CN106483187A - Chinese medicine based on ion mobility spectrometry and similarity easily mixes adulterant discrimination method with which - Google Patents
Chinese medicine based on ion mobility spectrometry and similarity easily mixes adulterant discrimination method with which Download PDFInfo
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- CN106483187A CN106483187A CN201610888887.1A CN201610888887A CN106483187A CN 106483187 A CN106483187 A CN 106483187A CN 201610888887 A CN201610888887 A CN 201610888887A CN 106483187 A CN106483187 A CN 106483187A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
Abstract
The invention discloses the Chinese medicine based on ion mobility spectrometry and similarity easily mixes adulterant discrimination method with which, the method is with Ion mobility spectrometry as analyzing means of testing, with Similarity Measure and quality control chart as quantization method, there is provided a kind of quantization method suitable for Chinese medicine quality control.Present invention achieves traditional Chinese medicine quality control differentiates the transformation based on qualitative analysis to quantitative evaluation, have quick, accurately, output can be quantified, new Evaluation example is provided for China's Chinese medicine quality safety, the control of centering quality of medicinal material ensures and clinical application is significant safely.
Description
Technical field
The invention belongs to Chinese medical herb technical field, more particularly to a kind of Chinese medicine easily mixes the discriminating side of adulterant with which
Method.
Background technology
Chinese medicine be Chinese medicine compound prescription play drug effect important foundation, adulterant be mixed into the performance that greatly have impact on drug effect,
Threaten the health of patient.By taking common Chinese medicine material Radix Codonopsis as an example, Radix Codonopsis (Radix Codonopsis) is Campanulaceae Radix Codonopsis platymiscium
Radix Codonopsis Codonopsis pilosula (Franch) Nannf, codonopsis pilosula var. modesta Codonopsis pilosula (Franch)
Nannf.Var.modesta (Nannf) L.T.shen or the dry root of radix codonpsis tangshen Codonpsis tangshen oliv, have
The effects such as tonifying middle-Jiao and Qi, strengthening spleen and tonifying lung, but as wild Radix Codonopsis quantity is more short, artificial culture product can not meet often
The market demand, causes mixed adulterant to happen occasionally.Radix stellariae dichotomae (Stellaria dichotoma L.var.lanceolata Bge)
It is more typical easily mixed adulterant, primary efficacy is clearind deficient heat, treating infantile malnutrition wide fever, and both effects are different, can not use with (Zhang Chaohui, practical
Traditional Chinese medicine magazine, 2011,27 (9), 636.), cuneate lespedeza herb with root (Lespedeza cuneate Dum Cours G.Don) is equally which
Relatively conventional adulterant, its effect are mainly liver-benefiting eye-brightening, heat clearing and blood circulation promoting, diuresis removing toxic substances, different from Radix Codonopsis drug effect, can not use with
(Huang Donglan, Sun Suqin, Xu Yongqun etc., modern instrument, 2008,5,22-25).
Common Chinese medicine quality control authentication technique mainly has two categories below strategy, and one is base chiller, proterties calibrating
With micro- calibrating, these calibration methods have certain artificial property (Qian Yunchuan, Zhou Lemin, Chinese medicine company, 2001,10 (5), 59. Lee
Dawn beautiful jade, Shao Aijuan, exhibition day dawn etc., CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2015,40 (7), 1271-1273.)；Two is that physics and chemistry calibrating and instrument divide
Analysis means, but the testing time is longer, expensive equipment (Zou Yuanfeng, Cao Chaosheng, Liu Jiang etc., Chinese herbal medicine, 2010,41 (3), attached 3- is attached
6).
Therefore, necessary on the basis of prior art, a kind of energy of research and design is quick, accurate, can quantify discriminating
The method that Chinese medicine easily mixes adulterant with which.
Content of the invention
In order to solve the technical problem of above-mentioned background technology proposition, the present invention is intended to provide based on ion mobility spectrometry and similar
The Chinese medicine of degree easily mixes adulterant discrimination method with which, can quickly, accurately, can quantify to differentiate Chinese medicine with its easy mixed adulterant.
In order to realize above-mentioned technical purpose, the technical scheme is that：
Chinese medicine based on ion mobility spectrometry and similarity easily mixes adulterant discrimination method with which, comprises the following steps：
(1) certain Chinese medicine and its easily mixed adulterant is selected, the Chinese medicine genuine piece sample and easy mixed adulterant is carried out respectively as follows
Pre-treatment：
Sample is processed using pulverizer and sample powder is obtained, 1-1000mg sample powder is accurately weighed, add 1-500mL to carry
Solvent is taken, magnetic agitation at 25-100 DEG C, 5min-24h is reacted under the conditions of condensate return, question response system is naturally cold
But, filtrate is collected by filtration, obtains sample solution；
(2) respectively the sample solution of Chinese medicine and the sample solution of easily mixed adulterant that step (1) is obtained is carried out by ion and is moved
Move modal data collection：
The sample solution that 5-50 μ L step (1) is obtained accurately is pipetted, is used for electrospray ion mobility spectrometry instrument, experiment ginseng
Number：Positive and negative ion pattern, ion source voltage are 1000-3000V, and migration tube voltage is 5000-10000V, and carrier gas temperature is
100-250 DEG C, migration tube temperature is 100-250 DEG C, and flow rate of carrier gas is 1-5L/min, and sample introduction speed is 1-5 μ L/min, gate voltage
For 1-100V, gatewidth is 5-300 μ s；
(3) the Chinese medicine Ion transfer modal data for being obtained according to step (2) and easily mixed adulterant Ion transfer modal data, calculate
Both similarity coefficient correlations and included angle cosine：
In above formula, xiAnd yiTwo ion mobility spectrometries intensity level i-th variable at is represented respectively,WithGeneration respectively
Mean value of two ion mobility spectrometries of table at i-th variable, n are the sum of variable in ion mobility spectrometry；
(4) gather at least 3 batches the Chinese medicine genuine piece sample, respectively according to step (1)-(3) described process at
Reason, obtains each batch Chinese medicine genuine piece and easily mixes coefficient correlation and the cosine angle of adulterant with which, and sets up and be based respectively on phase relation
Number and the quality control chart of cosine angle；
(5) traditional Chinese medicinal material samples to be detected are processed according to step (1)-(3) described process respectively, is obtained to be checked
The traditional Chinese medicinal material samples of survey easily mix the similar degrees of data of adulterant to which, and the similar degrees of data is substituted into the quality that step (4) is set up
In control figure, according to quality control chart judgment rule, so as to identify Chinese medicine to be detected with the presence or absence of adulterant risk.
Further, the quality control chart that step (4) is set up include average-range chart, mean-standard deviation figure, median-
Range chart and monodrome-moving range figure.
Further, the Extraction solvent in step (1) for methyl alcohol, ethanol, water or the former be mutually mixed solution.
Further, in step (1) pre-treatment design parameter：100mg sample powder is accurately weighed, adds 30mL methyl alcohol
In solution, magnetic agitation at 100 DEG C, 2h is reacted under the conditions of condensate return, question response system naturally cools to room temperature, filters
Filtrate is collected, obtains sample solution.
Further, the design parameter of step (2) intermediate ion migration modal data collection：25 μ L sample solution are accurately pipetted,
It is used for electrospray ion mobility spectrometry instrument, specific experiment condition：Negative ion mode, ion source voltage are 2000V, migration tube electricity
Press as 8000V, carrier gas temperature is 180 DEG C, migration tube temperature is 180 DEG C, flow rate of carrier gas is 1.00L/min, sample introduction speed is
1.20 μ L/min, gate voltage are 45V, and gatewidth is 65 μ s.
The beneficial effect that is brought using technique scheme：
The present invention with Ion mobility spectrometry as analyze means of testing, with similarity algorithm as quantization method, there is provided one
The quantization method suitable for Chinese medicine quality control is planted, with multiple advantages.1) amount of samples is few, it is only necessary to which several tens of microliters is extracted
Liquid, you can complete the collection of testing sample signal；2) analyze speed is fast, and loading was tested to the result output used time less than 1 minute；3)
Equipment volume is little, is easy to carry, and may be used to the analysis of live online experiment；4) result quantifies output, it is achieved that qualitative to quantitative
As a result differentiate and change；5) structure of quality control chart is realized, may be used to traditional Chinese medicinal material samples quality control.The present invention is expected to solve me
The unmanageable present Research of state's Chinese medicine supervision of quality safety.
Description of the drawings
Fig. 1 is the basic flow sheet of the present invention；
Fig. 2 is the ion transfer spectrogram of Radix Codonopsis；
Fig. 3 is the ion transfer spectrogram of radix stellariae dichotomae；
Fig. 4 is the ion transfer spectrogram of cuneate lespedeza herb with root；
Fig. 5 is the Radix Codonopsis mean-range chart based on coefficient correlation；
Fig. 6 is the Radix Codonopsis mean-range chart based on included angle cosine.
Specific embodiment
Below with reference to accompanying drawing, technical scheme is described in detail.
The present embodiment selects representative Radix Codonopsis as Chinese medicine, selects radix stellariae dichotomae and cuneate lespedeza herb with root as easily mixed puppet
Product, illustrate.
As shown in figure 1, the Chinese medicine based on ion mobility spectrometry and similarity easily mixes adulterant discrimination method with which, including following
Step：
Step 1：Select the Radix Codonopsis genuine piece sample of 8 batches, respectively the Radix Codonopsis genuine piece sample to this 8 batch and radix stellariae dichotomae and
Cuneate lespedeza herb with root sample carries out pre-treatment：
Above-mentioned sample is processed respectively using pulverizer (25000r/min, 3-10min) and corresponding powder is obtained, for every kind of sample
Product powder, accurately weighs 100mg sample powder, adds in 30mL methanol solution, magnetic agitation at 100 DEG C, condensate return
Under the conditions of react 2h, question response system naturally cools to room temperature, filtrate is collected by filtration, and obtains sample solution.
Step 2：The ion for gathering 8 batch Radix Codonopsis genuine piece sample solutions and radix stellariae dichotomae, the sample solution of cuneate lespedeza herb with root respectively is moved
Move modal data：
25 μ L sample solution are accurately pipetted, is used for electrospray ion mobility spectrometry instrument, specific experiment condition：Anion mould
Formula, ion source voltage are 2000V, and migration tube voltage is 8000V, and carrier gas temperature is 180 DEG C, and migration tube temperature is 180 DEG C, carrier gas
Flow velocity is 1.00L/min, and sample introduction speed is 1.20 μ L/min, and gate voltage is 45V, and gatewidth is 65 μ s.
As shown in Figure 2, Figure 3 and Figure 4, its abscissa is for during migration for the ion transfer spectrogram of Radix Codonopsis, radix stellariae dichotomae and cuneate lespedeza herb with root
Between, ordinate is intensity level.
Electrospray ion mobility spectrometry (Electrospray ion mobility spectrometry, ESI-IMS), is one
Plant the new technology that sample material ion analysis under atmospheric pressure are realized in electron spray soft ionization mode, with analysis
The advantages of speed is fast, amount of samples is few.Traditional Chinese medicine sample solution injects instrument through micro syringe, and it is anti-that electrospray mode introduces ionization
Area is answered, sample molecule occurs ionization to become ion in ionization source region, and ion is entered by ion fence gate under electric field action and moved
Area is moved, and detector is reached through migration area under electric field action.As the ion of different quality or structure is under electric field action
Mobility different, thus with different migration velocities, so each ion reaches detector successively, current generated through putting
Big and be converted into voltage signal output, the ionic strength of sample and the graph of a relation i.e. ion transfer spectrogram of transit time can be obtained.
This new Ion mobility spectrometry, can fast and accurately obtain the ion transfer spectrogram of experiment traditional Chinese medicinal material samples.
Step 3：The Ion transfer modal data of 8 batch Radix Codonopsis obtained according to step 2 and the ion of radix stellariae dichotomae, cuneate lespedeza herb with root are moved
Modal data is moved, and each batch Radix Codonopsis is calculated respectively to radix stellariae dichotomae, the similar degrees of data coefficient correlation of cuneate lespedeza herb with root and included angle cosine：
In above formula, xiAnd yiTwo ion mobility spectrometries intensity level i-th variable at is represented respectively,WithRepresent respectively
The mean value of two ion mobility spectrometries at i-th variable, n are the sum of variable in ion mobility spectrometry.
Analyze for Radix Codonopsis and its adulterant, solvent is methyl alcohol condition, before not considering transit time 9ms during Similarity Measure
Spectral peak information, with exclude solvent peak impact.In the present embodiment, taking 9 15ms Ion transfer modal data of transit time carries out phase
Calculate like degree.
Step 4：Similarity Measure result shows that, all 0.991 or so, included angle cosine value is all 0.997 for correlation coefficient value
Left and right, sets up the quality control chart based on coefficient correlation and the quality control chart based on included angle cosine accordingly.The quality control
Figure including average-range chart, mean-standard deviation figure, median-range chart and monodrome-moving range figure.
Step 5：Radix Codonopsis sample to be detected is processed according to process described in step 1-3 respectively, is obtained to be detected
Radix Codonopsis sample and radix stellariae dichotomae, the similar degrees of data of cuneate lespedeza herb with root, and the similar degrees of data is substituted into the quality control chart that step 4 is set up
In.Fig. 5 is the mean-range chart built based on correlation coefficient value, and Fig. 6 is the average-pole built based on included angle cosine value
, when sample value to be measured jumps out upper lower control limit scope, that is, can there is higher adulterant according to control figure judgment rule in difference control figure
Risk, can achieve Chinese medicine quality quantified controlling accordingly.In Fig. 4, Fig. 5, CL represents center line, and UCL represents upper control limit, LCL generation
Table lower control limit, Mean represent average, and Range represents extreme difference.
Above example technological thought only to illustrate the invention, it is impossible to which protection scope of the present invention is limited with this, every
According to technological thought proposed by the present invention, any change that is done on the basis of technical scheme, the scope of the present invention is each fallen within
Within.
Claims (5)
1. the Chinese medicine based on ion mobility spectrometry and similarity easily mixes adulterant discrimination method with which, it is characterised in that including following
Step：
(1) certain Chinese medicine and its easily mixed adulterant are selected, and to the Chinese medicine genuine piece sample and easily mixing adulterant carries out following front place respectively
Reason：
Sample is processed using pulverizer and sample powder is obtained, 1-1000mg sample powder is accurately weighed, adds 1-500mL to extract molten
Agent, magnetic agitation at 25-100 DEG C, 5min-24h, question response system natural cooling, mistake is reacted under the conditions of condensate return
Filtrate is collected in filter, obtains sample solution；
(2) respectively ion mobility spectrometry is carried out to the sample solution of Chinese medicine and the sample solution of easily mixed adulterant that step (1) is obtained
Data acquisition：
The sample solution that 5-50 μ L step (1) is obtained accurately is pipetted, is used for electrospray ion mobility spectrometry instrument, experiment condition：
Positive and negative ion pattern, ion source voltage are 1000-3000V, and migration tube voltage is 5000-10000V, and carrier gas temperature is 100-
250 DEG C, migration tube temperature is 100-250 DEG C, and flow rate of carrier gas is 1-5L/min, and sample introduction speed is 1-5 μ L/min, and gate voltage is 1-
100V, gatewidth are 5-300 μ s；
(3) the Chinese medicine Ion transfer modal data for being obtained according to step (2) and easily mixed adulterant Ion transfer modal data, calculate both
Similarity coefficient correlation and included angle cosine：
In above formula, xiAnd yiTwo ion mobility spectrometries intensity level i-th variable at is represented respectively,WithTwo are represented respectively
Mean value of the ion mobility spectrometry at i-th variable, n are the sum of variable in ion mobility spectrometry；
(4) the Chinese medicine genuine piece sample of at least 3 batches is gathered, is processed according to step (1)-(3) described process respectively, obtains
Coefficient correlation and the cosine angle of adulterant is easily mixed to each batch Chinese medicine genuine piece with which, and set up and be based respectively on coefficient correlation and remaining
The quality control chart of string angle；
(5) traditional Chinese medicinal material samples to be detected are processed according to step (1)-(3) described process respectively, is obtained to be detected
Traditional Chinese medicinal material samples easily mix the similar degrees of data of adulterant to which, and the similar degrees of data is substituted into the quality control that step (4) is set up
In figure, according to quality control chart judgment rule, so as to identify Chinese medicine to be detected with the presence or absence of adulterant risk.
2. Chinese medicine according to claim 1 based on ion mobility spectrometry and similarity easily mixes adulterant discrimination method with which, its
It is characterised by：The quality control chart that step (4) is set up include average-range chart, mean-standard deviation figure, median-range chart and
Monodrome-moving range figure.
3. Chinese medicine according to claim 1 based on ion mobility spectrometry and similarity easily mixes adulterant discrimination method with which, its
It is characterised by：Extraction solvent in step (1) for methyl alcohol, ethanol, water or the former be mutually mixed solution.
4. the Chinese medicine according to claim 1 or 3 based on ion mobility spectrometry and similarity easily mixes adulterant discrimination method with which,
It is characterized in that：The design parameter of pre-treatment in step (1)：
100mg sample powder is accurately weighed, is added in 30mL methanol solution, magnetic agitation at 100 DEG C, condensate return condition
Lower reaction 2h, question response system naturally cool to room temperature, filtrate are collected by filtration, and obtain sample solution.
5. Chinese medicine according to claim 1 based on ion mobility spectrometry and similarity easily mixes adulterant discrimination method with which, its
It is characterised by：The design parameter of step (2) intermediate ion migration modal data collection：
25 μ L sample solution are accurately pipetted, is used for electrospray ion mobility spectrometry instrument, specific experiment condition：Negative ion mode,
Ion source voltage is 2000V, and migration tube voltage is 8000V, and carrier gas temperature is 180 DEG C, and migration tube temperature is 180 DEG C, carrier gas stream
Speed is 1.00L/min, and sample introduction speed is 1.20 μ L/min, and gate voltage is 45V, and gatewidth is 65 μ s.
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Application publication date: 20170308 |