draws standard curve and result calculates：Standard working solution is introduced into gas chromatograph-mass spectrometer (GC-MS), with external standard method into
The concentration of the peak area of target compound quota ion and target compound is carried out linear regression analysis, obtained by row quantitative analysis
To standard curve equation of linear regression, related coefficient need to be not less than 0.999；The sample solution of preparation is measured, sample is measured
The peak area of target compound in product solution substitutes into unary linear regression equation and obtains containing for target compound in sample solution
Amount converts further according to following calculation formula (1) and calculation formula (2) and acquires containing for agalloch eaglewood tetrol and benzylacetone in sample respectively
Amount：
Calculation formula (1) is：
Calculation formula (2) is：
In formula：
X1--- the content of agalloch eaglewood tetrol in sample, unit %；
X2--- the content of benzylacetone in sample, unit mg/kg；
C --- the concentration of target compound in sample solution, unit are μ g/mL；
C0--- the concentration of target compound in blank solution, unit are μ g/mL；
V --- sample solution constant volume, unit mL；
The quality that M --- sample weighs, unit g.
2. method that is according to claim 1 a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood, feature exist
In the agalloch eaglewood include import agalloch eaglewood, Home-made occluder and using artificial Edgeworthia chrysantha technology production agalloch eaglewood.
3. method that is according to claim 1 a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood, feature exist
In step a, the condition of sample ultrasonic processing is frequency 40kHz, power 300W.
4. method that is according to claim 1 a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood, feature exist
In the standard working solution of preparation, the gradient concentration of agalloch eaglewood tetrol is 20 μ g/mL, 40 μ g/mL, 80 μ g/mL, 120 μ g/mL,
The gradient concentration of 160 μ g/mL, 200 μ g/mL, benzylacetone are 0.5 μ g/mL, 1 μ g/mL, 2 μ g/mL, 3 μ g/mL, 4 μ g/mL, 5 μ
g/mL。
5. method that is according to claim 4 a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood, feature exist
It is in the method for preparing standard working solution：
0.1g benzylacetone standard items accurately are weighed, 0.1mg is accurate to, is placed in 100mL volumetric flasks, is dissolved and determined with methanol
Hold, is configured to the benzylacetone standard reserving solution I of a concentration of 1mg/mL；0.02g agalloch eaglewood tetrol standard items accurately are weighed, are accurate to
0.1mg is placed in 10mL volumetric flasks, is dissolved with methanol, while accurately pipetting 0.500mL benzylacetones standard reserving solution I and being placed in
In same 10mL volumetric flasks and constant volume, it is configured to hybrid standard storing solution II, the wherein concentration of agalloch eaglewood tetrol and benzylacetone point
It Wei not 2mg/mL and 50 μ g/mL；The accurate hybrid standard deposit for pipetting 0.1mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL
Liquid II is respectively placed in 10mL volumetric flasks, with methanol constant volume to scale to get standard working solution.
6. method that is according to claim 1 a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood, feature exist
When selected ion monitoring scans, agalloch eaglewood tetrol and benzylacetone be quantitative, qualitative ion and its abundance ratio are：
Agalloch eaglewood tetrol, retention time 30.93min, quota ion 91, qualitative ion 282 assist qualitative ion 163, abundance ratio
100：40：3；
Benzylacetone, retention time 10.84min, quota ion 105, qualitative ion 91 assist qualitative ion 148, abundance ratio
100：87：57.
7. method that is according to claim 1 a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood, feature exist
In in drawing standard curve and result calculating, the selection ion chromatography peak of target compound and standard items is in identical guarantor in sample
It stays at the time and occurs, and the mass-to-charge ratio of corresponding mass spectrometric fragment ion is consistent with standard items, abundance ratio is answered compared with standard items
Meet：
When relative abundance ＞ 50%, allow ± 10% deviation；
When relative abundance 20%~50%, allow ± 15% deviation；
When relative abundance 10%~20%, allow ± 20% deviation；
When relative abundance≤10%, allow ± 50% deviation；Target analytes can be confirmed at this time.
8. method that is according to claim 1 a kind of while measuring agalloch eaglewood tetrol and benzylacetone in agalloch eaglewood, feature exist
When being calculated according to formula, with the average value of parallel determinations twice for final measurement result, as a result retain 3 effective three numbers
Word.
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| CN109406679A (en) * | 2018-12-11 | 2019-03-01 | 北京三和药业有限公司 | A kind of detection method of high effective liquid chromatography for measuring agalloch eaglewood quality |
| CN111272899A (en) * | 2020-03-11 | 2020-06-12 | 中国热带农业科学院热带生物技术研究所 | Method for identifying agilawood produced by thermal department No. 2 |
| CN114113411A (en) * | 2021-12-15 | 2022-03-01 | 深圳天祥质量技术服务有限公司 | Method for simultaneously detecting 5 harmful substances in consumer goods |
| CN117434170A (en) * | 2023-08-23 | 2024-01-23 | 珠海莱森博萃生物科技有限公司 | Method for simultaneously measuring benzyl acetone, adenosine and mangiferin in aquilaria sinensis leaves |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103630658A (en) * | 2013-11-06 | 2014-03-12 | 广东省微生物研究所 | Method for identifying natural agilawood and artificial agilawood |
| CN104597155A (en) * | 2015-01-09 | 2015-05-06 | 陕西科技大学 | Quality control method for agilawood |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2688855B2 (en) * | 1989-10-30 | 1997-12-10 | ダイセル化学工業株式会社 | Method for purifying benzylacetoxime |
-
2016
- 2016-06-28 CN CN201610506609.5A patent/CN106198791B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103630658A (en) * | 2013-11-06 | 2014-03-12 | 广东省微生物研究所 | Method for identifying natural agilawood and artificial agilawood |
| CN104597155A (en) * | 2015-01-09 | 2015-05-06 | 陕西科技大学 | Quality control method for agilawood |
Non-Patent Citations (6)
| Title |
|---|
| ANALYSIS OF HIGH QUALITY AGARWOOD OIL CHEMICAL COMPOUNDS BY MEANS OF SPME/GC-MS AND Z-SCORE TECHNIQUE;Nurlaila Ismail et al.;《Malaysian Journal of Analytical Sciences》;20131231;第17卷(第3期);第403-413页 * |
| GC-MS Analysis of Chemical Constituents from Various Agarwood;Kwhang-Ho Shin et al.;《Kor. J. Herbology》;20111231;第26卷(第1期);第7-12页 * |
| HPLC-DAD 测定沉香药材中沉香四醇的含量;顾宇凡等;《世界科学技术—中医药现代化》;20141231;第16卷(第12期);第2643-2646页 * |
| 一种新型人工结香沉香挥发性成分ＧＣ－ＭＳ分析;赵艳艳等;《亚太传统医药》;20130630;第9卷(第6期);第6-11页 * |
| 不同产区沉香药材挥发油成分GC-MS分析;郭晓玲等;《中药材》;20090930;第32卷(第9期);第1354-1358页 * |
| 国产沉香的固相微萃取-气相色谱-质谱分析;侯冬岩等;《鞍山师范学院学报》;20101231;第12卷(第6期);第44-46页 * |
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